27 CFR 7.22a - Voluntary disclosure of major food allergens.

Science.gov (United States)

2010-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Voluntary disclosure of major food allergens. 7.22a Section 7.22a Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX... Labeling Requirements for Malt Beverages § 7.22a Voluntary disclosure of major food allergens. (a...

Tropomyosin and Actin Identified as Major Allergens of the Carpet Clam (Paphia textile and the Effect of Cooking on Their Allergenicity

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Zailatul Hani Mohamad Yadzir

2015-01-01

Full Text Available Objectives. To identify the major allergenic proteins of clam (Paphia textile and to investigate the effect of different cooking methods on the allergenicity of these identified proteins. Methods. Clam protein extracts were separated by denaturing polyacrylamide gel electrophoresis. IgE reactive proteins were then analyzed by immunoblotting with sera from patients with positive skin prick tests (SPT to the raw clam extract. Mass spectrometry was used to identify the major allergenic proteins of this clam. Results. Raw extract showed 12 protein bands (18–150 kDa. In contrast, fewer protein bands were seen in the boiled extract; those ranging from 40 to 150 kDa were denatured. The protein profiles were similarly altered by frying or roasting. The immunoblots of raw and boiled extracts yielded 10 and 2 IgE-binding proteins, respectively. The fried and roasted extracts showed only a single IgE-binding protein at 37 kDa. Mass spectrometry analysis of the 37 and 42 kDa major allergens indicated that these spots were tropomyosin and actin, respectively. Conclusion. The two major allergens of Paphia textile were identified as the thermostable tropomyosin and a new thermolabile allergen actin.

27 CFR 4.32a - Voluntary disclosure of major food allergens.

Science.gov (United States)

2010-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Voluntary disclosure of major food allergens. 4.32a Section 4.32a Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX... Requirements for Wine § 4.32a Voluntary disclosure of major food allergens. (a) Definitions. For purposes of...

Use of gamma irradiation for microbial inactivation of buckwheat flour and buckwheat food products, 4

International Nuclear Information System (INIS)

Muramatsu, Nobuyuki; Oh-hinata, Hiroshi; Ohara, Tadahiko; Ohike, Terutake; Ito, Hitoshi.

1988-01-01

The effect of gamma irradiation for extension of shelf-life on buckwheat noodles was investigated from the number of microorganisms point of view by using noodles prepared by irradiated buckwheat flour or irradiated directly to fresh noodles in nylon pouch. The shelf-life of noodles prepared by irradiated buckwheat flour at 0.5 Mrad was extended 2∼2.5 times of compared with non-irradiated buckweat flour under the same condition of processing and storage temperature. Low temperature extended the shelf-life of noodles prepared by irradiated buckwheat flour at 0.5 Mrad for 7 to 8 days at 10deg C, which was extended 3∼5 times of shelf-life compared with storage at 25deg C. The shelf-life of direct-irradiated buckweat noodles at 1.0 Mrad was extended about 30 days at 25deg C. In the case of direct-irradiation to fresh buckwheat noodles at 0.5 Mrad, their shelf-life was extended about 30 days at 15deg C. (author)

Glycoproteomic analysis of seven major allergenic proteins reveals novel post-translational modifications

DEFF Research Database (Denmark)

Halim, Adnan; Carlsson, Michael C; Mathiesen, Caroline Benedicte K

2015-01-01

Allergenic proteins such as grass pollen and house dust mite (HDM) proteins are known to trigger hypersensitivity reactions of the immune system, leading to what is commonly known as allergy. Key allergenic proteins including sequence variants have been identified but characterization of their post...... allergens. Moreover, we identified more complex glycan structures than previously reported on the major grass pollen group 1 and 5 allergens, implicating important roles for carbohydrates in allergen recognition and response by the immune system. The new findings are important for understanding basic...

CYTOTOXIC AND ANTIOXIDANT ACTIVITY OF BUCKWHEAT HULL EXTRACTS

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Martina Danihelová

2013-02-01

Full Text Available Buckwheat contains many prophylactic compounds that are concentrated mainly in outer layers of buckwheat grain. The aim of this study was to prepare buckwheat hull extracts. Ten buckwheat cultivars were screened for their antioxidant and anticancer properties. Total polyphenol content was determined using Folin-Ciocalteau's reagent. Antioxidant activity was established by the method of binding free radical DPPH. Cytotoxic properties were measured on human cervical cancer cells HeLa using mitochondrial cytotoxic test (MTT. Total polyphenol content ranged from 166.67 to 635.31 mg GAE/100 g DW. The highest content displayed tartary buckwheat cultivar Madawaska (0.64% of hulls weight. Among common buckwheat the richest in polyphenols were cultivars Bamby and KASHO-2. The best free radical binding antioxidant activity was found for cultivars with highest polyphenol content. This relationship was not observed for cytotoxic action on human cervical cancer cells. The best growth inhibitory activity on HeLa cancer cells displayed common buckwheat cultivars Bamby and KASHO-2 (up to 50%, extract concentration 100 µg/ml. This was not found for tartary buckwheat cultivar Madawaska.

A clinically relevant major cross-reactive allergen from mesquite tree pollen.

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Dhyani, A; Singh, B P; Arora, N; Jain, V K; Sridhara, S

2008-10-01

Prosopis juliflora (mesquite) is one of the major sources of pollinosis in tropical and semi-arid countries of the world. The present study was undertaken to purify and characterize a major cross-reactive allergen from this tree species. Mesquite pollen extract was purified using reverse-phase chromatography. Allergen characterization was done by electrophoresis, enzyme-linked immunosorbent assay (ELISA) and Western blotting. Clinical relevance of the purified protein was analyzed by in vivo (skin tests) and in vitro experiments such as ELISA, histamine release, peripheral blood mononuclear cells (PBMC) proliferation and cytokine assays. Cross-reactivity of purified protein with allergenic tree species and lima bean (food) was assessed by inhibition assays. A 66-kDa protein was purified from mesquite pollen extract using octadecyl silica resin. Purified protein recognized 90% of mesquite-sensitized patients in skin test and ELISA. It induced significant histamine release in allergic patients' blood and interleukin-4 secretion in the PBMC culture supernatants. Inhibition assays suggested close allergenic relationship of this protein with Ailanthus excelsa, Cassia siamea, Salvadora persica pollen and Phaseolus lunatus (lima bean - an edible legume). A 66-kDa major cross-reactive allergen was isolated from mesquite pollen using single-step purification procedure. The protein seems relevant for clinical applications in allergic disorders.

Recent Progress in Perennial Buckwheat Development

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Qing-Fu Chen

2018-02-01

Full Text Available Grains in the genus Fagopyrum have benefits to human health and are an excellent gluten-free raw material. Of all cereal foods, this genus has the highest total content of amino-acid nutrients necessary for humans; nutrients that are resistant to digestion (protein and starch resulting in their sustained release; higher dietary fiber content than key cereals, and is rich in a special healthy ingredient (flavonoids. Fagopyrum includes 24 species of which five are perennial. Among them, golden buckwheat (F.cymosum complex is the most important perennial buckwheat, which is not only used in Chinese medicine, but also has great potential in healthy food crop. In order to provide some clues for perennial crop studies and their industry development, this paper presents the state of perennial buckwheat research in terms of taxonomy; natural chemical products and pharmacological and health functions; genetics and evolution; breeding; and product development and utilization. The great advances such as successful interspecific crossing and its subsequent new perennial buckwheat varieties will speed up the development of the perennial buckwheat industry.

A competitive solid-phase radioimmunoassay for quantitation of the major allergen of Parietaria pollen

International Nuclear Information System (INIS)

Corbi, A.L.; Ayuso, R.; Lombardero, M.; Duffort, O.; Carreira, J.

1985-01-01

A competitive solid-phase radioimmunoassay has been developed for quantitation of the major allergen of Parietaria judaica pollen. The assay is based on: (1) the ability of AC/1.1 monoclonal antibody to bind specifically to the P. judaica major allergen, and (2) the ability of crude pollen extracts or purified allergen to inhibit the binding of 125 I-labelled allergen to solid-phase-bound AC/1.1 monoclonal antibody. The assay is sensitive enough to detect as little as 10 ng of allergen. A good correlation is found when the results obtained are compared with those produced by RAST inhibition (r = 0.95; P < 0.001). Thus, this method can also be used for the estimation of the allergenic activity of P. judaica pollen extracts. The assay is easily completed in 2 h, allowing simultaneous analysis of a number of extracts. (Auth.)

Impact of buckwheat flour granulation and supplementation level on the quality of composite wheat/buckwheat ginger-nut-type biscuits

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Bojana Filipčev

2015-12-01

Full Text Available Effects of gradual wheat flour substitution with buckwheat flour in ginger-nut biscuit formulation were investigated regarding dough characteristics, physical and textural characteristics of finalproduct assessed after baking and 30 days of storage. Buckwheat flour was added at 30, 40, 50% levels and two granulations (fine and coarse. Addition of buckwheat flour significantly increaseddough hardness and decreased adhesiveness. Spread significantly increased in biscuits with 40% and 50% of coarse buckwheat flour. Biscuits containing coarse flour were harder and more fracturable than the control, whereas those with fine flour tended to be softer and less fracturable.Textural properties were significantly correlated to protein stability to heat and retrogradation tendency of starch in biscuit dough as well as moisture content.

Analysis of buckwheat production in the world and Serbia

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Popović Vera

2014-01-01

Full Text Available During the period 2010-2011 about 2.113 million ha of buckwheat was sown annually worldwide. Average yield of buckwheat during the monitored period was 913 kg ha-1. Areas and average yield have a rising tendency. The most significant producers of buckwheat in the world are: China, Russia and Ukraine. In Serbia buckwheat is produced on small areas. The paper presents results of testing of four buckwheat varieties, produced on plots of the Institute for crops and vegetables as follows: Novosadska, Godijevo, Bamby and Češka. Analysis of average yield has shown that Novosadska variety produced statistically significant higher yield (2626 kg ha-1 compared to the other varieties tested (p <0.05. From the results shown we can see that buckwheat yield in Serbia is significantly higher compared with the world average yield, which tells us that this plant can be successfully produced in our agro-ecological conditions of growing.

Complete Chloroplast Genome Sequence of Tartary Buckwheat (Fagopyrum tataricum and Comparative Analysis with Common Buckwheat (F. esculentum.

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Kwang-Soo Cho

Full Text Available We report the chloroplast (cp genome sequence of tartary buckwheat (Fagopyrum tataricum obtained by next-generation sequencing technology and compared this with the previously reported common buckwheat (F. esculentum ssp. ancestrale cp genome. The cp genome of F. tataricum has a total sequence length of 159,272 bp, which is 327 bp shorter than the common buckwheat cp genome. The cp gene content, order, and orientation are similar to those of common buckwheat, but with some structural variation at tandem and palindromic repeat frequencies and junction areas. A total of seven InDels (around 100 bp were found within the intergenic sequences and the ycf1 gene. Copy number variation of the 21-bp tandem repeat varied in F. tataricum (four repeats and F. esculentum (one repeat, and the InDel of the ycf1 gene was 63 bp long. Nucleotide and amino acid have highly conserved coding sequence with about 98% homology and four genes--rpoC2, ycf3, accD, and clpP--have high synonymous (Ks value. PCR based InDel markers were applied to diverse genetic resources of F. tataricum and F. esculentum, and the amplicon size was identical to that expected in silico. Therefore, these InDel markers are informative biomarkers to practically distinguish raw or processed buckwheat products derived from F. tataricum and F. esculentum.

Antioxidant activity of wheat and buckwheat flours

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Sedej Ivana J.

2010-01-01

Full Text Available Antioxidative activities of wheat flours (type 500 and wholegrain and buckwheat flours (light and wholegrain were tested using 1,1-diphenyl-2-picrylhydrazyl (DPPH·-scavenging activity, reducing power and chelating activity on Fe2+. Also, the content of the total phenolics of ethanolic extracts was estimated. Polyphenolics content (expressed as gallic acid equivalent, GAE in wheat flours varied between 37.1 and 137.2 μg GAE/g extract, while its content in buckwheat flour were at least four time higher and ranged between 476.3 and 618.9μg GAE/g extract. Ethanolic extracts of buckwheat flours exhibited higher antioxidant activities in all the assays, except for chelating activity. Regarding all the obtained results, it can be concluded that bakery products produced with buckwheat flour could be regarded as potential functional foods.

484 Allergen Standardisation in Allergens and Allergoids—Challenges and Considerations

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Skinner, Murray; Bullimore, Alan; Hewings, Simon; Swan, Nicola

2012-01-01

Background The range of therapeutics and dosing schedules for allergen preparations and allergoids produced and used clinically are considerable. Standardisation of allergy immunotherapies is considered a positive step; however there are difficulties in identifying universal metrics for standardisation. Many advocate the use of major allergen content whilst others advocate total allergenicity. Additionally as a compounding argument, where major allergen is used, many disagree on what the major allergen is for certain species. Methods Major allergen content measurement allows a consistent recognised measure, and IgE responses of a serum pool are often dominated by IgE against major allergens. However issues such as specificity of different assays toward isoforms and other variants of single allergens often results in diverging allergen contents that can cause unexpected and misleading disparity. Other aspects that increase complication are the relevance to modified allergens, use of adjuvants and differing dosing regimes. Results The major allergen content of key products in different therapeutic formats has been measured. Conclusions This has been performed in conjunction with techniques such as total allergenicity, as allergy treatments and therapeutics require careful characterisation to allow supply of consistent, safe and efficacious products.

Characterization of Ras k 1 a novel major allergen in Indian mackerel and identification of parvalbumin as the major fish allergen in 33 Asia-Pacific fish species.

Science.gov (United States)

Ruethers, T; Raith, M; Sharp, M F; Koeberl, M; Stephen, J N; Nugraha, R; Le, T T K; Quirce, S; Nguyen, H X M; Kamath, S D; Mehr, S S; Campbell, D E; Bridges, C R; Taki, A C; Swoboda, I; Lopata, A L

2018-04-01

Fish is a well-recognized cause of food allergy and anaphylaxis. The evolutionary and taxonomic diversity of the various consumed fish species pose a challenge in the identification and characterization of the major fish allergens critical for reliable diagnostics. Globally, fish is a rising cause of food allergy complicated by a large under-investigated variety of species as well as increasing global tourism and trade. This is the first comprehensive study on allergen profiles of heat-processed fish from Vietnam. The aim of this study was to identify the major heat-stable allergens from frequently exported Asia-Pacific freshwater and marine fish and to characterize the major allergen parvalbumin (PV) from one of the most consumed and exported fish species from Asia, the Indian mackerel (Rastrelliger kanagurta). Heated protein extracts from 33 fish species were separated by gel electrophoresis. PV isoforms were identified by immunoblotting utilizing 3 different PV-specific monoclonal and polyclonal antibodies and further characterized by mass spectrometry. IgE reactivity was investigated using sera from 21 patients with confirmed fish allergy. Heat-stable IgE-reactive PVs, with up to 5 isoforms per species, were identified in all 33 analysed fish species. In the Indian mackerel, 7 PV isoforms were identified by 2D-gel electrophoresis combined with mass spectrometric analyses. The amino acid sequence deduced from cDNA of the most expressed isoform showed a high identity (>90%) to PVs from 2 other mackerel species. Different PVs were identified as the major heat-stable allergens in all 33 analysed freshwater and marine fish species from Vietnam, many of which are exported world-wide and 21 species that have never been investigated before. The Indian mackerel PV represents a novel fish allergen, now officially registered as Ras k 1. Improved diagnostics for fish allergy against Asia-Pacific species should be developed with focus on PV. © 2017 John Wiley & Sons Ltd.

Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo

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Mojtaba Sankian

2013-05-01

Full Text Available Background: Several studies reported the clinical features of IgE-mediated hypersensitivity after ingestion of melon. Melon allergy is a common IgE-mediated fruit allergy in Iran. This prompted us to investigate immunochemical and molecular properties of the major allergen in melon fruit, to compare the IgE-binding capacity of the natural protein with the recombinant allergen, and to determine cross-reactivity of the major allergen with closely-related allergens from other plants displaying clinical cross-reactivity with melon. Methods: Identification and molecular characterization of the major melon allergen were performed using IgE immunoblotting, allergen-specific ELISA, affinity-based purifications, cross-inhibition assays, cloning, and expression of the allergen in Escherichia coli. Results: Melon profilin was identified and isolated as a major IgE-binding component and designated as Cuc m 2. Sequencing corresponding cDNA revealed an open reading frame of 363 bp coding for 131 amino acid residues and two fragments of 171 bp and 383 bps for the 5’and 3’ UTRs, respectively. Significant cross-reactivity was found between melon profilin and Cynodon dactylon, tomato, peach, and grape profilins in cross-inhibition assays. Although the highest degree of amino acid identity was revealed with watermelon profilin, there was no significant cross-reactivity between melon and watermelon profilins. Conclusion: Melon profilin is the major IgE-binding component in melon extract, and the recombinant and natural forms exhibited similar IgE-binding capacities. A part of the fruit-fruit and pollen-fruit cross-reactions could be explained by the presence of this conserved protein; however, sequence homology provides insufficient information to predict IgE cross-reactivity of profilins.

Component resolution reveals additional major allergens in patients with honeybee venom allergy.

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Köhler, Julian; Blank, Simon; Müller, Sabine; Bantleon, Frank; Frick, Marcel; Huss-Marp, Johannes; Lidholm, Jonas; Spillner, Edzard; Jakob, Thilo

2014-05-01

Detection of IgE to recombinant Hymenoptera venom allergens has been suggested to improve the diagnostic precision in Hymenoptera venom allergy. However, the frequency of sensitization to the only available recombinant honeybee venom (HBV) allergen, rApi m 1, in patients with HBV allergy is limited, suggesting that additional HBV allergens might be of relevance. We performed an analysis of sensitization profiles of patients with HBV allergy to a panel of HBV allergens. Diagnosis of HBV allergy (n = 144) was based on history, skin test results, and allergen-specific IgE levels to HBV. IgE reactivity to 6 HBV allergens devoid of cross-reactive carbohydrate determinants (CCD) was analyzed by ImmunoCAP. IgE reactivity to rApi m 1, rApi m 2, rApi m 3, nApi m 4, rApi m 5, and rApi m 10 was detected in 72.2%, 47.9%, 50.0%, 22.9%, 58.3%, and 61.8% of the patients with HBV allergy, respectively. Positive results to at least 1 HBV allergen were detected in 94.4%. IgE reactivity to Api m 3, Api m 10, or both was detected in 68.0% and represented the only HBV allergen-specific IgE in 5% of the patients. Limited inhibition of IgE binding by therapeutic HBV and limited induction of Api m 3- and Api m 10-specific IgG4 in patients obtaining immunotherapy supports recent reports on the underrepresentation of these allergens in therapeutic HBV preparations. Analysis of a panel of CCD-free HBV allergens improved diagnostic sensitivity compared with use of rApi m 1 alone, identified additional major allergens, and revealed sensitizations to allergens that have been reported to be absent or underrepresented in therapeutic HBV preparations. Copyright © 2014 The Authors. Published by Mosby, Inc. All rights reserved.

Identification of parvalbumin and two new thermolabile major allergens of Thunnus tonggol using a proteomics approach.

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Rosmilah, Misnan; Shahnaz, Murad; Meinir, Jones; Masita, Arip; Noormalin, Abdullah; Jamaluddin, Mohamed

2013-01-01

The longtail tuna (Thunnus tonggol) is widely consumed in Asia. Parvalbumin, the main major allergen of fish, has been well identified in multiple fish species, yet little is known about the allergenic proteins in T. tonggol. Thus, the aim of this study was to characterize the major allergens of T. tonggol using a proteomics approach. Raw and boiled extracts of the fish were prepared. Fish proteins were separated by means of SDS-PAGE and two-dimensional (2-DE) electrophoresis. 1-DE immunoblotting of raw extract was performed with sera from fish-allergic patients. Ten sera were further analysed by 2-DE immunoblotting. Selected major allergenic protein spots were excised, trypsin digested and analysed by means of mass spectrometry. SDS-PAGE of raw extract revealed 26 protein fractions, while boiled extract demonstrated fewer bands. The 2-DE gel profile of the raw extract further fractionated the protein bands to more than 100 distinct protein spots. 1-DE immunoblotting of raw extract exhibited two thermolabile protein fractions of 42 and 51 kDa as the major allergens, while the boiled extract only revealed a single IgE-binding band at 151 kDa. 2-DE immunoblotting of raw extract further detected numerous major IgE-reactive spots of 11-13, 42 and 51 kDa. Mass spectrometry analysis of the peptides generated from the 12, 42 and 51 kDa digested spots indicated that these spots were parvalbumin, creatine kinase and enolase, respectively. In addition to parvalbumin, two new thermolabile allergens were identified as major allergenic proteins of T. tonggol. This study proved that both thermostable and thermolabile proteins are important in local tuna allergy and should be included in diagnostic strategies.

Molecular cloning, expression and characterization of Pru a 1, the major cherry allergen.

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Scheurer, S; Metzner, K; Haustein, D; Vieths, S

1997-06-01

A high percentage of birch pollen allergic patients experiences food hypersensitivity reactions after ingestion of several fruits and vegetables. Previous work demonstrated common epitopes on an allergen of Mr 18,000 from sweet cherry (Prunus avium) and Bet v 1, the major allergen from birch pollen. N-terminal amino acid sequencing showed a sequence identity of 67% with Bet v 1. Here we report the cloning and cDNA sequencing of this cherry allergen. The entire deduced amino acid sequence described a protein of Mr 17,700 with 59.1% identity to Bet v 1. High degrees of identity in the range of 40 to 60% were also found with related allergens from other kinds of tree pollen and plant foods as well as with stress-induced proteins from food plants such as parsley, potato and soya. The coding DNA of the cherry protein was cloned into vector pET-16b and expressed in E. coli strain BL21(DE3) as a His-tag fusion protein. As shown by SDS-PAGE, the apparent molecular masses of the nonfusion protein and the natural allergen were identical. The fusion protein showed high IgE binding potency when sera from patients allergic to cherry were tested by immunoblotting and enzyme allergosorbent tests. Moreover, it cross-reacted strongly with IgE specific for the natural counterpart and for Bet v 1. The high biological activity of the recombinant fusion protein was further confirmed by the induction of a strong histamine release in basophils from cherry-allergic patients. Since sera from 17/19 of such patients contained IgE against this allergen it was classified as a major allergen and named Pru a 1. Recombinant Pru a 1 mimics most of the allergenic potency of cherry extract and hence could be a useful tool for studying the molecular and immunological properties of pollen related food allergens.

Ribonuclease activity of buckwheat plant (Fagopyrum esculentum cultivars with different sensitivities to buckwheat burn virus

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Y. R. Sindarovska

2014-06-01

Full Text Available Ribonucleases (RNases are present in base-level amounts in intact plants, but this level is able to increase greatly under stress conditions. The possible cause for such an increase is protection against plant RNA-virus attack. Buckwheat burn virus (BBV is a highly virulent pathogen that belongs to Rhabdoviridae family. In our study, we have analyzed the correlation between RNase activity and resistance of different buckwheat cultivars to BBV infection. Two cultivars, Kara-Dag and Roksolana, with different sensitivities to BBV have been used. Kara-Dag is a cultivar with medium sensitivity to virus and Roksolana is a tolerant cultivar. It has been shown that the base level of RNase activity in Roksolana cultivar was in most cases higher than the corresponding parameter in Kara-Dag cultivar. Both infected and uninfected plants of Roksolana cultivar demonstrated high RNase activity during two weeks. Whereas infected plants of Kara-Dag cultivar demonstrated unstable levels of RNase activity. Significant decline in RNase activity was detected on the 7th day post infection with subsequent gradual increase in RNase activity. Decline of the RNase activity during the first week could promote the virus replication and therefore more successful infection of upper leaves of plants. Unstable levels of RNase activity in infected buckwheat plants may be explained by insufficiency of virus-resistant mechanisms that determines the medium sensitivity of the cultivar to BBV. Thus, plants of buckwheat cultivar having less sensitivity to virus, displayed in general higher RNase activity.

Sensory analysis and aroma compounds of buckwheat containing products-a review.

Science.gov (United States)

Starowicz, Małgorzata; Koutsidis, Georgios; Zieliński, Henryk

2017-07-07

Buckwheat is a rich source of starch, proteins, minerals and antioxidants, and as such has become a popular functional ingredient incorporated in diverse recipes/products with particular use in the gluten free market. Due to the absence of gluten, application of buckwheat or buckwheat derived ingredients in this particular food sector has increased significantly over recent years with many buckwheat-based products appearing globally. Sensory analysis is an integral part of the development of products that fulfill consumer expectations. Therefore, investigations on the incorporation of health promoting functional ingredients such as buckwheat into traditional recipes are often complemented by the evaluation of appearance, aroma, taste and texture as well as overall quality through standardized procedures involving trained judges or consumer panels. Aroma is of particular importance in driving consumer preference and its sensory assessment is often complemented with analytical workflows aiming to isolate and determine the concentration of volatile compounds in food and understand the effect of food components on the overall aroma intensity and/or perception of the final product. The present manuscript provides a review of recent advances and knowledge on the sensory characteristics, consumer preference and volatile compound analysis of buckwheat and buckwheat based products.

Buckwheat-enriched wheat bread: National market placement possibilities

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Sakač Marijana B.

2015-01-01

Full Text Available Quality parameters and the possibility of successful placement of buckwheat-enriched wheat bread on the national market are presented in this paper. Analysis of the market position of buckwheat-enriched wheat bread includes demands, offer and competition. Elements that affect the overall retail price of buckwheat-enriched wheat bread are given in details, along with SWOT analysis and marketing plan including target market, market supply and product marketing mix. According to all performed analyses it could be concluded that this product should be positioned on the national market, especially for people with special needs and requirements.

Plant guide: Parsnipflower buckwheat: Eriogonum heracleoides Nutt

Science.gov (United States)

Derek Tilley; Dan Ogle; Loren St. John

2007-01-01

Parsnipflower buckwheat is a perennial forb to subshrub with a branching woody stem. Leaves are covered with dense white hairs making the herbage appear a light green to blue-grayish color. The flowers are a creamy-yellow color and have six petals which are borne in simple or compound umbels. Plants of parsnipflower buckwheat can be distinguished from other closely...

Rational design of hypoallergens applied to the major cat allergen Fel d 1.

Science.gov (United States)

Saarne, T; Kaiser, L; Grönlund, H; Rasool, O; Gafvelin, G; van Hage-Hamsten, M

2005-05-01

Allergen-specific immunotherapy is the only treatment for allergic disease providing long-lasting symptom relief. Currently, it is mainly based on the use of crude allergen extracts. The treatment may be improved by the use of genetically engineered allergens, hypoallergens, aiming at a more effective and safer therapy. The aim of this study was to provide a rational design of hypoallergen candidates for immunotherapy by using structural information and knowledge of B and T cell epitopes of an allergen. The three-dimensional structure of the major cat allergen Fel d 1 was systematically altered by duplication of selected T cell epitopes and disruption of disulphide bonds. Seven Fel d 1 derivatives were generated and screened for allergenic reactivity in comparison with recombinant Fel d 1 in competition-ELISA. The allergenicity was further evaluated in basophil activation experiments and T cell reactivity was assessed in a lymphoproliferation assay. Three out of seven Fel d 1 derivatives, with two duplicated T cell epitopes and one or two disulphide bonds disrupted, were carefully evaluated. The three derivatives displayed a strong reduction in allergenicity with 400-900 times lower IgE-binding capacity than recombinant Fel d 1. In addition, they induced a lower degree of basophil activation and similar or stronger T cell proliferation than recombinant Fel d 1. By a rational approach, we have constructed three Fel d 1 hypoallergens with reduced IgE-binding capacities and retained T cell reactivities. This strategy may be applied to any well-characterized allergen to improve immunotherapy for allergic patients.

Identification of hazelnut major allergens in sensitive patients with positive double-blind, placebo-controlled food challenge results

DEFF Research Database (Denmark)

Pastorello, Elide A; Vieths, Stefan; Pravettoni, Valerio

2002-01-01

The hazelnut major allergens identified to date are an 18-kd protein homologous to Bet v 1 and a 14-kd allergen homologous to Bet v 2. No studies have reported hazelnut allergens recognized in patients with positive double-blind, placebo-controlled food challenge (DBPCFC) results or in patients...

Purification, characterization and allergenicity assessment of 26kDa protein, a major allergen from Cicer arietinum.

Science.gov (United States)

Verma, Alok Kumar; Sharma, Akanksha; Kumar, Sandeep; Gupta, Rinkesh Kumar; Kumar, Dinesh; Gupta, Kriti; Giridhar, B H; Das, Mukul; Dwivedi, Premendra D

2016-06-01

Chickpea (CP), a legume of the family Fabaceae, is an important nutrient-rich food providing protein, essential amino acids, vitamins, dietary fibre, and minerals. Unfortunately, several IgE-binding proteins in CP have been detected that are responsible for allergic manifestations in sensitized population. Therefore, the prevalence of CP induced allergy prompted us towards purification, characterization and allergenicity assessment of a major ∼26kDa protein from chickpea crude protein extract (CP-CPE). Purification of CP 26kDa protein was done using a combination of fractionation and anion exchange chromatography. This protein was further characterized as "Chain A, crystal structure of a plant albumin" from Cicer arietinum with Mol wt 25.8kDa by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Further, allergenic potential of purified 25.8kDa protein was assessed using in vivo and in vitro model. Purified protein showed IgE-binding capacity with sensitized BALB/c mice and CP allergic patient's sera. Enhanced levels of specific and total IgE, MCP-1, MCPT-1, myeloperoxidase, histamine, prostaglandin D2, and cysteinyl leukotriene were found in sera of mice treated with CP ∼26kDa protein. Further, expressions of Th2 cytokines (i.e. IL-4, IL-5, IL-13), transcription factors (i.e. GATA-3, STAT-6, SOCS-3) and mast cell signaling proteins (Lyn, cFgr, Syk, PLC-γ2, PI-3K, PKC) were also found increased at mRNA and protein levels in the intestines of mice treated with CP ∼26kDa protein. In addition, enhanced release of β-hexosaminidase, histamine, cysteinyl leukotriene and prostaglandin D2 were observed in RBL2H3 cell line when treated (125μg) with CP 26kDa protein. Conclusively, in vivo and in vitro studies revealed the allergenic potential of purified CP 26kDa protein. Being a potential allergen, plant albumin may play a pivotal role in CP induced allergenicity. Current study will be helpful for better development of therapeutic approaches to

Purification and crystallization of Cor a 9, a major hazelnut allergen

International Nuclear Information System (INIS)

Guo, Feng; Kothary, Mahendra H.; Wang, Yang; Yu, Xiaoping; Howard, Andrew J.; Fu, Tong-Jen; Zhang, Yu-Zhu

2008-01-01

The major hazelnut allergen Cor a 9 was purified from the natural source and crystallized. Diffraction data were collected to 1.9 Å resolution using a synchrotron-radiation source. Hazelnut (Corylus avellana) is one of the food sources that induce allergic reaction in a subpopulation of people with food allergy. The 11S legumin-like seed-storage protein from hazelnut has been identified as one of the major hazelnut allergens and named Cor a 9. In this study, Cor a 9 was extracted from hazelnut kernels using a high-salt solution and was purified by desalting out and FPLC to a highly purified state. Diffraction-quality single crystals were obtained using the hanging-drop vapour-diffusion method. Diffraction data were collected and a structure solution has been obtained by molecular-replacement calculations. Further refinement of the structure is currently in progress

Identification of major rice allergen and their clinical significance in children

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You Hoon Jeon

2011-10-01

Full Text Available Purpose : Recently, an increase in the number of patients sensitized to rice allergen with or without clinical symptoms has been reported. This study was designed to determine the major allergens in rice and their clinical significance. Methods : Twenty-four children (15 boys and 9 girls; mean age, 16.3 months with allergic disease, who were sensitized to rice antigen (by UniCAP in the Pediatric Allergy Respiratory Center at Soonchunhyang University Hospital, were enrolled in this study. The allergenicity of various types of rice (raw, cooked, and heat-treated, simulated gastric fluid [SGF], and simulated intestinal fluid [SIF] was investigated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE and immunoglobulin E (IgE immunoblots. The patients’ medical records, including laboratory data and allergy symptoms after ingestion of rice were reviewed. Results : Patients were sensitized to an average of 13.5 food antigens and their mean total IgE was 6,888.7 kU/L. In SDS-PAGE, more than 16 protein bands were observed in the raw rice, whereas only 14-16 kDa and 31-35 kDa protein bands were observed in cooked rice. The common SDS-PAGE protein bands observed in SGF-, SIF-, and heattreated rice were 9, 14, and 31 kDa. In a heated-rice IgE immunoblot, protein bands of 9, 14, and 31-33 kDa were found in 27.8%, 38.9%, and 38.9% of all sera, respectively, and in 50%, 50%, and 75%, of ser a from the 4 symptomatic patients, respectively. Conclusion : The 9-, 14-, and 31-kDa protein bands appeared to be the major allergens responsible for rice allergy symptoms.

Physicochemical and functional properties of gamma irradiated buckwheat and potato starch

Science.gov (United States)

Verma, Ruchi; Jan, Shumaila; Rani, Savita; Jan, Kulsum; Swer, Tanya L.; Prakash, Kumar S.; Dar, M. Z.; Bashir, Khalid

2018-03-01

Starches isolated from buckwheat and potato were subject to different doses of irradiation at 0, 5, 10, 15 and 20 kGy. Native and irradiated starch samples were evaluated for their physicochemical and functional properties to assess the effect of gamma irradiation. Apparent amylose content decreased significantly from 26.84% to 22.12% and 27.01 to 16.11% for buckwheat and potato starch respectively as the dose increased. A significant decrease was observed in pH, swelling power and syneresis as the dose increased for both buckwheat and potato starch. pH decreased from 5.20 to 3.81 and 5.81 to 3.95 for buckwheat and potato starch, respectively. Carboxyl content, freeze thaw stability, water and oil absorption capacity and transmittance showed increasing trend with increasing irradiation dose. Carboxyl content increased from 0% to 0.23% and 0-0.22% for buckwheat and potato starch, respectively.

Allergenicity assessment of apple cultivars: hurdles in quantifying labile fruit allergens

NARCIS (Netherlands)

Zuidmeer, L.; van Leeuwen, W. A.; Kleine Budde, I.; Breiteneder, H.; Ma, Y.; Mills, C.; Sancho, A. I.; Meulenbroek, E. J.; van de Weg, E.; Gilissen, L.; Ferreira, F.; Hoffmann-Sommergruber, K.; van Ree, R.

2006-01-01

BACKGROUND: Assessment of allergenicity of foods is important for allergic consumers and regulators. Immunoassays to measure major food allergens are widely applied, often giving variable results. Using the major apple allergen Mal d 1 as a model, we aimed to establish at the molecular level why

Antioxidant properties of buckwheat flours and their contribution to functionality of bakery, pasta and confectionary products

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Sakač Marijana B.

2015-01-01

Full Text Available Buckwheat is grown primarily because of its grain which, after undergoing the processes of dehulling, grinding and sieving, is used to produce buckwheat flour which is characterized by a considerable content of antioxidants, especially polyphenols and tocopherols. Buckwheat polyphenols are represented by phenolic acids and flavonoids, mainly rutin, a proven potent antioxidant. The content of polyphenols and tocopherols in buckwheat grain primarily depends on the buckwheat species, growing area, climate and growing conditions. Common buckwheat (Fagopyrum esculentum Möench, which is often used for the production of light and wholegrain buckwheat flour, contains less polyphenols than tartary buckwheat. The content of polyphenols in common buckwheat grain varies depending on the grain part. As their largest amount is contained in the hull and the outer layers of the grain, the wholegrain buckwheat flour is superior in polyphenols than the light buckwheat flour. Therefore, the wholegrain buckwheat flour is characterized by a higher antioxidant capacity. Polyphenols in buckwheat flour exist in free and bound forms, where the contribution of free polyphenols ranges between 48-64%. Due to a relatively high content of antioxidants in light and wholegrain buckwheat flour, they are used for substitution of wheat or other cereal flours in bakery, pasta and confectionary formulations in order to create either added value or gluten-free products. The aim of a long-term consumption of buckwheat flours is to achieve health benefits and protect from many chronic diseases. Technological procedures and some treatments used during the food preparation influence polyphenol composition and content and consequently the functionality of food. Therefore, in order to minimize polyphenol losses and preserve the antioxidant capacity of the final products it is necessary to understand the thermal treatments and their mechanisms. [Projekat Ministarstva nauke Republike

Yield and Quality Features of Buckwheat-Soybean Mixtures in Organic Agricultural Conditions

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Mustafa Sürmen

2017-12-01

Full Text Available This study was carried out during the summer of 2014 to determine alternative quality forage sources that could be grown in the Aydın ecological conditions. In the study, effects of 3 different mixtures and 2 pure species (100% Buckwheat, 25% Buckwheat -75% Soybean, 50% Buckwheat -5 0% Soybean, 75% Buckwheat - 25% Soybean, 100% Soybean and 2 different harvesting times (50%-100% flowering/buckwheat on yield and quality characteristics were investigated. According to measurements, the highest average herbage yield was obtained from 75% Buckwheat-25% Soybean application (3100 kg/da at 2nd harvest time. When the average of hay yield was examined, the results were similar to herbage yield. When ADF and NDF were examined, the highest values were seen at the 2nd harvest. When the crude protein ratios were examined, it was found that they decreased at the 2nd harvesting time and the highest value was determined at 100% soybean application at the 1st harvesting time (21.08%. When Digestible Dry Matter (DDM and Relative Feed Value (RFV were examined, the highest values were determined in 100%Soybean applications at first harvest time and when the mixture applications were examined, the highest values were determined to be 75% Buckwheat - 25% Soybean application. As a result of the study, it was determined that the yields obtained at the 2nd harvest time were higher but the forage quality decreased. When the mixtures were examined, it was determined that the mixture having the highest roughage value was 75% buckwheat + 25% soybean application. In this study, the buckwheat which have short vegetation and good quality and the soybean, which is infront of with high quality, mixtures were examined. It has also been found that these mixtures may be important for obtaining high-quality forage in the short and intermediate periods.

A proteomic study of the major allergens from yellow jacket venoms.

Science.gov (United States)

Kolarich, Daniel; Loos, Andreas; Léonard, Renaud; Mach, Lukas; Marzban, Gorji; Hemmer, Wolfgang; Altmann, Friedrich

2007-05-01

The venoms of stinging insects belong to the most dangerous allergen sources and can cause fatal anaphylactic reactions. Reliable prediction of a patient's risk to anaphylactic reactions is vital, and diagnosis requires the knowledge of the relevant allergens. Recently, a new hyaluronidase -like glycoprotein from Vespula vulgaris (Ves v 2b) was identified. This led us to investigate hyaluronidases and also other major allergens from V. germanica and four additional Vespula species. By MALDI-Q-TOF-MS, the new hyaluronidase-like protein was shown to be the major component of the 43-kDa band in all Vespula species studied. LC-ESI-Q-TOF-MS/MS sequencing of Ves g 2a and Ves g 2b facilitated the cloning of their cDNA. Ves v 2b and Ves g 2b turned out to be essentially identical on protein level. Whereas the less abundant "a" form displayed enzymatic activity, the new "b" homologue did not. This is probably caused by amino acid exchanges in the active site, and it raises questions about the physiological role of this protein. Sequence comparisons by MS/MS of antigen 5 and phospholipases from V. vulgaris, germanica, maculifrons, pensylvanica, flavopilosa and squamosa revealed the latter as a taxonomic outlier and led to the discovery of several not previously reported amino acid differences.

Use of gamma irradiation for microbial inactivation of buckwheat flour and products, 8

International Nuclear Information System (INIS)

Muramatu, Nobuyuki; Ohinata, Hiroshi; Karasawa, Hideyuki; Oike, Terutake; Ito, Hitoshi; Ishigaki, Isao.

1991-01-01

Effects of irradiation at 3.0-7.0 kGy with 2 MeV electron beams were investigated on the number of microorganisms and quality of buckwheat flour. Electron beams and gamma-rays were compared in terms of the effects on the quality of buckwheat flour. The results were as follows. (1) Electron beams at 3 kGy reduced the number of microorganisms almost to the same level as gamma-rays. Oxygen content in buckwheat flour had no effect on inactivation of microorganisms by irradiation with electron beams and gamma-rays. (2) Peroxide-value (POV) of lipid in buckwheat flour increased with absorbed dose of gamma-rays and electron beams. The increase of POV was suppressed by the usage of oxygen absorber. The color change of buckwheat flour was suppressed by the usage of oxygen absorber as well. Acid-value (AV) of lipid in buckwheat flour was not changed by irradiation at high dose with gamma-rays or electron beams. (3) Maximum torque in Farinograph test of dough prepared from irradiated buckwheat flour decreased with increase of absorbed dose of electron beams. However, oxygen absorber suppressed the change of these properties induced by irradiation. (4) The usage of oxygen absorber resulted in a high sensory score of noodles from irradiated buckwheat flour with small changes of color, flavor and texture. (author)

Antioxidant and antidiabetic properties of tartary buckwheat rice flavonoids after in vitro digestion*

Science.gov (United States)

Bao, Tao; Wang, Ye; Li, Yu-ting; Gowd, Vemana; Niu, Xin-he; Yang, Hai-ying; Chen, Li-shui; Chen, Wei; Sun, Chong-de

2016-01-01

Oxidative stress and diabetes have a tendency to alter protein, lipid, and DNA moieties. One of the strategic methods used to reduce diabetes-associated oxidative stress is to inhibit the carbohydrate-digesting enzymes, thereby decreasing gastrointestinal glucose production. Plant-derived natural antioxidant molecules are considered a therapeutic tool in the treatment of oxidative stress and diabetes. The objective of this study was to identify tartary buckwheat rice flavonoids and evaluate the effect of in vitro digestion on their antioxidant and antidiabetic properties. High performance liquid chromatography (HPLC) analysis indicated the presence of rutin as a major component and quercitrin as a minor component of both digested and non-digested flavonoids. Both extracts showed a significant antioxidant capacity, but digested flavonoids showed reduced activity compared to non-digested. There were some decreases of the antioxidant activities (2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2-diphenyl-1-picrylhydrazy (DPPH) radical, and ferric reducing antioxidant power (FRAP)) of digested tartary buckwheat rice flavonoids compared with non-digested. Flavonoids from both groups significantly inhibited reactive oxygen species (ROS) production and α-glucosidase activity. Both digested and non-digested flavonoids markedly increased glucose consumption and glycogen content in HepG2 cells. Tartary buckwheat rice flavonoids showed appreciable antioxidant and antidiabetic properties, even after digestion. Tartary buckwheat rice appears to be a promising functional food with potent antioxidant and antidiabetic properties. PMID:27921399

Antioxidant and antidiabetic properties of tartary buckwheat rice flavonoids after in vitro digestion.

Science.gov (United States)

Bao, Tao; Wang, Ye; Li, Yu-Ting; Gowd, Vemana; Niu, Xin-He; Yang, Hai-Ying; Chen, Li-Shui; Chen, Wei; Sun, Chong-de

Oxidative stress and diabetes have a tendency to alter protein, lipid, and DNA moieties. One of the strategic methods used to reduce diabetes-associated oxidative stress is to inhibit the carbohydrate-digesting enzymes, thereby decreasing gastrointestinal glucose production. Plant-derived natural antioxidant molecules are considered a therapeutic tool in the treatment of oxidative stress and diabetes. The objective of this study was to identify tartary buckwheat rice flavonoids and evaluate the effect of in vitro digestion on their antioxidant and antidiabetic properties. High performance liquid chromatography (HPLC) analysis indicated the presence of rutin as a major component and quercitrin as a minor component of both digested and non-digested flavonoids. Both extracts showed a significant antioxidant capacity, but digested flavonoids showed reduced activity compared to non-digested. There were some decreases of the antioxidant activities (2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2-diphenyl-1-picrylhydrazy (DPPH) radical, and ferric reducing antioxidant power (FRAP)) of digested tartary buckwheat rice flavonoids compared with non-digested. Flavonoids from both groups significantly inhibited reactive oxygen species (ROS) production and α-glucosidase activity. Both digested and non-digested flavonoids markedly increased glucose consumption and glycogen content in HepG2 cells. Tartary buckwheat rice flavonoids showed appreciable antioxidant and antidiabetic properties, even after digestion. Tartary buckwheat rice appears to be a promising functional food with potent antioxidant and antidiabetic properties.

An Overview: Distribution, Production, and Diversity of Local Landraces of Buckwheat in Nepal

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Dol Raj Luitel

2017-01-01

Full Text Available Buckwheat is a sixth staple food crop after rice, wheat, maize, finger millet, and barley in Nepal. It is considered as an alternate cereal and poor man’s crop, representing an important food supply in remote places of Himalayas. It is the best crop in higher altitude in terms of adaptation to different climatic variables and easily fitted to different cropping patterns due to short duration. It is cultivated on marginal land in 61 out of 75 districts of Nepal from some 60 m to 4500 m asl, especially hilly and mountain districts like Rukum, Rolpa, Jajarkot, Dolpa, Humla, Jumla, Kalikot, Kavre, Dolakha, and Okhaldhunga. Sweet buckwheat varieties are generally grown in midhill and Terai but Tartary buckwheat varieties are grown in higher altitude. There are altogether 19 local landraces of sweat buckwheat and 37 for Tartary buckwheat listed from Nepal. The largest producers are China, USA, and Russia and Japan is principal user of global buckwheat grown in the world. In Nepal, it is cultivated in 10510 ha area with production of 10355 t/yr and yield of 0.983 t/ha. It has also medicinal value used in different forms including all its parts so the demand of buckwheat is increasing.

Hypolipidemic and antioxidant effects of buckwheat leaf and flower mixture in hyperlipidemic rats

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Maja Đurendić - Brenesel

2013-05-01

Full Text Available As a source of biologically active compounds, buckwheat has beneficial effects in nutrition due to its high content of flavonoids, particularly rutin. Aim of our study was to examine effects of buckwheat on plasma lipid status and phospholipids fatty acids composition, histological and parameters of oxidative stress in Wistar rats fed a high-fat diet. This study showed that buckwheat leaf and flower (BLF mixture supplementation significantly reduce weight gain, plasma lipid concentrations and atherogenic index in rats fed a high-fat diet. Treatment of the high-fat group of animals with buckwheat significantly increased percentage of n-6 fatty acids as well as eicosapentaenoic acid (EPA and decreased percentage of saturated fatty acids (SFA and oleic acid. Buckwheat antioxidant effects diminished negative influence of high-fat diet in hyperlipidemic rats, while pathohistological analysis of liver confirmed changes after high-fat consumption. Our results showed hypolipidemic, antiatherogenic and antioxidative features of buckwheat leaf and flower mixture, and these parts of the plant with the highest rutin content could be beneficial in prevention and curing of hyperlipidemia.

Lol p XI, a new major grass pollen allergen, is a member of a family of soybean trypsin inhibitor-related proteins.

Science.gov (United States)

van Ree, R; Hoffman, D R; van Dijk, W; Brodard, V; Mahieu, K; Koeleman, C A; Grande, M; van Leeuwen, W A; Aalberse, R C

1995-05-01

Monoclonal antibodies were obtained against an unknown allergen from Lolium perenne grass pollen. The allergen had an apparent molecular mass of 18 kd on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Earlier immunoblotting studies had shown that carbohydrate-specific IgG antibodies recognize an antigen of similar size. We sought to characterize the allergen biochemically and immunologically. The amino acid sequence of the allergen was determined by automated Edman degradation, and its monosaccharide composition was determined by gas chromatographic analysis. A panel of 270 grass pollen-positive sera was assessed in a RAST with the purified allergen. Protease digestion (proteinase K) and chemical deglycosylation (trifluoromethane sulfonic acid) were used to distinguish between carbohydrate and peptide epitopes for IgE antibodies. The allergen was shown to be a glycoprotein with a molecular mass of 16 kd, of which 8% is carbohydrate. Its amino acid sequence shares 32% homology with soybean trypsin inhibitor (Kunitz) but lacks its active site. No homology was found with known grass pollen allergens, hence it was designated Lol p XI. A high degree of homology (44%) was found with a tree pollen allergen, Ole e I, the major allergen of olive pollen. More than 65% of grass pollen-positive sera had IgE against Lol p XI. IgE reactivity was demonstrated both with the carbohydrate moiety and the peptide backbone. Lol p XI is a new major grass pollen allergen carrying an IgE-binding carbohydrate determinant. Lol p XI is structurally related to the major allergen from olive pollen.

The effect of baking temperature and buckwheat flour addition on the selected properties of wheat bread

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A. Selimović

2014-01-01

Full Text Available Wholegrain buckwheat flour was used to substitute 15 %, 30 % and 40 % of wheat flour to make buckwheat enriched wheat bread. Proximate composition, sensory evaluation, total phenols content and antioxidant activity of buckwheat enriched wheat breads were analysed and compared with wheat bread. Wholegrain buckwheat flour contained higher total phenols than wheat flour. The incorporation of buckwheat flour from 15 % to 40 % in bread samples increased the total phenols content from 0.25 (mg GA/g d.m. sample to 0.65 (mg GA/g d.m. sample, and antioxidant activity from 208.45 (µmol Fe2+/L extract to 354.45 (µmol Fe2+/L extract. Total phenols content decreased during the baking process, while the antioxidant activity increased. Bread samples with 15 %, 30 % and 40 % of wholegrain buckwheat flour showed lower lightness (L and whiteness index (WI values of crust and crumb colour compared to the wheat bread. Addition of buckwheat flour increased redness (a and yellowness (b colour values for crumb. Sensory results indicating that three breads with buckwheat flour were moderately acceptable. No differences were found in overall sensory attributes between buckwheat flour enriched bread samples with 15 % and wheat bread (control sample.

Food allergen digestibility: The influence on allergenicity

DEFF Research Database (Denmark)

Bøgh, Katrine Lindholm; Madsen, Charlotte Bernhard

potential exist. Resistance to digestion is therefore a test parameter included in the safety assessment of the allergenic potential of novel proteins in genetically modified foods. In recent years, the association between resistance to digestion and allergenic potential has been challenged. When reviewing......Food allergy is a major health problem in the Western countries, affecting 3-8% of the population. What makes a dietary protein a food allergen has not yet been established, though several characteristics have been proposed to be shared by the food allergens. One of the features believed...... existing data from digestibility studies on known food allergens, it becomes evident that food allergens do not necessarily resist digestion. However, the choice of assay conditions, the method used for detection of residual intact protein as well as fragments hereof greatly influences the outcome. Studies...

Component-resolved evaluation of the content of major allergens in therapeutic extracts for specific immunotherapy of honeybee venom allergy

DEFF Research Database (Denmark)

Blank, Simon; Etzold, Stefanie; Darsow, Ulf

2017-01-01

Allergen-specific immunotherapy is the only curative treatment of honeybee venom (HBV) allergy, which is able to protect against further anaphylactic sting reactions. Recent analyses on a molecular level have demonstrated that HBV represents a complex allergen source that contains more relevant...... major allergens than formerly anticipated. Moreover, allergic patients show very diverse sensitization profiles with the different allergens. HBV-specific immunotherapy is conducted with HBV extracts which are derived from pure venom. The allergen content of these therapeutic extracts might differ due...... to natural variations of the source material or different down-stream processing strategies of the manufacturers. Since variations of the allergen content of therapeutic HBV extracts might be associated with therapeutic failure, we adressed the component-resolved allergen composition of different therapeutic...

Molecular basis of IgE-recognition of Lol p 5, a major allergen of rye-grass pollen.

Science.gov (United States)

Suphioglu, C; Blaher, B; Rolland, J M; McCluskey, J; Schäppi, G; Kenrick, J; Singh, M B; Knox, R B

1998-04-01

Grass pollen, especially of rye-grass (Lolium perenne). represents an important cause of type I allergy. Identification of IgE-binding (allergenic) epitopes of major grass pollen allergens is essential for understanding the molecular basis of interaction between allergens and human IgE antibodies and therefore facilitates the devising of safer and more effective diagnostic and immunotherapy reagents. The aim of this study was to identify the allergenic epitopes of Lol p 5, a major allergen of rye-grass pollen, immunodissect these epitopes further so that the amino acid residues critical for antibody binding can be determined and investigate the conservation and nature of these epitopes within the context of the natural grass pollen allergens. Peptides, 12-13 amino acid residues long and overlapping each other by 4 amino acid residues, based on the entire deduced amino acid sequence of the coding region of Lol p 5, were synthesised and assayed for IgE-binding. Two strong IgE-binding epitopes (Lol p 5 (49-60) and (265-276), referred to as peptides 7 and 34, respectively) were identified. These epitopes were further resolved by truncated peptides and amino acid replacement studies and the amino acid residues critical for IgE-binding determined (Lol p 5 (49-60) residue Lys57 and (265-276) residue Lys275). Sequences of these epitopes were conserved in related allergens and may form the conserved allergenic domains responsible for the cross-reactivity observed between pollen allergens of taxonomically related grasses. Furthermore, due to its strong IgE-reactivity, synthetic peptide Lol p 5 (265-276) was used to affinity-purify specific IgE antibodies which recognised proteins of other clinically important grass pollens. further indicating presence of allergenic cross-reactivity at the level of allergenic epitope. Moreover, Lol p 5 (265 276) demonstrated a strong capacity to inhibit IgE-binding to natural rye-grass pollen proteins highlighting the antibody accessibility

Major grass pollen allergen Lol p 1 binds to diesel exhaust particles: implications for asthma and air pollution.

Science.gov (United States)

Knox, R B; Suphioglu, C; Taylor, P; Desai, R; Watson, H C; Peng, J L; Bursill, L A

1997-03-01

Grass pollen allergens are known to be present in the atmosphere in a range of particle sizes from whole pollen grains (approx. 20 to 55 microns in diameter) to smaller size fractions Lol p 1, immunogold labelling with specific monoclonal antibodies and a high voltage transmission electron-microscopic imaging technique. DECP are visualized as small carbon spheres, each 30-60 nm in diameter, forming fractal aggregates about 1-2 microns in diameter. Here we test our hypothesis and show by in vitro experiments that the major grass pollen allergen, Lol p 1, binds to one defined class of fine particles, DECP. DECP are in the respirable size range, can bind to the major grass pollen allergen Lol p 1 under in vitro conditions and represent a possible mechanism by which allergens can become concentrated in polluted air and thus trigger attacks of asthma.

Distribution of Radioactive Cesium during Milling and Cooking of Contaminated Buckwheat.

Science.gov (United States)

Hachinohe, Mayumi; Nihei, Naoto; Kawamoto, Shinichi; Hamamatsu, Shioka

2018-06-01

To clarify the behavior of radioactive cesium (Cs) in buckwheat grains during milling and cooking processes, parameters such as processing factor (Pf) and food processing retention factor (Fr) were evaluated in two lots of buckwheat grains, R1 and R2, with different concentrations of radioactive Cs. Three milling fractions, the husk, bran, and flour fractions, were obtained using a mill and electric sieve. The radioactive Cs ( 134 Cs + 137 Cs) concentrations in husk and bran were higher than that in grain, whereas the concentration in flour was lower than that in grain. Pf values for the flours of R1 and R2 were 0.60 and 0.80, respectively. Fr values for the flours of R1 and R2 were 0.28 and 0.53, respectively. Raw buckwheat noodles (soba) were prepared using a mixture of buckwheat flour and wheat flour according to the typical recipe and were cooked with boiling water for 0.5, 1, and 2 min, followed by rinsing with water. Pf values for the soba boiled for 2 min (optimal for eating) made with R1 and R2 were 0.34 and 0.40, respectively. Fr values for these R1 and R2 samples were 0.55 and 0.66, respectively. Pf and Fr values for soba boiled for different times for both R1 and R2 were less than 0.6 and 0.8, respectively. Thus, buckwheat flour and its product, soba, cooked by boiling, are considered acceptable for human consumption according to the standard limit for radioactive Cs in buckwheat grains.

Identification of IgE-binding proteins from Lepidoglyphus destructor and production of monoclonal antibodies to a major allergen.

Science.gov (United States)

Ventas, P; Carreira, J; Polo, F

1991-08-01

The allergen composition of one of the most important storage mites, Lepidoglyphus destructor, has been studied by immunodetection after SDS-PAGE with individual patient sera. An allergenic polypeptide of 14 kDa was identified with 95% of the sera. This major allergen was isolated in the supernatant of 60% ammonium sulfate salt precipitation of the whole extract, which was subsequently used to immunize BALB/c mice so as to produce monoclonal antibodies. Four mAbs recognizing molecules with IgE-binding ability were obtained. The specificity of the mAbs was assayed against different allergenic extracts, and the molecules recognized by them were characterized by immunoblotting. Two mAbs (Le5B5 and Le9E4) were directed to the 14-kDa allergen; the other two to several proteins of lesser allergenic significance.

Growth rate, carcass characteristics and meat quality of growing lambs fed buckwheat or maize silage

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Gurhan Keles

2018-04-01

Full Text Available Objective This study evaluated inclusion of buckwheat silage to the diet of growing lambs in terms of meat quality as compared to maize silage. Methods Buckwheat, rich in total phenols (TP, 33 g/kg dry matter [DM], was harvested at the end of the milk stage and ensiled in 40 kg plastic bags after wilting (294 g/kg silage DM. A total of 18 growing lambs (21.6±1.2 were individually fed isonitrogenous and isoenergetic total mixed rations (TMR for 75 d that either contained buckwheat or maize silage at DM proportions of 0.50. At the end of feeding trail all lambs were slaughtered to assess carcass characteristics and meat quality. Results Buckwheat silage increased (p0.05 on live weight gain and feed efficiency. Carcass weight, dressing percentage, meat pH, water holding capacity, cooking loss, shear force (kg/cm2, and total viable bacteria count of meat did not differ (p>0.05 between the treatments. However, TP content of meat increased (p<0.001 by feeding buckwheat TMR. Feeding buckwheat TMR also decreased (p<0.05 the b* values of meat. Conclusion The results provide that buckwheat silage is palatable and could successfully include TMR of growing lambs with no adverse effects on performance, carcass and meat quality. Additionally, feeding buckwheat silage to lambs offers increased TP in meat.

Crystal structure of prunin-1, a major component of the almond (Prunus dulcis) allergen amandin.

Science.gov (United States)

Jin, Tengchuan; Albillos, Silvia M; Guo, Feng; Howard, Andrew; Fu, Tong-Jen; Kothary, Mahendra H; Zhang, Yu-Zhu

2009-09-23

Seed storage proteins are accumulated during seed development and act as a reserve of nutrition for seed germination and young sprout growth. Plant seeds play an important role in human nutrition by providing a relatively inexpensive source of protein. However, many plant foods contain allergenic proteins, and the number of people suffering from food allergies has increased rapidly in recent years. The 11S globulins are the most widespread seed storage proteins, present in monocotyledonous and dicotyledonous seeds as well as in gymnosperms (conifers) and other spermatophytes. This family of proteins accounts for a number of known major food allergens. They are of interest to both the public and industry due to food safety concerns. Because of the interests in the structural basis of the allergenicity of food allergens, we sought to determine the crystal structure of Pru1, the major component of the 11 S storage protein from almonds. The structure was refined to 2.4 A, and the R/Rfree for the final refined structure is 17.2/22.9. Pru1 is a hexamer made of two trimers. Most of the back-to-back trimer-trimer association was contributed by monomer-monomer interactions. An alpha helix (helix 6) at the C-terminal end of the acidic domain of one of the interacting monomers lies at the cleft of the two protomers. The residues in this helix correspond to a flexible region in the peanut allergen Ara h 3 that encompasses a previously defined linear IgE epitope.

Crystal Structure of Prunin-1, a Major Component of the Almond (Prunus dulcis) Allergen Amandin

Energy Technology Data Exchange (ETDEWEB)

Jin, Tengchuan; Albillos, Silvia M.; Guo, Feng; Howard, Andrew; Fu, Tong-Jen; Kothary, Mahendra H.; Zhang, Yu-Zhu; (IIT); (US-FDA); (IT)

2010-10-28

Seed storage proteins are accumulated during seed development and act as a reserve of nutrition for seed germination and young sprout growth. Plant seeds play an important role in human nutrition by providing a relatively inexpensive source of protein. However, many plant foods contain allergenic proteins, and the number of people suffering from food allergies has increased rapidly in recent years. The 11S globulins are the most widespread seed storage proteins, present in monocotyledonous and dicotyledonous seeds as well as in gymnosperms (conifers) and other spermatophytes. This family of proteins accounts for a number of known major food allergens. They are of interest to both the public and industry due to food safety concerns. Because of the interests in the structural basis of the allergenicity of food allergens, we sought to determine the crystal structure of Pru1, the major component of the 11 S storage protein from almonds. The structure was refined to 2.4 {angstrom}, and the R/Rfree for the final refined structure is 17.2/22.9. Pru1 is a hexamer made of two trimers. Most of the back-to-back trimer-trimer association was contributed by monomer-monomer interactions. An {alpha} helix (helix 6) at the C-terminal end of the acidic domain of one of the interacting monomers lies at the cleft of the two protomers. The residues in this helix correspond to a flexible region in the peanut allergen Ara h 3 that encompasses a previously defined linear IgE epitope.

Characterization of cookies made from wheat flour blended with buckwheat flour and effect on antioxidant properties.

Science.gov (United States)

Jan, Ulfat; Gani, Adil; Ahmad, Mudasir; Shah, Umar; Baba, Waqas N; Masoodi, F A; Maqsood, Sajid; Gani, Asir; Wani, Idress Ahmed; Wani, S M

2015-10-01

Buckwheat flour was incorporated into wheat flour at different levels (0, 20, 40, 60, 80, and 100 %) and the physicochemical, functional and antioxidant properties of the blended flour were studied. This study also investigated the effect of buckwheat on the retention of antioxidant properties of cookies during baking. The results showed significant variation in physicochemical and functional properties of the blended flour. The addition of buckwheat flour into wheat flour also increased the antioxidant properties of blended flour proportionally, but metal chelating properties decreased. The incorporation of buckwheat in wheat flour helped in better retention of antioxidant potential of cookies during baking process as buckwheat cookies (100 % buckwheat) showed greater percentage increase in antioxidant properties than control (100 % wheat). Quality characteristics of cookies such as hardness and spread ratio decreased, while as non-enzymatic browning (NEB) increased significantly with increase in the proportion of buckwheat flour in wheat flour. The Overall acceptability of cookies by sensory analysis was highest at 40 % level of blending. This study concluded that addition of buckwheat in wheat flour, may not only improve the physico-chemical and functional properties of the blended flour but may also enhance the nutraceutical potential of the product prepared from it.

MALDI-TOF MS analysis of labile Lolium perenne major allergens in mixes.

Science.gov (United States)

Irañeta, S G; Acosta, D M; Duran, R; Apicella, C; Orlando, U D; Seoane, M A; Alonso, A; Duschak, V G

2008-08-01

It is well known that allergen extracts used for specific therapy of allergic disorders are commonly stored as mixtures, causing an alteration of its stability. The aim of this report is to identify pollen allergens susceptible to degradation during storage of mixtures containing different sources of proteases in the absence of glycerol as a preserving agent. Mixes containing Lolium perenne (Lol p) pollen extract with either Aspergillus fumigatus or Periplaneta americana extracts were prepared and co-incubated for 90 days at 4 degrees C. Samples were taken off at fixed times and comparatively tested by in vitro and in vivo assays with atopic patients. Selected pollinic allergens were subjected to MALDI-TOF MS analysis. ELISA inhibition evidenced the loss of potency from ryegrass extract, and immunoblotting assays showed the degradation of specific pollinic allergens during storage of mixtures containing protease-rich sources. An in vivo intradermal skin assay confirmed the gradual loss of the biological activity of L. perenne pollen extract co-incubated with non-related protease-rich extracts in comparison with that of the control pollen extract. MALDI-TOF MS analysis allowed us to determine that Lol p 1 and Lol p 5 are susceptible to proteolysis whereas Lol p 4 was found to be resistant to degradation during storage. Lol p 1 and Lol p 5 degradation is responsible for the loss of the biological activity of L. perenne pollen extract when co-incubated with protease-rich fungal and cockroach extracts in the same vial for months in the absence of glycerol as a preserving agent. The integrity of these major allergens must be preserved to increase the vaccine stability and to assure efficacy when mixes are used for immunotherapy.

Decreasing radioactive cesium in lodged buckwheat grain after harvest

Directory of Open Access Journals (Sweden)

Katashi Kubo

2016-01-01

Full Text Available This study assessed soil contamination with high radioactive cesium (R–Cs concentration in buckwheat grains by lodging, and assessed the possibility of R–Cs reduction in grain through post-harvest preparation. Analysis of buckwheat grain produced in farmers’ fields and reports from farmers indicated that grain from fields that had lodging showed higher R–Cs than grain from fields with no lodging. A field experiment demonstrated that R–Cs in grain after threshing and winnowing (TW was about six times higher in lodged plants than in nonlodged plants. In lodged plants, R–Cs in grain was decreased to about one-fourth by polishing, and was decreased to about one-seventh by ultrasonic cleaning, compared with R–Cs in grain after TW. These results demonstrate that R–Cs of buckwheat grain of lodged plants can be decreased by removing soil from the grain surface by polishing and winnowing.

Functional characterization of steam jet-cooked buckwheat flour as a fat replacer in cake-baking.

Science.gov (United States)

Min, Bockki; Lee, Seung Mi; Yoo, Sang-Ho; Inglett, George E; Lee, Suyong

2010-10-01

With rising consumer awareness of obesity, the food industry has a market-driven impetus to develop low-fat or fat-free foods with acceptable taste and texture. Fancy buckwheat flour was thus subjected to steam jet-cooking and the performance of the resulting product in cake-baking was evaluated as a fat replacer. Steam jet-cooking caused structural breakdown and starch gelatinization of buckwheat flour, thus increasing its water hydration properties. In the pasting measurements, steam jet-cooked buckwheat flour exhibited high initial viscosity, while no peak viscosity was observed. Also, the suspensions of steam jet-cooked buckwheat flour exhibited shear-thinning behaviors, which were well characterized by the power law model. When shortening in cakes was replaced with steam jet-cooked buckwheat gels, the specific gravity of cake batters significantly increased, consequently affecting cake volume after baking. However, shortening replacement with steam jet-cooked buckwheat up to 20% by weight appeared to be effective in producing cakes as soft as the control without volume loss. When buckwheat flour was thermomechanically modified by steam jet-cooking, it was successfully incorporated into cake formulations for shortening up to 20% by weight, producing low-fat cakes with comparable volume and textural properties to the control. Copyright © 2010 Society of Chemical Industry.

Crystallization and preliminary X-ray analysis of the major peanut allergen Ara h 1 core region

International Nuclear Information System (INIS)

Cabanos, Cerrone; Urabe, Hiroyuki; Masuda, Taro; Tandang-Silvas, Mary Rose; Utsumi, Shigeru; Mikami, Bunzo; Maruyama, Nobuyuki

2010-01-01

In this study, the core region of Ara h 1, one of the major peanut allergens, has been overexpressed in Escherichia coli, purified and crystallized. Diffraction data were collected to 2.25 Å resolution. Peanuts contain some of the most potent food allergens known to date. Ara h 1 is one of the three major peanut allergens. As a first step towards three-dimensional structure elucidation, recombinant Ara h 1 core region was cloned, expressed in Escherichia coli and purified to homogeneity. Crystals were obtained using 0.1 M sodium citrate pH 5.6, 0.1 M NaCl, 15% PEG 400 as precipitant. The crystals diffracted to 2.25 Å resolution using synchrotron radiation and belonged to the monoclinic space group C2, with unit-cell parameters a = 156.521, b = 88.991, c = 158.971 Å, β = 107.144°. Data were collected at the BL-38B1 station of SPring-8 (Hyogo, Japan)

Search for Allergens from the Pollen Proteome of Sunflower (Helianthus annuus L.): A Major Sensitizer for Respiratory Allergy Patients.

Science.gov (United States)

Ghosh, Nandini; Sircar, Gaurab; Saha, Bodhisattwa; Pandey, Naren; Gupta Bhattacharya, Swati

2015-01-01

Respiratory allergy triggered by pollen allergens is increasing at an alarming rate worldwide. Sunflower pollen is thought to be an important source of inhalant allergens. Present study aims to identify the prevalence of sunflower pollinosis among the Indian allergic population and characterizes the pollen allergens using immuno-proteomic tools. Clinico-immunological tests were performed to understand the prevalence of sensitivity towards sunflower pollen among the atopic population. Sera from selected sunflower positive patients were used as probe to detect the IgE-reactive proteins from the one and two dimensional electrophoretic separated proteome of sunflower pollen. The antigenic nature of the sugar moiety of the glycoallergens was studied by meta-periodate modification of IgE-immunoblot. Finally, these allergens were identified by mass-spectrometry. Prevalence of sunflower pollen sensitization was observed among 21% of the pollen allergic population and associated with elevated level of specific IgE and histamine in the sera of these patients. Immunoscreening of sunflower pollen proteome with patient sera detected seven IgE-reactive proteins with varying molecular weight and pI. Hierarchical clustering of 2D-immunoblot data highlighted three allergens characterized by a more frequent immuno-reactivity and increased levels of IgE antibodies in the sera of susceptible patients. These allergens were considered as the major allergens of sunflower pollen and were found to have their glycan moiety critical for inducing IgE response. Homology driven search of MS/MS data of these IgE-reactive proteins identified seven previously unreported allergens from sunflower pollen. Three major allergenic proteins were identified as two pectate lyases and a cysteine protease. Novelty of the present report is the identification of a panel of seven sunflower pollen allergens for the first time at immuno-biochemical and proteomic level, which substantiated the clinical evidence of

Antioxidant Capacity, Mineral Content and Sensory Properties of Gluten-Free Rice and Buckwheat Cookies

Directory of Open Access Journals (Sweden)

Mladenka Pestorić

2015-01-01

Full Text Available Light buckwheat flour was used to substitute rice fl our at the level of 10, 20 and 30 % to produce gluten-free cookies. The substitution of gluten-free cookie formulation with light buckwheat fl our contributed to the signifi cantly higher mineral content, especially magnesium, potassium, iron and copper, in comparison with the control rice cookies (p<0.05. Gluten-free cookies made with rice fl our and buckwheat fl our exhibited signifi cantly higher total phenolic and rutin content, scavenging activity against 1,1-diphenyl 2-picrylhydrazyl radicals (DPPH•, antioxidant activity and reducing power than the control cookies (p<0.05. Comparing all evaluated sensory properties, cookies containing 20 % of light buckwheat flour had the most acceptable sensory properties. The obtained results of principal component analysis showed that the cookies with 20 and 30 % buckwheat flour had better antioxidant and sensory properties in comparison with other two cookie samples.

Proteomic analysis of the major birch allergen Bet v 1 predicts allergenicity for 15 birch species

NARCIS (Netherlands)

Schenk, M.F.; Cordewener, J.H.G.; America, A.H.P.; Peters, J.; Smulders, M.J.M.; Gilissen, L.J.W.J.

2011-01-01

Pollen of the European and Asian white birch (Betula pendula and B. platyphylla) causes hay fever in humans. The allergenic potency of other birch species is largely unknown. To identify birch trees with a reduced allergenicity, we assessed the immunochemical characteristics of 15 species and two

Morphological characterization and estimation of genotype * environment interaction of indigenous buckwheat germplasm collected from Gilgit Baltistan, Pakistan

International Nuclear Information System (INIS)

Facho, Z.H.; Tarhatullah, A.; Khalil, I.H.; Ali, S.

2016-01-01

A two-year study (2011 and 2012) was conducted at two locations (Skardu and Ghanche) of Gilgit Baltistan, Pakistan to characterize and estimate genotype by environment interaction (GEI) for 20 buckwheat genotypes collected from 18 locations of Gilgit - Baltistan, Pakistan. The GE interaction was studied using a set of six quantitative descriptors. Wide range of variations was recorded for flowering, maturity, plant height, 1000 grain weight, grains plant-1 and grain yield ha-1. Significant differences in locations for all traits were recorded. Years * locations interactions was also significant (p= 0.05) for all traits except grains plant-1 and grain yield ha-1. Similarly, genotype * location * year interaction existed for most of the traits except plant height, grains plant-1 and grain yield ha-1. The genotype Sh-914 (29.91 g) excelled in 1000-grain weight, followed by Rd-915 (27.04 g). Maximum grains plant-1 were produced by Sh-914 (323.62) followed by the genotype Gh-918 (229.16). Maximum grain yield of 2010.27 kg ha-1 was produced by genotype Sh-914, followed by the genotype Gh-918 (1910.40 kg ha-1). Days to flowering, days to maturity, and 1000 grain weight were the major contributors towards genetic divergence among the buckwheat genotypes. Moderate to high estimates of broad-sense heritability and selection response were observed for traits at two test locations. Cluster analysis based on morphological and yield related traits classified buckwheat genotypes of two species (Fagopyrum esculentum and F. tataricum) into three main groups. The clustering pattern revealed that genotypes collected from the same location were grouped into different clusters. Data obtained on the basis of Dendrogram showed differences for various phenological and yield traits among buckwheat genotypes. Buckwheat genotypes Sh-914 and Gh-918 performed well at Skardu and Sh-914 and Rd-915 at Ghanche. The information on the existing genetic variability for morphological and yield

Mycopopulations of grain and flour of wheat, corn and buckwheat

Directory of Open Access Journals (Sweden)

Plavšić Dragana V.

2017-01-01

Full Text Available According to the nutritive characteristics, whole grain flour is a high quality product, due to its high vitamin, mineral, and dietary fiber content. However, the cereal grains are susceptible to the series of contamination during the ripening, harvesting, processing and storage. The aim of this work was to determine mold presence in grains and flour of wheat, corn and buckwheat. The determination of total number and identification of isolated genera and species of molds were the subject of this research. All samples were contaminated with the molds. The total number of molds per 100 cereal grains was between 60 cfu (wheat and 120 cfu (buckwheat. The total number of molds in the samples of flour ranged from 6.0x101 cfu/g in white wheat flour to 5.0 x102 cfu/g in buckwheat whole grain flour (DG18 medium. Eight fungal genera (Alternaria, Aspergillus, Cladosporium, Chrysonilia, Fusarium, Penicillium, Rhizopus and Scopulariopsis and fifteen species were isolated. The largest number of species of molds was isolated from the genus Aspergillus. About 66.7% of isolated fungi belonged to potentially toxigenic species. The results pointed out the necessity of grain surface treatment, preceding the milling of grains in wheat, corn and whole grain buckwheat flour production.

Targeting allergenic fungi in agricultural environments aids the identification of major sources and potential risks for human health.

Science.gov (United States)

Weikl, F; Radl, V; Munch, J C; Pritsch, K

2015-10-01

Fungi are, after pollen, the second most important producers of outdoor airborne allergens. To identify sources of airborne fungal allergens, a workflow for qPCR quantification from environmental samples was developed, thoroughly tested, and finally applied. We concentrated on determining the levels of allergenic fungi belonging to Alternaria, Cladosporium, Fusarium, and Trichoderma in plant and soil samples from agricultural fields in which cereals were grown. Our aims were to identify the major sources of allergenic fungi and factors potentially influencing their occurrence. Plant materials were the main source of the tested fungi at and after harvest. Amounts of A. alternata and C. cladosporioides varied significantly in fields under different management conditions, but absolute levels were very high in all cases. This finding suggests that high numbers of allergenic fungi may be an inevitable side effect of farming in several crops. Applied in large-scale studies, the concept described here may help to explain the high number of sensitization to airborne fungal allergens. Copyright © 2015 Elsevier B.V. All rights reserved.

27 CFR 4.32b - Petitions for exemption from major food allergen labeling.

Science.gov (United States)

2010-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Petitions for exemption from major food allergen labeling. 4.32b Section 4.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF WINE...

27 CFR 5.32b - Petitions for exemption from major food allergen labeling.

Science.gov (United States)

2010-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Petitions for exemption from major food allergen labeling. 5.32b Section 5.32b Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS LABELING AND ADVERTISING OF...

The influence of digestibility on the allergenicity of food allergens

DEFF Research Database (Denmark)

Bøgh, Katrine Lindholm; Madsen, Charlotte Bernhard

2012-01-01

potential exist. Resistance to digestion is therefore a test parameter included in the safety assessment of the allergenic potential of novel proteins in genetically modified foods. In recent years, the association between resistance to digestion and allergenic potential has been challenged. When reviewing......Food allergy is a major health problem in the Western countries, affecting 3-8% of the population. What makes a dietary protein a food allergen has not yet been established, though several characteristics have been proposed to be shared by the food allergens. One of the features believed...... existing data from digestibility studies on known food allergens, it becomes evident that food allergens do not necessarily resist digestion. However, the choice of assay conditions, the method used for detection of residual intact protein as well as fragments hereof greatly influences the outcome. Studies...

Textural properties of bread formulations based on buckwheat and rye flour

Directory of Open Access Journals (Sweden)

Petra Dvořáková

2012-01-01

Full Text Available Modern nutrition and nutritionists worldwide more and more require high nutritional quality foods including breads. Products based on rye (Secale cereale L. and other cereals such as buckwheat (Fagopyrum esculentum Moench provide nutritional benefits such as higher intake of fibre which has a positive effect on digestion and decreases a risk of obesity and heart disease, therefore current trend is to replace part of gluten breads with other cereal products. The main aim of this work was to observe changes in breads based on buckwheat and rye mixtures influenced by ratio of buckwheat and rye flour. Eleven ratios of buckwheat-rye flours were prepared. Dough and bread quality were tested in terms of dough machine workability, dough and pastry yield, baking loss, specific volume and texture analyses 24 and 72 hours after baking. The results were statistically evaluated and showed that rising amount of rye flour in mixtures did not affect dough machine workability but improved all of the investigated texture characteristics such as chewiness and gumminess, concerning specific volume of breads, no significant differences were found. All texture parameters deteriorated with staling time.

Cloning, expression, and mapping of allergenic determinants of alphaS1-casein, a major cow's milk allergen.

Science.gov (United States)

Schulmeister, Ulrike; Hochwallner, Heidrun; Swoboda, Ines; Focke-Tejkl, Margarete; Geller, Beate; Nystrand, Mats; Härlin, Annika; Thalhamer, Josef; Scheiblhofer, Sandra; Keller, Walter; Niggemann, Bodo; Quirce, Santiago; Ebner, Christoph; Mari, Adriano; Pauli, Gabrielle; Herz, Udo; Valenta, Rudolf; Spitzauer, Susanne

2009-06-01

Milk is one of the first components introduced into human diet. It also represents one of the first allergen sources, which induces IgE-mediated allergies in childhood ranging from gastrointestinal, skin, and respiratory manifestations to severe life-threatening manifestations, such as anaphylaxis. Here we isolated a cDNA coding for a major cow's milk allergen, alphaS1-casein, from a bovine mammary gland cDNA library with allergic patients' IgE Abs. Recombinant alphaS1-casein was expressed in Escherichia coli, purified, and characterized by circular dichroism as a folded protein. IgE epitopes of alphaS1-casein were determined with recombinant fragments and synthetic peptides spanning the alphaS1-casein sequence using microarrayed components and sera from 66 cow's milk-sensitized patients. The allergenic activity of ralphaS1-casein and the alphaS1-casein-derived peptides was determined using rat basophil leukemia cells transfected with human FcepsilonRI, which had been loaded with the patients' serum IgE. Our results demonstrate that ralphaS1-casein as well as alphaS1-casein-derived peptides exhibit IgE reactivity, but mainly the intact ralphaS1-casein induced strong basophil degranulation. These results suggest that primarily intact alphaS1-casein or larger IgE-reactive portions thereof are responsible for IgE-mediated symptoms of food allergy. Recombinant alphaS1-casein as well as alphaS1-casein-derived peptides may be used in clinical studies to further explore pathomechanisms of food allergy as well as for the development of new diagnostic and therapeutic strategies for milk allergy.

Use of gamma irradiation for microbial inactivation of buckwheat flour and buckwheat food products, 1

International Nuclear Information System (INIS)

Matsuhashi, Tetsujiro; Oh-hinata, Hiroshi; Muramatsu, Nobuyuki; O-hara, Tadahiko; Itoh, Hitoshi; Saito, Makoto.

1986-01-01

In an attempt to develop a feasible method for improving shelf-life extension of fresh 'soba' noodles, the usefulness of gamma irradiation was preliminarily investigated. Microorganism counts decreased to less than 10 2 per gram by irradiating gamma rays of 0.5 Mrad to buckwheat grains with hulls (Material I), industrially processed buckwheat flours (Material II), and uncooked 'soba' noodles packed in a plastic film bag (Material III). For Material II, the effective dose to decrease 90 % of the microorganism counts was 0.1 Mrad, and heat-resistant microorganism counts decreased to 10 or less per gram with gamma rays of 0.2 - 0.5 Mrad. Flavor in either Material II or Material III was not changed by irradiation; by contraries, enhanced 'soba'-like flavor was rarely observed. Although gamma-irradiation decreased the viscosity of Material II, it hardly exerted an effect on the process of making noodles. The texture of Material III became degraded by irradiation. The results suggest the feasibility of gamma irradiation in the use of inactivation of microorganisms. (Namekawa, K.)

Comparative Analysis of the Antioxidant Capacities and Phenolic Compounds of Oat and Buckwheat Vinegars During Production Processes.

Science.gov (United States)

Yu, Xiao; Yang, Mei; Dong, Jilin; Shen, Ruiling

2018-03-01

This study aimed to explore the dynamic changes in the antioxidant activities and phenolic acid profiles of oat and buckwheat vinegars during different production stages. The results showed that both oat and buckwheat vinegar products comparably attenuated D-galactose-induced oxidative damage in mice serum and liver, indicating no obvious dose dependence within the tested concentrations. However, oat vinegar product revealed more favorable in vitro antioxidant activities than those in buckwheat vinegar product as evaluated by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging abilities. Moreover, the alcoholic fermentation, acetic acid fermentation and fumigating induced successive increase in DPPH radical scavenging abilities and phenolic acid contents of the fermentation substrates of oat and buckwheat vinegars. Importantly, the different fermentation processes of oat and buckwheat vinegars were accompanied by the dynamic migration and transformation of specific phenolic acids across bound, esterified and free fractions. Thus, the antioxidant activities of oat and buckwheat vinegars could be improved through targeted modulation of the generation of specific phenolic acid fractions during production processes. We had evaluated the in vitro and in vivo antioxidant activities and phenolic acid contents of oat and buckwheat vinegars, and further explored the dynamic changes of bound, esterified and free phenolic acid fractions during successive fermentation processes of oat and buckwheat vinegars. This study provided the theoretical guidance for obtaining minor grain vinegar with the optimal antioxidant activities through targeted modulation of fermentation processes. © 2018 Institute of Food Technologists®.

A randomized, controlled, crossover study of appetite-related sensations after consuming snacks made from buckwheat.

Science.gov (United States)

Defries, Danielle M; Petkau, Jay C; Gregor, Terri; Blewett, Heather

2018-02-01

With the rising incidence of overweight and obesity in developed countries, there is an interest in developing food products that may aid in satiety and reduce energy intake. Buckwheat (Fagopyrum esculentum) is a gluten-free edible seed that has been previously shown to induce changes in postprandial concentrations of satiety hormones; however, subjective measures of appetite-related sensations and objective measures of energy intake at subsequent meals following buckwheat consumption have not been measured. Thirty-eight healthy adults were recruited to participate in a randomized, controlled, crossover trial with the main objective to determine if consuming snacks made from buckwheat would increase satiety and reduce energy intake compared with snacks comparable in serving size, physical characteristics, and nutrient composition. Water was included as a no-kilocalorie control. Participants received each of the treatments once separated by at least 7 days. Appetite related sensations were assessed using visual analog scales at fasting and after consuming the snack at 30-min intervals for 180 min. Lunch was provided at the clinic and the amount of food consumed was weighed. Participants recorded food intake for the rest of the day. Consuming buckwheat groats (32 g serving; 141 kcal) or pita bread made from buckwheat flour (50 g serving; 135 kcal) was not associated with changes in appetite related sensations or energy consumption compared with reference snack products made from corn or rice flour. Sensory questionnaires revealed that snacks made from buckwheat were liked to a similar degree or more as reference snack products, which shows commercial promise for developing buckwheat-containing snacks.

AGRO-ECONOMICAL ASPECTS OF GROWTH STIMULANTS AND MICROFERTILIZERS APPLICATION ON BUCKWHEAT

OpenAIRE

Kozlobaeva E. A.; Kozlobaev A. V.

2015-01-01

The article highlights the food value of buckwheat. It is said that the achievement of its biological potential under prevailing conditions of cultivation is impossible without the introduction of modern scientific and technical progress. The authors describe variants of integrated application of growth stimulants and microfertilizers in buckwheat production technology. Recommended technologies have been tested in conditions of the Central Chernozem region. It is revealed that the best varian...

Spectrum of allergens for Japanese cedar pollinosis and impact of component-resolved diagnosis on allergen-specific immunotherapy

Directory of Open Access Journals (Sweden)

Takashi Fujimura

2015-10-01

Full Text Available The high prevalence of Japanese cedar pollinosis in Japan is associated with a negative impact on the quality of life of patients, as well as significant loss of productivity among the workforce in early spring, thus representing a serious social problem. Furthermore, the prevalence is increasing, and has risen by more than 10% in this decade. Cry j 1 and Cry j 2 were identified as the major allergens in Japanese cedar pollen (JCP, and in 2004, the existence of other major and minor allergens were revealed by a combination of two-dimensional electrophoresis and immunoblotting analysis. Allergenome analysis identified a chitinase, a lipid transfer protein, a serine protease, and an aspartic protease as novel IgE-reactive allergens in patients with JCP allergy. Thaumatin-like protein (Cry j 3 was shown to be homologous to Jun a 3, a major allergen from mountain cedar pollen. Isoflavone reductase-like protein was also characterized in a study of a JCP cDNA library. The characterization of component allergens is required to clarify the sensitizer or cross-reactive elicitor allergens for component-resolved diagnosis (CRD. Increasing evidence from numerous clinical trials indicates that CRD can be used to design effective allergen-specific immunotherapy. In this review, we summarize the eight characterized JCP allergens and discuss the impact of CRD and characterization of novel allergens on allergen-specific immunotherapy.

Chemical modification of Art v 1, a major mugwort pollen allergen, by cis-aconitylation and citraconylation

Directory of Open Access Journals (Sweden)

DRAGANA STANIĆ

2009-04-01

Full Text Available Art v 1 is the major allergen of mugwort (Artemisia vulgaris pollen, a significant cause of hay fever all over Europe. Specific immunotherapy is the only treatment modality for allergic disease. Application of modified allergens makes the treatment safer and more efficient. In this work, two out of three (citraconic anhydride, cis-aconitic anhydride, 2,3-dimethylmaleic anhydride tested anhydrides were proven to be suitable for chemical modifications of allergens. Art v 1 was modified by cis-aconitylation and citraconylation in order to obtain derivatives of Art v 1 that may be suitable for further immunological testing. Acylation of Art v 1 gave derivatives (caaArt v 1 and citArt v 1 with about 80 % modified amino groups. The derivatives were in the monomeric form and had dramatically reduced pI values. Both derivatives were relatively stable at neutral pH values, while the acyl groups undergo hydrolysis under acidic conditions. Modification of allergens by cis-aconitylation and citraconylation could be a new tool for obtaining allergoids.

Isolation of cDNA encoding a newly identified major allergenic protein of rye-grass pollen: intracellular targeting to the amyloplast.

Science.gov (United States)

Singh, M B; Hough, T; Theerakulpisut, P; Avjioglu, A; Davies, S; Smith, P M; Taylor, P; Simpson, R J; Ward, L D; McCluskey, J

1991-01-01

We have identified a major allergenic protein from rye-grass pollen, tentatively designated Lol pIb of 31kDa and with pI 9.0. A cDNA clone encoding Lol pIb has been isolated, sequenced, and characterized. Lol pIb is located mainly in the starch granules. This is a distinct allergen from Lol pI, which is located in the cytosol. Lol pIb is synthesized in pollen as a pre-allergen with a transit peptide targeting the allergen to amyloplasts. Epitope mapping of the fusion protein localized the IgE binding determinant in the C-terminal domain. Images PMID:1671715

Food Allergens: Is There a Correlation between Stability to Digestion and Allergenicity?

DEFF Research Database (Denmark)

Bøgh, Katrine Lindholm; Madsen, Charlotte Bernhard

2016-01-01

and the method used for detection of residual intact protein as well as fragments hereof may greatly influence the outcome as well as the interpretation of results. The finding that digests from food allergens may retain allergenicity, stresses the importance of using immunological assays for evaluating......Food allergy is a major health problem in the Western countries, affecting 3-8% of the population. It has not yet been established what makes a dietary protein a food allergen. Several characteristics have been proposed to be shared by food allergens. One of these is resistance to digestion....... This paper reviews data from digestibility studies on purified food allergens and evaluates the predictive value of digestibility tests on the allergenic potential. We point out that food allergens do not necessarily resist digestion. We discuss how the choice of in vitro digestibility assay condition...

Purification and characterization of Lep d I, a major allergen from the mite Lepidoglyphus destructor.

Science.gov (United States)

Ventas, P; Carreira, J; Polo, F

1992-04-01

A major allergen of the storage mite Lepidoglyphus destructor (Lep d I) has been purified by affinity chromatography using an anti-Lep d I monoclonal antibody. The purity of the protein obtained by this procedure was assessed by reverse-phase HPLC. Lep d I displayed a molecular weight of 14 kD on SDS-PAGE under non-reducing conditions, and 16 kD in the presence of a reducing agent. Analytical IEF revealed a little charge microheterogeneity, showing three bands with pIs 7.6-7.8. Purified Lep d I retained IgE-binding ability, as proved by immunoblotting experiments after SDS-PAGE and RAST with individual sera from L. destructor-sensitive patients. Results from the latter technique demonstrated that 87% of L. destructor-allergic patients had specific IgE to Lep d I, and a good correlation between IgE reactivity with L. destructor extract and Lep d I was found. In addition, RAST inhibition experiments showed that IgE-binding sites on Lep d I are major L. destructor-allergenic determinants, since Lep d I could inhibit up to 75% the binding of specific IgE to L. destructor extract; on the other hand, Lep d I did not cross-react with D. pteronyssinus allergens.

[Different uses of Fagopyrum esculentum Moench (buckwheat) in Japan and China: what ancient medical documents reveal].

Science.gov (United States)

Tatsumi, Nami; Marui, Eiji

2012-03-01

The purpose of this thesis is to demonstrate that buckwheat has been recognized, both in Japan and China, as a crop that is useful in many ways: as an agricultural crop, and for the healing powers and properties that, according to traditional Chinese medicine, it has. A comparative study of ancient documents pertaining to medicine in these countries has made it clear that this is the case. Buckwheat, however, has been used quite differently in each country. As is shown in some ancient Chinese documents pertaining to medicine, China has treated buckwheat primarily as a medicine for clinical use rather than as an edible crop. Nowadays, buckwheat is eaten only in some regions of China. Although it came to Japan from China as a medicine, in Japan buckwheat gradually became a popular food crop. It has become an important component of traditional Japanese cuisine thanks in part to government support and the strong demand that developed in Japanese society.

CHOICE OF EFFICIENT METHOD OF ADDING FLOUR FROM BUCKWHEAT BRAN

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E. I. Ponomareva

2015-01-01

Full Text Available Nowadays the production of functional bakery products for therapeutic and preventive nutrition is of current importance. The problem of providing the population with functional products can be solved by enriching the recipes with vitamins, dietary fiber, micro and macronutrients. At the chair of "Technology of baking, confectionery, pasta and grain processing" of Voronezh State University of Engineering Technologies the recipe of no-salt bread from mixture of baker’s first grade wheat flour and whole-grain wheat with adding buckwheat bran flour, that increases the nutrition value of the product, has been developed. Flour from buckwheat bran is characterized by a high-scale balance of the content of essential amino acids, good digestibility, rich in vitamins (especially B vitamins, PP, dietary fiber, minerals (potassium, calcium, magnesium, iron. The use of buckwheat flour from the bran in bread production stimulates the creation of favorable intestinal microflora, normalizes blood sugar level, and excretes toxins, toxic salts and heavy metals from the body. We have conducted a research on how to make the rational choice of method of adding buckwheat bran flour enricher into dough from a mixture of first grade wheat flour and whole-grain wheat, compressed yeast and potable water, thus ensuring high and stable physical and chemical characteristics and the quality of the end product. Also, we have discovered that the bakery product prepared on tight sponge with addition of enricher has much better organoleptical properties in comparison with the others. Bread is characterized by a pleasant taste and flavour, elastic porous crumb. It has been found that the consumption of 100 g of a mixture of no-salt bread from first grade wheat flour and whole-grain with adding buckwheat bran flour will provide enough daily intake of protein 10.7%, fat 1.5%, carbohydrates 10.4% dietary fiber 16.3%, amino acids 2.5 14.0%. No-salt bread is recommended to people

Transcriptomic identification of salt-related genes and de novo assembly in common buckwheat (F. esculentum).

Science.gov (United States)

Lu, Qi-Huan; Wang, Ya-Qi; Song, Jin-Nan; Yang, Hong-Bing

2018-06-01

Common buckwheat (F. esculentum), annually herbaceous crop, is prevalent in people's daily life with the increasing development of economics. Compared with wheat, it is highly praised with high content of rutin and flavonoid. Common buckwheat is recognized as healthy food with good taste, and the product price of which such as noodles, flour, bread and so on are higher than wheat, and the seeds of which are bigger than that of tartary buckwheat, so if common buckwheat are planted more widely, people will spend less money on this healthy and delicious food. However, soil salinity has been a giant problem for agriculture production. The cultivation of salt tolerant crop varieties is an effective way to make full use of saline alkali land, and the highest salinity that the common buckwheat can sow is at 6.0%, so we chose 100 mM as the concentration of NaCl for treatment. Then we conducted transcriptome comparison between control and treatment groups. Potential regulatory genes related salt stress in common buckwheat were identified. A total of 29.36 million clean reads were produced via an illumina sequencing approach. We de novo assembled these reads into a transcriptome dataset containing 43,772 unigenes with N50 length of 1778 bp. A total of 26,672 unigenes could be found matches in public databases. GO, KEGG and Swiss-Prot classification suggested the enrichment of these unigenes in 47 sub-categories, 25 KOG and 129 pathways, respectively. We got 385 differentially expressed genes (DEGs) after comparing the transcriptome data between salt treatment and control groups. There are some genes encoded for responsing to stimulus, cell killing, metabolic process, signaling, multi-organism process, growth and cellular process might be relevant to salt stress in common buckwheat, which will provide a valuable references for the study on mechanism of salt tolerance and will be used as a genetic information for cultivating strong salt tolerant common buckwheat varieties in

Cloning and sequencing of Lol pI, the major allergenic protein of rye-grass pollen.

Science.gov (United States)

Griffith, I J; Smith, P M; Pollock, J; Theerakulpisut, P; Avjioglu, A; Davies, S; Hough, T; Singh, M B; Simpson, R J; Ward, L D

1991-02-25

We have isolated a full length cDNA clone encoding the major glycoprotein allergen Lol pI. The clone was selected using a combination of immunological screening of a cDNA expression library and PCR amplification of Lol pI-specific transcripts. Lol pI expressed in bacteria as a fusion protein shows recognition by specific IgE antibodies present in sera of grass pollen-allergic subjects. Northern analysis has shown that the Lol pI transcripts are expressed only in pollen of rye-grass. Molecular cloning of Lol pI provides a molecular genetic approach to study the structure-function relationship of allergens.

Impacts of Thermal Treatments on Major and Minor Allergens of Sea Snail, Cerithidea obtusa (Obtuse Horn Shell)

OpenAIRE

Rosmilah Misnan; Norazlin Salahudin Abd Aziz; Zailatul Hani Mohamad Yadzir; Faizal Bakhtiar; Noormalin Abdullah; Shahnaz Murad

2016-01-01

Snail is one of the worst causes of food allergy. Thus, the aim of this study was to identify the major and minor allergens of the local marine snail (Cerithidea obtusa) and subsequently to investigate the impacts of heat treatment on the IgE-binding activity of snail allergens. Proteins from raw and heat-treated snails (boiled, roasted and fried) were extracted and then resolved by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblotting of all extracts were then...

Immunological cross-reactivity of the major allergen from perennial ryegrass (Lolium perenne), Lol p I, and the cysteine proteinase, bromelain.

Science.gov (United States)

Pike, R N; Bagarozzi, D; Travis, J

1997-04-01

Antibodies prepared in rabbits against the major allergen from ryegrass (Lolium perenne), Lol p I, cross-reacted with the cysteine proteinase bromelain from pineapple and vice versa. Deglycosylation of the proteins showed that the cross-reaction was based on recognition of the carbohydrate moiety of the allergen, but for bromelain the cross-reaction was most likely due to a combination of factors. The results indicate that the carbohydrate residues from these allergens play an important role in cross-reactions found between them and possibly those from other species.

The effect of fermented buckwheat on producing l-carnitine- and γ-aminobutyric acid (GABA)-enriched designer eggs.

Science.gov (United States)

Park, Namhyeon; Lee, Tae-Kyung; Nguyen, Thi Thanh Hanh; An, Eun-Bae; Kim, Nahyun M; You, Young-Hyun; Park, Tae-Sub; Kim, Doman

2017-07-01

The potential of fermented buckwheat as a feed additive was studied to increase l-carnitine and γ-aminobutyric acid (GABA) in designer eggs. Buckwheat contains high levels of lysine, methionine and glutamate, which are precursors for the synthesis of l-carnitine and GABA. Rhizopus oligosporus was used for the fermentation of buckwheat to produce l-carnitine and GABA that exert positive effects such as enhanced metabolism, antioxidant activities, immunity and blood pressure control. A novel analytical method for simultaneously detecting l-carnitine and GABA was developed using liquid chromatography/mass spectrometry (LC/MS) and LC/MS/MS. The fermented buckwheat extract contained 4 and 34 times more l-carnitine and GABA respectively compared with normal buckwheat. Compared with the control, the fermented buckwheat extract-fed group showed enriched l-carnitine (13.6%) and GABA (8.4%) in the yolk, though only l-carnitine was significantly different (P < 0.05). Egg production (9.4%), albumen weight (2.1%) and shell weight (5.8%) were significantly increased (P < 0.05). There was no significant difference in yolk weight, and total cholesterol (1.9%) and triglyceride (4.9%) in the yolk were lowered (P < 0.05). Fermented buckwheat as a feed additive has the potential to produce l-carnitine- and GABA-enriched designer eggs with enhanced nutrition and homeostasis. These designer eggs pose significant potential to be utilized in superfood production and supplement industries. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Allergens of mites

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Emilia Siwak

2014-04-01

Full Text Available Mite allergens belong to the group of inhalant allergens and represent antigenic substances which are particutlarly important in the pathogenesis of respiratory system diseases and skin diseases. The most common diseases associated with chronic exposure to these aeroallergens include: allergic rhinitis, bronchial asthma and atopic dermatitis. Mite allergens are simple proteins or glycoproteins with different molecular structures and various biochemical functions. The sensitizing capacity of these proteins is connected from their physicochemical properties. Individual allergens perform, among others, the functions of structural proteins, act as enzymes, transport lipids, bind metal ions, and are capable of glycosylation. In addition, mite allergenic proteases degrade proteins of the skin epithelium-resulting in a weakening of its natural protective barrier-and induce the immune response. The proteases also induce the release of pro-inflammatory cytokines: interleukin-4 (IL-4, interleukin 6 (IL-6, interleukin 8 (IL-8, eotaxin, and granulocyte-macrophage colony-stimulating factor-GM-CSF. The article presents the tertiary structure of major and mid-range mite allergens and their classification. Based on literature reports concerning the chemical structure of allergenic proteins, it was emphasized that the structural differences between homologous proteins with allergenic pozoproperties relate to the distribution of amino acid residues on the surface of the molecule. IgE binding affinity and the similarities and differences in the amino acid sequence of the allergens were also the basis for determining cross-reactivity of allergenic proteins. The paper shows an example of this phenomenon, describing the existence of common allergens for various mite species.

Dose response and mutation induction by ion beam irradiation in buckwheat

International Nuclear Information System (INIS)

Morishita, T.; Yamaguchi, H.; Degi, K.; Shikazono, N.; Hase, Y.; Tanaka, A.; Abe, T.

2003-01-01

The biological effects of ion beams were investigated to pursue the development of a method for breeding by mutation in buckwheat. Common buckwheat (Botansoba, Bot) and tartary buckwheat (Rotundatiem, Rot) seeds were exposed to various ions in linear energy transfer (LET) at 9-630 keV/μm. The lethal dose 50 (LD 50 ) of ion beams were 10-300 Gy (Bot) and 30-500 Gy (Rot). It was indicated that a penetrating depth in excess of 1.7 mm is necessary to thoroughly saturate the target, and ions with a penetrating depth of less than 2.2 mm were affected by the presence of hulls. The maximum values of the relative biological effectiveness were 17.7 (Rot) and 22.5 (Bot) at 305 keV/μm. The effective cross sections increased with the LET, and the maximum values were 2.7 (Rot) and 3.0 μm 2 (Bot). The mutation induction effects of He and C ions were higher than those of gamma rays

Identification and characterization of an arginine kinase as a major allergen from silkworm (Bombyx mori) larvae.

Science.gov (United States)

Liu, Zhigang; Xia, Lixin; Wu, Yulan; Xia, Qingyou; Chen, Jiajie; Roux, Kenneth H

2009-01-01

The silkworm, Bombyx mori, is an important insect in the textile industry and its pupa are used in Chinese cuisine and traditional Chinese medicine. The silk, urine and dander of silkworms is often the cause of allergies in sericulture workers and the pupa has been found to be a food allergen in China. Recent studies have focused on reporting cases of silkworm allergies, but only a few studies have addressed the specific allergens present in the B. mori silkworm. We collected sera from 10 patients with a positive skin prick test to silkworm crude extract (SCE) and analyzed these samples by Western blot and ELISA. The cDNA of arginine kinase from the B. mori silkworm was also cloned and expressed in high yield in Escherichia coli. Allergenicity and cross-allergenicity of the recombinant B. mori arginine kinase (rBmAK) were investigated by ELISA inhibition assay. Collected sera all reacted to a 42-kDa protein in a Western blot with SCE as the antigen. Preincubation of sera with rBmAK eliminated the reactivity of the patients' sera to this 42-kDa band. All patient sera also exhibited positive reactivity to SCE in an ELISA assay. BmAK also demonstrated cross-reactivity with a recombinant AK from cockroach. Arginine kinase from the B. mori silkworm is a major allergen and crossreacts with cockroach AK. Copyright 2009 S. Karger AG, Basel.

Chemical composition of buckwheat plant parts and selected buckwheat products

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Petra Vojtíšková

2014-11-01

Full Text Available Chemical composition plant parts (roots, stalks, leaves, blossoms of common buckwheat (Fagopyrum esculentum Moench and selected products made from its seeds (peels, whole seed, wholemeal flour, broken seeds, crunchy products Natural and Cocoa, flour, and pasta was determined. Samples were dried and ground to a fine powder. All analyses were performed according to the Commission Regulation no. 152/2009, while rutin concentration was determined by the modified HPLC method. The lowest content of moisture was found in roots (4.3% and in peels (almost 8% and the highest moisture (nearly 11% was discovered in seeds. The lowest amount of crude protein (3.5% was found in peels, the highest crude protein amount (>13% in both flours and leaves (23%. The starch content (>50% in dry matter differs from one sample to another. Only in peels the content of starch was about 3.5%. From all examined samples, the lowest content of fat was found in crunchy products Cocoa, 1.7%. The lowest amount of histidine was determined in all studied samples, except peels, the highest content of glutamic acid was determined in almost all samples, except peels. Whole-meal flour is very rich source of Ca and Fe. The content of these elements was 1172 mg.kg-1 and 45.9 mg.kg-1, respectively. On the other hand, the highest content of Pb (>1 mg.kg-1 was found in broken seeds. The greatest concentration of rutin was determined in blossoms and leaves (83.6 and 69.9 mg.g-1, respectively. On the other hand, the lowest concentrations of rutin were found in buckwheat products (generally less then 1 mg.g-1, i.e. in wholemeal flour, 702 μg.kg-1, the lowest almost 10 μg.kg-1 in pasta.

The impact of nitration on the structure and immunogenicity of the major birch pollen allergen Bet v 1.0101.

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Chloé Ackaert

Full Text Available Allergy prevalence has increased in industrialized countries. One contributing factor could be pollution, which can cause nitration of allergens exogenously (in the air or endogenously (in inflamed lung tissue. We investigated the impact of nitration on both the structural and immunological behavior of the major birch pollen allergen Bet v 1.0101 to determine whether nitration might be a factor in the increased incidence of allergy. Bet v 1.0101 was nitrated with tetranitromethane. Immune effects were assessed by measuring the proliferation of specific T-cell lines (TCLs upon stimulation with different concentrations of nitrated and unmodified allergen, and by measurement of cytokine release of monocyte-derived dendritic cells (moDCs and primary DCs (primDCs stimulated with nitrated versus unmodified allergen. HPLC-MS, crystallography, gel electrophoresis, amino acid analysis, size exclusion chromatography and molecular dynamics simulation were performed to characterize structural changes after nitration of the allergen. The proliferation of specific TCLs was higher upon stimulation with the nitrated allergen in comparison to the unmodified allergen. An important structural consequence of nitration was oligomerization. Moreover, analysis of the crystal structure of nitrated Bet v 1.0101 showed that amino acid residue Y83, located in the hydrophobic cavity, was nitrated to 100%. Both moDCs and primDCs showed decreased production of TH1-priming cytokines, thus favoring a TH2 response. These results implicate that nitration of Bet v 1.0101 might be a contributing factor to the observed increase in birch pollen allergy, and emphasize the importance of protein modifications in understanding the molecular basis of allergenicity.

Buckwheat and CVD Risk Markers: A Systematic Review and Meta-Analysis

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Liangkui Li

2018-05-01

Full Text Available The effects of buckwheat intake on cardiovascular diseases (CVDs have not been systematically investigated. The aim of the present study was to comprehensively summarize studies in humans and animals, evaluating the impact of buckwheat consumption on CVD risk markers and to conduct a meta-analysis of relevant data. Thirteen randomized, controlled human studies, two cross-sectional human studies and twenty-one animal studies were identified. Using random-effects models, the weighted mean difference of post-intervention concentrations of blood glucose, total cholesterol and triglycerides were significantly decreased following buckwheat intervention compared with controls [differences in blood glucose: −0.85 mmol/L (95% CI: −1.31, −0.39, total cholesterol: 0.50 mmol/L (95% CI: −0.80, −0.20 and triglycerides: 0.25 mmol/L (95% CI: −0.49, −0.02]. Responses of a similar magnitude were seen in two cross-sectional studies. For animal studies, nineteen of twenty-one studies showed a significant reduction in total cholesterol of between 12% and 54%, and fourteen of twenty studies showed a significant reduction in triglycerides of between 2% and 74%. All exhibited high unexplained heterogeneity. There was inconsistency in HDL cholesterol outcomes in both human and animal studies. It remains unclear whether increased buckwheat intake significantly benefits other markers of CVD risk, such as weight, blood pressure, insulin, and LDL-cholesterol, and underlying mechanisms responsible for any effects are unclear.

Study on the high flavonoids mutants of tartarian buckwheat by radiation induced

International Nuclear Information System (INIS)

Li Guozhu; Shen Huifang

2011-01-01

Three different varieties of tartarian buckwheat seeds (Chuanqiao 1 # , Yu-621 and KP9920) were irradiated with various doses (100∼500 Gy) of 60 Co-γ ray and sowed. Ten, eight and six high flavonoid tartary buckwheat mutants were selected from three parent materials respectively. The flavonoid content of three parent materials were 8.33%, 10.18% and 9.80%. The range of flavonoid content of high flavonoids mutants for three parent materials 11.37%∼14.91%, 10.67%∼12.46% and 11.32%∼12.95% respectively. Cluster analysis was also carried out based on the agronomic traits and flavonoid content in the 27 tartary buckwheat materials (24 mutants and 3 parent material) The 27 materials were classified into four clusters by cluster analysis based on the agronomic traits and were classified into 5 groups based on flavonoid content. (authors)

Buckwheat seedling flavonoids do not undergo rapid turnover

International Nuclear Information System (INIS)

Margna, U.; Vainjaerv, T.

1981-01-01

Pulse-labelling with exogenous [1- 14 C]-L-phenylalanine showed that of all flavonoids of buckwheat seedling cotyledons only anthocyanins underwent a slow turnover (not more than 6-8 per cent a day) whilst the major flavonoids, rutin and four C-glycosylflavones, remained metabolically stable within at least 7 days. From excised hypocotyls pulse-labelled flavonoids (anthocyanins and rutin) began to disappear gradually beginning with the second day of incubation. As hypocotyls, however, during prolonged incubation soon began to autolyse a considerable part of that loss was obviously due to abnormal shifts in cell metabolic activities with a possibility that some microbial process was also involved. In hypocotyls of intact seedlings pulse-labelled rutin, similar to rutin in cotyledons, showed no signs of turnover. (author)

Quantification of major allergen parvalbumin in 22 species of fish by SDS-PAGE.

Science.gov (United States)

Kobayashi, Yukihiro; Yang, Tao; Yu, Cheng-Tao; Ume, Chiaki; Kubota, Hiroyuki; Shimakura, Kuniyoshi; Shiomi, Kazuo; Hamada-Sato, Naoko

2016-03-01

Fish is an important causative material of food allergy. Although the allergenicity of fish is considered to correlate with the content of parvalbumin, the major fish allergen, available information about the parvalbumin content in fish is limited. In this study, a simple and reliable quantification method for fish parvalbumin by SDS-PAGE was first established. Application of the SDS-PAGE method to 22 species of fish revealed a marked variation in parvalbumin content among fish. Furthermore, the parvalbumin content was found to be higher in dorsal white muscle than in ventral white muscle, in rostral part of white muscle than in caudal part of white muscle and in white muscle than in dark muscle. IgE reactivity of fish was roughly proportional to parvalbumin content. Interestingly, large-sized migratory fish, such as salmon, swordfish and tuna, were commonly very low in both parvalbumin content and IgE reactivity. Copyright © 2015 Elsevier Ltd. All rights reserved.

Development of hypo-allergenic apples: silencing of the major allergen Mal d 1 gene in "Elstar" apple and the effect of grafting

DEFF Research Database (Denmark)

Krath, Britta; Eriksen, Folmer Damsted; Pedersen, Bjarne H.

2009-01-01

Many people who are allergic to birch pollen are also allergic to apple fruit, due to cross-allergenicity. Since apples are the most extensively consumed fruit in Europe, it is highly relevant to develop a hypo-allergenic apple. Apples with significantly reduced levels of the allergen, Mal d 1, may...

The T Cell Response to Major Grass Allergens Is Regulated and Includes IL-10 Production in Atopic but Not in Non-Atopic Subjects

DEFF Research Database (Denmark)

Domdey, A.; Liu, A.; Millner, A.

2010-01-01

in allergen-specific responses. The aim was to determine whether major grass allergens induce production of suppressive cytokines in allergic and healthy subjects and to examine the inhibitory effect of these cytokines on allergic responses. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated...... from healthy and grass-allergic donors and stimulated with the major grass allergens Phl p 1 or Phl p 5. The effects of endogenous IL-10 and/or TGF-beta on proliferation and cytokine production were determined by use of blocking antibodies. In addition, the number of CD4(+)CD25(+) T cells...... in PBMCs in the two groups, but fewer cells from atopic donors were CD4(+)CD25(+)CCR4(+) and more cells were CD4(+)CD25(+)CLA(+) compared to healthy donors. Conclusion: Allergen-specific responses of grass allergic patients but not in non-atopic subjects are influenced by regulatory cytokines produced...

Buckwheat bran (Fagopyrum esculentum as partial replacement of corn and soybean meal in the laying hen diet

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Domenico Gatta

2012-01-01

Full Text Available The effect of partial substitution of corn (-20% and soybean meal (-10% with buckwheat bran (+30% (Fagopyrum esculentum Moench in the diet of ISA-Brown hens was investigated in sixteen 74-week old hens, housed in couple wire cages and submitted to a 16 h light:8 h dark photoperiod. The following traits were measured: body weight, egg production, egg mass, egg quality, feed intake, feed conversion, comparative palatability of ingredients and digestibility of diet. χ2 and non-parametric tests were used for production rate and yolk color score, respectively. ANOVA was used for all other parameters. Comparative choice of buckwheat, corn and soy was checked under different forms in 3 free choice tests. Results show that egg production rate (43.3% vs 50.5%; P<0.05 and feed intake (78.3±0.68 eggs/hen d vs 87.8±0.68 eggs/hen d; P<0.05 increased with the partial introduction of buckwheat bran in the diet. There was no difference in feed conversion between treatments. Nutrient balance confirmed that AMEn of diet was deeply lowered by the buckwheat bran use (6.5 MJ/kg vs 10.1 MJ/kg , due to the high fibre content of buckwheat bran (263 g/kg. Maize was always the most preferred ingredient, buckwheat bran was consumed more than expected in absence of any preference, and soybean was the food least chosen. Buckwheat bran can be used as an ingredient feed for low-producing laying hens; it induces a feed-intake increase, partially balanced by improved egg-production rates and a tendency to better albumen Haugh units.

Monoclonal antibody-based ELISA to quantify the major allergen of Cynodon dactylon (Bermuda grass) pollen, Cyn d 1.

Science.gov (United States)

Duffort, O; Calabozo, B; González, R; Carpizo, J A; Barber, D; Polo, F

2004-12-01

Pollen of Bermuda grass (Cynodon dactylon) is an important cause of pollinosis in many areas of the world. Most patients show sensitivity to the major allergen Cyn d 1, a glycoprotein composed of a number of isoforms with a molecular mass of 31-32 kDa. The aim of this work was to develop a monoclonal antibody (mAb)-based ELISA to quantify Cyn d 1, and to assess the correlation of the allergen content with the biological activity of C. dactylon pollen extracts. After fusion of myeloma cells with spleen cells from a BALB/c mouse immunized with C. dactylon pollen extract, Cyn d 1-specific mAbs secreting hybridomas were selected, and the antibodies characterized. One of them (4.4.1) was used as the capture antibody in an ELISA method for Cyn d 1 quantitation. An anti-Cyn d 1 rabbit serum was used as the second antibody. Cyn d 1 was purified by immunoaffinity chromatography with mAb 4.4.1, characterized, and used as the standard in the assay. The identity, purity and isoallergen composition of affinity-purified Cyn d 1 was confirmed by N-terminal amino acid sequencing, SDS-PAGE, Western blot and 2D electrophoresis. The Cyn d 1 ELISA is highly specific and sensitive, with a detection limit of 0.24 ng/ml and a linear range of 1.1-9.2 ng/ml. An excellent correlation was found when the content of Cyn d 1, measured in 16 different extracts, was compared with the allergenic activity of the same extracts determined by RAST inhibition. The results prove the usefulness of the Cyn d 1 ELISA for the standardization of C. dactylon-allergen products on the basis of major allergen content. 2004 S. Karger AG, Basel.

The Effect of Buckwheat Hull Extract on Lipid Oxidation in Frozen-Stored Meat Products.

Science.gov (United States)

Hęś, Marzanna; Szwengiel, Artur; Dziedzic, Krzysztof; Le Thanh-Blicharz, Joanna; Kmiecik, Dominik; Górecka, Danuta

2017-04-01

This study investigated the effect of antioxidants on lipid stability of frozen-stored meat products. Buckwheat hull extract was used to enrich fried meatballs made from ground pork. During 180-d storage of meat products, lipid oxidation (peroxide and 2-thiobarbituric acid reactive substances [TBARS] value) was periodically monitored. The results were compared with butylated hydroxytoluene (BHT). The addition of antioxidants decreased lipid oxidation in stored meatballs. The highest ability to control peroxide and TBARS values was demonstrated for buckwheat hull extract. Moreover, buckwheat hull extract showed a higher 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity as well as higher Fe(II) ion chelating ability, as compared with BHT. The total content of phenolic compounds are highly correlated to the individual polyphenols in extract of buckwheat hull, among which the following were assayed: 3,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, gallic acid, isovanillic acid and p-coumaric acid, and flavonoids: isoorientin, quercetin, quercetin 3-d-glucoside, rutin, and vitexin. These results indicate that plant extracts can be used to prolong shelf life of products by protecting them against lipid oxidation and deterioration of their nutritional quality. © 2017 Institute of Food Technologists®.

Combined effect of heat treatment and humidity on total polyphenol content of tartary buckwheat wholeflour

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Andrea BRUNORI

2016-12-01

Full Text Available Minor crops are gaining new interest due to the high content of bioactive compounds available in their grain and the consequent opportunity to be employed as ingredients for the production of healthy foodstuff. Tartary buckwheat (Fagopyrum tataricum Gaertn. grain is rich in flavonoids, the most important being represented by rutin, a compound possessing a high health value. When processing bakery products added with Tartary buckwheat whole flour, the key point is to prevent rutin from being hydrolysed to quercetin. In this view, a combination of heat treatment and controlled humidity level was applied for different lengths of time, in the attempt to deactivate the enzymes catalysing the reaction. Tartary buckwheat grain contains other polyphenols also capable to confer health properties. This class of compounds has been associated with the prevention of cardiovascular diseases, cancers, neurodegenerative diseases, diabetes, and osteoporosis. In this study it was observed how the physical treatments meant to preserve rutin would influence the overall content of polyphenols in Tartary buckwheat whole flour and dough.

PROTEIN COMPLEX OF WHEAT, BUCKWHEAT AND MAIZE IN RELATION TO CELIAC DISEASE

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Milan Chňapek

2014-02-01

Full Text Available Cereals are the most wide spread and very important plants utilized as a food source for mankind and for animals where they play role in energetical metabolism and proteosynthesis. Cereals contain proteins with unique properties. These properties allow us to produce leavened bread. Technological characteristic of cereal grain is determined by quantity and quality of storage proteins which represent alcohol soluble prolamins and glutenins soluble in acids and basis solutions. Celiac disease is one of the most frequent food intolerance caused by cereal storage proteins. Therapy consists of strict diet without consumptions of cereals or gluten. Pseudocereals are very perspective groups of plants in gluten free diet, due to absence of celiac active proteins, but on the other hand, flour from pseudocereals is not very suitable for baking. There are a lot of analytical methods applicable for detection of celiac active proteins in cereal and pseudocereal grain. Electrophoretical and immunochemical methods are the most utilized. Genotypes of wheat and maize were homogeneous and singlelined in contrast with genotypes of buckwheat. Average content of HMW-GS was highest in genotypes of bread wheat and lowest in buckwheat varieties. A celiac active fraction of storage proteins (LMW-GS and gliadins was detected at the highest content level in wheat genotypes. Genotypes of buckwheat and maize showed similar low content of this protein fraction. Presence of residual albumins and globulins in buckwheat varieties showed the highest value.

Study on the high flavonoids mutants of tartarian buckwheat by radiation induced mutation

International Nuclear Information System (INIS)

Li Guozhu; Shen Huifang

2012-01-01

Three different varieties of tartarian buckwheat seeds (Chuanqiao 1#, Yu-621 and KP9920) were irradiated with various doses (100∼500 Gy) of 60 Co-γ ray and sowed. Ten, eight and six high flavonoid tartary buckwheat mutants were selected from three parent materials respectively. The flavonoid content of three parent materials were 8.33%, 10.18% and 9.80%. The range of flavonoid content of high flavonoids mutants for three parent materials 11.37%∼14.91%, 10.67%∼12.46% and 11.32%∼12.95% respectively. Cluster analysis was also carried out based on the agronomic traits and flavonoid content in the 27 tartary buckwheat materials (24 mutants and 3 parent material) The 27 materials were classified into four clusters by cluster analysis based on the agronomic traits and were classified into 5 groups based on flavonoid content. (authors)

Analysis of major paralogs encoding the Fra a 1 allergen based on their organ-specificity in Fragaria × ananassa.

Science.gov (United States)

Ishibashi, Misaki; Nabe, Takeshi; Nitta, Yoko; Tsuruta, Hiroki; Iduhara, Miho; Uno, Yuichi

2018-03-01

Fra a 1 protein in strawberry causes oral allergic syndrome. Over 39 Fra a 1 paralogs have been identified in strawberry genome. Fra a 1.01 is major accumulating protein in edible organs. Strawberry fruits contain allergenic proteins that cause oral allergic syndrome. The hypothesized major allergen is Fra a 1, an ortholog of the birch pollen allergen protein Bet v 1. We organized Fra a 1 genes and analyzed their localizations at the transcriptional and translational levels. In total, 15 new Fra a 1 proteins were identified from the genomic database, increasing the total number of Fra a 1 to 30 proteins encoded by 39 genes. Fra a 1.02 was mostly expressed in receptacles, and Fra a 1.01 in achenes, when analyzed by RNA sequencing. Immunoblotting showed that the Fra a 1.01 protein was broadly accumulated in strawberry organs, while the Fra a 1.02 protein was mostly expressed in receptacles. Recombinant Fra a 1.01 strongly reacted with human IgE. The mRNA and protein expression levels of Fra a 1 did not correlate, indicating the importance of protein levels when evaluating the abundance of allergens in strawberry. Based on the localizations, accumulation levels and reactivity to human IgE, we determined that Fra a 1.01 was the most important allergen, followed by Fra a 1.02, and then other Fra a 1 proteins. The information obtained here will be useful for selecting the target Fra a 1 paralogs when breeding hypoallergenic strawberry.

Rabbit IgG directed to a synthetic C-terminal peptide of the major grass pollen allergen Lol p I inhibits human basophil histamine release induced by natural Lol p I

NARCIS (Netherlands)

van Ree, R.; Aalberse, R. C.

1995-01-01

The potential role of allergen-specific IgG antibodies as 'blocking' antibodies in allergen-induced human basophil histamine release was investigated. This was studied in a model with the major grass pollen allergen Lol p I and polyclonal rabbit antisera directed against this allergen and against a

HYMENOPTERA ALLERGENS: FROM VENOM TO VENOME

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Edzard eSpillner

2014-02-01

Full Text Available In Western Europe hymenoptera venom allergy primarily relates to venoms of the honeybee and the common yellow jacket. In contrast to other allergen sources, only a few major components of hymenoptera venoms had been characterized until recently. Improved expression systems and proteomic detection strategies have allowed the identification and characterization of a wide range of additional allergens. The field of hymenoptera venom allergy research has moved rapidly from focusing on venom extract and single major allergens to a molecular understanding of the entire venome as a system of unique and characteristic components. An increasing number of such components has been identified, characterized regarding function and assessed for allergenic potential. Moreover, advanced expression strategies for recombinant production of venom allergens allow selective modification of molecules and provide insight into different types of IgE reactivities and sensitization patterns. The obtained information contributes to an increased diagnostic precision in hymenoptera venom allergy and may serve for monitoring, reevaluation and improvement of current therapeutic strategies.

Responsiveness of the major birch allergen Bet v 1 scaffold to the gastric environment: Impact on structure and allergenic activity

DEFF Research Database (Denmark)

Sancho, Ana I; Wangorsch, Andrea; Jensen, Bettina M

2011-01-01

Four Bet v 1 homologous food allergens from celeriac (rApi g 1), apple (rMal d 1), peach (rPru p 1) and hazelnut (rCor a 1), were used to probe the structural responsiveness of the Bet v 1 scaffold to gastric digestion conditions and its impact on allergenicity....

Analysis of U.S. Food and Drug Administration food allergen recalls after implementation of the food allergen labeling and consumer protection act.

Science.gov (United States)

Gendel, Steven M; Zhu, Jianmei

2013-11-01

To avoid potentially life-threatening reactions, food allergic consumers rely on information on food labels to help them avoid exposure to a food or ingredient that could trigger a reaction. To help consumers in the United States obtain the information that they need, the Food Allergen Labeling and Consumer Protection Act of 2004 defined a major food allergen as being one of eight foods or food groups and any ingredient that contains protein from one of these foods or food groups. A food that contains an undeclared major food allergen is misbranded under the U.S. Food, Drug, and Cosmetic Act and is subject to recall. Food allergen labeling problems are the most common cause of recalls for U.S. Food and Drug Administration (FDA)-regulated food products. To help understand why food allergen recalls continue to occur at a high rate, information on each food allergen recall that occurred in fiscal years 2007 through 2012 was obtained from the FDA recall database. This information was analyzed to identify the food, allergen, root cause, and mode of discovery for each food allergen recall. Bakery products were the most frequently recalled food type, and milk was the most frequently undeclared major food allergen. Use of the wrong package or label was the most frequent problem leading to food allergen recalls. These data are the first reported that indicate the importance of label and package controls as public health measures.

Effects of high hydrostatic pressure on the structure and potential allergenicity of the major allergen bovine β-lactoglobulin.

Science.gov (United States)

Meng, Xuanyi; Bai, Yuxin; Gao, Jinyan; Li, Xin; Chen, Hongbing

2017-03-15

Bovine β-lactoglobulin (β-Lg) is recognized as a significant milk allergen in several countries. In this study, β-Lg was isolated and treated with high hydrostatic pressure (HHP) at 100, 200, 300, 400, and 500MPa. The allergenic properties of the HHP-treated β-Lg were characterized by indirect competitive enzyme-linked immunosorbent assay with anti-β-Lg rabbit antibody and the sera of patients allergic to cows' milk. The conformation of the HHP-treated β-Lg was examined with ultraviolet absorption spectroscopy, endogenous fluorescence spectroscopy, exogenous fluorescence spectroscopy, and circular dichroism spectroscopy analyses. The results indicated that IgG binding increased with treatment pressure, and IgE binding was lowest at 200MPa and highest at 400MPa. The tertiary structure of β-Lg changed significantly after HHP, whereas the primary and secondary structures remained stable. Overall, this study suggests that the conformational changes in HHP-treated β-Lg contribute to its altered allergenicity. Copyright © 2016 Elsevier Ltd. All rights reserved.

The current state of recombinant allergens for immunotherapy

DEFF Research Database (Denmark)

Pauli, Gabrielle; Malling, H-J

2010-01-01

Subcutaneous immunotherapy is a well documented treatment of allergic rhinitis and asthma. The majority of the disadvantages of the treatment are related to the poor quality of the natural allergen extracts which can contain varying amounts of individual allergens including allergens to which...

Solution structure of the major fish allergen parvalbumin Sco j 1 derived from the Pacific mackerel.

Science.gov (United States)

Kumeta, Hiroyuki; Nakayama, Haruka; Ogura, Kenji

2017-12-07

Although fish is an important part of the human diet, it is also a common source of food allergy. The major allergen in fish is parvalbumin, a well-conserved Ca 2+ -binding protein found in the white muscle of many fish species. Here, we studied the solution structure of the parvalbumin Sco j 1, derived from the Pacific mackerel, using nuclear magnetic resonance spectroscopy. We mapped the IgE-binding epitope proposed in a recent study onto the present structure. Interestingly, three of four residues, which were elucidated as key residues of the IgE-binding epitope, were exposed to solvent, whereas one residue faced the inside of the molecule. We expect that this solution structure can be used in future studies attempting to analyze the various IgE-binding modes of these allergens.

Identification of sole parvalbumin as a major allergen: study of cross-reactivity between parvalbumins in a Spanish fish-allergic population.

Science.gov (United States)

Perez-Gordo, M; Cuesta-Herranz, J; Maroto, A S; Cases, B; Ibáñez, M D; Vivanco, F; Pastor-Vargas, C

2011-05-01

Fish allergy is becoming an important health problem in Spain, a country with the third highest level of fish consumption after Japan and Portugal. The most common fish allergens are parvalbumins. In our area, the most widely consumed fish species are lean, such as whiff (Lepidorhombus whiffiagonis) and sole (Solea solea). Adverse reactions to fish are usually related to these species, a fact that is largely unknown to allergists in other countries. The aim of this study was to identify and purify the major allergen implicated in allergic response to sole and evaluate the IgE cross-reactivity of purified parvalbumins from whiff and sole, which are phylogenetically close, and more distant species (i.e. cod and salmon). Eighteen Spanish fish-allergic patients with a positive history of type I allergy to fish were recruited from the clinic. Total protein extracts and purified parvalbumins from whiff and sole were tested for their IgE-binding properties by combining two-dimensional Western blotting and mass spectrometry. The extent of cross-reactivity between these parvalbumins along with cod and salmon parvalbumins was investigated by IgE ELISA inhibition assay. An IgE-binding spot of approximately 14 kDa was identified as parvalbumin and confirmed as a major allergen in sole extract, which is recognized by almost 70% of the patients. Whiff parvalbumin was recognized by 83.4% of the patients. High cross-reactivity was determined for all purified parvalbumins by IgE inhibition assay. Sole and whiff parvalbumin were confirmed as major allergens. The parvalbumins of sole, whiff, cod and salmon were highly cross-reactive, thus suggesting a high amino acid sequence identity between them. © 2011 Blackwell Publishing Ltd.

The high molecular weight dipeptidyl peptidase IV Pol d 3 is a major allergen of Polistes dominula venom.

Science.gov (United States)

Schiener, Maximilian; Hilger, Christiane; Eberlein, Bernadette; Pascal, Mariona; Kuehn, Annette; Revets, Dominique; Planchon, Sébastien; Pietsch, Gunilla; Serrano, Pilar; Moreno-Aguilar, Carmen; de la Roca, Federico; Biedermann, Tilo; Darsow, Ulf; Schmidt-Weber, Carsten B; Ollert, Markus; Blank, Simon

2018-01-22

Hymenoptera venom allergy can cause severe anaphylaxis in untreated patients. Polistes dominula is an important elicitor of venom allergy in Southern Europe as well as in the United States. Due to its increased spreading to more moderate climate zones, Polistes venom allergy is likely to gain importance also in these areas. So far, only few allergens of Polistes dominula venom were identified as basis for component-resolved diagnostics. Therefore, this study aimed to broaden the available panel of important Polistes venom allergens. The 100 kDa allergen Pol d 3 was identified by mass spectrometry and found to be a dipeptidyl peptidase IV. Recombinantly produced Pol d 3 exhibited sIgE-reactivity with approximately 66% of Polistes venom-sensitized patients. Moreover, its clinical relevance was supported by the potent activation of basophils from allergic patients. Cross-reactivity with the dipeptidyl peptidases IV from honeybee and yellow jacket venom suggests the presence of exclusive as well as conserved IgE epitopes. The obtained data suggest a pivotal role of Pol d 3 as sensitizing component of Polistes venom, thus supporting its status as a major allergen of clinical relevance. Therefore, Pol d 3 might become a key element for proper diagnosis of Polistes venom allergy.

Purification, crystallization and preliminary X-ray characterization of prunin-1, a major component of the almond (Prunus dulcis) allergen amandin.

Science.gov (United States)

Albillos, Silvia M; Jin, Tengchuan; Howard, Andrew; Zhang, Yuzhu; Kothary, Mahendra H; Fu, Tong-Jen

2008-07-09

The 11S globulins from plant seeds account for a number of major food allergens. Because of the interest in the structural basis underlying the allergenicity of food allergens, we sought to crystallize the main 11S seed storage protein from almond ( Prunus dulcis). Prunin-1 (Pru1) was purified from defatted almond flour by water extraction, cryoprecipitation, followed by sequential anion exchange, hydrophobic interaction, and size exclusion chromatography. Single crystals of Pru1 were obtained in a screening with a crystal screen kit, using the hanging-drop vapor diffusion method. Diffraction quality crystals were grown after optimization. The Pru1 crystals diffracted to at least 3.0 A and belong to the tetragonal space group P4(1)22, with unit cell parameters of a = b = 150.912 A, c = 165.248 A. Self-rotation functions and molecular replacement calculations showed that there are three molecules in the asymmetry unit with water content of 51.41%. The three Pru1 protomers are related by a noncrystallographic 3-fold axis and they form a doughnut-shaped trimer. Two prunin trimers form a homohexamer. Elucidation of prunin structure will allow further characterization of the allergenic features of the 11S protein allergens at the molecular level.

Purification, Crystallization and Preliminary X-ray Characterization of Prunin-1, a Major Component of the Almond (Prunus dulcis) Allergen Amandin

Energy Technology Data Exchange (ETDEWEB)

Albillos, Silvia M.; Jin, Tengchuan; Howard, Andrew; Zhang, Yuzhu; Kothary, Mahendra H.; Fu, Tong-Jen (IIT); (US-FDA)

2008-08-04

The 11S globulins from plant seeds account for a number of major food allergens. Because of the interest in the structural basis underlying the allergenicity of food allergens, we sought to crystallize the main 11S seed storage protein from almond (Prunus dulcis). Prunin-1 (Pru1) was purified from defatted almond flour by water extraction, cryoprecipitation, followed by sequential anion exchange, hydrophobic interaction, and size exclusion chromatography. Single crystals of Pru1 were obtained in a screening with a crystal screen kit, using the hanging-drop vapor diffusion method. Diffraction quality crystals were grown after optimization. The Pru1 crystals diffracted to at least 3.0 {angstrom} and belong to the tetragonal space group P4{sub 1}22, with unit cell parameters of a = b = 150.912 {angstrom}, c = 165.248 {angstrom}. Self-rotation functions and molecular replacement calculations showed that there are three molecules in the asymmetry unit with water content of 51.41%. The three Pru1 protomers are related by a noncrystallographic 3-fold axis and they form a doughnut-shaped trimer. Two prunin trimers form a homohexamer. Elucidation of prunin structure will allow further characterization of the allergenic features of the 11S protein allergens at the molecular level.

Rheological properties and mineral content of buckwheat enriched wholegrain wheat pasta

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Nedeljković Nataša

2014-01-01

Full Text Available Light buckwheat flour (LBF was used to substitute 20% of whole wheat flour (WWF in the formulation of wholegrain wheat pasta. Wholegrain wheat pasta (WWP and buckwheat enriched wholegrain wheat pasta (BWWP were produced on an industrial scale. Substitution level of buckwheat flour (20% was based on previously conducted rheological tests on LBF/WWF blends which were performed using 10, 20 and 30% of LBF. The obtained Mixolab profiles have indicated that wheat blend containing 20% LBF expressed the most similar rheological parameters to WWF. Proximate composition, cooking quality and mineral content of BWWP were analyzed and compared with those of WWP. The substitution of WWF with LBF in the pasta formulation resulted in significantly increased (P < 0.05 contents of P, Mg, K and Zn compared to WWP in dry pasta. The reduction in mineral content of BWWP during cooking was significantly higher (P < 0.05 compared to WWP. The content of P, Mg and K were at same level in both type of pasta after cooking. The obtained results suggest that enrichment of WWP with LBF at the level of 20% did not improve the mineral content of cooked pasta, although increase in minerals was observed in dry pasta. [Projekat Ministarstva nauke Republike Srbije, br. TR31029

Determination of allergen specificity by heavy chains in grass pollen allergen-specific IgE antibodies.

Science.gov (United States)

Gadermaier, Elisabeth; Flicker, Sabine; Lupinek, Christian; Steinberger, Peter; Valenta, Rudolf

2013-04-01

Affinity and clonality of allergen-specific IgE antibodies are important determinants for the magnitude of IgE-mediated allergic inflammation. We sought to analyze the contribution of heavy and light chains of human allergen-specific IgE antibodies for allergen specificity and to test whether promiscuous pairing of heavy and light chains with different allergen specificity allows binding and might affect affinity. Ten IgE Fabs specific for 3 non-cross-reactive major timothy grass pollen allergens (Phl p 1, Phl p 2, and Phl p 5) obtained by means of combinatorial cloning from patients with grass pollen allergy were used to construct stable recombinant single chain variable fragments (ScFvs) representing the original Fabs and shuffled ScFvs in which heavy chains were recombined with light chains from IgE Fabs with specificity for other allergens by using the pCANTAB 5 E expression system. Possible ancestor genes for the heavy chain and light chain variable region-encoding genes were determined by using sequence comparison with the ImMunoGeneTics database, and their chromosomal locations were determined. Recombinant ScFvs were tested for allergen specificity and epitope recognition by means of direct and sandwich ELISA, and affinity by using surface plasmon resonance experiments. The shuffling experiments demonstrate that promiscuous pairing of heavy and light chains is possible and maintains allergen specificity, which is mainly determined by the heavy chains. ScFvs consisting of different heavy and light chains exhibited different affinities and even epitope specificity for the corresponding allergen. Our results indicate that allergen specificity of allergen-specific IgE is mainly determined by the heavy chains. Different heavy and light chain pairings in allergen-specific IgE antibodies affect affinity and epitope specificity and thus might influence clinical reactivity to allergens. Copyright © 2012 American Academy of Allergy, Asthma & Immunology. Published by

Buckwheat and quinoa seeds as supplements in wheat bread production

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Demin Mirjana A.

2013-01-01

Full Text Available The aim of this work was to compare the nutritional characteristics of wheat bread with the bread produced of wheat flour supplemented with quinoa and buckwheat seeds. Bread making properties of these blends were analyzed in order to investigate their ability to make moulded bread. Quinoa (Chenopodium quinoa Will. and buckwheat seeds were grown in the vicinity of Belgrade, Serbia. The addition of pseudocereal seeds (at levels of 30% and 40% and a selected technological process, which included hydrothermal preparation of supplements, resulted with a valuable effect on nutritive value of breads. In comparison with the wheat bread that was used as control sample, the protein increase of 2% and the increase of crude fiber content at around 0.5% in 30% supplemented breads were registered. Furthermore, the incorporation of both seeds mixture at the level of 40%, increased the content of protein for 2.5% and fiber content for 0.4%. In regard to the starch, fat, and ash contents there were no major differences. The investigated breads were nutritionally superior to the wheat bread. Chemical composition of the selected seeds was also investigated. The results showed that the blends containing either 30% or 40% of selected seeds expressed high potential for the production of molded breads, as new baking products with enhanced nutritional composition. The applied technological procedure was modified in such way that for all blended combination of supplements it changed rheological properties of dough. Furthermore, it resulted in a good volume of breads with excellent sensory properties of aroma-odor and taste.

Animal Allergens and Their Presence in the Environment

Science.gov (United States)

Zahradnik, Eva; Raulf, Monika

2014-01-01

Exposure to animal allergens is a major risk factor for sensitization and allergic diseases. Besides mites and cockroaches, the most important animal allergens are derived from mammals. Cat and dog allergies affect the general population; whereas, allergies to rodents or cattle is an occupational problem. Exposure to animal allergens is not limited to direct contact to animals. Based on their aerodynamic properties, mammalian allergens easily become airborne, attach to clothing and hair, and can be spread from one environment to another. For example, the major cat allergen Fel d 1 was frequently found in homes without pets and in public buildings, including schools, day-care centers, and hospitals. Allergen concentrations in a particular environment showed high variability depending on numerous factors. Assessment of allergen exposure levels is a stepwise process that involves dust collection, allergen quantification, and data analysis. Whereas a number of different dust sampling strategies are used, ELISA assays have prevailed in the last years as the standard technique for quantification of allergen concentrations. This review focuses on allergens arising from domestic, farm, and laboratory animals and describes the ubiquity of mammalian allergens in the human environment. It includes an overview of exposure assessment studies carried out in different indoor settings (homes, schools, workplaces) using numerous sampling and analytical methods and summarizes significant factors influencing exposure levels. However, methodological differences among studies have contributed to the variability of the findings and make comparisons between studies difficult. Therefore, a general standardization of methods is needed and recommended. PMID:24624129

Animal allergens and their presence in the environment

Directory of Open Access Journals (Sweden)

Eva eZahradnik

2014-03-01

Full Text Available Exposure to animal allergens is a major risk factor for sensitization and allergic diseases. Besides mites and cockroaches, the most important animal allergens are derived from mammals. Cat and dog allergies affect the general population; whereas, allergies to rodents or cattle is an occupational problem. Exposure to animal allergens is not limited to direct contact to animals. Based on their aerodynamic properties, mammalian allergens easily become airborne, attach to clothing and hair, and can be spread from one environment to another. For example, the major cat allergen Fel d 1 was frequently found in homes without pets and in public buildings, including schools, day care centers and hospitals. Allergen concentrations in a particular environment showed high variability depending on numerous factors.Assessment of allergen exposure levels is a stepwise process that involves dust collection, allergen quantification and data analysis. Whereas a number of different dust sampling strategies are used, ELISA assays have prevailed in the last years as the standard technique for quantification of allergen concentrations. This review focuses on allergens arising from domestic, farm and laboratory animals and describes the ubiquity of mammalian allergens in the human environment. It includes an overview of exposure assessment studies carried out in different indoor settings (homes, schools, workplaces using numerous sampling and analytical methods and summarizes significant factors influencing exposure levels. However, methodological differences among studies have contributed to the variability of the findings and make comparisons between studies difficult. Therefore, a general standardization of methods is needed and recommended.

Anti-Fatigue Properties of Tartary Buckwheat Extracts in Mice

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Ping Wei

2011-07-01

Full Text Available Anti-fatigue properties of tartary buckwheat extracts (TBE was investigated in male Kunming mice. The animals were divided into four groups. The first group, designated as the control group (control, was administered with distilled water by gavage every day for 28 days. The other three groups, designated as TBE treatment groups, were administered with TBE of 60, 120 and 240 mg/kg body weight, respectively, by gavage every day for 28 days. Exhaustive swimming time, blood lactic acid (BLA, blood urea nitrogen (BUN, tissue glycogen, glutathione peroxidase (GPx and superoxide dismutase (SOD of mice after swimming were determined. The results showed that tartary buckwheat extracts had anti-fatigue properties, which extended the exhaustive swimming time of mice, effectively inhibiting the increase of BLA, decreasing the level of BUN, increasing the tissue glycogen content and the activities of SOD and GPx of mice. However, further study is needed to elucidate the exact mechanism of the effect of TBE on fatigue.

Specific B-cell Epitope of Per a 1: A Major Allergen of American Cockroach (Periplaneta americana) and Anatomical Localization.

Science.gov (United States)

Sookrung, Nitat; Khetsuphan, Thanyathon; Chaisri, Urai; Indrawattana, Nitaya; Reamtong, Onrapak; Chaicumpa, Wanpen; Tungtrongchitr, Anchalee

2014-07-01

Cockroach (CR) is a common source of indoor allergens, and Per a 1 is a major American CR (Periplaneta americana) allergen; however, several attributes of this protein remain unknown. This study identifies a novel specific B cell epitope and anatomical locations of Per a 1.0105. Recombinant Per a 1.0105 (rPer a 1.0105) was used as BALB/c mouse immunogen for the production of monoclonal antibodies (MAb). The MAb specific B cell epitope was identified by determining phage mimotopic peptides and pair-wise alignment of the peptides with the rPer a 1.0105 amino acid sequence. Locations of the Per a 1.0105 in P. americana were investigated by immunohistochemical staining. The rPer a 1.0105 (~13 kDa) had 100%, 98% and ≥90% identity to Per a 1.0105, Per a 1.0101, and Cr-PII, respectively. The B-cell epitope of the Per a 1.0105 specific-MAb was located at residues(99) QDLLLQLRDKGV(110) contained in all 5 Per a 1.01 isoforms and Per a 1.02. The epitope was analogous to the Bla g 1.02 epitope; however, this B-cell epitope was not an IgE inducer. Per a 1.0105 was found in the midgut and intestinal content of American CR but not in the other organs. The amount of the Per a 1 was ~544 ℃g per gram of feces. The novel Per a 1 B-cell epitope described in this study is a useful target for allergen quantification in samples; however, the specific MAb can be used as an allergen detection reagent. The MAb based-affinity resin can be made for allergen purification, and the so-purified protein can serve as a standard and diagnostic allergen as well as a therapeutic vaccine component. The finding that the Per a 1 is contained in the midgut and feces is useful to increase yield and purity when preparing this allergen.

Optimization of bioactive compounds in buckwheat sprouts and their effect on blood cholesterol in hamsters.

Science.gov (United States)

Lin, Li-Yun; Peng, Chiung-Chi; Yang, Ya-Lu; Peng, Robert Y

2008-02-27

Nutrient levels in buckwheats that were maximized in day 8 sprouts (D8SP) included total phenolics, quercetin, and l-ascorbic acid, whereas those of oxalic, malic, tartaric, and citric acids, rutin, and gamma-aminobutyric acid (GABA) were found to reach maximum levels on day 10. Ethanolic extract of D8SP (2.5 mg/mL) revealed potent free-radical scavenging (FRS) and antioxidative (ANO) capabilities. However, its Fe2+-chelating capability was only moderate. To further study the hypolipidemic activity of D8SP, 36 Syrian hamsters were grouped into six groups and fed for 28 days, respectively, with (i) control meal, (ii) high fat plus high cholesterol meal, (iii) high fat plus high cholesterol plus 2.5% of buckwheat seeds, (iv) high fat plus high cholesterol plus 25% of buckwheat seeds, (v) high fat plus high cholesterol plus 2.5% of D8SP, and (vi) high fat plus high cholesterol plus 25% of D8SP. High seed meal prominently enhanced body weight gain, whereas high sprout meal exhibited the highest feed efficiency. Ratios of liver/body weight (L/B) were significantly lowered by all BS meals. Although low seed meal reduced serum total cholesterol (TC) levels (pcholesterol (LDL-C) were significantly suppressed by all buckwheat meals (pcholesterol (HDL-C) levels were increased, however, insignificantly. Nutraceutically more meaningful is that both LDL-C/HDL-C and TC/HDL-C ratios were significantly lowered (p<0.01). Apparently, hepatic TC levels were significantly reduced, whereas hepatic TG levels were totally unaffected. Conclusively, sprouting triggers a variety of nutritional changes in buckwheats. Day 8 sprouts, consisting of high polyphenolic and moderate quercetin contents, are nutraceutically maximized when hypocholesterolemic, hypotriglyceridemic, and antioxidative activities are concerned.

Characterization of parvalbumin, the major allergen in Alaska pollack, and comparison with codfish Allergen M.

Science.gov (United States)

Van Do, Thien; Hordvik, Ivar; Endresen, Curt; Elsayed, Said

2005-02-01

Increased fish consumption has led to frequent reporting of fish allergy and adverse reactions. Alaska pollack (Theragra chalcogramma) is a globally important commercial fish species, belonging to the Gadidae family. This family of fish also includes cod whose parvalbumin, Allergen M (Gad c 1), has been thoroughly studied and considered as a reference to sensitization in fish allergy. In the present study, parvalbumin from Alaska pollack, designated The c 1, was purified by use of anion exchange chromatography. To demonstrate the homogeneity of the purified protein, reverse phase high performance liquid chromatography was performed and showed two distinct fractions which had similar IgG and IgE binding capacities. Accordingly, cDNA cloning revealed two isotypic parvalbumin transcripts in pollack muscle. Recombinant parvalbumins of pollack exhibited low IgG and IgE binding capacities, in contrast to the native counterparts, which were almost as potent as cod Gad c 1. The allergenicity of The c 1 was assayed by ELISA inhibition, and compared to cod, the concentration required for obtaining 50% ELISA inhibition (C 50%) was only 18% higher for The c 1.

QUALITY OF GLUTEN-FREE BUCKWHEAT-RICE BREAD

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Petra Dvořáková

2013-02-01

Full Text Available In case of celiac disease the products containing gluten must be excluded from the nutrition. The offer of gluten-free products (especially pastry is low and in addition the gluten-free breads are typical of dry crust and crumb and higher firmness in comparison with wheat bread. This work deals with gluten-free mixtures prepared from buckwheat and rice flour and the effect of rising amount of these flours on bread quality, crumb hardness, elasticity, chewiness and gumminess. With rising portion of buckwheat flour in the mixture the bread volume, dough and bread yield increased. The biggest improvement was found for mean bread volume (30% between the samples FO 1090 (166.7 cm3 and FO 9010 (216.7 cm3. The texture analysis showed positive effect of rice flour on hardness, chewiness and gumminess. Hardness decreased from 114.5 N (F 100 to 91.3 N (FO 1090. Very similar results showed chewiness and gumminess. Chewiness of F 100 (314.0 was reduced by 32.5% to 212.2 at the sample FO 1090. Gumminess was improved almost linearly through the samples, the biggest difference (44.3% was found between the check sample F 100 (88.3 and FO 1090 (49.7.

Biologically Active Substances in Buckwheat ( Fagopyrum tataricum L. Cul- tivated in Mongolia

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S. Badamtsetseg

2006-06-01

Full Text Available From thoroughly air-dried samples of buckwheat plant, we revealed a biochemical composition of 14 components. By thin layer chromatography and quantitative analysis methods we showed that buck- wheat has 7 kinds of alkaloids with one dominating alkaloid, and the total weight of all alkaloids equals 0.05%. We also determined the aboveground parts of buckwheat contain the following substances: rutin- 3.14%, fat-0.91%, protein-8.23%, carbohydrate-18.52%, monosaccharide-0.37%, disaccharides-1.11%, vitamin C-0.02%, ash-10.57%, acidity-0.05, carotine-15.6mg, cellulose-40.8%, tannin-1.70%, soluble pectins-0.266%, insoluble pectins-0.507%, total amount of alkaloids-0.05%.

Effects of Aeration Treatment on γ-Aminobutyric Acid Accumulation in Germinated Tartary Buckwheat (Fagopyrum tataricum

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Yuanxin Guo

2016-01-01

Full Text Available To explore the optimum condition of γ-aminobutyric acid (GABA accumulation in germinated tartary buckwheat, effects of some factors including aeration treatment, physiological indexes, air flow rate, culture temperature, and pH value of cultivating solution under hypoxia on GABA in germinated tartary buckwheat were investigated. The results showed that the dark cultures with distilled water at 30°C, 2 days, and aeration stress with 1.0 L/min air flow rate at 30°C were optimal for GABA accumulation. Under these conditions, the predicted content of GABA was up to 371.98 μg/g DW. The analysis of correlation indicated that there was a significant correlation (P<0.01 between GABA accumulation and physiological indexes. Box-Behnken experimental analysis revealed that optimal conditions with aeration treatment for GABA accumulation in germinated tartary buckwheat were air flow rate of 1.04 L/min, culture temperature of 31.25°C, and a pH value of 4.21. Under these conditions, the GABA content was predicted as high as 386.20 μg/g DW, which was close to the measured value (379.00±9.30 μg/g DW. The variance analysis and validation test suggested that this established regression model could predict GABA accumulation in tartary buckwheat during germination.

Mountain cedar allergens found in nonpollen tree parts.

Science.gov (United States)

Goetz, D W; Goetz, M A; Whisman, B A

1995-09-01

Mountain cedar (Juniperus ashei) pollen is the principal aeroallergen in south central Texas from late December through February. The major mountain cedar allergen is a 40-kD glycoprotein, gp40. To identify allergens in mountain cedar wood, leaves, and berries and to detect mountain cedar allergen in smoke from burning male or female trees. SDS-PAGE plus mountain cedar human sIgE and monoclonal antibody immunoblots identified mountain cedar allergens within pollen and nonpollen tree part extracts. IgE immunoblots identified a single wood allergen at 36 kD and three berry allergens at 36, 26-27, and 21 kD, in addition to known pollen allergens. Mountain cedar monoclonal antibody bound an allergen epitope present not only on 40, 33, and 28-kD pollen allergens, but also on 36 and 32-kD wood allergens, and the 26-27-kD berry allergen. Immunoblot studies detected no mountain cedar allergen in leaves and no allergen in smoke from burning male and female trees. Allergens constituted a much smaller percentage of extractable protein in wood and berries than in pollen. Mountain cedar berry allergen content is too small to give credence to the ingestion of berries as a folk medicine treatment of mountain cedar pollinosis. In addition, while smoke from burning mountain cedar trees may be irritating, it contains no allergens that could cause allergic rhinoconjunctivitis.

Risk assessment of buckwheat flour contaminated by thorn-apple (Datura stramonium L.) alkaloids: a case study from Slovenia.

Science.gov (United States)

Perharič, Lucija; Koželj, Gordana; Družina, Branko; Stanovnik, Lovro

2013-01-01

In Slovenia, a mass poisoning incident involving 73 consumers with symptoms such as dry mouth, hot red skin, blurred vision, tachycardia, urinary retention, ataxia, speech disturbance, disorientation and visual hallucinations occurred in 2003. In all cases, consumers had eaten buckwheat flour food products within the last few hours. Investigations by responsible authorities identified the contamination of a range of buckwheat food products with thorn-apple (Datura stramonium L.) seeds containing toxic alkaloids, atropine and scopolamine. To ensure the safe consumption of buckwheat food products, we carried out risk characterisation and proposed provisional maximum residue levels (MRLs) of atropine and scopolamine mixture in buckwheat flour. In the absence of critical "no observed adverse effect levels" for atropine and scopolamine, we based our estimation of the acute reference doses on the lowest recommended therapeutic doses. Taking into account the additive effect of the two alkaloids, we calculated acute reference doses of the mixture, that is 0.05 µg/kg of body mass for atropine and 0.03 µg/kg of body mass for scopolamine. MRLs for atropine and scopolamine mixture in buckwheat flour were estimated in a worst-case scenario, that is consumption of 100 g of flour by a child weighing 10 kg and taking into account a range of atropine/scopolamine ratio in implicated food products, that is 0.85-3.3. We proposed the national MRLs for atropine/scopolamine mixture in buckwheat food products: 4.0 µg/kg (atropine) and 2.0 µg/kg(scopolamine). However, in view of the large variability in the alkaloid content, depending on the origin of the Datura, we propose that risk assessment should be carried out on a case-by-case basis, taking into account the ratio between atropine and scopolamine content in a particular sample.

Formation of disulfide bonds and homodimers of the major cat allergen Fel d 1 equivalent to the natural allergen by expression in Escherichia coli.

Science.gov (United States)

Grönlund, Hans; Bergman, Tomas; Sandström, Kristofer; Alvelius, Gunvor; Reininger, Renate; Verdino, Petra; Hauswirth, Alexander; Liderot, Karin; Valent, Peter; Spitzauer, Susanne; Keller, Walter; Valenta, Rudolf; van Hage-Hamsten, Marianne

2003-10-10

Dander from the domestic cat (Felis domesticus) is one of the most common causes of IgE-mediated allergy. Attempts to produce tetrameric folded major allergen Fel d 1 by recombinant methods with structural features similar to the natural allergen have been only partially successful. In this study, a recombinant folded Fel d 1 with molecular and biological properties similar to the natural counterpart was produced. A synthetic gene coding for direct fusion of the Fel d 1 chain 2 N-terminally to chain 1 was constructed by overlapping oligonucleotides in PCR. Escherichia coli expression resulted in a non-covalently associated homodimer with an apparent molecular mass of 30 kDa defined by size exclusion chromatography. Furthermore, each 19,177-Da subunit displayed a disulfide pattern identical to that found in the natural Fel d 1, i.e. Cys3(1) Cys73(2), Cys44(1)-Cys48(2), Cys70(1)-Cys7(2), as determined by electrospray mass spectrometry after tryptic digestion. Circular dichroism analysis showed identical folds of natural and recombinant Fel d 1. Furthermore, recombinant Fel d l reacted specifically with serum IgE, inducing expression of CD203c on basophils and lymphoproliferative responses in cat-allergic patients. The results show that the overall fold and immunological properties of the recombinant Fel d 1 are very similar to those of natural Fel d 1. Moreover, the recombinant Fel d 1 construct provides a tool for defining the three-dimensional structure of Fel d 1 and represents a reagent for diagnosis and allergen-specific immunotherapy of cat allergy.

Potential of sunflower, castor bean, common buckwheat and vetiver as lead phytoaccumulators

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Jailson do C. Alves

2016-03-01

Full Text Available ABSTRACT Studies concerning the tolerance, absorption and distribution of heavy metals in plants are essential for the success of phytoremediation programs. The present study was carried out in order to evaluate the potential of the sunflower, castor bean, common buckwheat and vetiver as lead phytoaccumulators. The species were grown in nutrient solution containing increasing doses of Pb (0, 50, 100, 200 and 400 mg L-1 during a 30-day exposure period. A completely randomized split-plot design was used, with a 4 x 5 factorial and three replicates. Significant reductions of dry matter of the root, shoot and whole plant were found in the all species under study as a function of the increased Pb doses. Vetiver showed higher tolerance to Pb contamination; sunflower and castor bean had intermediate tolerance and the common buckwheat proved to be the most sensitive species. The concentration and total content of Pb in plant compartments were significantly affected by the increased Pb doses in solution, and higher accumulation of this element was observed, in general, in the roots of the studied species. Common buckwheat proved to be not much promising for Pb-phytoremediation programs; sunflower showed potential for Pb phytoextraction and castor bean and vetiver were the most appropriate for Pb phytostabilization.

Characterization of a monoclonal antibody (P40) against the 68 kD major allergen of Penicillium notatum.

Science.gov (United States)

Shen, H D; Choo, K B; Chen, J H; Lin, W L; Chang, Z N; Han, S H

1992-04-01

A monoclonal antibody (MoAb P40) against the 68 kD major allergen of Penicillium notatum (P. notatum) was obtained by immunizing the mouse with a crude extract of P. notatum. Analysed by two-dimensional gel electrophoresis and immunoblotting, P40 reacted with two different isoforms of the 68 kD component of P. notatum with pIs of 5.4 and 5.5. In addition to P. notatum, P40 showed positive ELISA activity to Aspergillus fumigatus (A. fumigatus) but not to components of six other fungi including Alternaria porri, Cladosporium cladosporoides, Aureobasidium pullulans, Fusarium solani, Rhizopus arrhizus and Candida albicans. Analysed by ELISA, MoAb P40 also showed positive activity to two (P. frequentans and P. roseopurpureum) of the 10 other Penicillium species and two (A. terreus and A. flavus) of the four other Aspergillus species tested. SDS-PAGE and immunoblotting studies demonstrated P40 positive reactivity to components with MW of about 67 kD in all these Penicillium and Aspergillus species with positive ELISA activity to P40. Furthermore, immunoblotting activity of MoAb P40 to the 67 kD component of A. niger was also observed. The epitope of the 68 kD allergen of P. notatum recognized by MoAb P40 was resistant to treatment of periodate oxidation with concentration of NaIO4 up to 20 mM. This MoAb may thus be useful in the characterization and purification of the 68 kD allergen from crude extracts, and in the molecular cloning of allergen genes.

Widespread IgE-mediated hypersensitivity in the Sudan to the 'green nimitti' midge, Cladotanytarsus lewisi (Diptera: Chironomidae) II. Identification of a major allergen.

Science.gov (United States)

Gad El Rab, M O; Thatcher, D R; Kay, A B

1980-01-01

A major allergen has been identified in an aqueous extract of the 'green nimitti' midge, Cladotanytarsus lewisi (Diptera: Chironomidae). Following chromatography on Sephadex G-100 allergenic activity, as assessed by skin ('prick') testing, eluted as two closely related peaks (pools I and II) at about 50% bed volume. When these pools were applied separately to columns of CM-cellulose, activity in each eluted with 0 . 05 M NaCl. Isoelectric focusing of the unfractionated allergen gave a single peak of activity at pI 4 . 3. By SDS--PAGE, biological activity in the whole 'green nimitti' extract and the material eluting from both pools I and II of the Sephadex G-100 column migrated to the same positions and were associated with a molecular size of 15,000--20,000 daltons. Skin test reactivity of the unfractionated material and the Sephadex G-100 pool I and II eluates were all destroyed following incubation with trypsin, chymotrypsin, thermolysin and neuraminidase. These experiments indicate that a major allergen derived from the 'green nimitti' midge, a cause of widespread and severe immediate-type allergy in the Sudan, is an acidic glycoprotein of 15,000--20,000 molecular weight. Images Fig. 3 PMID:7438559

Combined Effects of Blue and Ultraviolet Lights on the Accumulation of Flavonoids in Tartary Buckwheat Sprouts

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Ji Hongbin

2016-06-01

Full Text Available The effects of blue and UV-A (365 nm/UV-C (254 nm or their combinations on the levels of total flavonoids, rutin, quercetin, phenylalanine ammonialyase (PAL, chalcone isomerase (CHI, rutin degrading enzymes (RDEs and 1,1-diphenyl-2-picrylhydrazyl (DPPH radical scavenging activity in tartary buckwheat sprouts were investigated in this study. The total flavonoids content in the tartary buckwheat sprouts irradiated with blue light followed by UV-C (BL+UV-C raised by 10%, compared with the opposite combination sequence (UV-C+BL. However, blue light did not show the same results when combined with UV-A, and their combinations on the accumulation of total flavonoids were still lower than that of UV-A/UV-C. Key enzymes (PAL, CHI and RDEs revealed a significant correlation with total flavonoids in tartary buckwheat sprouts.

Evaluation of In Vitro Inhibitory Activity of Rye-Buckwheat Ginger Cakes with Rutin on the Formation of Advanced Glycation End-Products (AGEs)

OpenAIRE

Przygodzka Małgorzata; Zieliński Henryk

2015-01-01

In this study, the relationship between the inhibitory effects of extracts from rye-buckwheat ginger cakes supplemented with low and high rutin dosage baked without or with dough fermentation step on the formation of fluorescent advanced glycation end-products (AGEs), and phenolic compounds, rutin, D-chiro-inositol and antioxidant capacity were addressed. The cakes were based on rye flour substituted by light buckwheat flour or flour from roasted buckwheat groats at 30% level, and were produc...

Human CD4+ T cell responses to the dog major allergen Can f 1 and its human homologue tear lipocalin resemble each other.

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Aino L K Liukko

Full Text Available Lipocalin allergens form a notable group of proteins, as they contain most of the significant respiratory allergens from mammals. The basis for the allergenic capacity of allergens in the lipocalin family, that is, the development of T-helper type 2 immunity against them, is still unresolved. As immunogenicity has been proposed to be a decisive feature of allergens, the purpose of this work was to examine human CD4+ T cell responses to the major dog allergen Can f 1 and to compare them with those to its human homologue, tear lipocalin (TL. For this, specific T cell lines were induced in vitro from the peripheral blood mononuclear cells of Can f 1-allergic and healthy dog dust-exposed subjects with peptides containing the immunodominant T cell epitopes of Can f 1 and the corresponding TL peptides. We found that the frequency of Can f 1 and TL-specific T cells in both subject groups was low and close to each other, the difference being about two-fold. Importantly, we found that the proliferative responses of both Can f 1 and TL-specific T cell lines from allergic subjects were stronger than those from healthy subjects, but that the strength of the responses within the subject groups did not differ between these two antigens. Moreover, the phenotype of the Can f 1 and TL-specific T cell lines, determined by cytokine production and expression of cell surface markers, resembled each other. The HLA system appeared to have a minimal role in explaining the allergenicity of Can f 1, as the allergic and healthy subjects' HLA background did not differ, and HLA binding was very similar between Can f 1 and TL peptides. Along with existing data on lipocalin allergens, we conclude that strong antigenicity is not decisive for the allergenicity of Can f 1.

Airborne seafood allergens as a cause of occupational allergy and asthma.

Science.gov (United States)

Lopata, Andreas L; Jeebhay, Mohamed F

2013-06-01

Occupational allergy and asthma is a serious adverse health outcome affecting seafood-processing workers. Allergic reactions are directed to two major seafood groups: fish and shellfish, with the latter group comprising crustaceans and molluscs. Several allergenic proteins have been identified in these different groups, but few have been characterised on a molecular level. Parvalbumin appears to be the major fish allergen, while tropomyosin the major crustacean allergen. Other IgE-binding proteins have also been identified in molluscs and other seafood-associated agents (e.g. Anisakis sp), although their molecular nature has not been characterised. Aerosolised allergens can be identified and quantified using immunological and chemical approaches, detecting levels as low as 10 ng/m(3). This contemporary review discusses interesting and recent findings in the area of occupational seafood allergy including high-risk occupations, environmental risk factors for airborne exposures, major and minor allergens implicated and innovative approaches in diagnosing and managing occupational allergy and asthma associated with seafood processing.

Effect of presowing gamma irradiation of buckwheat seeds on anatomic structure of adult plants

Energy Technology Data Exchange (ETDEWEB)

Kiseleva, N S [Gomel' skij Gosudarstvennyj Univ. (Byelorussian SSR)

1975-01-01

Changes in the structure of the leaves and stems of buckwheat resulting from presowing irradiation of seeds in doses of 5, 10, 20, and 30 kR were studied. It was shown that small doses of radiation caused an increase in cell dimensions and significant development of the stem parenchyma. With higher radiation doses xeromorphic changes occurred. The number of epidermal cells and orifices per unit surface area of the leaf, a decrease in the sizes of the perenchyma cells, and considerable development of mechanical tissues in the stem were observed. A 30-kR dose was lethal for buckwheat.

Purification, reactivity with IgE and cDNA cloning of parvalbumin as the major allergen of mackerels.

Science.gov (United States)

Hamada, Y; Tanaka, H; Ishizaki, S; Ishida, M; Nagashima, Y; Shiomi, K

2003-08-01

Three species of mackerels (Scomber japonicus, S. australasicus and S. scombrus) are widely consumed and considered to be most frequently involved in incidents of IgE-mediated fish allergy in Japan. In this study, parvalbumin, a possible candidate for the major allergen, was purified from the white muscle of three species of mackerels by gel filtration on Sephadex G-75 and reverse-phase HPLC on TSKgel ODS-120T. All the purified preparations from three species gave a single band of about 11 kDa and were clearly identified as parvalbumins by analyses of their partial amino acid sequences. In ELISA experiments, four of five sera from fish-allergic patients reacted to all the purified parvalbumins, demonstrating that parvalbumin is the major allergen in common with the mackerels. Antigenic cross-reactivity among the mackerel parvalbumins was also established by ELISA inhibition experiments. A cDNA library was constructed from the white muscle of S. japonicus and the cDNA encoding parvalbumin was cloned. The amino acid sequence translated from the nucleotide sequence revealed that the S. japonicus parvalbumin is composed of 108 residues, being a member of beta-type parvalbumins.

Household Arthropod Allergens in Korea

Science.gov (United States)

Jeong, Kyoung Yong

2009-01-01

Arthropods are important in human health, which can transmit pathogens to humans, parasitize, or produce important allergens. Allergy prevalence becomes higher in Korea recently as well as other developed countries in contrast to a decrease of infectious diseases. Allergic diseases caused by household arthropods have increased dramatically during the last few decades since human beings spend more their time for indoor activities in modernized life style. Household arthropods are one of the most common causes of allergic diseases. Biological characterization of household arthropods and researches on their allergens will provide better understanding of the pathogenesis of allergic diseases and suggest new therapeutic ways. Therefore, studies on arthropods of allergenic importance can be considered one of the major research areas in medical arthropodology and parasitology. Here, the biology of several household arthropods, including house dust mites and cockroaches, the 2 most well known arthropods living indoor together with humans worldwide, and characteristics of their allergens, especially the research activities on these allergens performed in Korea, are summarized. PMID:19885330

Correlation between airborne Olea europaea pollen concentrations and levels of the major allergen Ole e 1 in Córdoba, Spain, 2012-2014

Science.gov (United States)

Plaza, M. P.; Alcázar, P.; Galán, C.

2016-12-01

Olea europaea L. pollen is the second-largest cause of pollinosis in the southern Iberian Peninsula. Airborne-pollen monitoring networks provide essential data on pollen dynamics over a given study area. Recent research, however, has shown that airborne pollen levels alone do not always provide a clear indicator of actual exposure to aeroallergens. This study sought to evaluate correlations between airborne concentrations of olive pollen and Ole e 1 allergen levels in Córdoba (southern Spain), in order to determine whether atmospheric pollen concentrations alone are sufficient to chart changes in hay fever symptoms. The influence of major weather-related variables on local airborne pollen and allergen levels was also examined. Monitoring was carried out from 2012 to 2014. Pollen sampling was performed using a Hirst-type sampler, following the protocol recommended by the Spanish Aerobiology Network. A multi-vial cyclone sampler was used to collect aeroallergens, and allergenic particles were quantified by ELISA assay. Significant positive correlations were found between daily airborne allergen levels and atmospheric pollen concentrations, although there were occasions when allergen was detected before and after the pollen season and in the absence of airborne pollen. The correlation between the two was irregular, and pollen potency displayed year-on-year variations and did not necessarily match pollen-season-intensity.

The effects of gastric digestion on codfish allergenicity

DEFF Research Database (Denmark)

Untersmayr, Eva; Poulsen, Lars K.; Platzer, Michael H

2005-01-01

In a recent murine study, we showed that impaired gastric digestion supports the induction of fish allergy by protecting the digestion-sensitive major allergen parvalbumin and thus enhancing its sensitizing properties.......In a recent murine study, we showed that impaired gastric digestion supports the induction of fish allergy by protecting the digestion-sensitive major allergen parvalbumin and thus enhancing its sensitizing properties....

Food allergen analysis for processed food using a novel extraction method to eliminate harmful reagents for both ELISA and lateral-flow tests.

Science.gov (United States)

Ito, Kaori; Yamamoto, Takayuki; Oyama, Yuriko; Tsuruma, Rieko; Saito, Eriko; Saito, Yoshikazu; Ozu, Takeshi; Honjoh, Tsutomu; Adachi, Reiko; Sakai, Shinobu; Akiyama, Hiroshi; Shoji, Masahiro

2016-09-01

Enzyme-linked immunosorbent assay (ELISA) is commonly used to determine food allergens in food products. However, a significant number of ELISAs give an erroneous result, especially when applied to highly processed food. Accordingly, an improved ELISA, which utilizes an extraction solution comprising the surfactant sodium lauryl sulfate (SDS) and reductant 2-mercaptoethanol (2-ME), has been specially developed to analyze food allergens in highly processed food by enhancing analyte protein extraction. Recently, however, the use of 2-ME has become undesirable. In the present study, a new extraction solution containing a human- and eco-friendly reductant, which is convenient to use at the food manufacturing site, has been established. Among three chemicals with different reducing properties, sodium sulfite, tris(3-hydroxypropyl)phosphine, and mercaptoethylamine sodium sulfite was selected as a 2-ME substitute. The protein extraction ability of SDS/0.1 M sodium sulfite solution was comparable to that of SDS/2-ME solution. Next, the ELISA performance for egg, milk, wheat, peanut, and buckwheat was evaluated by using model-processed foods and commercially available food products. The data showed that the SDS/0.1 M sulfite ELISA significantly correlated with the SDS/2-ME ELISA for all food allergens examined (p analysis of food allergens in processed food, showing consistency with the SDS/0.1 M sulfite ELISA results. Accordingly, a harmonized analysis system for processed food comprising a screening LF test and a quantitative ELISA with identical extraction solution has been established. The ELISA based on the SDS/0.1 M sulfite extraction solution has now been authorized as the revised official method for food allergen analysis in Japan.

IgE-Binding Epitope Mapping and Tissue Localization of the Major American Cockroach Allergen Per a 2.

Science.gov (United States)

Lee, Mey Fann; Chang, Chia Wei; Song, Pei Pong; Hwang, Guang Yuh; Lin, Shyh Jye; Chen, Yi Hsing

2015-07-01

Cockroaches are the second leading allergen in Taiwan. Sensitization to Per a 2, the major American cockroach allergen, correlates with clinical severity among patients with airway allergy, but there is limited information on IgE epitopes and tissue localization of Per a 2. This study aimed to identify Per a 2 linear IgE-binding epitopes and its distribution in the body of a cockroach. The cDNA of Per a 2 was used as a template and combined with oligonucleotide primers specific to the target areas with appropriate restriction enzyme sites. Eleven overlapping fragments of Per a 2 covering the whole allergen molecule, except 20 residues of signal peptide, were generated by PCR. Mature Per a 2 and overlapping deletion mutants were affinity-purified and assayed for IgE reactivity by immunoblotting. Three synthetic peptides comprising the B cell epitopes were evaluated by direct binding ELISA. Rabbit anti-Per a 2 antibody was used for immunohistochemistry. Human linear IgE-binding epitopes of Per a 2 were located at the amino acid sequences 57-86, 200-211, and 299-309. There was positive IgE binding to 10 tested Per a 2-allergic sera in 3 synthetic peptides, but none in the controls. Immunostaining revealed that Per a 2 was localized partly in the mouth and midgut of the cockroach, with the most intense staining observed in the hindgut, suggesting that the Per a 2 allergen might be excreted through the feces. Information on the IgE-binding epitope of Per a 2 may be used for designing more specific diagnostic and therapeutic approaches to cockroach allergy.

Comparison of Anti-Inflammatory Effects of Flavonoid-Rich Common and Tartary Buckwheat Sprout Extracts in Lipopolysaccharide-Stimulated RAW 264.7 and Peritoneal Macrophages

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Tae Gyu Nam

2017-01-01

Full Text Available Buckwheat sprouts have been widely consumed all around world due to their great abundance of bioactive compounds. In this study, the anti-inflammatory effects of flavonoid-rich common buckwheat sprout (CBS and tartary buckwheat sprout (TBS extracts were evaluated in lipopolysaccharide- (LPS- stimulated RAW 264.7 murine macrophages and primary peritoneal macrophages from male BALB/c mice. Based on the reversed-phase HPLC analysis, the major flavonoids in CBS were determined to be C-glycosylflavones (orientin, isoorientin, vitexin, and isovitexin, quercetin-3-O-robinobioside, and rutin, whereas TBS contained only high amounts of rutin. The TBS extract exhibited higher inhibitory activity as assessed by the production of proinflammatory mediators such as nitric oxide and cytokines including tumor necrosis factor-α, interleukin- (IL- 6, and IL-12 in LPS-stimulated RAW 264.7 macrophages than CBS extract. In addition, TBS extract suppressed nuclear factor-kappa B activation by preventing inhibitor kappa B-alpha degradation and mitogen-activated protein kinase phosphorylation in LPS-stimulated RAW 264.7 macrophages. Moreover, the TBS extract markedly reduced LPS-induced cytokine production in peritoneal macrophages. Taken together, these findings suggest that TBS extract can be a potential source of anti-inflammatory agents that may influence macrophage-mediated inflammatory disorders.

Complementary DNA cloning of the major allergen Phl p I from timothy grass (Phleum pratense); recombinant Phl p I inhibits IgE binding to group I allergens from eight different grass species

NARCIS (Netherlands)

Laffer, S.; Valenta, R.; Vrtala, S.; Susani, M.; van Ree, R.; Kraft, D.; Scheiner, O.; Duchêne, M.

1994-01-01

BACKGROUND: Grass pollens, such as pollen from timothy grass (Phleum pratense), represent a major cause of type I allergy. OBJECTIVE: In this report we attempted to determine how cross-reactive allergenic components of grass pollens from different species can be represented by a minimum number of

Evaluation of different glycoforms of honeybee venom major allergen phospholipase A2 (Api m 1) produced in insect cells

DEFF Research Database (Denmark)

Blank, Simon; Seismann, Henning; Plum, Melanie

2011-01-01

Allergic reactions to hymenoptera stings are one of the major reasons for IgE-mediated anaphylaxis. However, proper diagnosis using venom extracts is severely affected by molecular cross-reactivity. In this study recombinant honeybee venom major allergen phospholipase A2 (Api m 1) was produced......-derived recombinant Api m 1 with defined CCD phenotypes might provide further insights into hymenoptera venom IgE reactivities and contribute to an improved diagnosis of hymenoptera venom allergy....

BUCKWHEAT AS A GLUTEN-FREE CEREAL IN COMBINATION WITH MAIZE FLOUR

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Stanislav Kráčmar

2012-02-01

Full Text Available Celiac disease is an autoimmune disorder of the small intestine that occurs in genetically predisposed people of all ages. Symptoms include chronic diarrhoea, and fatigue. The only treatment is long life diet with absence of gluten. Many researches concerning gluten-free nutrition have been done but it is still a big challenge. The main aim of this work was to observe changes in gluten-free breads quality made from maize-buckwheat mixtures depending on ratio of maize and buckwheat flour. To obtain samples, bread baking test was applied and these were provided to analyses (dough and pastry yield, baking loss, specific volume and texture analysis. The results showed that rising amount of maize flour in mixtures improved texture characteristics such as chewiness and gumminess, concerning specific volume of breads no significant differences were found and it was proved, that all texture parameters deteriorate with staling time.

Heat processing of peanut seed enhances the sensitization potential of the major peanut allergen Ara h 6.

Science.gov (United States)

Guillon, Blanche; Bernard, Hervé; Drumare, Marie-Françoise; Hazebrouck, Stéphane; Adel-Patient, Karine

2016-12-01

Processing of food has been shown to impact IgE binding and functionality of food allergens. In the present study, we investigated the impact of heat processing on the sensitization capacity of Ara h 6, a major peanut allergen and one of the most potent elicitors of the allergic reaction. Peanut extracts obtained from raw or heat-processed peanut and some fractions thereof were biochemically and immunochemically characterized. These extracts/fractions, purified Ara h 6, or recombinant Ara h 6 including Ara h 6 mutants lacking disulfide bridges were used in in vitro digestion tests and mouse models of experimental sensitization. Peanut roasting led to the formation of complexes of high molecular weight, notably between Ara h 6 and Ara h 1, which supported the induction of IgE specific to native Ara h 6. On the contrary, a fraction containing free monomeric 2S albumins or purified native Ara h 6 displayed no intrinsic allergenicity. In addition to complex formation, heat denaturation and/or partial destabilization enhanced Ara h 6 immunogenicity and increased its sensitivity to digestion. These results suggest that sensitization potency and IgE binding capacity can be supported by different structures, modified and/or produced during food processing in interaction with other food constituents. © 2016 The Authors. Molecular Nutrition & Food Research published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Sensitization to fungal allergens: Resolved and unresolved issues

Directory of Open Access Journals (Sweden)

Yuma Fukutomi

2015-10-01

Despite its importance in the management of allergic diseases, precise recognition of species-specific IgE sensitization to fungal allergens is often challenging because the majority of fungal extracts exhibit broad cross-reactivity with taxonomically unrelated fungi. Recent progress in gene technology has contributed to the identification of specific and cross-reactive allergen components from different fungal sources. However, data demonstrating the clinical relevance of IgE reactivity to these allergen components are still insufficient.

Molecular cloning and immunochemical characterization of a novel major Japanese cedar pollen allergen belonging to the aspartic protease family.

Science.gov (United States)

Ibrahim, Ahmed Ragaa Nour; Kawamoto, Seiji; Aki, Tsunehiro; Shimada, Yayoi; Rikimaru, Satoshi; Onishi, Nobukazu; Babiker, Elfadil Elfadl; Oiso, Isao; Hashimoto, Kunihiko; Hayashi, Takaharu; Ono, Kazuhisa

2010-01-01

Japanese cedar (Cryptomeria japonica) pollen is a major cause of seasonal pollinosis in Japan. Protease activity in the pollen grains may trigger pro-allergic responses but no such proteases have yet been identified as pollen allergens. We report the molecular cloning and immunochemical characterization of a novel C. japonica pollen allergen belonging to the aspartic protease family. We focused on the C. japonica pollen allergen spot No. 63 (CPA63, 47.5% IgE binding frequency) on our 2-dimensional IgE immunoblot map. The internal amino acid sequences were determined using time-of-flight mass spectrometry. Full-length cpa63 cDNA was cloned by rapid amplification of cDNA ends (RACE)-PCR. Recombinant CPA63 (r-CPA63) was expressed using the baculovirus-insect cell culture system and its IgE binding capacity was analyzed by enzyme-linked immunosorbent assay (ELISA). The proteolytic activity of r-CPA63 was also assessed using a putative mature enzyme produced upon autolysis. cpa63 cDNA encoded a 472 amino acid polypeptide showing about 40% sequence identity to members of the plant atypical aspartic protease family. ELISA showed that r-CPA63 was recognized by IgE antibodies in the serum of 58% (18/31) of Japanese cedar pollinosis patients. We also demonstrated an aspartic protease-like enzyme activity of the putative mature r-CPA63. We have identified the first plant aspartic protease allergen from Japanese cedar pollen. The availability of the CPA63 sequence and its recombinant allergen production system are useful not only for pharmaceutical applications but also for further examination of the role of protease activity in the pathogenesis of cedar pollinosis. 2010 S. Karger AG, Basel.

The major Alternaria alternata allergen, Alt a 1: A reliable and specific marker of fungal contamination in citrus fruits.

Science.gov (United States)

Gabriel, M F; Uriel, N; Teifoori, F; Postigo, I; Suñén, E; Martínez, J

2017-09-18

The ubiquitously present spores of Alternaria alternata can spoil a wide variety of foodstuffs, including a variety of fruits belonging to the Citrus genus. The major allergenic protein of A. alternata, Alt a 1, is a species-specific molecular marker that has been strongly associated with allergenicity and phytopathogenicity of this fungal species. This study aimed to evaluate the potential of the detection of Alt a 1 as a reliable indicator of A. alternata contamination in citrus fruits. To accomplish this aim, sixty oranges were artificially infected with a spore suspension of A. alternata. Internal fruit material was collected at different incubation times (one, two and three weeks after the fungal inoculation) and used for both total RNA extraction and protein extraction. Alt a 1 detection was then performed by polymerase chain reaction (PCR) amplification using Alt a 1 specific primers and by enzyme-linked immunosorbent assay (ELISA). The experimental model presented in this work was effective to simulate the typical Alternaria black rot phenotype and its progression. Although both PCR and ELISA techniques have been successfully carried out for detecting Alt a 1 allergen in A. alternata infected oranges, the PCR method was found to be more sensitive than ELISA. Nevertheless, ELISA results were highly valuable to demonstrate that considerable amounts of Alt a 1 are produced during A. alternata fruit infection process, corroborating the recently proposed hypothesis that this protein plays a role in the pathogenicity and virulence of Alternaria species. Such evidence suggests that the detection of Alt a 1 by PCR-based assay may be used as a specific indicator of the presence of pathogenic and allergenic fungal species, A. alternata, in fruits. This knowledge can be employed to control the fungal infection and mitigate agricultural losses as well as human exposure to A. alternata allergens and toxins. Copyright © 2017 Elsevier B.V. All rights reserved.

Development of gluten-free bread using tartary buckwheat and chia flour rich in flavonoids and omega-3 fatty acids as ingredients.

Science.gov (United States)

Costantini, Lara; Lukšič, Lea; Molinari, Romina; Kreft, Ivan; Bonafaccia, Giovanni; Manzi, Laura; Merendino, Nicolò

2014-12-15

In this study, chia seed flour, which is rich in omega-3 alpha-linolenic acid, and common and tartary buckwheat flour, which has a high antioxidant activity, were integrated into different types of bread with the aim of improving their nutritional value and healthy features. Our results indicate that bread made with chia and tartary buckwheat flour was more acceptable in many nutritional aspects compared to the control (common wheat bread); it contained a higher amount of protein (20%), insoluble dietary fibres (74%), ash (51%), and alpha-linolenic acid (67.4%). Moreover, this bread possessed lower energy (14%) and carbohydrate contents (24%) compared to the control. Tartary buckwheat also improved the total antioxidant capacity of the bread (about 75%) and provided a considerable amount of flavonoids, which are healthy non-nutritional compounds. Overall, chia and tartary buckwheat represent excellent raw materials for the formulation of gluten-free bread with high nutritional value. Copyright © 2014 Elsevier Ltd. All rights reserved.

Air-conditioner filters enriching dust mites allergen.

Science.gov (United States)

Zhan, Xiaodong; Li, Chaopin; Xu, Haifeng; Xu, Pengfei; Zhu, Haibin; Diao, Jidong; Li, Na; Zhao, Beibei

2015-01-01

We detected the concentration of dust mites allergen (Der f1 & Der p1) in the air of different places before and after the starting of air-conditioners in Wuhu City, Anhui, China, and to discuss the relation between the dust mites allergen in air-conditioner filters and the asthma attack. The dust samples were collected from the air-conditioner filters in dining rooms, shopping malls, hotels and households respectively. Concentrations of dust mites major group allergen 1 (Der f 1, Der p1) were detected with enzyme linked immunosorbent assay (ELISA), and the dust mite immune activities were determined by dot-ELISA. The concentration of Der f1 in dining rooms, shopping malls, hotels and households was 1.52 μg/g, 1.24 μg/g, 1.31 μg/g and 1.46 μg/g respectively, and the concentration of Der p1 in above-mentioned places was 1.23 μg/g, 1.12 μg/g, 1.16 μg/g and 1.18 μg/g respectively. The concentration of Der f1 & Der p1 in air was higher after the air-conditioners starting one hours later, and the difference was significant (Pair-conditioner filters can enrich dust mites major group allergen, and the allergens can induce asthma. The air-conditioner filters shall be cleaned or replaced regularly to prevent or reduce accumulation of the dust mites and its allergens.

Unique and cross-reactive T cell epitope peptides of the major Bahia grass pollen allergen, Pas n 1.

Science.gov (United States)

Etto, Tamara; de Boer, Carmela; Prickett, Sara; Gardner, Leanne M; Voskamp, Astrid; Davies, Janet M; O'Hehir, Robyn E; Rolland, Jennifer M

2012-01-01

Bahia grass pollen (BaGP) is a major cause of allergic rhinitis. Subcutaneous allergen-specific immunotherapy is effective for grass pollen allergy, but is unsuitable for patients with moderate to severe asthma due to the risk of anaphylaxis. T cell-reactive but IgE nonreactive peptides provide a safer treatment option. This study aimed to identify and characterize dominant CD4(+) T cell epitope peptides of the major BaGP allergen, Pas n 1. Pas n 1-specific T cell lines generated from the peripheral blood of BaGP-allergic subjects were tested for proliferative and cytokine response to overlapping 20-mer Pas n 1 peptides. Cross-reactivity to homologous peptides from Lol p 1 and Cyn d 1 of Ryegrass and Bermuda grass pollen, respectively, was assessed using Pas n 1 peptide-specific T cell clones. MHC class II restriction of Pas n 1 peptide T cell recognition was determined by HLA blocking assays and peptide IgE reactivity tested by dot blotting. Three Pas n 1 peptides showed dominant T cell reactivity; 15 of 18 (83%) patients responded to one or more of these peptides. T cell clones specific for dominant Pas n 1 peptides showed evidence of species-specific T cell reactivity as well as cross-reactivity with other group 1 grass pollen allergens. The dominant Pas n 1 T cell epitope peptides showed HLA binding diversity and were non-IgE reactive. The immunodominant T cell-reactive Pas n 1 peptides are candidates for safe immunotherapy for individuals, including those with asthma, who are allergic to Bahia and possibly other grass pollens. Copyright © 2012 S. Karger AG, Basel.

Rabbit IgG directed to a synthetic C-terminal peptide of the major grass pollen allergen Lol p I inhibits human basophil histamine release induced by natural Lol p I.

Science.gov (United States)

van Ree, R; Aalberse, R C

1995-03-01

The potential role of allergen-specific IgG antibodies as 'blocking' antibodies in allergen-induced human basophil histamine release was investigated. This was studied in a model with the major grass pollen allergen Lol p I and polyclonal rabbit antisera directed against this allergen and against a synthetic peptide of its C terminus. When allergen and antibodies were allowed to preincubate, Lol p I induced histamine release was inhibited up to 85% by the antiserum against Lol p I. By omitting preincubation, and thereby more closely mimicking an in vivo situation, up to 55% inhibition was realized. This indicates that allergen-specific IgG can act as 'blocking' antibody without preincubation. Immunization of rabbits with a synthetic C-terminal peptide of Lol p I resulted in antibodies reactive with natural Lol p I. Despite their 100-fold lower avidity for Lol p I (as compared with antinatural Lol p I), these antibodies had the capacity to inhibit Lol p I induced histamine release for > 90% (up to 50% without preincubation). This indicates that it is possible to block histamine release induced by a major allergen with low-avidity IgG antibodies directed against a minor proportion of the allergen (25 amino acids). IgE antibodies from the donors studied were unreactive with this synthetic peptide, indicating that for blocking activity identical epitope specificity of IgE and IgG is not essential. This opens interesting perspectives for application of synthetic peptides in immunotherapy, distinct from their effects on T cell reactivity.

Evaluation of In Vitro Inhibitory Activity of Rye-Buckwheat Ginger Cakes with Rutin on the Formation of Advanced Glycation End-Products (AGEs

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Przygodzka Małgorzata

2015-09-01

Full Text Available In this study, the relationship between the inhibitory effects of extracts from rye-buckwheat ginger cakes supplemented with low and high rutin dosage baked without or with dough fermentation step on the formation of fluorescent advanced glycation end-products (AGEs, and phenolic compounds, rutin, D-chiro-inositol and antioxidant capacity were addressed. The cakes were based on rye flour substituted by light buckwheat flour or flour from roasted buckwheat groats at 30% level, and were produced with or without dough fermentation step. The inhibitory effect against AGEs formation was studied in bovine serum albumin (BSA-glucose and BSA-methylglyoxal (MGO systems. The antioxidant capacity was measured by 2,2-diphenyl- -1-picrylhydrazyl (DPPH and cyclic voltammetry (CV, rutin and D-chiro-inositol contents by HPLC and total phenolics (TPC by spectrophotometric assays. The study showed the inhibitory effects of extracts from rye-buckwheat ginger cakes supplemented with low and high rutin dosage. The results of the inhibitory activity were highly correlated in two applied model systems. Enrichment of rye-buckwheat ginger cakes with rutin improved their antioxidant properties. The correlation studies showed that the inhibitory effects of rye-buckwheat ginger cakes produced with dough fermentation step and enhanced with rutin against formation of AGEs were highly correlated with TPC, rutin and D-chiro-inositol contents, and antioxidant capacity. Moreover, the effect of rutin enrichment was clearly seen in cakes obtained with dough fermentation step, even the inhibitory activity was slightly lower as compared to the cakes produced without dough fermentation.

Sensitization to common allergens among patients with allergies in major Iranian cities: a systematic review and meta-analysis.

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Moghtaderi, Mozhgan; Hosseini Teshnizi, Saeed; Farjadian, Shirin

2017-01-01

Various allergens are implicated in the pathogenesis of allergic diseases in different regions. This study attempted to identify the most common allergens among patients with allergies based on the results of skin prick tests in different parts of Iran. Relevant studies conducted from 2000 to 2016 were identified from the MEDLINE database. Six common groups of allergen types, including animal, cockroach, food, fungus, house dust mite, and pollen were considered. Subgroup analysis was performed to determine the prevalence of each type of allergen. The Egger test was used to assess publication bias. We included 44 studies in this meta-analysis. The overall prevalence of positive skin test results for at least one allergen was estimated to be 59% in patients with allergies in various parts of Iran. The number of patients was 11,646 (56% male and 44% female), with a mean age of 17.46±11.12 years. The most common allergen sources were pollen (47.0%), mites (35.2%), and food (15.3%). The prevalence of sensitization to food and cockroach allergens among children was greater than among adults. Pollen is the most common allergen sensitization in cities of Iran with a warm and dry climate; however, sensitization to house dust mites is predominant in northern and southern coastal areas of Iran.

What do we know about plant food allergens?

DEFF Research Database (Denmark)

Jenkins, J. A.; Sancho, A. I.; Madsen, Charlotte Bernhard

2005-01-01

databases has allowed their classification into families. This has shown that plant food allergens fall into four main families, with the prolamin superfamily (including the 2S albumins, nonspecific lipid transfer proteins and cc-amylase inhibitors) predominating, followed by the family of allergens related...... to the major birch pollen allergen, Bet v 1, and the cupin superfamily, including the I IS and 7S seed storage globulins. Future studies will be required to allow us to begin understand what it is about these protein families - whether it be their abundance, stability or some as yet unidentified factor...... - that is predisposing certain family members to becoming allergens....

Mutational epitope analysis of Pru av 1 and Api g 1, the major allergens of cherry (Prunus avium) and celery (Apium graveolens): correlating IgE reactivity with three-dimensional structure.

Science.gov (United States)

Neudecker, Philipp; Lehmann, Katrin; Nerkamp, Jörg; Haase, Tanja; Wangorsch, Andrea; Fötisch, Kay; Hoffmann, Silke; Rösch, Paul; Vieths, Stefan; Scheurer, Stephan

2003-01-01

Birch pollinosis is often accompanied by adverse reactions to food due to pollen-allergen specific IgE cross-reacting with homologous food allergens. The tertiary structure of Pru av 1, the major cherry (Prunus avium) allergen, for example, is nearly identical with Bet v 1, the major birch (Betula verrucosa) pollen allergen. In order to define cross-reactive IgE epitopes, we generated and analysed mutants of Pru av 1 and Api g 1.0101, the major celery (Apium graveolens) allergen, by immunoblotting, EAST (enzyme allergosorbent test), CD and NMR spectroscopy. The mutation of Glu45 to Trp45 in the P-loop region, a known IgE epitope of Bet v 1, significantly reduced IgE binding to Pru av 1 in a subgroup of cherry-allergic patients. The backbone conformation of Pru av 1 wild-type is conserved in the three-dimensional structure of Pru av 1 Trp45, demonstrating that the side chain of Glu45 is involved in a cross-reactive IgE epitope. Accordingly, for a subgroup of celery-allergic patients, IgE binding to the homologous celery allergen Api g 1.0101 was enhanced by the mutation of Lys44 to Glu. The almost complete loss of IgE reactivity to the Pru av 1 Pro112 mutant is due to disruption of its tertiary structure. Neither the mutation Ala112 nor deletion of the C-terminal residues 155-159 influenced IgE binding to Pru av 1. In conclusion, the structure of the P-loop partially explains the cross-reactivity pattern, and modulation of IgE-binding by site-directed mutagenesis is a promising approach to develop hypo-allergenic variants for patient-tailored specific immunotherapy. PMID:12943529

Assessment of the Allergenic Potential of Transgenic Wheat (Triticum aestivum) with Reduced Levels of ω5-Gliadins, the Major Sensitizing Allergen in Wheat-Dependent Exercise-Induced Anaphylaxis.

Science.gov (United States)

Altenbach, Susan B; Tanaka, Charlene K; Pineau, Florence; Lupi, Roberta; Drouet, Martine; Beaudouin, Etienne; Morisset, Martine; Denery-Papini, Sandra

2015-10-28

The ω5-gliadins are the major sensitizing allergens in wheat-dependent exercise-induced anaphylaxis (WDEIA). In this study, two-dimensional immunoblot analysis was used to assess the allergenic potential of two transgenic wheat lines in which ω5-gliadin genes were silenced by RNA interference. Sera from 7 of 11 WDEIA patients showed greatly reduced levels of immunoglobulin E (IgE) reactivity to ω5-gliadins in both transgenic lines. However, these sera also showed low levels of reactivity to other gluten proteins. Sera from three patients showed the greatest reactivity to proteins other than ω5-gliadins, either high-molecular-weight glutenin subunits (HMW-GSs), α-gliadins, or non-gluten proteins. The complexity of immunological responses among these patients suggests that flour from the transgenic lines would not be suitable for individuals already diagnosed with WDEIA. However, the introduction of wheat lacking ω5-gliadins could reduce the number of people sensitized to these proteins and thereby decrease the overall incidence of this serious food allergy.

Solution structure of the major fish allergen parvalbumin Sco j 1 derived from the Pacific mackerel

OpenAIRE

Kumeta, Hiroyuki; Nakayama, Haruka; Ogura, Kenji

2017-01-01

Although fish is an important part of the human diet, it is also a common source of food allergy. The major allergen in fish is parvalbumin, a well-conserved Ca2+-binding protein found in the white muscle of many fish species. Here, we studied the solution structure of the parvalbumin Sco j 1, derived from the Pacific mackerel, using nuclear magnetic resonance spectroscopy. We mapped the IgE-binding epitope proposed in a recent study onto the present structure. Interestingly, three of four re...

Herbal complex of green buckwheat in the production technology of cakes

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L. E. Glagoleva

2016-01-01

Full Text Available The possibility of using vegetable complex of green buckwheat in production technology cheesecakes. The unique chemical, rich mineral and vitamin composition buckwheat green indicates desirability of not only its use, but also th e possibility of using for cooking preventive and functional orientation. Using green buckwheat plant complex due to the hi gh water-holding ability to reduce the time of the process, eliminate some technological preparatory operations, such as pressing off. It is found that the maximum degree of swelling (2.5 g/g was observed at 40 oC and pH 4, 2. The time to reach an equilibrium state is reached after 30 minutes. On the basis of the research has been defined in a way to make plant complex curd basis, in an amount of from 10 to 70 % based on standardized physical-chemical and organoleptic characteristics of harmonious order to minimize-if-operation of flour to make the product functional and preventive properties. Technology of production of a pre -from adequate use of shock freezing, thus preserving the shape, color and flavor of the product. Microbiologists -cal studies analyzed frozen semi-finished products indicates that the microbiological-cal indicators remain stable throughout the shelf life. The use of herbal ingredients in the development of technology and the adjustment of the nutritional value of cheese bioproducts will solve the issue of their care-enrichment in essential micronutrients and natural way to improve the taste and increase the life -sti Useful developed products that currently is a modern and topical focus of research.

Immune response to allergens in sheep sensitized to house dust mite

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Velden Joanne

2008-10-01

Full Text Available Abstract Background House dust mite (HDM allergens are a major cause of allergic asthma. Most studies using animal models of allergic asthma have used rodents sensitized with the 'un-natural' allergen ovalbumin. It has only recently been recognized that the use of animal models based on HDM provide a more relevant insight into the allergen-induced mechanisms that underpin human allergic disease. We have previously described a sheep model of human allergic asthma that uses Dermatophagoides pteronyssinus HDM. The present study extends our understanding of the immune effects of HDM and the allergens Der p 1 and Der p 2 in the sheep model of asthma. Methods Peripheral blood sera from non-sensitized (control sheep and sheep sensitized to HDM was collected to determine immunoglobulin (Ig reactivities to HDM, Der p 1 and Der p 2 by ELISA. Bronchoalveolar lavage (BAL fluid collected following allergen challenge was also assessed for the presence of HDM-specific antibodies. To examine the cellular immune response to HDM allergens, T cell proliferation and cutaneous responses were assessed in sensitized and control sheep. Results Strong HDM- and Der p 1-specific IgE, IgG1, IgG2 and IgA serum responses were observed in sensitized sheep, while detectable levels of HDM-specific IgG1 and IgA were seen in BAL fluid of allergen-challenged lungs. In contrast, minimal antibody reactivity was observed to Der p 2. Marked T cell proliferation and late phase cutaneous responses, accompanied by the recruitment of eosinophils, indicates the induction of a cellular and delayed-type hypersensitivity (DTH type II response by HDM and Der p 1 allergen, but not Der p 2. Conclusion This work characterizes the humoral and cellular immune effects of HDM extract and its major constituent allergens in sheep sensitized to HDM. The effects of allergen in HDM-sensitized sheep were detectable both locally and systemically, and probably mediated via enzymatic and immune actions of the

Solution structure, copper binding and backbone dynamics of recombinant Ber e 1-the major allergen from Brazil nut.

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Louise Rundqvist

Full Text Available BACKGROUND: The 2S albumin Ber e 1 is the major allergen in Brazil nuts. Previous findings indicated that the protein alone does not cause an allergenic response in mice, but the addition of components from a Brazil nut lipid fraction were required. Structural details of Ber e 1 may contribute to the understanding of the allergenic properties of the protein and its potential interaction partners. METHODOLOGY/PRINCIPAL FINDINGS: The solution structure of recombinant Ber e 1 was solved using NMR spectroscopy and measurements of the protein back bone dynamics at a residue-specific level were extracted using (15N-spin relaxation. A hydrophobic cavity was identified in the structure of Ber e 1. Using the paramagnetic relaxation enhancement property of Cu(2+ in conjunction with NMR, it was shown that Ber e 1 is able to specifically interact with the divalent copper ion and the binding site was modeled into the structure. The IgE binding region as well as the copper binding site show increased dynamics on both fast ps-ns timescale as well as slower µs-ms timescale. CONCLUSIONS/SIGNIFICANCE: The overall fold of Ber e 1 is similar to other 2S albumins, but the hydrophobic cavity resembles that of a homologous non-specific lipid transfer protein. Ber e 1 is the first 2S albumin shown to interact with Cu(2+ ions. This Cu(2+ binding has minimal effect on the electrostatic potential on the surface of the protein, but the charge distribution within the hydrophobic cavity is significantly altered. As the hydrophobic cavity is likely to be involved in a putative lipid interaction the Cu(2+ can in turn affect the interaction that is essential to provoke an allergenic response.

Enhancement of Phenolic Production and Antioxidant Activity from Buckwheat Leaves by Subcritical Water Extraction.

Science.gov (United States)

Kim, Dong-Shin; Kim, Mi-Bo; Lim, Sang-Bin

2017-12-01

To enhance the production of phenolic compounds with high antioxidant activity and reduce the level of phototoxic fagopyrin, buckwheat leaves were extracted with subcritical water (SW) at 100~220°C for 10~50 min. The major phenolic compounds were quercetin, gallic acid, and protocatechuic acid. The cumulative amount of individual phenolic compounds increased with increasing extraction temperature from 100°C to 180°C and did not change significantly at 200°C and 220°C. The highest yield of individual phenolic compounds was 1,632.2 μg/g dry sample at 180°C, which was 4.7-fold higher than that (348.4 μg/g dry sample) at 100°C. Total phenolic content and total flavonoid content increased with increasing extraction temperature and decreased with increasing extraction time, and peaked at 41.1 mg gallic acid equivalents/g and 26.9 mg quercetin equivalents/g at 180°C/10 min, respectively. 2,2-Diphenyl-1-picrylhydrazyl free radical scavenging activity and ferric reducing ability of plasma reached 46.4 mg ascorbic acid equivalents/g and 72.3 mmol Fe 2+ /100 g at 180°C/10 min, respectively. The fagopyrin contents were reduced by 92.5~95.7%. Color values L * and b * decreased, and a * increased with increasing extraction temperature. SW extraction enhanced the yield of phenolic compounds with high antioxidant activity and reduced the fagopyrin content from buckwheat leaves.

Metabolomic analysis and phenylpropanoid biosynthesis in hairy root culture of tartary buckwheat cultivars.

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Aye Aye Thwe

Full Text Available Buckwheat, Fagopyrum tataricum Gaertn., is an important medicinal plant, which contains several phenolic compounds, including one of the highest content of rutin, a phenolic compound with anti-inflammatory properties. An experiment was conducted to investigate the level of expression of various genes in the phenylpropanoid biosynthetic pathway to analyze in vitro production of anthocyanin and phenolic compounds from hairy root cultures derived from 2 cultivars of tartary buckwheat (Hokkai T8 and T10. A total of 47 metabolites were identified by gas chromatography-time-of-flight mass spectrometry (GC-TOFMS and subjected to principal component analysis (PCA in order to fully distinguish between Hokkai T8 and T10 hairy roots. The expression levels of phenylpropanoid biosynthetic pathway genes, through qRT-PCR, showed higher expression for almost all the genes in T10 than T8 hairy root except for FtF3'H-2 and FtFLS-2. Rutin, quercetin, gallic acid, caffeic acid, ferulic acid, 4-hydroxybenzoic acid, and 2 anthocyanin compounds were identified in Hokkai T8 and T10 hairy roots. The concentration of rutin and anthocyanin in Hokkai T10 hairy roots of tartary buckwheat was several-fold higher compared with that obtained from Hokkai T8 hairy root. This study provides useful information on the molecular and physiological dynamic processes that are correlated with phenylpropanoid biosynthetic gene expression and phenolic compound content in F. tataricum species.

The major birch pollen allergen Bet v 1 induces different responses in dendritic cells of birch pollen allergic and healthy individuals.

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Ursula Smole

Full Text Available Dendritic cells play a fundamental role in shaping the immune response to allergens. The events that lead to allergic sensitization or tolerance induction during the interaction of the major birch pollen allergen Bet v 1 and dendritic cells are not very well studied. Here, we analyzed the uptake of Bet v 1 and the cross-reactive celery allergen Api g 1 by immature monocyte-derived dendritic cells (iMoDCs of allergic and normal donors. In addition, we characterized the allergen-triggered intracellular signaling and transcriptional events. Uptake kinetics, competitive binding, and internalization pathways of labeled allergens by iMoDCs were visualized by live-cell imaging. Surface-bound IgE was detected by immunofluorescence microscopy and flow cytometry. Allergen- and IgE-induced gene expression of early growth response genes and Th1 and Th2 related cytokines and chemokines were analyzed by real-time PCR. Phosporylation of signaling kinases was analyzed by Western blot. Internalization of Bet v 1 by iMoDCs of both donor groups, likely by receptor-mediated caveolar endocytosis, followed similar kinetics. Bet v 1 outcompeted Api g 1 in cell surface binding and uptake. MoDCs of allergic and healthy donors displayed surface-bound IgE and showed a pronounced upregulation of Th2 cytokine- and NFκB-dependent genes upon non-specific Fcε receptor cross-linking. In contrast to these IgE-mediated responses, Bet v 1-stimulation increased transcript levels of the Th2 cytokines IL-4 and IL-13 but not of NFκB-related genes in MoDCs of BP allergic donors. Cells of healthy donors were either unresponsive or showed elevated mRNA levels of Th1-promoting chemokines. Moreover, Bet v 1 was able to induce Erk1/2 and p38 MAPK activation in BP allergics but only a slight p38 activation in normal donors. In conclusion, our data indicate that Bet v 1 favors the activation of a Th2 program only in DCs of BP allergic individuals.

Effect of microbiological fertilizer and soil additive on yield of buckwheat (Fagopyrum esculentum Moenchunder high altitude conditions

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Oljača Snežana

2012-01-01

Full Text Available Effect of microbiological fertilizer (Slavol and soil additives (zeolite and hydrogel on buckwheat (Fagopyrum esculentum Moench yield was investigated in this paper. Trial was set up in the village of Radijevići, Serbia in agroecological conditions of mountain Zlatar (altitude 1,065 m during a two-year period 2009 and 2010. A randomized complete block design with four replications was set up. In organic cropping system three combinations of microbiological fertilizer (Slavol with zeolite and hydrogel were used prior to sowing. Different combinations of the microbiological fertilizer and the soil additives gave positive results especially in the second year of the trial. The best combination in organic cropping system was Slavol+hydrogel with foliar application of the microbiological fertilizer, which resulted in the greatest yield of buckwheat and this treatment can be recommended to producers. Buckwheat performed very well under limited conditions of acidic soil on high altitude in organic cropping system and it can be recommended as a very suitable crop for organic producers.

Quality requirements for allergen extracts and allergoids for allergen immunotherapy.

Science.gov (United States)

Zimmer, J; Bonertz, A; Vieths, S

2017-12-01

All allergen products for allergen immunotherapy currently marketed in the European Union are pharmaceutical preparations derived from allergen-containing source materials like pollens, mites and moulds. Especially this natural origin results in particular demands for the regulatory requirements governing allergen products. Furthermore, the development of regulatory requirements is complicated by the so far missing universal link between certain quality parameters, in particular biological potency, on the one hand and clinical efficacy on the other hand. As a consequence, each allergen product for specific immunotherapy has to be assessed individually for its quality, safety and efficacy. At the same time, biological potency of allergen products is most commonly determined using IgE inhibition assays based on human sera relative to product-specific in house references, ruling out full comparability of products from different manufacturers. This review article aims to summarize the current quality requirements for allergen products including the special requirements implemented for control of chemically modified allergen extracts (allergoids). Copyright © 2017 SEICAP. Published by Elsevier España, S.L.U. All rights reserved.

Analysis of glabrous canary seeds by ELISA, mass spectrometry, and Western blotting for the absence of cross-reactivity with major plant food allergens.

Science.gov (United States)

Boye, Joyce Irene; Achouri, Allaoua; Raymond, Nancy; Cleroux, Chantal; Weber, Dorcas; Koerner, Terence B; Hucl, Pierre; Patterson, Carol Ann

2013-06-26

Glabrous (hairless) canary seed belongs to the Poaceae (Gramineae) family and could serve as an alternative source of gluten-free cereal grain. In this study, allergenic cross-reactivities between hairless, dehulled canary seeds (Phalaris canariensis) and major allergenic proteins from gluten, soy, peanuts, tree nuts, sesame, and mustard were studied using commercial enzyme-linked immune sorbent assay (ELISA) kits specific for these target allergens. Mass spectrometry (MS) and immunoblotting were further used to assess for the presence of gluten-specific protein fragments. MS results revealed the likely presence of proteins homologous with rice, oat, corn, carrot, tomato, radish, beet, and chickpea. However, no presence of celiac-related gluten fragments from wheat, rye, barley, or their derivatives was found. Immunoblotting studies yielded negative results, further confirming the absence of gluten in the canary seed samples tested. No cross-reactivities were detected between canary seeds and almond, hazelnut, mustard, peanut, sesame, soy, walnut, and gluten using ELISA.

Distribution of peanut allergen in the environment.

Science.gov (United States)

Perry, Tamara T; Conover-Walker, Mary Kay; Pomés, Anna; Chapman, Martin D; Wood, Robert A

2004-05-01

Patients with peanut allergy can have serious reactions to very small quantities of peanut allergen and often go to extreme measures to avoid potential contact with this allergen. The purpose of this study was to detect peanut allergen under various environmental conditions and examine the effectiveness of cleaning agents for allergen removal. A monoclonal-based ELISA for Arachis hypogaea allergen 1 (Ara h 1; range of detection, 30-2000 ng/mL) was used to assess peanut contamination on cafeteria tables and other surfaces in schools, the presence of residual peanut protein after using various cleaning products on hands and tabletops, and airborne peanut allergen during the consumption of several forms of peanut. After hand washing with liquid soap, bar soap, or commercial wipes, Ara h 1 was undetectable. Plain water and antibacterial hand sanitizer left detectable Ara h 1 on 3 of 12 and 6 of 12 hands, respectively. Common household cleaning agents removed peanut allergen from tabletops, except dishwashing liquid, which left Ara h 1 on 4 of 12 tables. Of the 6 area preschools and schools evaluated, Ara h 1 was found on 1 of 13 water fountains, 0 of 22 desks, and 0 of 36 cafeteria tables. Airborne Ara h 1 was undetectable in simulated real-life situations when participants consumed peanut butter, shelled peanuts, and unshelled peanuts. The major peanut allergen, Ara h 1, is relatively easily cleaned from hands and tabletops with common cleaning agents and does not appear to be widely distributed in preschools and schools. We were not able to detect airborne allergen in many simulated environments.

Food allergen law and the Food Allergen Labeling and Consumer Protection Act of 2004: falling short of true protection for food allergy sufferers.

Science.gov (United States)

Roses, Jonathan B

2011-01-01

In 2004, Congress mandated labeling of food allergens on packaged foods for the first time by passing the Food Allergen Labeling and Consumer Protection Act (FALCPA). FALCPA requires that manufacturers of foods containing one of the eight major allergens responsible for 90 percent of food allergies either state on the food's packaging that the food contains the allergen, or refers to the allergen by a name easily understandable by consumers in the ingredients listing. Despite this important first step in protecting consumers with food allergies, FALCPA left unregulated the use of conditional precautionary statements (e.g., "may contain [allergen]"), which many manufacturers have used as a low-cost shield to liability. Further, FALCPA applies only to packaged foods, and does not mandate listing of food allergen ingredients in restaurants. This article discusses the history of food allergen litigation in the United States, highlighting the problems plaintiffs have faced in seeking recovery for allergic reactions to a defendants' food product, and some of the practical difficulties still extant due to the lack of regulation of precautionary statements. Also presented is a review of the Massachusetts Food Allergy Awareness Act, the first state legislation requiring restaurants to take an active role in educating employees and consumers about the presence and dangers of food allergens.

Studies on `allergoids' prepared from naturally occurring allergens

Science.gov (United States)

Marsh, D. G.; Lichtenstein, L. M.; Campbell, D. H.

1970-01-01

The highly purified major allergenic component of rye grass pollen (Group I) was used to investigate the possibility of destroying selectively the allergenic properties of an antigen, while largely retaining its original immunizing capacities. The allergen was treated under mild conditions with formalin alone or formalin plus a reactive low molecular weight additive. Certain derivatives (allergoids) showed well over 99 per cent reduction in allergenicity, determined by the histamine released from allergic human leucocytes in vitro, but were still able to combine with rabbit antibody against native antigen. Furthermore, the allergoids stimulated production (in guinea-pigs) of appreciable amounts of antibody able to inhibit native allergen-mediated human allergic histamine release in vitro and to cross-react with native antigen by PCA tests in normal guinea-pigs. Residual allergenicity and cross-immunogenicity (by the inhibition assay) of the different formalinized derivatives varied appreciably according to the additive used in formalinization, but the cross-reactivities of the different preparations in quantitative precipitin analysis against rabbit anti-native antigen serum were similar. The residual allergenicities of individual derivatives varied by up to 1000-fold in different cell preparations, suggesting a heterogeneity of allergenic determinants. Allergoid derivatives showed no hapten-like activity in that they were unable to inhibit allergen-mediated histamine release from leucocytes. The theoretical and practical application of allergoids is discussed, including their potential usefulness in improving the immunotheraphy of atopic humans. ImagesFIG. 2 PMID:4192674

Rapid genotyping with DNA micro-arrays for high-density linkage mapping and QTL mapping in common buckwheat (Fagopyrum esculentum Moench)

Science.gov (United States)

Yabe, Shiori; Hara, Takashi; Ueno, Mariko; Enoki, Hiroyuki; Kimura, Tatsuro; Nishimura, Satoru; Yasui, Yasuo; Ohsawa, Ryo; Iwata, Hiroyoshi

2014-01-01

For genetic studies and genomics-assisted breeding, particularly of minor crops, a genotyping system that does not require a priori genomic information is preferable. Here, we demonstrated the potential of a novel array-based genotyping system for the rapid construction of high-density linkage map and quantitative trait loci (QTL) mapping. By using the system, we successfully constructed an accurate, high-density linkage map for common buckwheat (Fagopyrum esculentum Moench); the map was composed of 756 loci and included 8,884 markers. The number of linkage groups converged to eight, which is the basic number of chromosomes in common buckwheat. The sizes of the linkage groups of the P1 and P2 maps were 773.8 and 800.4 cM, respectively. The average interval between adjacent loci was 2.13 cM. The linkage map constructed here will be useful for the analysis of other common buckwheat populations. We also performed QTL mapping for main stem length and detected four QTL. It took 37 days to process 178 samples from DNA extraction to genotyping, indicating the system enables genotyping of genome-wide markers for a few hundred buckwheat plants before the plants mature. The novel system will be useful for genomics-assisted breeding in minor crops without a priori genomic information. PMID:25914583

Allergenic fragments of ryegrass (Lolium perenne) pollen allergen Lol p IV.

Science.gov (United States)

Jaggi, K S; Ekramoddoullah, A K; Kisil, F T

1989-01-01

To facilitate studies on establishing the nature of structure/function relationships of allergens, ryegrass pollen allergen, Lol p IV, was cleaved into smaller fragments by cyanogen bromide (CNBr) and the resulting peptides were further digested with trypsin. The resulting peptides were then fractionated by high performance liquid chromatography (HPLC) on a C-18 reverse phase column. The allergenic activity of the HPLC fractions was evaluated in terms of their ability to inhibit the binding of 125I-Lol p IV to serum IgE antibodies of a grass-allergic patient. Many of these fractions inhibited the binding between the native allergen and IgE antibodies in a dose-dependent manner. The inhibitions were specific, i.e., the fractions did not inhibit the binding between 125I-Lol p I (a group-I ryegrass pollen allergen) and the IgE antibodies present in the allergic human serum. The possibility that the allergenic peptide fractions were contaminated by the native undegraded allergen, which might have accounted for the observed inhibition, was ruled out by the fact that the native allergen could not be detected by SDS-PAGE and the elution profiles of allergenically active peptides did not coincide with that of native allergen. One of the allergenic sites recognized by monoclonal antibody (Mab) 90, i.e., site A, was located in HPLC fractions 90-100 while another allergenic site B (recognized by Mab 12) appeared to be lost following the sequential digestion of Lol p IV with CNBr and trypsin.

New species of Braggia (Hemiptera: Aphididae) on buckwheat in western North America

Science.gov (United States)

K. S. Pike; G. Graf; R. G. Foottit; H. E. L. Maw; P. Stary; R. Hammon; D. G. Miller

2009-01-01

Species of Braggia Gillette and Palmer (Hemiptera: Aphididae: Aphidinae: Aphidini) feed on various buckwheat, Eriogonum Michx. (Polygonaceae), species in western North America. Two new species, Braggia columbiana Pike n. sp. from Washington and Oregon and Braggia longicauda Pike n. sp. from Washington, Oregon, and northern California, are proposed. Descriptions,...

Effectiveness of allergen-specific immunotherapy with pollen allergens in children from the viewpoint of molecular allergology

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S.M. Nedelska

2017-03-01

, allergic rhinitis is complicated by sinusitis in 13 % of cases. Children received 3 to 4 courses of preseason subcutaneous ASIT (December to May according to classic scheme. The vast majority of children got mixed extracts of ragweed, cyclachaena, mugwort, quinoa, sunflower (90 %. One allergen (ragweed or sunflower has been used only in 6 children (10 %. We detected that 1/5 children had no clinical effect, the symptoms of hay fever were not diminished or changed their temporal frames. 82 % of patients without effect of ASIT had cross plant-food allergy (oral allergy syndrome. Conclusion. Before the beginning of АSІТ in children with hay fever, the doctor must define a high-risk group in relation to its possible ineffectiveness. We suggest taking into account children with sensitization to 4 and more pollen allergens, with the presence of oral allergic syndrome, concomitant pathology of ENT organs, in particular, adenoid hypertrophy, chronic sinusitis. Such children are recommended to make a profile of molecular allergy diagnostics to determine the specific immunoglobulines Е to the major and minor allergens. Component resolved molecular allergy diagnostics maybe also helpful in estimation the risk of systemic (anaphylactic reactions in children with cross reactivity (pollen-food allergy and in improvement of preventive measures, in particular, in patients with oral allergy syndrome.

Insect Sting Reactions and Specific IgE to Venom and Major Allergens in a General Population

DEFF Research Database (Denmark)

Mosbech, H; Tang, L; Linneberg, A

2016-01-01

BACKGROUND: Insect sting reactions are frequently reported, but population studies documenting the frequency and the relation to IgE-sensitization and serum tryptase are scarce. METHODS: Questionnaire data and results from measurements of specific IgE against venom, major allergens and cross...... or wasp. IgE to CCDs occurred in only 0.7%, but 80% of these were DS. Finally, 36% with IgE to CCDs had had symptoms, mostly local. Serum tryptase was not associated with a history of sting reactions. CONCLUSIONS: In a temperate climate, self-reported insect sting reactions and sensitization to venom...

Modified Allergens for Immunotherapy.

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Satitsuksanoa, Pattraporn; Głobińska, Anna; Jansen, Kirstin; van de Veen, Willem; Akdis, Mübeccel

2018-02-16

During the past few decades, modified allergens have been developed for use in allergen-specific immunotherapy (AIT) with the aim to improve efficacy and reduce adverse effects. This review aims to provide an overview of the different types of modified allergens, their mechanism of action and their potential for improving AIT. In-depth research in the field of allergen modifications as well as the advance of recombinant DNA technology have paved the way for improved diagnosis and research on human allergic diseases. A wide range of structurally modified allergens has been generated including allergen peptides, chemically altered allergoids, adjuvant-coupled allergens, and nanoparticle-based allergy vaccines. These modified allergens show promise for the development of AIT regimens with improved safety and long-term efficacy. Certain modifications ensure reduced IgE reactivity and retained T cell reactivity, which facilities induction of immune tolerance to the allergen. To date, multiple clinical trials have been performed using modified allergens. Promising results were obtained for the modified cat, grass and birch pollen, and house dust mite allergens. The use of modified allergens holds promise for improving AIT efficacy and safety. There is however a need for larger clinical studies to reliably assess the added benefit for the patient of using modified allergens for AIT.

Identification of T-cell epitopes of Lol p 9, a major allergen of ryegrass (Lolium perenne) pollen.

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Blaher, B; Suphioglu, C; Knox, R B; Singh, M B; McCluskey, J; Rolland, J M

1996-07-01

T-cell recognition of Lol p 9, a major allergen of ryegrass pollen, was investigated by using a T-cell line and T-cell clones generated from the peripheral blood of an atopic donor. The T-cell line reacted with purified Lol p 9, as well as with crude ryegrass pollen extract, but failed to cross-react with Bermuda grass pollen extract. All of six T-cell clones generated from this line proliferated in response to Lol p 9. Epitope mapping was carried out with a panel of 34 overlapping synthetic peptides, which spanned the entire sequence of the Lol p 9 12R isoform. The T-cell line responded to two of the peptides, Lol p 9 (105-116) and Lol p 9 (193-204), whereas reactivity with one or other of these peptides was shown by five T-cell clones. These two peptides contained sequences consistent with motifs previously reported for major histocompatibility complex class II-restricted peptides. HLA antibody blocking studies showed that presentation of peptide Lol p 9 (105-116) to one T-cell clone was HLA-DR-restricted; this clone expressed a T helper cell phenotype (CD3+, CD4+) and the T-cell receptor alpha beta. The identification of immunodominant T-cell epitope(s) on allergens is essential for devising safer and more effective immunotherapy strategies, which can interrupt the chain of events leading to allergic disease.

Purification and characterization of parvalbumins, the major allergens in red stingray (Dasyatis akajei).

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Cai, Qiu-Feng; Liu, Guang-Ming; Li, Teng; Hara, Kenji; Wang, Xi-Chang; Su, Wen-Jin; Cao, Min-Jie

2010-12-22

Fish has received increasing attention because it induces IgE-mediated food allergy. Parvalbumin (PV) represents the major allergen of fish, and IgE cross-reactivity to PV in various teleost fish species has been shown, while little information is available about allergens in elasmobranch fish. In this study, two PV isoforms (named as PV-I and PV-II) from red stingray (Dasyatis akajei) were purified to homogeneity by a series of procedures including ammonium sulfate precipitation and column chromatographies of DEAE-Sepharose and Sephacryl S-200. Purified PVs revealed a single band on tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular masses of PV-I and PV-II were 12.29 and 11.95 kDa, respectively, as determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot using antifrog PV monoclonal antibody (PARV-19) showed positive reactions to the two proteins, confirming that they were PVs, although their immunological reactivities were weaker than those of PV from silver carp. The N-terminal amino acid sequence of PV-I was determined, and comparison with PVs from other fish species showed low homology between teleost and elasmobranch fish. The isoelectric points of PV-I and PV-II were 5.4 and 5.0, respectively, as determined by two-dimensional electrophoresis (2-DE), suggesting that both isoforms belong to the α-group. IgE immunoblotting analysis showed that sera from fish-allergic patients reacted to both PV-I and PV-II from red stingray. Thermal stability revealed that PV-I easily formed oligomers than PV-II, which might contribute to the maintenance of its allerginicity during heat processing.

Environmental allergens in patients with allergic rhinitis

International Nuclear Information System (INIS)

Anwar, M.S.; Bokhari, S.R.

2002-01-01

Objective: to find out the common environmental allergens responsible for sensitivity in patients with allergic rhinitis. Design: Descriptive cross sectional study. Place and Duration of Study: A local allergy clinic in an urban area of Lahore during the year 2000-2001. Subjects and Methods: Eighty patients with allergic rhinitis irrespective of age and sex were studied. These cases were selected on the basis of symptoms like sneezing, itching, watery nasal discharge and eosinophilia in nasal secretions. Forty matched healthy subjects as controls were also studied. Allergy test was performed on all the subjects by skin prick test to determine sensitivity to common environmental allergens using Bencard (England) allergy kit. Results: common environmental allergens responsible for sensitivity in allergic rhinitis patients were house dust (82.5 %), house dust mites (73.7%), mixed threshing (80%), straw dust (58.7%, hay dust (63.7%), mixed feathers (45%), cat fur (57.5%), cotton flock (56.2%), tree pollens (45%) and grass pollens (48.7%). Sensitivity to these allergens was observed in significantly higher (P<0.01) percentage of allergic rhinitis patients as compared with control subjects. Sensitivity to house dust, house dust mites and cat fur was of severe degree in majority of allergic rhinitis patients. While sensitivity to mixed threshing, straw dust, hay dust and mixed feathers was of moderate to severe degree in majority of these patients. Conclusion: Skin prick tests provide an effective and definitive mean to find out sensitivity to different allergens in cases with allergic rhinitis. Based on these findings, the physician can manage these patients in better way. (author)

Identification, cloning, and characterization of a major cat flea salivary allergen (Cte f 1).

Science.gov (United States)

McDermott, M J; Weber, E; Hunter, S; Stedman, K E; Best, E; Frank, G R; Wang, R; Escudero, J; Kuner, J; McCall, C

2000-05-01

An 18 kDa protein isolated from saliva of the cat flea, Ctenocephalides felis, elicits a positive intradermal skin test (IDST) in 100 and 80% of experimental and clinical flea allergic dogs, respectively. Using solid-phase enzyme-linked immuno assay (ELISA), this protein detected IgE in 100 and 80% of experimental and clinical flea allergic dogs, respectively. A cDNA (pFSI) encoding a full-length Cte f 1 protein was isolated from a C. felis salivary gland cDNA library, using a combination of PCR and hybridization screening. This cDNA is 658 bp in length, and contains an open reading frame of 528 bp. The open reading frame encodes a protein of 176 amino acids, consisting of an 18 amino acid signal sequence and a 158 amino acid mature protein. The calculated molecular weight and pI of the mature protein are 18106 Da and 9.3, respectively. The protein, named Cte f 1, is the first novel major allergen described for canine flea allergy. Recombinant Cte f 1 (rCte f 1) was expressed in Escherichia coli, Pichia pastoris and baculovirus infected Trichoplusia ni cells. Approximately, 90% of the rCte f 1 expressed in E. coli accumulated in insoluble inclusion bodies, which could be refolded to a soluble mixture of disulfide isomers with partial IgE binding activity. Small quantities of an apparently correctly refolded form of rCte f 1, which had IgE binding activity equal to the native antigen, was isolated from the soluble fraction of E. coli cells. However, P. pastoris and baculovirus infected insect cells expressed and secreted a fully processed, correctly refolded and fully active form of rCte f 1. Mass spectrometry analysis of the active forms of rCte f 1confirmed that eight intact disulfide bonds were present, matching the number observed in the native allergen. The relative ability of rCte f 1 to bind IgE in the serum of flea allergic animals, produced in these three expression systems, matched that of the native allergen. Competition ELISA demonstrated that

Challenges in testing genetically modified crops for potential increases in endogenous allergen expression for safety.

Science.gov (United States)

Panda, R; Ariyarathna, H; Amnuaycheewa, P; Tetteh, A; Pramod, S N; Taylor, S L; Ballmer-Weber, B K; Goodman, R E

2013-02-01

Premarket, genetically modified (GM) plants are assessed for potential risks of food allergy. The major risk would be transfer of a gene encoding an allergen or protein nearly identical to an allergen into a different food source, which can be assessed by specific serum testing. The potential that a newly expressed protein might become an allergen is evaluated based on resistance to digestion in pepsin and abundance in food fractions. If the modified plant is a common allergenic source (e.g. soybean), regulatory guidelines suggest testing for increases in the expression of endogenous allergens. Some regulators request evaluating endogenous allergens for rarely allergenic plants (e.g. maize and rice). Since allergic individuals must avoid foods containing their allergen (e.g. peanut, soybean, maize, or rice), the relevance of the tests is unclear. Furthermore, no acceptance criteria are established and little is known about the natural variation in allergen concentrations in these crops. Our results demonstrate a 15-fold difference in the major maize allergen, lipid transfer protein between nine varieties, and complex variation in IgE binding to various soybean varieties. We question the value of evaluating endogenous allergens in GM plants unless the intent of the modification was production of a hypoallergenic crop. © 2012 John Wiley & Sons A/S.

Effect of thermal treatment on rutin content in selected buckwheat products using calcium as an internal tracer

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Eliška Krejzová

2017-01-01

Full Text Available Reversed-phase high-performance liquid chromatography (RP-HPLC was used for rutin (quercetin-3-rutinoside determination in selected buckwheat products (whole meal flour, broken seeds, seed hulls, herbs and baked cereal breads. The effect of various thermal procedures on content of rutin was evaluated using calcium as an internal tracker to correct changes in mass and composition of the buckwheat products. These factors are very seldom taken into account. The results show non-significant changes in rutin levels obtained in whole meal flour and broken seed samples after thermal treatment up to 150°C. Higher temperature already caused sudden fall in the observed rutin concentrations. The evaporation of some volatile compounds and degradation products can decrease the mass of the samples and formally increase the content of rutin (35.5 ±4.7 mg per 100 g for whole meal flour and 10.2 ±0.4 mg per100 g for broken seeds at 150°C. Serious decrease of rutin contents at elevated temperatures (>150°C can be explained by its degradation (by breaking off the C-C bond in quercetin-3-rutinoside moiety and/or evaporation (24.3 ±1.4 mg per 100 g for whole meal flour and 3.06 ±0.3 mg per100 g for broken seeds at 180°C. In case of baked cereal breads the level of rutin changed in dependence on the ratio between buckwheat and corn flour. Longer time leaching and higher temperature implicate higher rutin content in infusions prepared from buckwheat seed hulls and herbs.

Identification of Aspergillus (A. flavus and A. niger) Allergens and Heterogeneity of Allergic Patients' IgE Response.

Science.gov (United States)

Vermani, Maansi; Vijayan, Vannan Kandi; Agarwal, Mahendra Kumar

2015-08-01

Aspergillus species (A. flavus and A. niger) are important sources of inhalant allergens. Current diagnostic modalities employ crude Aspergillus extracts which only indicate the source to which the patient has been sensitized, without identifying the number and type of allergens in crude extracts. We report a study on the identification of major and minor allergens of the two common airborne Aspergillus species and heterogeneity of patients' IgE response to them. Skin prick tests were performed on 300 patients of bronchial asthma and/or allergic rhinitis and 20 healthy volunteers. Allergen specific IgE in patients' sera was estimated by enzyme allergosorbent test (EAST). Immunoblots were performed to identify major/minor allergens of Aspergillus extracts and to study heterogeneity of patients'IgE response to them. Positive cutaneous responses were observed in 17% and 14.7% of patients with A. flavus and A. niger extracts, respectively. Corresponding EAST positivity was 69.2% and 68.7%. In immunoblots, 5 allergenic proteins were identified in A. niger extract, major allergens being 49, 55.4 and 81.5 kDa. Twelve proteins bound patients' IgE in A. flavus extract, three being major allergens (13.3, 34 and 37 kDa). The position and slopes of EAST binding and inhibition curves obtained with individual sera varied from patient to patient. The number and molecular weight of IgE-binding proteins in both the Aspergillus extracts varied among patients. These results gave evidence of heterogeneity of patients' IgE response to major/minor Aspergillus allergens. This approach will be helpful to identify disease eliciting molecules in the individual patients (component resolved diagnosis) and may improve allergen-specific immunotherapy.

Mutants of the major ryegrass pollen allergen, Lol p 5, with reduced IgE-binding capacity: candidates for grass pollen-specific immunotherapy.

Science.gov (United States)

Swoboda, Ines; De Weerd, Nicole; Bhalla, Prem L; Niederberger, Verena; Sperr, W R; Valent, Peter; Kahlert, Helga; Fiebig, Helmut; Verdino, Petra; Keller, Walter; Ebner, Christof; Spitzauer, Susanne; Valenta, Rudolf; Singh, Mohan B

2002-01-01

More than 400 million individuals are sensitized to grass pollen allergens. Group 5 allergens represent the most potent grass pollen allergens recognized by more than 80 % of grass pollen allergic patients. The aim of our study was to reduce the allergenic activity of group 5 allergens for specific immunotherapy of grass pollen allergy. Based on B- and T-cell epitope mapping studies and on sequence comparison of group 5 allergens from different grasses, point mutations were introduced by site-directed mutagenesis in highly conserved sequence domains of Lol p 5, the group 5 allergen from ryegrass. We obtained Lol p 5 mutants with low IgE-binding capacity and reduced allergenic activity as determined by basophil histamine release and by skin prick testing in allergic patients. Circular dichroism analysis showed that these mutants exhibited an overall structural fold similar to the recombinant Lol p 5 wild-type allergen. In addition, Lol p 5 mutants retained the ability to induce proliferation of group 5 allergen-specific T cell lines and clones. Our results demonstrate that a few point mutations in the Lol p 5 sequence yield mutants with reduced allergenic activity that represent potential vaccine candidates for immunotherapy of grass pollen allergy.

Quantifying Dustiness, Specific Allergens, and Endotoxin in Bulk Soya Imports

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Howard J. Mason

2017-11-01

Full Text Available Soya is an important bulk agricultural product often transported by sea as chipped beans and/or the bean husks after pelletisation. There are proven allergens in both forms. Bulk handling of soya imports can generate air pollution containing dust, allergens, and pyrogens, posing health risks to dockside workers and surrounding populations. Using an International Organization for Standardization (ISO standardised rotating drum dustiness test in seven imported soya bulks, we compared the generated levels of dust and two major soya allergens in three particle sizes related to respiratory health. Extractable levels of allergen and endotoxin from the bulks showed 30–60 fold differences, with levels of one allergen (hydrophobic seed protein and endotoxin higher in husk. The generated levels of dust and allergens in the three particle sizes also showed very wide variations between bulks, with aerolysed levels of allergen influenced by both the inherent dustiness and the extractable allergen in each bulk. Percentage allergen aerolysed from pelletized husk—often assumed to be of low dustiness—after transportation was not lower than that from chipped beans. Thus, not all soya bulks pose the same inhalation health risk and reinforces the importance of controlling dust generation from handling all soya bulk to as low as reasonably practicable.

Teff, buckwheat, quinoa and amaranth: Ancient whole grain gluten-free egg-free pasta

Science.gov (United States)

This report demonstrates innovative ancient whole grains, gluten-free, egg-free pasta (no chemicals added) made using a kitchen counter-top appliance. Whole grain, fusilli pasta was prepared with teff, buckwheat, quinoa and amaranth flours. These ancient grains are called “Super Foods” due to thei...

Elemental analysis of edible grains by micro-PIXE: Common buckwheat case study

Energy Technology Data Exchange (ETDEWEB)

Vogel-Mikus, K. [Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, SI-1000 Ljubljana (Slovenia); Pelicon, P.; Vavpetic, P. [Jozef Stefan Institute, Jamova 39, SI-1000 Ljubljana (Slovenia); Kreft, I. [Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, SI-1000 Ljubljana (Slovenia)], E-mail: ivan.kreft@guest.arnes.si; Regvar, M. [Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, SI-1000 Ljubljana (Slovenia)

2009-09-01

The aim of this study was the adaptation of the micro-PIXE method for analysis of nutritionally relevant heavy elements in different tissues of the grain of common buckwheat (Fagopyrum esculentum), as a representative nutritionally interesting grain food source. At 57% of the buckwheat grain biomass, the endosperm was a modest nutrient source when compared to the cotyledons, at 17% of the biomass. These latter contained high concentrations of trace elements, representing 91% of the total grain Zn, 87% for P, 70% for S, 62% for Mg, 60% for K, 54% for Cu, 53% for Mn and 35% for Fe. The husk provided storage for 85% of the total Ca, 84% for Al, 83% for Si, 76% for Cl, 69% for Ti and 46% for Fe. Knowledge on these preferential elemental constitutions of the different grain tissues makes the possibility of designing target products with nutritionally optimal constitution more feasible. These data represent a basis for a more targeted approach to nutritional improvement of grains intended for human consumption.

Elemental analysis of edible grains by micro-PIXE: Common buckwheat case study

International Nuclear Information System (INIS)

Vogel-Mikus, K.; Pelicon, P.; Vavpetic, P.; Kreft, I.; Regvar, M.

2009-01-01

The aim of this study was the adaptation of the micro-PIXE method for analysis of nutritionally relevant heavy elements in different tissues of the grain of common buckwheat (Fagopyrum esculentum), as a representative nutritionally interesting grain food source. At 57% of the buckwheat grain biomass, the endosperm was a modest nutrient source when compared to the cotyledons, at 17% of the biomass. These latter contained high concentrations of trace elements, representing 91% of the total grain Zn, 87% for P, 70% for S, 62% for Mg, 60% for K, 54% for Cu, 53% for Mn and 35% for Fe. The husk provided storage for 85% of the total Ca, 84% for Al, 83% for Si, 76% for Cl, 69% for Ti and 46% for Fe. Knowledge on these preferential elemental constitutions of the different grain tissues makes the possibility of designing target products with nutritionally optimal constitution more feasible. These data represent a basis for a more targeted approach to nutritional improvement of grains intended for human consumption.

Optimization of corn, rice and buckwheat formulations for gluten-free wafer production.

Science.gov (United States)

Dogan, Ismail Sait; Yildiz, Onder; Meral, Raciye

2016-07-01

Gluten-free baked products for celiac sufferers are essential for healthy living. Cereals having gluten such as wheat and rye must be removed from the diet for the clinical and histological improvement. The variety of gluten-free foods should be offered for the sufferers. In the study, gluten-free wafer formulas were optimized using corn, rice and buckwheat flours, xanthan and guar gum blend as an alternative product for celiac sufferers. Wafer sheet attributes and textural properties were investigated. Considering all wafer sheet properties in gluten-free formulas, better results were obtained by using 163.5% water, 0.5% guar and 0.1% xanthan in corn formula; 173.3% water, 0.45% guar and 0.15% xanthan gum in rice formula; 176% water, 0.1% guar and 0.5% xanthan gum in buckwheat formula. Average desirability values in gluten-free formulas were between 0.86 and 0.91 indicating they had similar visual and textural profiles to control sheet made with wheat flour. © The Author(s) 2015.

Prevalence of food and airborne allergens in allergic patients in Kerman

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Hamed Fouladseresht

2014-07-01

Full Text Available Background: Detection of various environmental allergens is the major challenge in allergic diseases and the only treatment is avoiding these allergens. The aim of this study was to determine the prevalence of food and airborne allergens in allergic patients using Skin Prick Test (SPT. Methods: A cross-sectional study was done on clinically confirmed patients of atopic-dermatitis (n=54, allergenic-rhinitis (n=64 and chronic-urticaria (n=39 who referred to asthma and allergy clinic at Afzali-Pour hospital in Kerman during 2008-2010. Skin prick test was done using allergen extracts to determine the patients' sensitivity to food and airborne antigens. Results: Fifty-nine percent of patients responded to at least one allergen. Allergy to airborne and food allergens was 55.9 % and 21.7%, respectively. Chenopodiaceae (22.9% and egg white (10.2% were most prevalent airborne and food allergens. Allergy to cockroach, egg white, egg yolk and tomato was significantly higher in males than in females (P<0.05. Conclusion: The results indicated that allergy to food and airborne allergens is different depending on the nutrition and environmental conditions.

Physicochemical characterization of allergens: quantity, identity, purity, aggregation and conformation.

Science.gov (United States)

Koppelman, Stef J; Luykx, Dion M A M; de Jongh, Harmen H J; Veldhuizen, Willem Jan

2009-01-01

Allergens and allergoids can be characterized by means of physicochemical methods, resulting in a description of the protein on different structural levels. Several techniques are available and their suitability depends on the composition of the particular sample. Current European legislation on allergen products demands characterization of final products in particular focusing on identity, degree of modification (for allergoids) and stability of the composition. Structural parameters of allergens may be used to investigate the stability of an allergen product. The challenge is to identify and optimize techniques that allow determination of protein structure in the context of a formulated pharmaceutical product. As the majority of the products currently marketed are formulated with aluminium hydroxide or aluminium phosphate as a depot, most of the methods and techniques used for protein characterization in solution are not applicable. An additional hurdle is that allergen products are based on natural extracts, comprising a mixture of proteins, both allergens and non-allergens, sometimes in the presence of other uncharacterized components from the raw material. This paper describes which methods are suitable for the different stages of allergen product manufacturing, and how these relate to the current regulatory requirements. Some of the techniques are demonstrated using a model allergen, cod parvalbumin, and a chemically modified form thereof. We conclude that a variety of methods is available for characterization of proteins in solution, and that a limited number of techniques appear to be suitable for modified allergens (allergoids). Adaptation of existing methods, e.g. mass spectroscopy and infrared spectroscopy may be helpful to obtain protein parameters of allergens in a formulated allergen product. By choosing a combination of techniques, including those additional to physicochemical approaches, relevant parameters of allergens in formulated allergen

pH and Heat Resistance of the Major Celery Allergen Api g 1.

Science.gov (United States)

Rib-Schmidt, Carina; Riedl, Philipp; Meisinger, Veronika; Schwaben, Luisa; Schulenborg, Thomas; Reuter, Andreas; Schiller, Dirk; Seutter von Loetzen, Christian; Rösch, Paul

2018-05-25

The major celery allergen Api g 1 is a member of the pathogenesis-related 10 class protein family. Here we aimed to investigate the impact of heat and pH on the native protein conformation required for Immunoglobulin E (IgE) recognition. Spectroscopic methods, MS and IgE binding analyses were used to study the effects of pH and thermal treatment on Api g 1.0101. Heat processing results in a loss of the native protein fold via denaturation, oligomerisation and precipitation along with a subsequent reduction of IgE recognition. The induced effects and timescales are strongly pH depended. While Api g 1 refolds partially into an IgE-binding conformation at physiological pH, acidic pH treatment leads to the formation of structurally heat resistant, IgE-reactive oligomers. Thermal processing in the presence of a celery matrix or at pH conditions close to the isoelectric point (pI = 4.63) of Api g 1.0101 results in almost instant precipitation. Our data demonstrate that Api g 1.0101 is not intrinsically susceptible to heat treatment in vitro. However, the pH and the celery matrix strongly influence the stability of Api g 1.0101 and might be the main reasons for the observed temperature lability of this important food allergen. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Bioavailability of House Dust Mite Allergens in Sublingual Allergy Tablets Is Highly Dependent on the Formulation.

Science.gov (United States)

Ohashi-Doi, Katsuyo; Kito, Hirokazu; Du, Weibin; Nakazawa, Hiroshi; Ipsen, Henrik; Gudmann, Pernille; Lund, Kaare

2017-01-01

In sublingual immunotherapy (SLIT), the immune system is addressed by solubilized allergen that interacts with immunocompetent cells of the oral mucosa, the efficiency of which is governed by 2 main factors of SLIT allergen bioavailability: the allergen concentration and the mucosal contact time. Recently, 3 house dust mite (HDM) SLIT tablets were developed that differ with regard to allergen content, nominal strength (maintenance doses: 6 SQ-HDM/10,000 Japanese Allergen Units [JAU], 12 SQ-HDM/ 20,000 JAU, and 300 IR/57,000 JAU), and formulation (freeze-dried/compressed). Here, the importance of the SLIT tablet formulation for HDM major allergen bioavailability is examined. The HDM major allergen content, tablet disintegration times, and allergen release kinetics were determined. Dissolution kinetics (allergen concentration vs. time) of Der f 1, Der p 1, and Der 2 were measured. Area under the curve (AUC) was used as a surrogate parameter for allergen bioavailability. The release of HDM major allergens from the freeze-dried tablets was complete after 30 s, while only partial release was achieved with the compressed tablets, even after prolonged dissolution. At 1 min, i.e., the recommended sublingual holding time for the freeze-dried tablets, the allergen bioavailability (AUC) of the compressed 300 IR/57,000 JAU tablet was 4.7-fold (Der f 1), 10.8-fold (Der p 1), and 23.6-fold (Der 2) lower than that of the freeze-dried 12 SQ-HDM/20,000 JAU tablet and similar to (Der f 1) and 5.3-fold (Der p 1) and 12.5-fold (Der 2) lower than that of the freeze-dried 6 SQ-HDM/10,000 JAU tablet. SLIT tablet allergen bioavailability depends highly on the tablet formulation. Only the fast-dissolving freeze-dried tablets provide maximal delivery of soluble allergens and achieve allergen concentrations that reflect the nominal tablet strengths within the recommended sublingual holding time. © 2017 S. Karger AG, Basel.

Effect of selected spices on chemical and sensory markers in fortified rye-buckwheat cakes.

Science.gov (United States)

Przygodzka, Małgorzata; Zieliński, Henryk; Ciesarová, Zuzana; Kukurová, Kristina; Lamparski, Grzegorz

2016-07-01

The aim of this study was to find out the effect of selected spices on chemical and sensorial markers in cakes formulated on rye and light buckwheat flour fortified with spices. Among collection of spices, rye-buckwheat cakes fortified individually with cloves, nutmeg, allspice, cinnamon, vanilla, and spice mix revealed the highest sensory characteristics and overall quality. Cakes fortified with cloves, allspice, and spice mix showed the highest antioxidant capacity, total phenolics, rutin, and almost threefold higher available lysine contents. The reduced furosine content as well as free and total fluorescent intermediatory compounds were observed as compared to nonfortified cakes. The FAST index was significantly lowered in all cakes enriched with spices, especially with cloves, allspice, and mix. In contrast, browning index increased in compare to cakes without spices. It can be suggested that clove, allspice, vanilla, and spice mix should be used for production of safety and good quality cakes.

Fractionation and analysis of allergenicity of allergens from Prosopis juliflora pollen.

Science.gov (United States)

Thakur, I S

1989-01-01

Prosopis juliflora pollen allergen extract was prepared, and its crude allergen extract was fractionated by Sephadex G-100 gel filtration. Six different fractions were obtained which was confirmed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Protein and carbohydrate content of each fraction were estimated. Fraction E (MW 20,000) showed a 25% carbohydrate concentration. The amino acid analysis indicated that this fraction was rich in glutamic acid and alanine. Antigenicity or allergenicity of fractionated allergens were checked by gel diffusion test, rocket immunoelectrophoresis, skin prick test, and radioallergosorbent test. All these test indicate that fraction E consisted mainly of allergenic molecules (MW 20,000) of P. juliflora pollen.

Crude protein, fibre and phytic acid in vitro digestibility of selected legume and buckwheat samples

Directory of Open Access Journals (Sweden)

Petra Vojtíšková

2013-01-01

Full Text Available The aim of this study was to determine crude protein, fibre and phytic acid in vitro digestibility of selected legumes and buckwheat products. All analyses except the phytic acid contents were performed in the line with the Commission Regulation (EC No. 152/2009. A modified version of Holt’s Method was used for phytic acid (phytate determination. None of all samples contained more than 11% of moisture. Soybeans are rich in crude protein; they contain nearly 40% of this compound. The content of crude protein in buckwheat flours was about 14%. The highest amount of phytate was found in common beans and soybeans-about 2 g/100 g of dry matter. On the other hand, the lowest phytate content was observed in buckwheat pasta (F. esculentum groats was 1.9 g per 100 g of dry matter. In vitro digestibility was determined using an incubator Daisy and pepsin enzymes and the combination of pepsin and pancreatin. The highest coefficient of crude protein digestibility was discovered to be in peels and wholemeal flour. The greatest fibre digestibility coefficients were obtained for peels, which contain about 65% of fibre in their dry matter. When pepsin was used, a higher phytic acid digestibility coefficient for G. max, Ph. vulgaris, peels, flour, groats and broken groats was observed; while when the combination of pepsin and pancreatin was used, higher phytic acid digestibility coefficients for peas, lentil and wholemeal flour were observed.

From Allergen Back to Antigen:. a Rational Approach to New Forms of Immunotherapy

Science.gov (United States)

Colombo, Paolo; Trapani, Antonino; Geraci, Domenico; Golino, Massimiliano; Gianguzza, Fabrizio; Bonura, Angela

2007-12-01

Mapping an epitope on a protein by gene fragmentation and/or point mutations is often expensive and time consuming. Analysis of a 3D model can be utilized to detect the amino acids residues which are exposed to the solvent surface and thus represent potential epitope residues. Parj1 and Parj2 are the two major allergens of the Parietaria judaica pollen belonging to the Lipid Transfer Protein family. Using their three-dimensional structures as a guide, a head to tail dimer expressing disulphide bond variants of the major allergens was generated by means of DNA recombinant technology. The hybrid was expressed in E.coli and its immunological activity studied in vivo and in vitro. Our results demonstrate that a hybrid polypeptide expressing disulphide bond variants of the major allergens of the Parietaria pollen displayed reduced allergenicity and enhanced T cell reactivity for induction of protective antibodies able to block human IgE induced during the natural course of sensitization against the Parietaria pollen.

Economic Factors Impacting Food Allergen Management: Perspectives from the Food Industry.

Science.gov (United States)

Gupta, Ruchi S; Taylor, Steve L; Baumert, Joseph L; Kao, Lauren M; Schuster, Erik; Smith, Bridget M

2017-10-01

Food allergies affect up to 8% of children in the United States and may occasionally lead to severe life-threatening reactions. Because there is currently no cure for food allergies, strict avoidance of the allergen-containing foods is the only means of preventing an allergic reaction. Consumers rely on food manufacturers to reliably track and declare the presence of food allergens in products. Over the past 10 to 20 years, the food industry has increasingly adopted allergen control approaches in its processing facilities. However, the major industry costs related to food allergen management have not been fully described. The objective of this study was to characterize the factors that contribute to the economic impact of food allergen control practices on the food industry. A focus group (n = 100) was conducted with food industry professionals to identify key areas of cost for food allergen management. A survey based on the domains identified was then developed and disseminated to a convenience sample (n = 50) of quality control food industry specialists with knowledge of their company's food allergen management practices. Nearly all companies (92%) produced food products containing one or more of the top eight allergenic foods recognized by the U.S. Food and Drug Administration or sesame seeds. Cleaning procedures, employee training, and the potential for a recall due to allergen cross-contact were most frequently rated as the important factors in food allergen management. Recalls due to food allergen cross-contact, cleaning procedures, equipment and premises design, and employee training were ranked as the greatest allergen management expenses. Although 96% of companies had a food allergen control plan in place, nearly half (42%) had at least one food allergen-related recall within the past 5 years. The industry appears to endorse a willingness to unify precautionary allergen labeling to communicate a clear message more effectively to consumers.

Generation and epitope analysis of human monoclonal antibody isotypes with specificity for the timothy grass major allergen Phl p 5a

DEFF Research Database (Denmark)

Hecker, J.; Diethers, A.; Seismann, H.

2011-01-01

The scarcity of monoclonal human IgE antibodies with specificity for defined allergens is a bottleneck for the molecular characterisation of allergens and their epitopes. Insights into the characteristics of such antibodies may allow for analyses of the molecular basis underlying allergenicity an...

Ancient whole grain Gluten-free egg-free Teff, Buckwheat, Quinoa and Amaranth pasta (abstract)

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This report demonstrates innovative ancient whole grain, gluten-free, egg-free pasta (no chemicals added) made using a kitchen counter-top appliance. Whole grain, fusilli pasta was prepared with teff, buckwheat, quinoa and amaranth flours. These ancient grains are called “Super Foods” due to thei...

Effective Allergen Management : Precautionary (may contain) allergen labeling; when to apply?

NARCIS (Netherlands)

Zandvoort. M.M.J. van

2013-01-01

When do you label food products as having been possibly cross contaminated by allergens? TNO can help you to develop a quantitative risk management guidance for food allergens, based on a unique method that quantifies the risk of food allergen traces in products and validated data on thresholds.

Epitope mapping of the major allergen from Atlantic cod in Spanish population reveals different IgE-binding patterns.

Science.gov (United States)

Perez-Gordo, Marina; Pastor-Vargas, Carlos; Lin, Jing; Bardina, Ludmilla; Cases, Barbara; Ibáñez, Maria Dolores; Vivanco, Fernando; Cuesta-Herranz, Javier; Sampson, Hugh A

2013-07-01

IgE-epitope mapping of allergens reveal important information about antigen components involved in allergic reactions. The peptide-based microarray immunoassay has been used to map epitopes of some food allergens. We developed a peptide microarray immunoassay to map allergenic epitopes in parvalbumin from Atlantic cod (Gad m 1), the most consumed cod species in Spain. Sera from 13 fish-allergic patients with specific IgE to cod parvalbumin were used. A library of overlapping peptides was synthesized, representing the primary sequence of Gad m 1. Peptides were used to analyze allergen-specific IgE antibodies in patient sera. 100% of the patients recognized one antigenic region of 15 amino acids in length in Gad m 1. This region only partially correlated with one of the three antigenic determinants of Gad c 1 (Allergen M), parvalbumin from Baltic cod (Gadus callarias). In the 3D model of the protein, this region was located on the surface of the protein. We have identified a relevant antigenic region in Gad m 1. This epitope could be considered as a severity marker and provides additional information to improve fish allergy diagnosis and the design of safe immunotherapeutic tools. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Influence of the 3D-conformation, glycan component and microheterogeneity on the epitope structure of Ole e 1, the major olive allergen - Use of recombinant isoforms and specific monoclonal antibodies as immunological tools

NARCIS (Netherlands)

González, Eva M.; Villalba, Mayte; Lombardero, Manuel; Aalbers, Marja; van Ree, Ronald; Rodríguez, Rosalía

2002-01-01

Ole e 1 is the main allergen of olive pollen, which is a major cause of pollinosis in countries of the Mediterranean area. Nine Ole e 1-specific murine monoclonal antibodies (mAbs), as well as two Ole e 1-isoforms and two Ole e 1-like allergens from lilac and privet, all of them obtained in Pichia

Standardization of RP-HPLC methods for the detection of the major peanut allergens Ara h 1, Ara h 2 and Ara h 3

Science.gov (United States)

Crude peanut extract (CPE) was analyzed for three major allergens (Ara h 1, h 2, and h 3) using a C12 and a C18 column at two wavelengths (280 and 220 nm) and under different solvent conditions. HPLC profiles were compared for retention time, resolution, and peak heights. CPE samples were spiked wit...

Protective effect of the DNA vaccine encoding the major house dust mite allergens on allergic inflammation in the murine model of house dust mite allergy

Directory of Open Access Journals (Sweden)

Lee Jaechun

2006-02-01

Full Text Available Abstract Background Vaccination with naked DNA encoding antigen induces cellular and humoral immunity characterized by the activation of specific Th1 cells. Objective To evaluate the effects of vaccination with mixed naked DNA plasmids encoding Der p 1, Der p 2, Der p 3, Der f 1, Der f 2, and Der f 3, the major house dust mite allergens on the allergic inflammation to the whole house dust mites (HDM crude extract. Methods Three hundred micrograms of these gene mixtures were injected into muscle of BALB/c mice. Control mice were injected with the pcDNA 3.1 blank vector. After 3 weeks, the mice were actively sensitized and inhaled with the whole house dust mite extract intranasally. Results The vaccinated mice showed a significantly decreased synthesis of total and HDM-specific IgE compared with controls. Analysis of the cytokine profile of lymphocytes after challenge with HDM crude extract revealed that mRNA expression of interferon-γ was higher in the vaccinated mice than in the controls. Reduced infiltration of inflammatory cells and the prominent infiltration of CD8+ T cells were observed in histology of lung tissue from the vaccinated mice. Conclusion Vaccination with DNA encoding the major house dust mite allergens provides a promising approach for treating allergic responses to whole house dust mite allergens.

Effect of reagins and allergen extracts on radioallergosorbent assays for mite allergen

International Nuclear Information System (INIS)

Tovey, E.R.; Vandenberg, R.A.

1978-01-01

The reproducibility of the radioallergosorbent (RAST) inhibition and direct binding assays with mite allergen were investigated in the presence of heterogeneous extracts and non-mite sensitive atopic sera. Both contain components similar to potential contaminants which would occur in the assay of mite allergen and dust allergen extracts. The standardized inhibition and direct binding assays employed had a day to day (n = 4) coefficient of variation [(s.d. x 100)/mean] of 15% and 24% respectively. The inhibition assay for mite allergen was reproducible in the presence of protein concentrations of added plant, fungal, arthropod and animal extracts in excess of the protein concentrations that occur under the operational mite assay conditions. The mite inhibition assay was also reproducible in the presence of non-mite allergen extracts, with and without additional sera containing IgE specific for the non0mite allergens. The binding of a constant quantity of mite allergen to the activated solid phase in the direct binding assay was reproducible in the presence of added bovine serum albumin, and of a fungal or arthropod extract, representing the heterogeneous components of an allergen extract at the concentrations of total protein known to occur in the direct binding assay of mite extracts. (author)

Food allergens in mattress dust in Norwegian homes - a potentially important source of allergen exposure.

Science.gov (United States)

Bertelsen, R J; Faeste, C K; Granum, B; Egaas, E; London, S J; Carlsen, K-H; Lødrup Carlsen, K C; Løvik, M

2014-01-01

Sensitization to food allergens and food allergic reactions are mostly caused by ingesting the allergen, but can also occur from exposure via the respiratory tract or the skin. Little is known about exposure to food allergens in the home environment. The objective of this study was firstly to describe the frequency of detection of allergens from fish, egg, milk, and peanut in mattress dust collected from homes of 13-year-old adolescents and secondly to identify home characteristics associated with the presence of food allergen contamination in dust. Food allergens were measured by dot blot analysis in mattress dust from 143 homes in Oslo, Norway. We analysed associations between home characteristics (collected by parental questionnaires and study technicians) and food allergens by multivariate regression models. Fish allergen was detected in 46%, peanut in 41%, milk in 39%, and egg allergen in 22% of the mattress dust samples; only three samples contained none of these allergens. All four food allergens were more frequently detected in mattresses in small dwellings (Food allergens occurred frequently in beds in Norwegian homes, with dwelling size and proximity of kitchen and bedroom as the most important determinants. Due to the amount of time children spent in the bedroom, mattress dust may be an important source of exposure to food allergens. © 2013 John Wiley & Sons Ltd.

Food allergens of plant origin - their molecular and evolutionary relationships

DEFF Research Database (Denmark)

Mills, E. N. C.; Madsen, Charlotte Bernhard; Shewry, P. R.

2003-01-01

Along with other forms of allergic disease, food allergies appear to be on the increase, with childhood allergies to foods such as peanuts being of particular concern. Around 7-10 foods are responsible for the majority of allergies, including several of plant origin, notably peanut. Allergies...... are usually triggered by the protein components in a food, which are also known as allergens. However, not all the proteins in an allergenic food like peanut are allergens. Why should this be? This question has been addressed by an EU-funded inter-disciplinary network of clinicians, food chemists and plant...

Food processing and allergenicity.

Science.gov (United States)

Verhoeckx, Kitty C M; Vissers, Yvonne M; Baumert, Joseph L; Faludi, Roland; Feys, Marcel; Flanagan, Simon; Herouet-Guicheney, Corinne; Holzhauser, Thomas; Shimojo, Ryo; van der Bolt, Nieke; Wichers, Harry; Kimber, Ian

2015-06-01

Food processing can have many beneficial effects. However, processing may also alter the allergenic properties of food proteins. A wide variety of processing methods is available and their use depends largely on the food to be processed. In this review the impact of processing (heat and non-heat treatment) on the allergenic potential of proteins, and on the antigenic (IgG-binding) and allergenic (IgE-binding) properties of proteins has been considered. A variety of allergenic foods (peanuts, tree nuts, cows' milk, hens' eggs, soy, wheat and mustard) have been reviewed. The overall conclusion drawn is that processing does not completely abolish the allergenic potential of allergens. Currently, only fermentation and hydrolysis may have potential to reduce allergenicity to such an extent that symptoms will not be elicited, while other methods might be promising but need more data. Literature on the effect of processing on allergenic potential and the ability to induce sensitisation is scarce. This is an important issue since processing may impact on the ability of proteins to cause the acquisition of allergic sensitisation, and the subject should be a focus of future research. Also, there remains a need to develop robust and integrated methods for the risk assessment of food allergenicity. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Characterization of the human T cell response to rye grass pollen allergens Lol p 1 and Lol p 5.

Science.gov (United States)

Burton, M D; Papalia, L; Eusebius, N P; O'Hehir, R E; Rolland, J M

2002-12-01

Knowledge of dominant T cell epitopes of major allergens recognized by allergic individuals is required to improve efficacy and safety of allergen immunotherapy. Rye grass pollen (RGP) is the most important source of seasonal aeroallergens in temperate climates and Lol p 1 and Lol p 5 are the two major IgE-reactive allergens. This study aimed to characterize the T cell response to these allergens using a large panel of RGP-sensitive individuals. Short-term RGP-specific T cell lines (TCL) were generated from 38 RGP-sensitive subjects and stimulated with Lol p 1 and/or Lol p 5 allergens and synthetic 20-mer peptides. Proliferative responses were determined by 3H-thymidine uptake and IL-5 and IFN-gamma in culture supernatants analysed by ELISA. Of 17 subjects tested for reactivity to both allergens 16 (94%) responded to Lol p 1 and/or Lol p 5, establishing these as major T cell-reactive allergens. Sites of T cell reactivity were spread throughout the allergen molecules but regions of high reactivity were found. For Lol p 1 these spanned residues 19-38, 109-128, 154-173, 190-209, and for Lol p 5 37-56, 100-119, 145-164, 154-173, 190-209, 217-236 and 226-245. IL-5 and IFN-gamma were produced by T cells cultured with proliferation-inducing peptides. T cell responses to RGP major allergens have been extensively characterized, providing fundamental information for developing T cell-targeted immunotherapy for RGP allergy.

Modulation of allergic immune responses by mucosal application of recombinant lactic acid bacteria producing the major birch pollen allergen Bet v 1.

Science.gov (United States)

Daniel, C; Repa, A; Wild, C; Pollak, A; Pot, B; Breiteneder, H; Wiedermann, U; Mercenier, A

2006-07-01

Probiotic lactic acid bacteria (LAB) are able to modulate the host immune system and clinical trials have demonstrated that specific strains have the capacity to reduce allergic symptoms. Therefore, we aimed to evaluate the potential of recombinant LAB producing the major birch pollen allergen Bet v 1 for mucosal vaccination against birch pollen allergy. Recombinant Bet v 1-producing Lactobacillus plantarum and Lactococcus lactis strains were constructed. Their immunogenicity was compared with purified Bet v 1 by subcutaneous immunization of mice. Intranasal application of the live recombinant strains was performed to test their immunomodulatory potency in a mouse model of birch pollen allergy. Bet v 1 produced by the LAB was recognized by monoclonal anti-Bet v 1 and IgE antibodies from birch pollen-allergic patients. Systemic immunization with the recombinant strains induced significantly lower IgG1/IgG2a ratios compared with purified Bet v 1. Intranasal pretreatment led to reduced allergen-specific IgE vs enhanced IgG2a levels and reduced interleukin (IL)-5 production of splenocytes in vitro, indicating a shift towards non-allergic T-helper-1 (Th1) responses. Airway inflammation, i.e. eosinophils and IL-5 in lung lavages, was reduced using either Bet v 1-producing or control strains. Allergen-specific secretory IgA responses were enhanced in lungs and intestines after pretreatment with only the Bet v 1-producing strains. Mucosal vaccination with live recombinant LAB, leading to a shift towards non-allergic immune responses along with enhanced allergen-specific mucosal IgA levels offers a promising approach to prevent systemic and local allergic immune responses.

Bioanalytical methods for food allergy diagnosis, allergen detection and new allergen discovery.

Science.gov (United States)

Gasilova, Natalia; Girault, Hubert H

2015-01-01

For effective monitoring and prevention of the food allergy, one of the emerging health problems nowadays, existing diagnostic procedures and allergen detection techniques are constantly improved. Meanwhile, new methods are also developed, and more and more putative allergens are discovered. This review describes traditional methods and summarizes recent advances in the fast evolving field of the in vitro food allergy diagnosis, allergen detection in food products and discovery of the new allergenic molecules. A special attention is paid to the new diagnostic methods under laboratory development like various immuno- and aptamer-based assays, including immunoaffinity capillary electrophoresis. The latter technique shows the importance of MS application not only for the allergen detection but also for the allergy diagnosis.

Comparison of buckwheat, red clover, and purple tansy as potential surrogate plants for use in semi-field pesticide risk assessments with Bombus impatiens

Directory of Open Access Journals (Sweden)

Angela E. Gradish

2016-07-01

Full Text Available Background. Bumble bees (Bombus spp. are important wild and managed pollinators. There is increased interest in incorporating data on bumble bees into risk assessments for pesticides, but standardized methods for assessing hazards of pesticides in semi-field and field settings have not yet been established for bumble bees. During semi-field studies, colonies are caged with pesticide-treated flowering surrogate plants, which must be attractive to foragers to ensure colony exposure to the test compound, and must produce an ample nectar and pollen to sustain colonies during testing. However, it is not known which plant(s are suitable for use in semi-field studies with bumble bees. Materials and Methods. We compared B. impatiens foraging activity and colony development on small plots of flowering buckwheat (Fagopyrum esculentum, var. common, red clover (Trifolium pratense, and purple tansy (Phacelia tanacetifolia under semi-field conditions to assess their suitability as surrogate plants for pesticide risk assessment studies with bumble bees. We also compared the growth characteristics and input requirements of each plant type. Results. All three plant types generally established and grew well. Red clover and purple tansy experienced significant weed pressure and/or insect pest damage. In contrast, pest pressure was extremely low in buckwheat. Overall, B. impatiens foraging activity was significantly greater on buckwheat plots than red clover or purple tansy, but plant type had no effect on number of individuals produced per colony or colony weight. Discussion. Because of the consistently high foraging activity and successful colony development observed on buckwheat plots, combined with its favourable growth characteristics and low maintenance requirements, we recommend buckwheat as a surrogate plant for use in semi-field pesticide toxicity assessments with B. impatiens.

Depigmented and polymerised house dust mite allergoid: allergen content, induction of IgG4 and clinical response.

Science.gov (United States)

Gallego, M T; Iraola, V; Himly, M; Robinson, D S; Badiola, C; García-Robaina, J C; Briza, P; Carnés, J

2010-01-01

Polymerised allergenic extracts (allergoids) are commonly used in allergen immunotherapy. Clinical efficacy and safety of these extracts have been demonstrated. Recently, allergen sequences have been identified by mass spectrometry in depigmented and polymerised (Dpg-Pol) extracts. The objectives of this study were to investigate the presence of allergens in Dpg-Pol extracts of house dust mite and to analyze the immunological changes induced by these extracts in asthmatic patients enrolled in a double-blind, placebo-controlled study. Dpg-Pol extracts were manufactured and vaccines with a composition of 50% Dermatophagoides pteronyssinus and 50% D. farinae (100 HEPL/ml) were prepared. Allergen composition was analyzed by mass spectrometry. Patients with asthma and rhinoconjunctivitis were treated in a 1-year, double-blind, placebo-controlled, parallel-group study with 6 up-dosing and monthly maintenance injections. Specific IgE and IgG4 titres to D. pteronyssinus, Der p 1 and Der p 2 were measured in patients' sera using the CAP system and direct ELISA experiments. Sequences from the major allergens Der p 1 and Der p 2 and from other allergens were identified in native and Dpg-Pol extracts. There was a statistically significant increase in specific IgG4, a decrease in the ratio of IgE/IgG4 to D. pteronyssinus and a significant increase in specific IgG4 to Der p 1 and Der p 2 in the patients allotted to active treatment. The detection of allergen sequences suggests preservation of major and minor allergens in Dpg-Pol allergoids from house dust mites. Efficacy in asthma treatment and the increase in specific IgG4 seem to be associated with the presence of major allergens in Dpg-Pol allergen extracts. Copyright (c) 2010 S. Karger AG, Basel.

Low concentration of sodium bicarbonate improves the bioactive compound levels and antioxidant and α-glucosidase inhibitory activities of tartary buckwheat sprouts.

Science.gov (United States)

Qin, Peiyou; Wei, Aichun; Zhao, Degang; Yao, Yang; Yang, Xiushi; Dun, Baoqing; Ren, Guixing

2017-06-01

This study aimed to investigate the effects of different concentrations of sodium bicarbonate (NaHCO 3 ) on the accumulation of flavonoids, total phenolics and d-chiro-inositol (DCI), as well as the antioxidant and α-glucosidase inhibitory activities, in tartary buckwheat sprouts. Treatment with low concentrations of NaHCO 3 (0.05, 0.1, and 0.2%) resulted in an increase in flavonoids, total phenolic compounds and DCI concentrations, and improved DPPH radical-scavenging and α-glucosidase inhibition activities compared with the control (0%). The highest levels of total flavonoids (26.69mg/g DW), individual flavonoids (rutin, isoquercitrin, quercetin, and kaempferol), total phenolic compounds (29.31mg/g DW), DCI (12.56mg/g DW), as well as antioxidant and α-glucosidase inhibition activities, were observed in tartary buckwheat sprouts treated with 0.05% NaHCO 3 for 96h. These results indicated that appropriate treatment with NaHCO 3 could improve the healthy benefits of tartary buckwheat sprouts. Copyright © 2016 Elsevier Ltd. All rights reserved.

Cat is a major allergen in patiens with asthma from west Siberia, Russia

Czech Academy of Sciences Publication Activity Database

Gusareva, Elena; Bragina, E.J.; Deeva, E.V.; Kazakevich, N.V.; Puzyrev, V.P.; Ogorodova, L.M.; Lipoldová, Marie

2006-01-01

Roč. 61, č. 4 (2006), s. 509-510 ISSN 0105-4538 R&D Projects: GA ČR(CZ) GD310/03/H147 Institutional research plan: CEZ:AV0Z50520514 Keywords : asthma patients * allergens * sensitization Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.334, year: 2006

Immunogold electron microscopic localization of timothy grass (Phleum pratense) pollen major allergens Phl p I and Phl p V after anhydrous fixation in acrolein vapor

NARCIS (Netherlands)

Grote, M.; Dolecek, C.; van Ree, R.; Valenta, R.

1994-01-01

We used the vapor phase of acrolein as an anhydrous fixative for timothy grass pollen in an immunogold double-labeling localization study of two different major allergens, Phl p I and Phl p V. More than 48 hr of fixation were needed for the subcellular pollen structures to be satisfactorily

Current challenges facing the assessment of the allergenic capacity of food allergens in animal models.

Science.gov (United States)

Bøgh, Katrine Lindholm; van Bilsen, Jolanda; Głogowski, Robert; López-Expósito, Iván; Bouchaud, Grégory; Blanchard, Carine; Bodinier, Marie; Smit, Joost; Pieters, Raymond; Bastiaan-Net, Shanna; de Wit, Nicole; Untersmayr, Eva; Adel-Patient, Karine; Knippels, Leon; Epstein, Michelle M; Noti, Mario; Nygaard, Unni Cecilie; Kimber, Ian; Verhoeckx, Kitty; O'Mahony, Liam

2016-01-01

Food allergy is a major health problem of increasing concern. The insufficiency of protein sources for human nutrition in a world with a growing population is also a significant problem. The introduction of new protein sources into the diet, such as newly developed innovative foods or foods produced using new technologies and production processes, insects, algae, duckweed, or agricultural products from third countries, creates the opportunity for development of new food allergies, and this in turn has driven the need to develop test methods capable of characterizing the allergenic potential of novel food proteins. There is no doubt that robust and reliable animal models for the identification and characterization of food allergens would be valuable tools for safety assessment. However, although various animal models have been proposed for this purpose, to date, none have been formally validated as predictive and none are currently suitable to test the allergenic potential of new foods. Here, the design of various animal models are reviewed, including among others considerations of species and strain, diet, route of administration, dose and formulation of the test protein, relevant controls and endpoints measured.

Expression and purification of a major allergen, Pla a 1, from Platanus acerifolia pollen and the preparation of its monoclonal antibody.

Science.gov (United States)

Ni, Wei-Wei; Huang, Wen; Wu, De-Qin; Zhou, Yan-Jun; Ji, Chun-Mei; Cao, Meng-Da; Guo, Miao; Sun, Jin-Lu; Wei, Ji-Fu

2017-09-01

Platanus acerifolia pollen is considered an important source of airborne allergens in numerous cities. Pla a 1 is a major allergen from P. acerifolia pollen. The present study aimed to express and purify Pla a 1, and to prepare its monoclonal antibody. In the present study, the Pla a 1 gene was subcloned into a pET‑28a vector and transformed into the ArcticExpress™ (DE3) RP Escherichia coli host strain. The purified Pla a 1 was then used to immunize BALB/c mice. When serum detection was positive, spleen cells were isolated from the mice and fused with SP2/0 myeloma cells at a ratio of 10:1. Hybridoma cells were screened by indirect ELISA and limiting dilution. Positive cells were used to induce the formation of antibody‑containing ascites fluid, and the antibodies were purified using protein A‑agarose. The results of the present study demonstrated that recombinant Pla a 1 was successfully expressed and purified, and exhibited positive immunoglobulin E‑binding to serum from patients allergic to P. acerifolia. A total of 11 hybridomas that steadily secreted anti‑Pla a 1 antibody were obtained and an immunoblotting analysis indicated that all of these monoclonal antibodies specifically recognized the Pla a 1 protein. These results suggested that specific anti‑Pla a 1 antibodies may be obtained, which can be used for the rapid detection of Pla a 1 allergens and in the preparation of vaccines against P. acerifolia pollen.

Production of recombinant peanut allergen Ara h 2 using Lactococcus lactis

DEFF Research Database (Denmark)

Glenting, J.; Poulsen, Lars K.; Kato, K.

2007-01-01

lactis is an attractive microorganism for use in the production of protein therapeutics. L. lactis is considered food grade, free of endotoxins, and is able to secrete the heterologous product together with few other native proteins. Hypersensitivity to peanut represents a serious allergic problem. Some...... of the major allergens in peanut have been described. However, for therapeutic usage more information about the individual allergenic components is needed. In this paper we report recombinant production of the Ara h 2 peanut allergen using L. lactis. Results: A synthetic ara h 2 gene was cloned into an L...

Legumin allergens from peanuts and soybeans: Effects of denaturation and aggregation on allergenicity

NARCIS (Netherlands)

Boxtel, E.L. van; Broek, L.A.M. van den; Koppelman, S.J.; Gruppen, H.

2008-01-01

Legumin proteins Ara h 3 from peanuts and glycinin from soybeans are increasingly described as important allergens. The stability of an allergen's IgE binding capacity towards heating and digestion is considered an important characteristic for food allergens. We investigated the effects of heating

Legumin allergens from peanuts and soybeans : Effects of denaturation and aggregation on allergenicity

NARCIS (Netherlands)

Boxtel, van E.L.; Broek, van den L.A.M.; Koppelman, S.J.; Gruppen, H.

2008-01-01

Legumin proteins Ara h 3 from peanuts and glycinin from soybeans are increasingly described as important allergens. The stability of an allergen's IgE binding capacity towards heating and digestion is considered an important characteristic for food allergens. We investigated the effects of heating

Comparing Proteolytic Fingerprints of Antigen-Presenting Cells during Allergen Processing.

Science.gov (United States)

Hofer, Heidi; Weidinger, Tamara; Briza, Peter; Asam, Claudia; Wolf, Martin; Twaroch, Teresa E; Stolz, Frank; Neubauer, Angela; Dall, Elfriede; Hammerl, Peter; Jacquet, Alain; Wallner, Michael

2017-06-08

Endolysosomal processing has a critical influence on immunogenicity as well as immune polarization of protein antigens. In industrialized countries, allergies affect around 25% of the population. For the rational design of protein-based allergy therapeutics for immunotherapy, a good knowledge of T cell-reactive regions on allergens is required. Thus, we sought to analyze endolysosomal degradation patterns of inhalant allergens. Four major allergens from ragweed, birch, as well as house dust mites were produced as recombinant proteins. Endolysosomal proteases were purified by differential centrifugation from dendritic cells, macrophages, and B cells, and combined with allergens for proteolytic processing. Thereafter, endolysosomal proteolysis was monitored by protein gel electrophoresis and mass spectrometry. We found that the overall proteolytic activity of specific endolysosomal fractions differed substantially, whereas the degradation patterns of the four model allergens obtained with the different proteases were extremely similar. Moreover, previously identified T cell epitopes were assigned to endolysosomal peptides and indeed showed a good overlap with known T cell epitopes for all four candidate allergens. Thus, we propose that the degradome assay can be used as a predictor to determine antigenic peptides as potential T cell epitopes, which will help in the rational design of protein-based allergy vaccine candidates.

Endogenous allergens and compositional analysis in the allergenicity assessment of genetically modified plants.

Science.gov (United States)

Fernandez, A; Mills, E N C; Lovik, M; Spoek, A; Germini, A; Mikalsen, A; Wal, J M

2013-12-01

Allergenicity assessment of genetically modified (GM) plants is one of the key pillars in the safety assessment process of these products. As part of this evaluation, one of the concerns is to assess that unintended effects (e.g. over-expression of endogenous allergens) relevant for the food safety have not occurred due to the genetic modification. Novel technologies are now available and could be used as complementary and/or alternative methods to those based on human sera for the assessment of endogenous allergenicity. In view of these developments and as a step forward in the allergenicity assessment of GM plants, it is recommended that known endogenous allergens are included in the compositional analysis as additional parameters to be measured. Copyright © 2013 Elsevier Ltd. All rights reserved.

New routes of allergen immunotherapy.

Science.gov (United States)

Aricigil, Mitat; Muluk, Nuray Bayar; Sakarya, Engin Umut; Sakalar, Emine Güven; Senturk, Mehmet; Reisacher, William R; Cingi, Cemal

2016-11-01

Allergen immunotherapy is the only cure for immunoglobulin E mediated type I respiratory allergies. Subcutaneous immunotherapy (SCIT) and sublingual immunotherapy (SLIT) are the most common treatments. In this article, we reviewed new routes of allergen immunotherapy. Data on alternative routes to allow intralymphatic immunotherapy (ILIT), epicutaneous immunotherapy (EPIT), local nasal immunotherapy (LNIT), oral immunotherapy (OIT), and oral mucosal immunotherapy (OMIT) were gathered from the literature and were discussed. ILIT features direct injection of allergens into lymph nodes. ILIT may be clinically effective after only a few injections and induces allergen-specific immunoglobulin G, similarly to SCIT. A limitation of ILIT is that intralymphatic injections are required. EPIT features allergen administration by using patches mounted on the skin. EPIT seeks to target epidermal antigen-presenting Langerhans cells rather than mast cells or the vasculature; this should reduce both local and systemic adverse effects. LNIT involves the spraying of allergen extracts into the nasal cavity. Natural or chemically modified allergens (the latter, termed allergoids, lack immunoglobulin E reactivity) are prepared in a soluble form. OIT involves the regular administration of small amounts of a food allergen by mouth and commences with low oral doses, which are then increased as tolerance develops. OMIT seeks to deliver allergenic proteins to an expanded population of Langerhans cells in the mucosa of the oral cavity. ILIT, EPIT, LNIT, OIT, and OMIT are new routes for allergen immunotherapy. They are safe and effective.

Expanded bed adsorption as a fast technique for the large-scale purification of the complete isoform pool of Ber e 1, the major allergen from Brazil nuts.

NARCIS (Netherlands)

Boxtel, van E.L.; Koningsveld, van G.A.; Koppelman, S.J.; Broek, van den L.A.M.; Voragen, A.G.J.; Gruppen, H.

2006-01-01

A new, fast, large-scale purification method for Ber e 1, the major allergen from Brazil nuts, using expanded bed adsorption (EBA) chromatography, is presented. Using EBA, crude extracts can be applied to a fluidized column, which allows the unhindered passage of particulate impurities, thereby

Responses of human birch pollen allergen-reactive T cells to chemically modified allergens (allergoids).

Science.gov (United States)

Dormann, D; Ebner, C; Jarman, E R; Montermann, E; Kraft, D; Reske-Kunz, A B

1998-11-01

Allergoids are widely used in specific immunotherapy for the treatment of IgE-mediated allergic diseases. The aim of this study was to analyse whether a modification of birch pollen allergens with formaldehyde affects the availability of T-cell epitopes. Efficient modification of the allergens was verified by determining IgE and IgG binding activity using ELISA inhibition tests. T-cell responses to birch pollen allergoids were analysed in polyclonal systems, using peripheral blood mononuclear cells (PBMC) of five birch pollen-allergic individuals, as well as birch pollen extract-reactive T-cell lines (TCL), established from the peripheral blood of 14 birch pollen-allergic donors. To determine whether the modification of natural (n)Bet v 1 with formaldehyde or maleic anhydride results in epitope-specific changes in T-cell reactivities, 22 Bet v 1-specific T-cell clones (TCC), established from nine additional birch pollen-allergic individuals, were tested for their reactivity with these products. The majority of PBMC and TCL showed a reduced response to the birch pollen extract allergoid. Bet v 1-specific TCC could be divided into allergoid-reactive and -non-reactive TCC. No simple correlation between possible modification sites of formaldehyde in the respective T-cell epitopes and the stimulatory potential of the allergoid was observed. Mechanisms of suppression or of anergy induction were excluded as an explanation for the non-reactivity of representative TCC. All TCC could be stimulated by maleylated and unmodified nBet v 1 to a similar extent. These results demonstrate differences in the availability of T-cell epitopes between allergoids and unmodified allergens, which are most likely due to structural changes within the allergen molecule.

Physicochemical and Antioxidant Properties of Buckwheat Protein Isolates with Different Polyphenolic Content Modified by Limited Hydrolysis with Trypsin

Directory of Open Access Journals (Sweden)

Xiao-Yan Wang

2012-01-01

Full Text Available Effects of limited hydrolysis with trypsin on the physicochemical and antioxidant properties of buckwheat protein isolates (BPIs obtained with untreated and 2-propanol-extracted meal have been investigated and compared. The dephenolization treatment significantly improved the hydrolysis of BPI, which resulted in the gradual decrease in total and protein-bound polyphenolic content, but an increase in the free polyphenolic content. The hydrolysis of globulins was much easier than that of the albumins. The removal of polyphenols improved the hydrolysis of the albumin fraction. The modified BPIs with high polyphenolic content exhibited much higher DPPH radical scavenging activity and reducing power, but poorer ferrous ion chelating ability than those with low polyphenolic content. These results suggest that the limited hydrolysis is suitable for modification of the properties of buckwheat proteins.

[Evaluation of the total biological activity and allergenic composition of allergenic extracts].

Science.gov (United States)

Lombardero, M; González, R; Duffort, O; Juan, F; Ayuso, R; Ventas, P; Cortés, C; Carreira, J

1986-01-01

In the present study, a complete procedure is presented in order to standardize allergenic extracts, the meaning of which is the measurement of the total allergenic activity and the determination of the allergenic composition. The measurement of the biological activity comprises 2 steps: Preparation of Reference Extracts and determination of their "in vivo" activity. Evaluation of the total allergenic activity of extracts for clinical use. Reference extracts were prepared from the main allergens and their "in vivo" biological activity was determined by a quantitative skin prick test in a sample of at least 30 allergic patients. By definition, the protein concentration of Reference Extract that produces, in the allergic population, a geometric mean wheal of 75 mm.2 has an activity of 100 biological units (BUs). The determination of the biological activity of a problem extract is made by RAST inhibition. The sample is compared with the corresponding Reference Extract by this technique and, from this comparison, it is possible to quantify the activity of the problem extract in biologic units (BUs) with clinical significance. Likewise, different techniques have been used to determine the allergenic composition of extracts. These techniques comprise 2 steps: Separation of the components of the extract. Identification of the components that bind specific human IgE. The separation of the components of the extract has been carried out by isoelectric focusing (IEF) and electrophoresis in the presence of sodium dodecyl sulphate (SDS-PAGE). In order to identify the allergenic components, an immunoblotting technique has been employed. The separated components in the IEF gel or SDS-PAGE gel are transferred to a nitrocellulose sheet and later on, this membrane is overlaid with a serum pool from allergic patients and a mouse monoclonal anti-human IgE, labelled with 125I. Finally, the autoradiography of the nitrocellulose membrane is obtained. In this way it is possible to compare

Food-cooking processes modulate allergenic properties of hen's egg white proteins.

Science.gov (United States)

Liu, Xiaoyu; Feng, Bai-Sui; Kong, Xiaoli; Xu, Hong; Li, Xiumin; Yang, Ping-Chang; Liu, Zhigang

2013-01-01

Reducing the allergenicity of food allergens can suppress the clinical symptoms of food allergy. The objective of the present study was to investigate the effects of processing on the allergenic properties of hen's egg white proteins. Eggs were processed by traditional Chinese cooking, including steaming, water boiling, frying, spicing and tea boiling. The contents of processed egg protein were assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis; the allergenicity was evaluated by Western blotting, enzyme-linked immunosorbent assay and enzyme allergosorbent test inhibition. Circular dichroism spectrum analysis of four major egg allergens from various egg products was performed as well. A mouse model of food allergy was developed to test the allergenicity of processed egg protein in vivo. Protein degradation was significant following tea boiling and spiced-tea boiling. The total allergenic potential of water-boiled egg and fried egg was relatively higher than that of steamed egg, spiced egg and tea-boiled egg. Challenge with proteins from raw egg, water-boiled egg and fried egg induced skewed T-helper 2 pattern responses (Th2 responses) in the intestine of mice sensitized to egg proteins; however, when the mice sensitized to egg proteins were challenged with proteins from steamed egg, spiced egg and tea-boiled egg, respectively, only weak Th2 responses were induced in their intestine. Processing by steaming, spicing, or tea boiling can weaken the allergenicity of egg proteins. Copyright © 2012 S. Karger AG, Basel.

A rapid and efficient two-step gel electrophoresis method for the purification of major rye grass pollen allergens.

Science.gov (United States)

Levy, D; Davies, J; O'Hehir, R; Suphioglu, C

2001-06-01

Purified proteins are mandatory for molecular, immunological and cellular studies. However, purification of proteins from complex mixtures requires specialised chromatography methods (i.e., gel filtration, ion exchange, etc.) using fast protein liquid chromatography (FPLC) or high-performance liquid chromatography (HPLC) systems. Such systems are expensive and certain proteins require two or more different steps for sufficient purity and generally result in low recovery. The aim of this study was to develop a rapid, inexpensive and efficient gel-electrophoresis-based protein purification method using basic and readily available laboratory equipment. We have used crude rye grass pollen extract to purify the major allergens Lol p 1 and Lol p 5 as the model protein candidates. Total proteins were resolved on large primary gel and Coomassie Brilliant Blue (CBB)-stained Lol p 1/5 allergens were excised and purified on a secondary "mini"-gel. Purified proteins were extracted from unstained separating gels and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analyses. Silver-stained SDS-PAGE gels resolved pure proteins (i.e., 875 microg of Lol p 1 recovered from a 8 mg crude starting material) while immunoblot analysis confirmed immunological reactivity of the purified proteins. Such a purification method is rapid, inexpensive, and efficient in generating proteins of sufficient purity for use in monoclonal antibody (mAb) production, protein sequencing and general molecular, immunological, and cellular studies.

8 Allergenic Composition of Polymerized Allergen Extracts of Betula verrucosa, Dermatophagoides Pteronyssinus and Phleum Pratense

OpenAIRE

Fernandez-Caldas, Enrique; Cases, Barbara; Tudela, Jose Ignacio; Fernandez, Eva Abel; Casanovas, Miguel; Subiza, Jose Luis

2012-01-01

Background Allergoids have been successfully used in the treatment of respiratory allergic diseases. They are modified allergen extracts that allow the administration of high allergen doses, due to their reduced IgE binding capacity.They maintain allergen-specific T-cell recognition. Since they are native allergen extracts that have been polymerized with glutaraldehyde, identification of the allergenic molecules requires more complicated methods. The aim of the study was to determine the qual...

Transcripts of Anthocyanidin Reductase and Leucoanthocyanidin Reductase and Measurement of Catechin and Epicatechin in Tartary Buckwheat

Directory of Open Access Journals (Sweden)

Yeon Bok Kim

2014-01-01

Full Text Available Anthocyanidin reductase (ANR and leucoanthocyanidin reductase (LAR play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions.

Allergens from fish and egg

DEFF Research Database (Denmark)

Poulsen, L.K.; Hansen, Tine Kjær; Norgaard, A.

2001-01-01

Allergens from fish and egg belong to some of the most frequent causes of food allergic reactions reported in the literature. Egg allergens have been described in both white and yolk, and the egg white proteins ovomucoid, ovalbumin, ovotransferrin and lysozyme have been adopted in the allergen...... nomenclature as Gal d1-d4. The most reported allergen from egg yolk seems to be alpha-livitin. In fish, the dominating allergen is the homologues of Gad c1 from cod, formerly described as protein M. A close cross-reactivity exists within different species of fish between this calcium-binding protein family...

Chemical modification of birch allergen extract leads to a reduction in allergenicity as well as immunogenicity.

Science.gov (United States)

Würtzen, Peter Adler; Lund, Lise; Lund, Gitte; Holm, Jens; Millner, Anders; Henmar, Helene

2007-01-01

In Europe, specific immunotherapy is currently conducted with vaccines containing allergen preparations based on intact extracts. In addition to this, chemically modified allergen extracts (allergoids) are used for specific allergy treatment. Reduced allergenicity and thereby reduced risk of side effects in combination with retained ability to activate T cells and induce protective allergen-specific antibody responses has been claimed for allergoids. In the current study, we compared intact allergen extracts and allergoids with respect to allergenicity and immunogenicity. The immunological response to birch allergen extract, alum-adsorbed extract, birch allergoid and alum-adsorbed allergoid was investigated in vitro in human basophil histamine release assay and by stimulation of human allergen-specific T cell lines. In vivo, Bet v 1-specific IgG titers in mice were determined after repetitive immunizations. In all patients tested (n = 8), allergoid stimulations led to reduced histamine release compared to the intact allergen extract. However, the allergoid preparations were not recognized by Bet v 1-specific T cell lines (n = 7), which responded strongly to the intact allergen extract. Mouse immunizations showed a clearly reduced IgG induction by allergoids and a strongly potentiating effect of the alum adjuvant. Optimal IgG titers were obtained after 3 immunizations with intact allergen extracts, while 5 immunizations were needed to obtain maximal response to the allergoid. The reduced histamine release observed for allergoid preparations may be at the expense of immunological efficacy because the chemical modifications lead to a clear reduction in T cell activation and the ability to induce allergen-specific IgG antibody responses. Copyright 2007 S. Karger AG, Basel.

Allergenicity, immunogenicity and dose-relationship of three intact allergen vaccines and four allergoid vaccines for subcutaneous grass pollen immunotherapy.

Science.gov (United States)

Henmar, H; Lund, G; Lund, L; Petersen, A; Würtzen, P A

2008-09-01

Different vaccines containing intact allergens or chemically modified allergoids as active ingredients are commercially available for specific immunotherapy. Allergoids are claimed to have decreased allergenicity without loss of immunogenicity and this is stated to allow administration of high allergoid doses. We compared the allergenicity and immunogenicity of four commercially available chemically modified grass pollen allergoid products with three commercially available intact grass pollen allergen vaccines. The allergenicity was investigated with immunoglobulin (Ig)E-inhibition and basophil activation assays. Human T cell proliferation and specific IgG-titres following mouse immunizations were used to address immunogenicity. Furthermore, intact allergen vaccines with different contents of active ingredients were selected to study the influence of the allergen dose. In general, a lower allergenicity for allergen vaccines was clearly linked to a reduced immunogenicity. Compared with the vaccine with the highest amount of intact allergen, the allergoids caused reduced basophil activation as well as diminished immunogenicity demonstrated by reduced T cell activation and/or reduced induction of murine grass-specific IgG antibodies. Interestingly, intact allergen vaccines with lower content of active ingredient exhibited similarly reduced allergenicity, while immunogenicity was still higher or equal to that of allergoids. The low allergenicity observed for some allergoids was inherently linked to a significantly lower immunogenic response questioning the rationale behind the chemical modification into allergoids. In addition, the linkage between allergenicity, immunogenicity and dose found for intact allergen vaccines and the immunogen as well as allergenic immune responses observed for allergoids suggest that the modified allergen vaccines do not contain high doses of immunologically active ingredients.

Suspension-cultured BY-2 tobacco cells produce and mature immunologically active house dust mite allergens

NARCIS (Netherlands)

Lienard, David; Tran Dinh, Olivia; van Oort, Erika; van Overtvelt, Laurence; Bonneau, Caroline; Wambre, Eric; Bardor, Muriel; Cosette, Pascal; Didier-Laurent, Alain; de Borne, François Dorlhac; Delon, Rene; van Ree, Ronald; Moingeon, Philippe; Faye, Loïc; Gomord, Véronique

2007-01-01

The replacement of crude allergen extracts by selected allergens currently represents a major goal for the improvement of allergy diagnosis and immunotherapy. Indeed, the development of molecularly defined vaccines would facilitate both standardization and enhance batch-to-batch reproducibility as

The distribution of dust mite allergen in the houses of patients with asthma

International Nuclear Information System (INIS)

Tovey, E.R.; Chapman, M.D.; Wells, C.W.; Platts-Mills, T.A.

1981-01-01

Using an inhibition radioimmunoassay for the major allergen from Dermatophagoides pteronyssinus (antigen P1), we studied the distribution of this dust allergen in the houses of patients with asthma. Both bed and floor dust samples contained a wide range of antigen P1, 100 to 100,000 ng/g of fine dust, and this concentration correlated well with the number of mite bodies (r . 0.81, p less than 0.001). We were unable to detect antigen P1 in the air of undisturbed rooms. However, during domestic activity, between 1 and 30 ng were collected on a filter than sampled air for 45 min at 17 L/min. Using a cascade impactor it was shown that greater than 80% of the airborne antigen P1 was associated with particles greater than 10 mu in diameter. Some of the particles containing allergen could be identified because they formed precipitin rings when impacted onto agarose containing rabbit antimite antiserum. These particles had the physical appearance of mite feces, which are the major source of antigen P1 in mite cultures. The results suggested that natural exposure to this dust allergen allows occasional fecal particles to enter the lungs and that these particles contain very concentrated allergen

Time-course study and effects of drying method on concentrations of gamma-aminobutyric acid, flavonoids, anthocyanin, and 2''-hydroxynicotianamine in leaves of buckwheats.

Science.gov (United States)

Suzuki, Tatsuro; Watanabe, Masami; Iki, Makiko; Aoyagi, Yasuo; Kim, Sun-Ju; Mukasa, Yuji; Yokota, Satoshi; Takigawa, Shigenobu; Hashimoto, Naoto; Noda, Takahiro; Yamauchi, Hiroaki; Matsuura-Endo, Chie

2009-01-14

Concentrations of gamma-aminobutyric acid (GABA), rutin, minor flavonoids (such as orientin), anthocyanin, and 2''-hydroxynicotianamine (2HN) were quantified in the leaves of common and tartary buckwheat (Fagopyrum esculentum Moench and Fagopyrum tataricum Gaertn., respectively), at 14, 28, and 42 days after sowing (DAS). GABA and rutin concentrations peaked at 42 DAS, whereas anthocyain, 2HN, and minor flavonoid concentrations declined with the age of the plants. However, at 42 DAS, anthocyanin concentrations in the leaves of tartary buckwheat Hokkai T10 leaves were at least 10-fold greater than in the other buckwheats tested. In addition, the effects on target compound concentrations and 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of three different drying methods (20 h at 40 degrees C, 7 h at 70 degrees C, or lyophilization) were investigated. In general, the drying method had no significant effect on the parameters tested. These results indicate that, in terms of GABA, rutin, and anthocyanin concentrations, leaf powder from 42 day old Hokkai T10 has the potential to be a useful food ingredient, such as Ao-jiru juice.

Bioanalytical methods for food allergy diagnosis, allergen detection and new allergen discovery

OpenAIRE

Gasilova, Natalia; Girault, Hubert H

2015-01-01

For effective monitoring and prevention of the food allergy, one of the emerging health problems nowadays, existing diagnostic procedures and allergen detection techniques are constantly improved. Meanwhile, new methods are also developed, and more and more putative allergens are discovered. This review describes traditional methods and summarizes recent advances in the fast evolving field of the in vitro food allergy diagnosis, allergen detection in food products and discovery of the new all...

Allergen relative abundance in several wheat varieties as revealed via a targeted quantitative approach using MS.

Science.gov (United States)

Rogniaux, Hélène; Pavlovic, Marija; Lupi, Roberta; Lollier, Virginie; Joint, Mathilde; Mameri, Hamza; Denery, Sandra; Larré, Colette

2015-05-01

Food allergy has become a major health issue in developed countries, therefore there is an urgent need to develop analytical methods able to detect and quantify with a good sensitivity and reliability some specific allergens in complex food matrices. In this paper, we present a targeted MS/MS approach to compare the relative abundance of the major recognized wheat allergens in the salt-soluble (albumin/globulin) fraction of wheat grains. Twelve allergens were quantified in seven wheat varieties, selected from three Triticum species: T. aestivum (bread wheat), T. durum (durum wheat), and T. monococcum. The allergens were monitored from one or two proteotypic peptides and their relative abundance was deduced from the intensity of one fragment measured in MS/MS. Whereas the abundance of some of the targeted allergens was quite stable across the genotypes, others like alpha-amylase inhibitors showed clear differences according to the wheat species, in accordance with the results of earlier functional studies. This study enriches the scarce knowledge available on allergens content in wheat genotypes, and brings new perspectives for food safety and plant breeding. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Assessment of the Olea pollen and its major allergen Ole e 1 concentrations in the bioearosol of two biogeographical areas

Science.gov (United States)

Moreno-Grau, S.; Aira, M. J.; Elvira-Rendueles, B.; Fernández-González, M.; Fernández-González, D.; García-Sánchez, A.; Martínez-García, M. J.; Moreno, J. M.; Negral, L.; Vara, A.; Rodríguez-Rajo, F. J.

2016-11-01

The Olea pollen is currently an important allergy source. In some regions of Southern Spain, olive pollen is the main cause of allergic sensitization exceeding 40% of the sensitized individuals. Due to the scarce presence of olive trees in Northern Spain, limited to some cultivated fields in the South of the Galicia region where they also grow wild, only 8% of the sensitized individuals showed positive results for Olea pollen. The aim of the paper was to assess the behaviour pattern of the Olea pollen and its aeroallergens in the atmosphere, as this information could help us to improve the understanding and prevention of clinical symptoms. Airborne Olea pollen and Ole e 1 allergens were quantified in Cartagena (South-eastern Spain) and Ourense (North-western Spain). A volumetric pollen trap and a Burkard Cyclone sampler were used for pollen and allergen quantification. The Olea flowering took place in April or May in both biometeorological sampling areas. The higher concentrations were registered in the Southern area of Spain, for both pollen and Ole e 1, with values 8 times higher for pollen concentrations and 40 times higher for allergens. An alternate bearing pattern could be observed, characterized by years with high pollen values and low allergen concentrations and vice versa. Moreover, during some flowering seasons the allergen concentrations did not correspond to the atmospheric pollen values. Variations in weather conditions or Long Distance Transport (LDT) processes could explain the discordance. The back trajectory analysis shows that the most important contributions of pollen and allergens in the atmosphere are coincident with air masses passing through potential source areas. The exposure to olive pollen may not be synonym of antigen exposure.

Isolation, cloning, and characterization of the 2S albumin: A new allergen from hazelnut

NARCIS (Netherlands)

Garino, Cristiano; Zuidmeer, Laurian; Marsh, Justin; Lovegrove, Alison; Morati, Maria; Versteeg, Serge; Schilte, Piet; Shewry, Peter; Arlorio, Marco; van Ree, Ronald

2010-01-01

Scope: 2S albumins are the major allergens involved in severe food allergy to nuts, seeds, and legumes. We aimed to isolate, clone, and express 2S albumin from hazelnut and determine its allergenicity. Methods: 2S albumin from hazelnut extract was purified using size exclusion chromatography and

Highlight on Bottlenecks in Food Allergen Analysis: Detection and Quantification by Mass Spectrometry.

Science.gov (United States)

Planque, Mélanie; Arnould, Thierry; Renard, Patricia; Delahaut, Philippe; Dieu, Marc; Gillard, Nathlie

2017-07-01

Food laboratories have developed methods for testing allergens in foods. The efficiency of qualitative and quantitative methods is of prime importance in protecting allergic populations. Unfortunately, food laboratories encounter barriers to developing efficient methods. Bottlenecks include the lack of regulatory thresholds, delays in the emergence of reference materials and guidelines, and the need to detect processed allergens. In this study, ultra-HPLC coupled to tandem MS was used to illustrate difficulties encountered in determining method performances. We measured the major influences of both processing and matrix effects on the detection of egg, milk, soy, and peanut allergens in foodstuffs. The main goals of this work were to identify difficulties that food laboratories still encounter in detecting and quantifying allergens and to sensitize researchers to them.

Safety of allergen immunotherapy: a review of premedication and dose adjustment.

Science.gov (United States)

Morris, A Erika; Marshall, Gailen D

2012-03-01

From the first allergen immunotherapy proposed in the early 1900s to the present day, numerous studies have proven the efficacy of allergen immunotherapy for the treatment of allergic rhinitis, allergic conjunctivitis, allergic asthma and stinging insect hypersensitivity. The major risk, however small, with allergen immunotherapy is anaphylaxis. There has been considerable interest and debate regarding risk factors for immunotherapy reactions (local and systemic) and interventions to reduce the occurrence of these reactions. One of these interventions that is especially debated regards dose adjustment for various reasons, but in particular for local reactions. In this review, we discuss the safety of immunotherapy and provide a comprehensive review of the literature regarding immunotherapy schedules and doses.

Lol p XI, a new major grass pollen allergen, is a member of a family of soybean trypsin inhibitor-related proteins

NARCIS (Netherlands)

van Ree, R.; Hoffman, D. R.; van Dijk, W.; Brodard, V.; Mahieu, K.; Koeleman, C. A.; Grande, M.; van Leeuwen, W. A.; Aalberse, R. C.

1995-01-01

BACKGROUND: Monoclonal antibodies were obtained against an unknown allergen from Lolium perenne grass pollen. The allergen had an apparent molecular mass of 18 kd on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Earlier immunoblotting studies had shown that carbohydrate-specific IgG

Effects of nasal corticosteroids on boosts of systemic allergen-specific IgE production induced by nasal allergen exposure.

Directory of Open Access Journals (Sweden)

Cornelia Egger

Full Text Available Allergen exposure via the respiratory tract and in particular via the nasal mucosa boosts systemic allergen-specific IgE production. Intranasal corticosteroids (INCS represent a first line treatment of allergic rhinitis but their effects on this boost of allergen-specific IgE production are unclear.Here we aimed to determine in a double-blind, placebo-controlled study whether therapeutic doses of an INCS preparation, i.e., nasal fluticasone propionate, have effects on boosts of allergen-specific IgE following nasal allergen exposure.Subjects (n = 48 suffering from grass and birch pollen allergy were treated with daily fluticasone propionate or placebo nasal spray for four weeks. After two weeks of treatment, subjects underwent nasal provocation with either birch pollen allergen Bet v 1 or grass pollen allergen Phl p 5. Bet v 1 and Phl p 5-specific IgE, IgG1-4, IgM and IgA levels were measured in serum samples obtained at the time of provocation and one, two, four, six and eight weeks thereafter.Nasal allergen provocation induced a median increase to 141.1% of serum IgE levels to allergens used for provocation but not to control allergens 4 weeks after provocation. There were no significant differences regarding the boosts of allergen-specific IgE between INCS- and placebo-treated subjects.In conclusion, the application of fluticasone propionate had no significant effects on the boosts of systemic allergen-specific IgE production following nasal allergen exposure.http://clinicaltrials.gov/NCT00755066.

Beneficial Influence of Short-Term Germination on Decreasing Allergenicity of Peanut Proteins.

Science.gov (United States)

Li, Yingchao; Sun, Xiulan; Ma, Zhezhe; Cui, Yan; Du, Chao; Xia, Xiuhua; Qian, He

2016-01-01

Most allergenic storage proteins in peanuts are degraded during seed germination. By altering this natural physiological process, it might be possible to reduce peanut protein allergenicity. However, little is known about the change in allergenic proteins and their corresponding immunocreactivity, and the effects of major environmental conditions on their allergenicity during germination. In this study, the influence of different germination conditions (temperature and light) on the degradation of Ara h1 and allergenicity changes of peanut seeds was evaluated by ELISA and Western blotting. The results showed that the 40- and 65-kDa proteins in peanut seeds degraded rapidly during the time course, beginning at 60 (at 25 °C) and 108 h (at 20 °C), and the corresponding immunocreactivity of Ara h1 decreased approximately one-third after 5 to 7 d of germination. Compared with the cotyledons, the embryonic axes had a higher proportion of Ara h1, which was then degraded relatively faster during germination, resulting in a significant reduction in its allergenicity. Although a higher temperature improved the seed germination rate, it affected sprout quality (as did light); therefore, 25 °C and dark surroundings were suitable conditions under which peanut sprouts were processed; neither factor significantly affected the allergenicity of Ara h1. These results provided a theoretical basis for studies using biological methods to reduce peanut allergenicity. © 2015 Institute of Food Technologists®

Diversity and pollination value of insects visiting the flowers of a rare buckwheat (Eriogonum pelinophilum: Polygonaceae in disturbed and “natural” areas

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Griswold, T. L.

2011-06-01

Full Text Available We compared flower-visitors of the endangered plant Eriogonum pelinophilum, at relatively undisturbed and highly disturbed sites. We found no difference between sites in flower visitation rate or species richness of flower-visitors; species diversity of flower-visitors was higher at disturbed than at undisturbed sites but there was no difference in equitability. We found significant differences in total E. pelinophilum pollen carried on the body among 14 abundant bee species; eight abundant wasp species; and 12 abundant fly species. Both bee and wasp species carried significantly more pollen on the ventral compared to dorsal segments of the body; pollen on the body of fly species was more equally distributed across body surfaces. Total pollen carried on flower-visitor bodies was significantly related to visitor length, suggesting that larger visitors were more effective pollinators. Total Pollination Value, a measure combining both visitor abundance and body pollen was greater at the disturbed site than the undisturbed site, further suggesting that pollination in fragments of this rare species is not a major concern. We conclude that the high diversity of insect flower-visitors and the generalized nature of E. pelinophilum flowers make a special management programme to conserve pollinators unnecessary. Conservation of this buckwheat is best achieved by simple habitat preservation, together with a program to enlist private citizens to include buckwheat plants in their backyard gardens.

Molecular Allergen-Specific IgE Assays as a Complement to Allergen Extract-Based Sensitization Assessment

NARCIS (Netherlands)

Aalberse, Rob C.; Aalberse, Joost A.

2015-01-01

Molecular allergen-based component-resolved diagnostic IgE antibody tests have emerged in the form of singleplex assays and multiplex arrays. They use both native and recombinant allergen molecules, sometimes in combination with each other, to supplement allergen extract-based IgE antibody analyses.

Effect of roasting on the allergenicity of major peanut allergens Ara h 1 and Ara h 2/6: the necessity of degranulation assays

DEFF Research Database (Denmark)

Vissers, Y. M.; Iwan, M.; Adel-Patient, K.

2011-01-01

process on the allergenicity of Ara h 1 and a mix of 2S albumins from peanut (Ara h 2/6). Methods Ara h 1 and Ara h 2/6 were purified from raw peanuts and heated in a dry form for 20 min at 145 degrees C in the presence (R + g) or absence (R - g) of glucose, and soluble proteins were then extracted. Sera...... obtained from 12 well-characterized peanut-allergic patients were used to assess the IgE binding and degranulation capacities of the allergens. Results Extensive heating at low moisture resulted in the hydrolysis of both Ara h 1 and Ara h 2/6. However, in contrast to Ara h 2/6, soluble R + g Ara h 1 formed...... large aggregates. Although the IgE-binding capacity of R + g and R - g Ara h 1 was decreased 9000- and 3.6-fold, respectively, compared with native Ara h 1, their capacity to elicit mediator release was increased. Conversely, both the IgE-binding capacity and the degranulation capacity of R - g Ara h 2...

Specific IgE response to different grass pollen allergen components in children undergoing sublingual immunotherapy

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Marcucci Francesco

2012-06-01

Full Text Available Abstract Background Grass pollen is a major cause of respiratory allergy worldwide and contain a number of allergens, some of theme (Phl p 1, Phl p 2, Phl p 5, and Phl 6 from Phleum pratense, and their homologous in other grasses are known as major allergens. The administration of grass pollen extracts by immunotherapy generally induces an initial rise in specific immunoglobulin E (sIgE production followed by a progressive decline during the treatment. Some studies reported that immunotherapy is able to induce a de novo sensitisation to allergen component previously unrecognized. Methods We investigated in 30 children (19 males and 11 females, mean age 11.3 years, 19 treated with sublingual immunotherapy (SLIT by a 5-grass extract and 11 untreated, the sIgE and sIgG4 response to the different allergen components. Results Significant increases (p  Conclusions These findings confirm that the initial phase of SLIT with a grass pollen extract enhances the sIgE synthesis and show that the sIgE response concerns the same allergen components which induce IgE reactivity during natural exposure.

Characterisation of purified parvalbumin from five fish species and nucleotide sequencing of this major allergen from Pacific pilchard, Sardinops sagax.

Science.gov (United States)

Beale, Janine E; Jeebhay, Mohamed F; Lopata, Andreas L

2009-09-01

IgE-mediated allergic reaction to seafood is a common cause of food allergy including anaphylactic reactions. Parvalbumin, the major fish allergen, has been shown to display IgE cross-reactivity among fish species consumed predominantly in Europe and the Far East. However, cross-reactivity studies of parvalbumin from fish species widely consumed in the Southern hemisphere are limited as is data relating to immunological and molecular characterisation. In this study, antigenic cross-reactivity and the presence of oligomers and isomers of parvalbumin from five highly consumed fish species in Southern Africa were assessed by immunoblotting using purified parvalbumin and crude fish extracts. Pilchard (Sardinops sagax) parvalbumin was found to display the strongest IgE reactivity among 10 fish-allergic consumers. The cDNA sequence of the beta-form of pilchard parvalbumin was determined and designated Sar sa 1.0101 (accession number FM177701 EMBL/GenBank/DDBJ databases). Oligomeric forms of parvalbumin were observed in all fish species using a monoclonal anti-parvalbumin antibody and subject's sera. Isoforms varied between approximately 10-13 kDa. A highly cross-reactive allergenic isoform of parvalbumin was identified and sequenced, providing a successful primary step towards the generation of a recombinant form that could be used for diagnostic and potential therapeutic use in allergic individuals.

Allergen management in the food industry

National Research Council Canada - National Science Library

Boye, Joyce I; Godefroy, Samuel Benrejeb

2010-01-01

.... Starting with an introduction to food allergens, the book follows with sections on food allergen management during production and processing, guidelines for the processing of specific allergen-free...

Application of recombinant latex allergens in diagnostics of occupational latex allergy

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Ewa Nowakowska-Świrta

2015-02-01

Full Text Available Over many years, allergy to natural rubber latex has been a major problem among health care workers (HCW. The diagnosis of occupational allergy requires methods of high diagnostic accuracy in view of certification implications (e.g., a sick worker quits a job. With the development of molecular methods, the frequency of application of recombinant allergens in the diagnostics of allergic diseases continues to increase. This paper reviews the applicability of laboratory tests which use recombinant allergens in the diagnostics of occupational allergy. The diagnosis of latex allergy is based on the presence of clinical symptoms linked with exposure to latex allergens, positive skin prick tests and detection of specific IgE antibodies to latex in serum. Moreover, in some cases specific challenge tests are conducted. The analysis of literature indicates that applying the panel of recombinant latex allergens in diagnostic tests, cross-reactivity can very likely be excluded and/or sensitization can be confirmed without the need for specific challenge tests, which in case of latex allergens carries a potential risk of generalized reactions. Med Pr 2015;66(1:85–97

Allergenic characterization of a novel allergen, homologous to chymotrypsin, from german cockroach.

Science.gov (United States)

Jeong, Kyoung Yong; Son, Mina; Lee, Jae Hyun; Hong, Chein Soo; Park, Jung Won

2015-05-01

Cockroach feces are known to be rich in IgE-reactive components. Various protease allergens were identified by proteomic analysis of German cockroach fecal extract in a previous study. In this study, we characterized a novel allergen, a chymotrypsin-like serine protease. A cDNA sequence homologous to chymotrypsin was obtained by analysis of German cockroach expressed sequence tag (EST) clones. The recombinant chymotrypsins from the German cockroach and house dust mite (Der f 6) were expressed in Escherichia coli using the pEXP5NT/TOPO vector system, and their allergenicity was investigated by ELISA. The deduced amino acid sequence of German cockroach chymotrypsin showed 32.7 to 43.1% identity with mite group 3 (trypsin) and group 6 (chymotrypsin) allergens. Sera from 8 of 28 German cockroach allergy subjects (28.6%) showed IgE binding to the recombinant protein. IgE binding to the recombinant cockroach chymotrypsin was inhibited by house dust mite chymotrypsin Der f 6, while it minimally inhibited the German cockroach whole body extract. A novel allergen homologous to chymotrypsin was identified from the German cockroach and was cross-reactive with Der f 6.

Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis

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Frøkiær Hanne

2007-08-01

Full Text Available Abstract Background Natural allergen sources can supply large quantities of authentic allergen mixtures for use as immunotherapeutics. However, such extracts are complex, difficult to define, vary from batch to batch, which may lead to unpredictable efficacy and/or unacceptable levels of side effects. The use of recombinant expression systems for allergen production can alleviate some of these issues. Several allergens have been tested in high-level expression systems and in most cases show immunereactivity comparable to their natural counterparts. The gram positive lactic acid bacterium Lactococcus lactis is an attractive microorganism for use in the production of protein therapeutics. L. lactis is considered food grade, free of endotoxins, and is able to secrete the heterologous product together with few other native proteins. Hypersensitivity to peanut represents a serious allergic problem. Some of the major allergens in peanut have been described. However, for therapeutic usage more information about the individual allergenic components is needed. In this paper we report recombinant production of the Ara h 2 peanut allergen using L. lactis. Results A synthetic ara h 2 gene was cloned into an L. lactis expression plasmid containing the P170 promoter and the SP310mut2 signal sequence. Flask cultures grown overnight showed secretion of the 17 kDa Ara h 2 protein. A batch fermentation resulted in 40 mg/L recombinant Ara h 2. Purification of Ara h 2 from the culture supernatant was done by hydrophobic exclusion and size separation. Mass spectrometry and N-terminal analysis showed a recombinant Ara h 2 of full length and correctly processed by the signal peptidase. The immunological activity of recombinant Ara h 2 was analysed by ELISA using antibodies specific for native Ara h 2. The recombinant Ara h 2 showed comparable immunereactivity to that of native Ara h 2. Conclusion Recombinant production of Ara h 2 using L. lactis can offer high yields

AllergenOnline: A peer-reviewed, curated allergen database to assess novel food proteins for potential cross-reactivity.

Science.gov (United States)

Goodman, Richard E; Ebisawa, Motohiro; Ferreira, Fatima; Sampson, Hugh A; van Ree, Ronald; Vieths, Stefan; Baumert, Joseph L; Bohle, Barbara; Lalithambika, Sreedevi; Wise, John; Taylor, Steve L

2016-05-01

Increasingly regulators are demanding evaluation of potential allergenicity of foods prior to marketing. Primary risks are the transfer of allergens or potentially cross-reactive proteins into new foods. AllergenOnline was developed in 2005 as a peer-reviewed bioinformatics platform to evaluate risks of new dietary proteins in genetically modified organisms (GMO) and novel foods. The process used to identify suspected allergens and evaluate the evidence of allergenicity was refined between 2010 and 2015. Candidate proteins are identified from the NCBI database using keyword searches, the WHO/IUIS nomenclature database and peer reviewed publications. Criteria to classify proteins as allergens are described. Characteristics of the protein, the source and human subjects, test methods and results are evaluated by our expert panel and archived. Food, inhalant, salivary, venom, and contact allergens are included. Users access allergen sequences through links to the NCBI database and relevant references are listed online. Version 16 includes 1956 sequences from 778 taxonomic-protein groups that are accepted with evidence of allergic serum IgE-binding and/or biological activity. AllergenOnline provides a useful peer-reviewed tool for identifying the primary potential risks of allergy for GMOs and novel foods based on criteria described by the Codex Alimentarius Commission (2003). © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

AllerHunter: a SVM-pairwise system for assessment of allergenicity and allergic cross-reactivity in proteins.

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Hon Cheng Muh

Full Text Available Allergy is a major health problem in industrialized countries. The number of transgenic food crops is growing rapidly creating the need for allergenicity assessment before they are introduced into human food chain. While existing bioinformatic methods have achieved good accuracies for highly conserved sequences, the discrimination of allergens and non-allergens from allergen-like non-allergen sequences remains difficult. We describe AllerHunter, a web-based computational system for the assessment of potential allergenicity and allergic cross-reactivity in proteins. It combines an iterative pairwise sequence similarity encoding scheme with SVM as the discriminating engine. The pairwise vectorization framework allows the system to model essential features in allergens that are involved in cross-reactivity, but not limited to distinct sets of physicochemical properties. The system was rigorously trained and tested using 1,356 known allergen and 13,449 putative non-allergen sequences. Extensive testing was performed for validation of the prediction models. The system is effective for distinguishing allergens and non-allergens from allergen-like non-allergen sequences. Testing results showed that AllerHunter, with a sensitivity of 83.4% and specificity of 96.4% (accuracy = 95.3%, area under the receiver operating characteristic curve AROC = 0.928+/-0.004 and Matthew's correlation coefficient MCC = 0.738, performs significantly better than a number of existing methods using an independent dataset of 1443 protein sequences. AllerHunter is available at (http://tiger.dbs.nus.edu.sg/AllerHunter.

Quality characteristics of gluten-free cookies made of buckwheat, corn, and rice flour with/without transglutaminase.

Science.gov (United States)

Altındağ, Gülçin; Certel, Muharrem; Erem, Fundagül; İlknur Konak, Ülgen

2015-04-01

Buckwheat is one of the most valuable pseudo-cereals in terms of its nutritional composition, and it is suitable for celiac patients because of its gluten-free characteristic. However, gluten is the main structure-forming protein responsible for the development of structure in baked products. Therefore, it is a challenge to produce high-quality gluten-free products. Transglutaminase addition is a relatively common application used in the production of gluten-free baked goods. The objective of this study was to investigate the combination of buckwheat flour with rice and corn flour at different levels in gluten-free cookie formulations and the impact of transglutaminase on the quality of cookies. Quality parameters evaluated were proximal chemical composition, spread ratio, color, and textural parameters (hardness and fracturability). Spread ratio, protein, crude fiber, ash content, and also b* and hardness values were significantly (p flour combinations. Further, addition of transglutaminase resulted in increased moisture content, spread ratio, and fracturability but decreased hardness values. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

DISTRIBUTION OF ALLERGENIC PLANTS IN RUSSIA

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Tatiana V. Dikareva

2015-01-01

Full Text Available We analyzed, for the first time ever, the geographical distribution of the main allergenic plants in Russia. All materials were organized as database and attached to the map in GIS Mapinfo. For each region of Russian Federation, two indices were calculated: the total number of allergenic plants in the region and the “allergenic index”. A series of maps wascompiled: the number of spring-flowering species, the number of summer-flowering species,the total number of species flowering during the whole year, the overall allergen danger during spring and summer seasons, respectively, and the overall allergen danger during the whole year. In terms of the number of allergenic species and by the “allergenic index,” the most dangerous regions appeared to be the Ryazan and Voronezh Oblasts, while the less dangerous – the Chukotka Autonomous Okrug, and the Magadan Oblast. The maps may serve as a reference source for allergologists and allergy sufferers.

Application of chemical mutagens and radiation in breeding buckwheat for larger seeds

International Nuclear Information System (INIS)

Alekseeva, E.S.

1988-01-01

Full text: In 1974, seeds of the Viktoriya variety of buckwheat were treated with 20-30 krad gamma radiation and chemical mutagens in the Biophysics Department of the Kishinev Agricultural Institute. For the chemical mutagen treatment, we used N-ethylnitroso-urea NEH (0.025 and 0.012%), N-methylnitroso-urea NMH (0.01 and 0.005%), ethylenimine EI (0.01 and 0.005%), dimethyl sulphate DMS (0.01 and 0.005%) and 1.4-bis-diazoacetyl butane DAB (0.01 and 0.05%). Since some investigators think that different results are produced by changing the order of the treatment, we treated seeds with chemical mutagens before and after irradiation and this was followed by drying. A total of 2400 seeds were treated. Selection started with M 2 seeds produced by M 1 plants. The thousand seed weight of the best ones ranged from 40.7 to 47.8 g, which was 11.9-18.7 g heavier than the control. The large seed size thus selected was heritable. Since larger seeds are very important for the creation of high yielding varieties buckwheat, only families with these characteristics were selected for further work. We observed even some further increase in seed weight in the next generation. It was observed that when planting large seeds, after six days of growth the cotyledons were significantly larger than in the control plants. This characteristic was used in selecting for a high yielding large-seed variety of buckwheat. The plants were selected twice: once for development of large cotyledon leaves and the second time for plant yield. In the fourth generation, the families thus obtained continued to be studied in greenhouse experiments and the same time be propagated under field conditions. The seeds of these families were then combined and under the name Podolyanka in 1976 were subjected to competitive variety testing. Following the competitive variety testing the mutant variety Podolyanka was released in 1984. It is high yielding (2950 kg/ha), has a short vegetation period (matures 17-18 days

Insect (food) allergy and allergens.

Science.gov (United States)

de Gier, Steffie; Verhoeckx, Kitty

2018-05-03

Insects represent an alternative for meat and fish in satisfying the increasing demand for sustainable sources of nutrition. Approximately two billion people globally consume insects. They are particularly popular in Asia, Latin America, and Africa. Most research on insect allergy has focussed on occupational or inhalation allergy. Research on insect food safety, including allergenicity, is therefore of great importance. The objective of this review is to provide an overview of cases reporting allergy following insect ingestion, studies on food allergy to insects, proteins involved in insect allergy including cross-reactive proteins, and the possibility to alter the allergenic potential of insects by food processing and digestion. Food allergy to insects has been described for silkworm, mealworm, caterpillars, Bruchus lentis, sago worm, locust, grasshopper, cicada, bee, Clanis bilineata, and the food additive carmine, which is derived from female Dactylopius coccus insects. For cockroaches, which are also edible insects, only studies on inhalation allergy have been described. Various insect allergens have been identified including tropomyosin and arginine kinase, which are both pan-allergens known for their cross-reactivity with homologous proteins in crustaceans and house dust mite. Cross-reactivity and/or co-sensitization of insect tropomyosin and arginine kinase has been demonstrated in house dust mite and seafood (e.g. prawn, shrimp) allergic patients. In addition, many other (allergenic) species (various non-edible insects, arachnids, mites, seafoods, mammals, nematoda, trematoda, plants, and fungi) have been identified with sequence alignment analysis to show potential cross-reactivity with allergens of edible insects. It was also shown that thermal processing and digestion did not eliminate insect protein allergenicity. Although purified natural allergens are scarce and yields are low, recombinant allergens from cockroach, silkworm, and Indian mealmoth are

Quadrupole time-of-flight mass spectometry : a method to study the actual expression of allergen isoforms identified by PCR cloning

NARCIS (Netherlands)

Helsper, J.P.F.G.; Gilissen, L.J.W.J.; Ree, van R.; America, A.H.P.; Cordewener, J.H.G.; Bosch, D.

2002-01-01

Background: Over the past 2 decades, molecular biology has shown that most major allergens exist in multiple isoforms. Very little is known about the relevance of allergen isoforms at the level of expressed protein (ie, actual allergen exposure). Objective: The aim of this study was to evaluate the

Quadrupole time-of-flight mass spectrometry: a method to study the actual expression of allergen isoforms identified by PCR cloning

NARCIS (Netherlands)

Helsper, Johannes P. F. G.; Gilissen, Luud J. W. J.; van Ree, Ronald; America, Antoine H. P.; Cordewener, Jan H. G.; Bosch, Dirk

2002-01-01

BACKGROUND: Over the past 2 decades, molecular biology has shown that most major allergens exist in multiple isoforms. Very little is known about the relevance of allergen isoforms at the level of expressed protein (ie, actual allergen exposure). OBJECTIVE: The aim of this study was to evaluate the

[Purification, characterization and partial primary structure analysis of rutin-degrading enzyme in tartary buckwheat seeds].

Science.gov (United States)

Zhang, Yuwei; Li, Jie; Yuan, Yong; Gu, Jijuan; Chen, Peng

2017-05-25

Rutin-degrading enzymes (RDE) can degrade rutin into poorly water soluble compound, quercetin, and cause the bitter taste in tartary buckwheat. In the present study RDE from Yu 6-21 tartary buckwheat seeds was purified by ammonium sulphate precipitation, followed by hydrophobic interaction chromatography on Phenyl Sepharose CL-4B, ion exchange chromatography on CM-Cellulose and gel filtration chromatography on Sephadex G-150. Purified RDE showed single band with molecular weight of 66 kDa on SDS-PAGE. The optimum pH and temperature of RDE were 5.0 and 50 ℃ respectively. The Km was 0.27 mmol/L, and the Vmax was 39.68 U/mg. The RDE activity could be inhibited by Cu²⁺, Zn²⁺, Mn²⁺ and EDTA, and showed tolerance to 50% methanol (V/V). The N terminal sequence (TVSRSSFPDGFLFGL) was obtained by Edman degradation method and 15 internal peptide sequences were determined by MALDI-TOF-MS (matrix-assisted laser desorption ionization time of flight mass spectrometry). These results established the foundations for identification of the candidate gene of RDE via transcriptome data and further studying RDE biological function.

Comparison of allergenicity and immunogenicity of an intact allergen vaccine and commercially available allergoid products for birch pollen immunotherapy.

Science.gov (United States)

Lund, L; Henmar, H; Würtzen, P A; Lund, G; Hjortskov, N; Larsen, J N

2007-04-01

Specific immunotherapy with intact allergen vaccine is a well-documented treatment for allergic diseases. Different vaccine formulations are currently commercially available, the active ingredient either being intact allergens or chemically modified allergoids. The rationale behind allergoids is to decrease allergenicity while maintaining immunogenicity. However, data from the German health authorities based on reporting of adverse events over a 10-year period did not indicate increased safety of allergoids over intact allergens. The objective of this study was to investigate the effect of chemical modification on allergenicity and immunogenicity comparing four commercial allergoid products for birch pollen immunotherapy with an intact allergen vaccine. Solid-phase IgE inhibition and histamine release assays were selected as model systems for allergenicity, and a combination of human T cell proliferation and IgG titres following mouse immunizations were used to address the immunogenicity of the intact allergen vaccine and the four allergoids. In all assays, the products were normalized with respect to the manufacturer's recommended maintenance dose. IgE inhibition experiments showed a change in epitope composition comparing intact allergen vaccine with allergoid. One allergoid product induced enhanced histamine release compared to the intact allergens, while the other three allergoids showed reduced release. Standard T cell stimulation assays using lines from allergic patients showed a reduced response for all allergoids compared with the intact allergen vaccine regardless of the cell type used for antigen presentation. All allergoids showed reduced capacity to induce allergen-specific IgG responses in mice. While some allergoids were associated with reduced allergenicity, a clear reduction in immunogenicity was observed for all allergoid products compared with the intact allergen vaccine, and the commercial allergoids tested therefore do not fulfil the allergoid

The prebiotic and protective effects of buckwheat flour and oat bran on Lactobacillus acidophilus

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Aida VASILE

2016-12-01

Full Text Available During conservation, the probiotic bacteria currently used in food industry have been shown to have less viability in the matter of fermented products microbiota and also to present lower levels of colonization and survival in vivo. This study describes a new way of improving the behavior and functionality of Lactobacillus acidophilus - LA 5® commercial strain by using buckwheat flour (Fagopyrum esculentum and oat bran (Avena sativa as fermentation ingredients, in view of their high content of bioactive compounds that have a great impact both on fermentation microorganisms and consumers’ health. The effect of these two raw vegetal products on LA 5® strain was studied by cultivation on MRS broth and on milk. The supplementation of the fermentation medium with 4.0% - 6.0% of buckwheat flour or oat bran, respectively, had a positive effect on metabolic activity and viability of lactic acid bacteria. Thus, the rapid decrease of the pH and the increase of the multiplication rate were observed after 6 hours of lactic acid fermentation. Furthermore, the presence of the vegetal substrates substantially improved the cells survival during the storage of the fermented products for 28 days at 4 °C, comparing to samples without supplementation used as control.

Contents of fragrance allergens in children's cosmetics and cosmetic-toys

DEFF Research Database (Denmark)

Rastogi, S C; Johansen, Jeanne Duus; Menné, T

1999-01-01

was present in a maximum concentration of 0.07%. In one cosmetic-toy, cinnamic alcohol was present at 3.7% which exceeds the current industry guideline for safe products by a factor of 5. In all types of products other fragrance allergens were frequently found. In conclusion, children are already exposed......Fragrances are one of the major causes of allergic contact dermatitis from use of cosmetics. The aim of the current study was to assess the possible exposure of infants and children to fragrance allergens from cosmetic products and "toy-cosmetics". 25 children's cosmetics or toy-cosmetic products...... at an early age to well-known allergens, sometimes at concentrations which are considered to be unsafe. As contact allergy usually persists for life, manufacturers of children's cosmetics should be aware of their special responsibility and apply the highest possible safety standards....

LINE'S OF MANUFACTURE OF KISSELS ON THE BASIS OF BUCKWHEAT AND BARLEY STARCH

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J. J. Ermolaev

2012-01-01

Full Text Available This work is devoted to development of a production line of kissels on the basis of grechishny and barley крахмалов, allowing to use their structural and mechanical and chemical properties, investigating rheological characteristics of solutions buckwheat and barley крахмалов, to receive the dry granulated drinks on the basis of vegetative raw materials which will partially solve problems of equation of a food.

ALLERGEN-SPECIFIC IMMUNOTHERAPY IN CHILDREN WITH POLLINOSIS

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R. M. Torshkhoeva

2014-01-01

Full Text Available Aim: to compare clinical efficacy and safety of sublingual and parenteral allergen-specific immunotherapy in children with pollinosis. Patients and methods: 143 patients with pollinosis aged from 5 to 16 years old were included into the study. They were divided into 4 groups and received allergen-specific immunotherapy. Patients of the groups I and III were administered water-salt mixtures of extracts of tree pollen allergens. Patients from the II group received standardized adjuvant mixture of extracts of tree pollen allergens. Patients from the IV group were administered standardized extract of birch pollen allergens. Prophylaxis with water-salt solutions was performed before seasons of increased allergy risk during 3 years in autumns and winters. Prophylaxis with standardized extracts of allergens was performed uninterruptedly for 3 years. Results: allergen-specific immunotherapy prevents increase of sensitization and enlargement of allergen spectrum of elevated organism perceptibility, as well as prevents aggravation of disease course and conversion to more severe forms. It also decreases requirements of anti-allergic drugs and therefore elongates the duration of remission. Conclusions: allergen-specific immunotherapy with the use of standardized allergens is the most effective method of treatment of pollen sensitization in children. In order to increase its efficacy not less than 3 courses of immunotherapy are needed.

Allergens from fish and egg

DEFF Research Database (Denmark)

Poulsen, Lars K.; Hansen, T K; Nørgaard, A

2001-01-01

, denominated the parvalbumins. This cross-reactivity has been indicated to be of clinical relevance for several species, since patients with a positive double-blind, placebo-controlled food challenge to cod will also react with other fish species, such as herring, plaice and mackerel. In spite......Allergens from fish and egg belong to some of the most frequent causes of food allergic reactions reported in the literature. Egg allergens have been described in both white and yolk, and the egg white proteins ovomucoid, ovalbumin, ovotransferrin and lysozyme have been adopted in the allergen...... nomenclature as Gal d1-d4. The most reported allergen from egg yolk seems to be alpha-livitin. In fish, the dominating allergen is the homologues of Gad c1 from cod, formerly described as protein M. A close cross-reactivity exists within different species of fish between this calcium-binding protein family...

Structural stability of Amandin, a major allergen from almond (Prunus dulcis), and its acidic and basic polypeptides.

Science.gov (United States)

Albillos, Silvia M; Menhart, Nicholas; Fu, Tong-Jen

2009-06-10

Information relating to the resistance of food allergens to thermal and/or chemical denaturation is critical if a reduction in protein allergenicity is to be achieved through food-processing means. This study examined the changes in the secondary structure of an almond allergen, amandin, and its acidic and basic polypeptides as a result of thermal and chemical denaturation. Amandin ( approximately 370 kDa) was purified by cryoprecipitation followed by gel filtration chromatography and subjected to thermal (13-96 degrees C) and chemical (urea and dithiothreitol) treatments. Changes in the secondary structure of the protein were followed using circular dichroism spectroscopy. The secondary structure of the hexameric amandin did not undergo remarkable changes at temperatures up to 90 degrees C, although protein aggregation was observed. In the presence of a reducing agent, irreversible denaturation occurred with the following experimental values: T(m) = 72.53 degrees C (transition temperature), DeltaH = 87.40 kcal/mol (unfolding enthalpy), and C(p) = 2.48 kcal/(mol degrees C) (heat capacity). The concentration of urea needed to achieve 50% denaturation was 2.59 M, and the Gibbs free energy of chemical denaturation was calculated to be DeltaG = 3.82 kcal/mol. The basic and acidic polypeptides of amandin had lower thermal stabilities than the multimeric protein.

Comparison of allergenicity and allergens between fish white and dark muscles.

Science.gov (United States)

Kobayashi, A; Tanaka, H; Hamada, Y; Ishizaki, S; Nagashima, Y; Shiomi, K

2006-03-01

Fish is one of the most frequent causes of immunoglobulin E (IgE)-mediated food allergy. Although the fish dark muscle is often ingested with the white muscle, no information about its allergenicity and allergens is available. Heated extracts were prepared from both white and dark muscles of five species of fish and examined for reactivity with IgE in fish-allergic patients by enzyme-linked immunosorbent assay (ELISA) and for allergens by immunoblotting. Cloning of cDNAs encoding parvalbumins was performed by rapid amplification cDNA ends. Parvalbumin contents in both white and dark muscles were determined by ELISA using antiserum against mackerel parvalbumin. Patient sera were less reactive to the heated extract from the dark muscle than to that from the white muscle. A prominent IgE-reactive protein of 12 kDa, which was detected in both white and dark muscles, was identified as parvalbumin. Molecular cloning experiments revealed that the same parvalbumin molecule is contained in both white and dark muscles of either horse mackerel or Pacific mackerel. Parvalbumin contents were four to eight times lower in the dark muscle than in the white muscle. The fish dark muscle is less allergenic than the white muscle, because the same allergen molecule (parvalbumin) is contained at much lower levels in the dark muscle than in the white muscle. Thus, the dark muscle is less implicated in fish allergy than the white muscle.

Identification of novel allergen in edible insect, Gryllus bimaculatus and its cross-reactivity with Macrobrachium spp. allergens.

Science.gov (United States)

Srinroch, Chutima; Srisomsap, Chantragan; Chokchaichamnankit, Daranee; Punyarit, Phaibul; Phiriyangkul, Pharima

2015-10-01

Edible insects have recently been promoted as a source of protein and have a high nutrition value. Identification of allergens and cross-reactivity between Macrobrachium spp. and the field cricket (Gryllus bimaculatus) is necessary for food safety control and to assist in the diagnosis and therapy of allergy symptoms. Denaturing polyacrylamide gel electrophoresis (SDS-PAGE) was used to separate proteins. Allergens were determined and identified by IgE-immunoblotting with pooled sera from prawn-allergic patients (n=16) and LC-MS/MS. Arginine kinase (AK) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were determined as the important allergens in muscle of Macrobrachium rosenbergii whereas, hemocyanin (HC) was identified as an allergen in Macrobrachium spp. The allergens in Macrobrachium lanchesteri were identified as AK and HC. In addition, hexamerin1B (HEX1B) was identified as a novel and specific allergen in G. bimaculatus. The important allergen in G. bimaculatus and Macrobrachium spp. is AK and was found to cross-react between both species. Copyright © 2015 Elsevier Ltd. All rights reserved.

Allergen specific responses in cord and adult blood are differentially modulated in the presence of endotoxins

DEFF Research Database (Denmark)

Eiwegger, T.; Mayer, E.; Pedersen, Susanne Brix

2008-01-01

Background Endotoxins are common contaminants in allergen preparations and affect antigen-specific cellular responses. Distinct effects of endotoxin on cells in human umbilical cord and adult blood are poorly defined. Objectives To examine the effect of endotoxins in allergen preparations...... on cellular responses in human cord and peripheral blood (PB). Methods The endotoxin content in beta lactoglobulin (BLG), the peanut allergen Ara h 1 and the major birch pollen allergen Bet v 1 was assessed. Proliferation and cytokine response of mononuclear cells towards contaminated and lipopolysaccharide....... Results The proliferative response of cord blood (CB)-derived mononuclear cells towards allergen-preparations at day 3 was related to the level of LPS contamination. At day 7, proliferation was also detected in the absence of endotoxin. Cytokine production in CB was strongly affected by the content...

Identification of novel allergens of Aspergillus fumigatus using immunoproteomics approach.

Science.gov (United States)

Gautam, P; Sundaram, C S; Madan, T; Gade, W N; Shah, A; Sirdeshmukh, R; Sarma, P U

2007-08-01

Approximately 20% of the world's asthmatics are suffering from Aspergillus fumigatus (Afu)-induced allergies. The characterization of specific IgE-inducing allergens in allergic aspergillosis patients is fundamental for clinical diagnosis and for immunotherapy. Immunoproteomics combined with mass spectrometric analysis was used to identify proteins of third-week culture filtrate (3wcf) potentially responsible for Afu-specific IgE immunoreactivity, using pooled sera from Afu-sensitized asthmatics. Their allergenic potential was also tested against patients with allergic bronchopulmonary aspergillosis (ABPA), by two-dimensional (2-D) gel electrophoresis immunoblotting of 3wcf proteins with individual sera from such patients. This helped us to establish a set of candidate allergens, which could be explored further for diagnostic application in allergic aspergillosis asthmatics including ABPA. Peptide mass fingerprint using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and/or de novo sequencing by MS/MS analysis of the protein spots from 2-D gels led to the identification of a total of 16 allergens of Afu. Eleven of them are being reported as allergens for the first time and five had been reported earlier. Putative isoforms of the proteins Asp f 13 and chitosanase have been observed for the first time. When studied for reactivity of these proteins among patients with ABPA using their individual sera, these patients exhibited sensitization although the pattern was varying. Taken together, these proteins could thus be considered as potential allergens even among patients with ABPA. Three of these proteins viz. the hypothetical protein (# spot no. 5), extracellular arabinase (# spot no. 6) and chitosanase (# spot no. 11) could be major allergens with specific IgE immunoreactivity with six out of eight patients' sera. The immunoproteomic approach applied to the analysis of culture filtrate proteins resulted in the

8 Allergenic Composition of Polymerized Allergen Extracts of Betula verrucosa, Dermatophagoides Pteronyssinus and Phleum Pratense

Science.gov (United States)

Fernandez-Caldas, Enrique; Cases, Barbara; Tudela, Jose Ignacio; Fernandez, Eva Abel; Casanovas, Miguel; Subiza, Jose Luis

2012-01-01

Background Allergoids have been successfully used in the treatment of respiratory allergic diseases. They are modified allergen extracts that allow the administration of high allergen doses, due to their reduced IgE binding capacity.They maintain allergen-specific T-cell recognition. Since they are native allergen extracts that have been polymerized with glutaraldehyde, identification of the allergenic molecules requires more complicated methods. The aim of the study was to determine the qualitative composition of different polymerized extracts and investigate the presence of defined allergenic molecules using Mass spectrometry. Methods Proteomic analysis was carried out at the Proteomics Facility of the Hospital Nacional de Parapléjicos (Toledo, Spain). After reduction and alkylation, proteins were digested with trypsin and the resulting peptides were cleaned using C18 SpinTips Sample Prep Kit; peptides were separated on an Ultimate nano-LC system using a Monolithic C18 column in combination with a precolumn for salt removal. Fractionation of the peptides was performed with a Probot microfraction collector and MS and MS/MS analysis of offline spotted peptide samples were performed using the Applied Biosystems 4800 plus MALDI TOF/TOF Analyzer mass spectrometer. ProteinPilot Software V 2.0.1 and the Paragon algorithm were used for the identification of the proteins. Each MS/MS spectrum was searched against the SwissProt 2010_10 database, Uniprot-Viridiplantae database and Uniprot_Betula database. Results Analysis of the peptides revealed the presence of native allergens in the polymerized extracts: Der p 1, Der p 2, Der p 3, Der p 8 and Der p 11 in D. pteronyssinus; Bet v 2, Bet v 6, Bet v 7 and several Bet v 1 isoforms in B. verrucosa and Phl p 1, Phl p 3, Phl p 5, Phl p 11 and Phl p 12 in P. pratense allergoids. In all cases, potential allergenic proteins were also identified, including ubiquitin, actin, Eenolase, fructose-bisphosphate aldolase, luminal

Allergenicity assessment strategy for novel food proteins and protein sources.

Science.gov (United States)

Verhoeckx, Kitty; Broekman, Henrike; Knulst, André; Houben, Geert

2016-08-01

To solve the future food insecurity problem, alternative and sustainable protein sources (e.g. insects, rapeseed, fava bean and algae) are now being explored for the production of food and feed. To approve these novel protein sources for future food a comprehensive risk assessment is needed according to the European food legislation. Allergenicity risk assessment might pose some major difficulties, since detailed guidance on how to assess the allergenic potential of novel foods is not available. At present, the approach relies mostly on the guidance of allergenicity assessment for genetically modified (GM) plant foods. The most recent one was proposed by EFSA (2010 and 2011); "weight-of-evidence approach". However this guidance is difficult to interpret, not completely applicable or validated for novel foods and therefore needs some adjustments. In this paper we propose a conceptual strategy which is based on the "weight-of-evidence approach" for food derived from GM plants and other strategies that were previously published in the literature. This strategy will give more guidance on how to assess the allergenicity of novel food proteins and protein sources. Copyright © 2016 Elsevier Inc. All rights reserved.

Effects of daily food processing on allergenicity.

Science.gov (United States)

Cabanillas, Beatriz; Novak, Natalija

2017-08-11

Daily food processing has the potential to alter the allergenicity of foods due to modification of the physico-chemical properties of proteins. The degree of such modifications depends on factors such as processing conditions, type of food considered, allergenic content, etc. The impact of daily food processing like boiling, roasting, frying or baking on food allergenicity have been extensively studied. The influence of other thermal treatments such as microwave heating or pressure cooking on allergenicity has also been analyzed. Non-thermal treatment such as peeling impacts on the allergenic content of certain foods such as fruits. In this review, we give an updated overview of the effects of daily processing treatments on the allergenicity of a wide variety of foods. The different variables that contribute to the modification of food allergenicity due to processing are also reviewed and discussed.

Mechanisms of allergen-specific immunotherapy

Directory of Open Access Journals (Sweden)

Fujita Hiroyuki

2012-01-01

Full Text Available Abstract Allergen-specific immunotherapy (allergen-SIT is a potentially curative treatment approach in allergic diseases. It has been used for almost 100 years as a desensitizing therapy. The induction of peripheral T cell tolerance and promotion of the formation of regulatory T-cells are key mechanisms in allergen-SIT. Both FOXP3+CD4+CD25+ regulatory T (Treg cells and inducible IL-10- and TGF-β-producing type 1 Treg (Tr1 cells may prevent the development of allergic diseases and play a role in successful allergen-SIT and healthy immune response via several mechanisms. The mechanisms of suppression of different pro-inflammatory cells, such as eosinophils, mast cells and basophils and the development of allergen tolerance also directly or indirectly involves Treg cells. Furthermore, the formation of non-inflammatory antibodies particularly IgG4 is induced by IL-10. Knowledge of these molecular basis is crucial in the understanding the regulation of immune responses and their possible therapeutic targets in allergic diseases.

AllergenOnline: A peer-reviewed, curated allergen database to assess novel food proteins for potential cross-reactivity

NARCIS (Netherlands)

Goodman, Richard E.; Ebisawa, Motohiro; Ferreira, Fatima; Sampson, Hugh A.; van Ree, Ronald; Vieths, Stefan; Baumert, Joseph L.; Bohle, Barbara; Lalithambika, Sreedevi; Wise, John; Taylor, Steve L.

2016-01-01

Increasingly regulators are demanding evaluation of potential allergenicity of foods prior to marketing. Primary risks are the transfer of allergens or potentially cross-reactive proteins into new foods. AllergenOnline was developed in 2005 as a peer-reviewed bioinformatics platform to evaluate

Mapping of Lol p I allergenic epitopes by using murine monoclonal antibodies.

Science.gov (United States)

Mourad, W; Bernier, D; Jobin, M; Hébert, J

1989-11-01

Murine monoclonal antibodies (MAbs) against three non-overlapping epitopes of Lol p I allergen were previously produced and subsequently used for purification of the allergen. In the present study, these MAbs were further characterized, and the biological activity of the purified allergen assessed. The three MAbs were of the IgG isotype and carried a kappa light chain. Their affinity constants were in the range of 7.4-15.1 x 10(-9) mol/l. Purified Lol p I kept its biological activity, as shown by its ability to induce histamine release by basophils of Lol p I-sensitive patients. The profiles of histamine release induced by either Lol p I or crude Lolium perenne extracts were comparable. This observation suggests that human IgE bound to basophils are polyspecific which has been confirmed by immunoblot and inhibition assay. Our data indicated also that Lol p I possesses a major allergenic epitope recognized by all human serum IgE tested. This epitope seems to be partially shared by those recognized by the three MAbs. Finally, preincubation of Lol p I with either one of the Mabs did not affect significantly the basophil-histamine release induced by the purified allergen. This suggests that Lol p I possesses allergenic sites other than the one shared by MAbs and IgE Abs.

Buckwheat yield and its quality as affected by laser biostimulation of its seeds

International Nuclear Information System (INIS)

Koper, R.; Mikos-Bielak, M.

2003-01-01

The influence of various doses of laser radiation applied to buckwheat seed bio stimulation on the yield, and changes of chemical composition was analysed. A 12-25 percent increase of yield was observed in bio stimulated plants. The most positive effects were achieved after seed triple radiation using a laser of 30 mW power for 0.1 s. Bio stimulation caused a slight increase of protein, fat and fiber content a large increase of soluble and reducing sugars and a decrease of starch level

Allergen extracts and recombinant proteins: comparison of efficiency of in vitro allergy diagnostics using multiplex assay on a biological microchip.

Science.gov (United States)

Smoldovskaya, Olga; Feyzkhanova, Guzel; Arefieva, Alla; Voloshin, Sergei; Ivashkina, Olga; Reznikov, Yuriy; Rubina, Alla

2016-01-01

Immunological test systems for diagnostics of type I hypersensitivity involve the following types of antigens: whole allergen extracts, individual highly purified proteins and their recombinant analogues. The goal of this study was to compare the results obtained with whole allergen extracts (birch pollen, cat dander, and timothy grass pollen) and their respective recombinant proteins in biochip-based immunoassay. Multiplex fluorescent immunoassay of 139 patients' blood serum samples was carried out using biological microchips (biochips). sIgE concentrations for the chosen allergens and their recombinant components were measured. ROC analysis was used for comparison of the results and determination of diagnostic accuracy. The results for the birch pollen extract and its recombinant allergens have shown that the diagnostic accuracy of the methods utilizing the whole allergen extract, its major component Bet v 1 and the combination of major and minor components (Bet v 1 and Bet v 2) was the same. Values for diagnostic accuracy for the cat dander extract and its major recombinant component Fel d 1 were equal. In contrast with birch pollen and cat dander allergens, using of recombinant components of timothy grass pollen (Phl p 1, Phl p 5, Phl p 7 and Phl p 12) did not allow reaching the diagnostic accuracy of using natural extract. Multiplex analysis of samples obtained from patients with allergy to birch pollen and cat dander using biological microchips has shown that comparable accuracy was observed for the assay with natural extracts and recombinant allergens. In the case of timothy grass allergen, using the recombinant components may be insufficient.

Sublingual allergen immunotherapy

DEFF Research Database (Denmark)

Calderón, M A; Simons, F E R; Malling, Hans-Jørgen

2012-01-01

To cite this article: Calderón MA, Simons FER, Malling H-J, Lockey RF, Moingeon P, Demoly P. Sublingual allergen immunotherapy: mode of action and its relationship with the safety profile. Allergy 2012; 67: 302-311. ABSTRACT: Allergen immunotherapy reorients inappropriate immune responses......-presenting cells (mostly Langerhans and myeloid dendritic cells) exhibit a tolerogenic phenotype, despite constant exposure to danger signals from food and microbes. This reduces the induction of pro-inflammatory immune responses leading to systemic allergic reactions. Oral tissues contain relatively few mast...... cells and eosinophils (mostly located in submucosal areas) and, in comparison with subcutaneous tissue, are less likely to give rise to anaphylactic reactions. SLIT-associated immune responses include the induction of circulating, allergen-specific Th1 and regulatory CD4+ T cells, leading to clinical...

Evaluation of different glycoforms of honeybee venom major allergen phospholipase A2 (Api m 1) produced in insect cells

DEFF Research Database (Denmark)

Blank, Simon; Seismann, Henning; Plum, Melanie

2011-01-01

for the first time in insect cells. Using baculovirus infection of different insect cell lines allergen versions providing a varying degree of cross-reactive carbohydrate determinants as well as a non glycosylated variant could be obtained as secreted soluble proteins in high yields. The resulting molecules...... were analyzed for their glycosylation and proved to show advantageous properties regarding cross-reactivity in sIgE-based assays. Additionally, in contrast to the enzymatically active native protein the inactivated allergen did not induce IgE-independent effector cell activation. Thus, insect cell...

Cloning and characterization of an 11S legumin, Car i 4, a major allergen in pecan.

Science.gov (United States)

Sharma, Girdhari M; Irsigler, Andre; Dhanarajan, Pushparani; Ayuso, Rosalia; Bardina, Luda; Sampson, Hugh A; Roux, Kenneth H; Sathe, Shridhar K

2011-09-14

Among tree nut allergens, pecan allergens remain to be identified and characterized. The objective was to demonstrate the IgE-binding ability of pecan 11S legumin and characterize its sequential IgE-binding epitopes. The 11S legumin gene was amplified from a pecan cDNA library and expressed as a fusion protein in Escherichia coli. The native 11S legumin in pecan extract was identified by mass spectrometry/mass spectrometry (MS/MS). Sequential epitopes were determined by probing the overlapping peptides with three serum pools prepared from different patients' sera. A three-dimensional model was generated using almond legumin as a template and compared with known sequential epitopes on other allergenic tree nut homologues. Of 28 patients tested by dot blot, 16 (57%) bound to 11S legumin, designated Car i 4. MS/MS sequencing of native 11S legumin identified 33 kDa acidic and 20-22 kDa basic subunits. Both pecan and walnut seed protein extracts inhibited IgE binding to recombinant Car i 4, suggesting cross-reactivity with Jug r 4. Sequential epitope mapping results of Car i 4 revealed weak, moderate, and strong reactivity of serum pools against 10, 5, and 4 peptides, respectively. Seven peptides were recognized by all three serum pools, of which two were strongly reactive. The strongly reactive peptides were located in three discrete regions of the Car i 4 acidic subunit sequence (residues 118-132, 208-219, and 238-249). Homology modeling of Car i 4 revealed significant overlapping regions shared in common with other tree nut legumins.

The Effect of Water Stress on the Gas Exchange Parameters, Productivity and Seed Health of Buckwheat (Fagopyrum esculentum Moench

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Agnieszka Pszczółkowska

2012-12-01

Full Text Available The present pot experiment studied the effect of different soil moisture contents (60 - 70% CWC (capillary water capacity - control; 30 - 35% CWC - water stress on buckwheat productivity, the gas exchange parameters and health of buckwheat nuts. It was found that water deficit affected adversely certain biometric features investigated (plant height, number of nuts per cluster and caused a decrease in seed weight per plant. It was also shown that water stress reduced the values of the investigated gas exchange parameters (photosynthesis rate, transpiration rate, intercellular-space CO2 concentration, and stomatal conductance relative to the control treatment. Different soil moisture contents did not have a clear effect on fungal colonization of seeds. The multiplex PCR assays did not enable the detection of the genes responsible for mycotoxin synthesis. Under water deficit conditions, an increase was found in the content of albumin and globulin fractions as well as of glutelin fractions.

Public protection - reliable allergen risk management

Science.gov (United States)

Janković, V.; Popov Raljić, J.; Đorđević, V.

2017-09-01

Consumers with potentially fatal food allergies are dependent on correct product labelling to protect their health. The food industry is responsible for providing every detail consumers need to make informed decisions. Considering public health, food suppliers have to monitor the presence of allergens, prevent cross-contamination and label products accurately. Allergen labelling of food products, drinks and non pre-packed food and drink products is clearly defined with legal regulations. To achieve this, a complete understanding of each product’s allergenic ingredients is needed and cross-contamination of food with allergens must be avoided. Raw materials need to be checked, every ingredient must be verified and every single allergen has to be stipulated. A mislabeled product could be recalled at potential cost, financially damaging business and at the same time, negatively impacting brand and reputation.

Immunoproteomic tools are used to identify masked allergens: Ole e 12, an allergenic isoflavone reductase from olive (Olea europaea) pollen.

Science.gov (United States)

Castro, Lourdes; Crespo, Jesús F; Rodríguez, Julia; Rodríguez, Rosalía; Villalba, Mayte

2015-12-01

Proteins performing important biochemical activities in the olive tree (Olea europaea) pollen have been identified as allergens. One novel 37-kDa protein seems to be associated to the IgE-binding profile of a group of patients suffering allergy to peach and olive pollen. Three previously described olive pollen allergens exhibit very similar molecular mass. Our objective was to identify this allergen by using immunoproteomic approaches. After 2D-electrophoresis and mass spectrometry, peptide sequences from several IgE-binding spots, allowed identifying this new allergen, as well as cloning and DNA sequencing of the corresponding gene. The allergen, named Ole e 12, is a polymorphic isoflavone reductase-like protein of 308 amino acids showing 80% and 74% identity with birch and pear allergens, Bet v 6 and Pyr c 5, respectively. A prevalence of 33% in the selected population is in contrast to 4%-10% in groups of subjects suffering from pollinosis. Recombinant allergen was produced in Escherichia coli, and deeply characterised. Immunoblotting and ELISA detection as well as inhibition experiments were performed with polyclonal antisera and allergic patients' sera. The recombinant allergen retains the IgE reactivity of its natural counterpart. Close structural and immunological relationships between members of this protein family were supported by their IgG recognition in vegetable species. In summary, Ole e 12 is a minor olive pollen allergen, which gains relevance in patients allergic to peach with olive pollinosis. Proteomic approaches used to analyse this allergen provide useful tools to identify hidden allergens, relevant for several allergic populations and thus complete allergenic panels. Copyright © 2015 Elsevier B.V. All rights reserved.

AllerML: markup language for allergens.

Science.gov (United States)

Ivanciuc, Ovidiu; Gendel, Steven M; Power, Trevor D; Schein, Catherine H; Braun, Werner

2011-06-01

Many concerns have been raised about the potential allergenicity of novel, recombinant proteins into food crops. Guidelines, proposed by WHO/FAO and EFSA, include the use of bioinformatics screening to assess the risk of potential allergenicity or cross-reactivities of all proteins introduced, for example, to improve nutritional value or promote crop resistance. However, there are no universally accepted standards that can be used to encode data on the biology of allergens to facilitate using data from multiple databases in this screening. Therefore, we developed AllerML a markup language for allergens to assist in the automated exchange of information between databases and in the integration of the bioinformatics tools that are used to investigate allergenicity and cross-reactivity. As proof of concept, AllerML was implemented using the Structural Database of Allergenic Proteins (SDAP; http://fermi.utmb.edu/SDAP/) database. General implementation of AllerML will promote automatic flow of validated data that will aid in allergy research and regulatory analysis. Copyright © 2011 Elsevier Inc. All rights reserved.

Recombinant allergen Lol p II: expression, purification and characterization.

Science.gov (United States)

Tamborini, E; Brandazza, A; De Lalla, C; Musco, G; Siccardi, A G; Arosio, P; Sidoli, A

1995-05-01

Pollen from perennial rye grass (Lolium perenne) is a major cause of type I allergies worldwide. It contains complex mixtures of proteins, among which Lol p II is a major allergen. Previously, we have reported the cloning and sequencing of Lol p II and its expression in fusion with the heavy chain of human ferritin as carrier polypeptide (Sidoli et al., 1993, J. biol. Chem. 268, 21819-21825). Here, we describe the expression, purification and characterization of a recombinant Lol p II overproduced as a non-fusion protein in the periplasm of E. coli. The recombinant allergen was expressed in high yields and was easily purified in milligram amounts. It competed with the natural Lol p II for binding to specific IgE, and it induced allergic responses in skin prick tests, indicating to be immunologically analogous to the natural protein. Biochemical analyses indicate that recombinant Lol p II is a highly stable and soluble monomeric molecule which behaves like a small globular protein.

Modeling of allergen proteins found in sea food products

Directory of Open Access Journals (Sweden)

Nataly Galán-Freyle

2012-06-01

Full Text Available Shellfish are a source of food allergens, and their consumption is the cause of severe allergic reactions in humans. Tropomyosins, a family of muscle proteins, have been identified as the major allergens in shellfish and mollusks species. Nevertheless, few experimentally determined three-dimensional structures are available in the Protein Data Base (PDB. In this study, 3D models of several homologous of tropomyosins present in marine shellfish and mollusk species (Chaf 1, Met e1, Hom a1, Per v1, and Pen a1 were constructed, validated, and their immunoglobulin E binding epitopes were identified using bioinformatics tools. All protein models for these allergens consisted of long alpha-helices. Chaf 1, Met e1, and Hom a1 had six conserved regions with sequence similarities to known epitopes, whereas Per v1 and Pen a1 contained only one. Lipophilic potentials of identified epitopes revealed a high propensity of hydrophobic amino acids in the immunoglobulin E binding site. This information could be useful to design tropomyosin-specific immunotherapy for sea food allergies.

Evaluation of molecular basis of cross reactivity between rye and Bermuda grass pollen allergens.

Science.gov (United States)

Tiwari, Ruby; Bhalla, Prem L; Singh, Mohan B

2009-12-01

Allergenic cross reactivity between the members of the Pooids (Lolium perenne, Phleum pratense, and Poa pratensis) and Chloridoids (Cynodon dactylon and Paspalum notatum) is well established. Studies using crude extracts in the past have demonstrated limited cross reactivity between the Pooids and the Chloridoids suggesting separate diagnosis and therapy. However, little is known regarding the molecular basis for the limited cross reactivity observed between the 2 groups of grasses. The present study was undertaken to gain insights into the molecular basis of cross allergenicity between the major allergens from rye and Bermuda grass pollens. Immunoblot inhibition tests were carried out to determine the specificity of the proteins involved in cross reactivity. Crude pollen extract and bacterially expressed and purified recombinant Lol p 1and Lol p 5 from rye grass were subjected to cross inhibition experiments with crude and purified recombinant Cyn d 1 from Bermuda grass using sera from patients allergic to rye grass pollen. The immunoblot inhibition studies revealed a high degree of cross inhibition between the group 1 allergens. In contrast, a complete lack of inhibition was observed between Bermuda grass group 1 allergen rCyn d 1, and rye grass group 5 allergen rLol p 5. Crude rye grass extract strongly inhibited IgE reactivity to Bermuda grass, whereas crude Bermuda grass pollen extract showed a weaker inhibition. Our data suggests that a possible explanation for the limited cross reactivity between the Pooids and Chloridoids may, in part, be due to the absence of group 5 allergen from Chloridoid grasses. This approach of using purified proteins may be applied to better characterize the cross allergenicity patterns between different grass pollen allergens.

Specific IgE for Fag e 3 Predicts Oral Buckwheat Food Challenge Test Results and Anaphylaxis: A Pilot Study.

Science.gov (United States)

Yanagida, Noriyuki; Sato, Sakura; Maruyama, Nobuyuki; Takahashi, Kyohei; Nagakura, Ken-Ichi; Ogura, Kiyotake; Asaumi, Tomoyuki; Ebisawa, Motohiro

2018-01-01

Buckwheat (BW) is the source of a life-threatening allergen. Fag e 3-specific serum IgE (sIgE) is more useful than BW-sIgE for diagnosis; however, it is unknown whether Fag e 3-sIgE can predict oral food challenge (OFC) results and anaphylaxis. This study aimed to clarify the efficacy of Fag e 3-sIgE in predicting OFC results and anaphylaxis. We conducted a retrospective review of BW- and Fag e 3-sIgE data obtained using the ImmunoCAP® assay system and fluorescent enzyme-linked immunosorbent assay from children who underwent OFC using 3,072 mg of BW protein between July 2006 and March 2014 at Sagamihara National Hospital, Kanagawa, Japan. We analyzed 60 patients aged 1.9-13.4 years (median 6.0 years); 20 (33%) showed objective symptoms upon BW OFC. The patients without symptoms had significantly lower Fag e 3-sIgE than those with non-anaphylactic (p tested factor that significantly predicted positive OFC results (odds ratio 8.93, 95% confidence interval 3.10-25.73, p < 0.001) and OFC-induced anaphylaxis (2.67, 1.12-6.35, p = 0.027). We suggest that a threshold Fag e 3-sIgE level of 18.0 kUE/L has 95% probability of provoking a positive reaction to BW. Fag e 3-sIgE predicted OFC results and OFC-induced anaphylaxis. We further emphasize paying careful attention to the risk of BW OFC-induced anaphylaxis. © 2018 The Author(s) Published by S. Karger AG, Basel.

Hyposensitization therapy with whole pollen extract or purified allergens monitored by immunoblotting

DEFF Research Database (Denmark)

Jarolim, E; Matthiesen, F; Skov, P S

1990-01-01

. Inhibition experiments using allergenic components isolated by preparative sodium dodecyl sulphate-polyacrylamide gel electrophoresis indicated that all antigenic components of timothy grass pollen detected in immunoblot dispose of private and cross-reactive determinants for binding of human IgE. The worse......Patients allergic to grass pollen were hyposensitized with two major allergenic components or whole extract of timothy grass pollen. Specific IgE, IgG1, and IgG4 formed during immunotherapy were analyzed by immunoblotting. Similar antibody-binding patterns were observed in both patient groups...

Biochemical and immunological characterization of recombinant allergen Lol p 1.

Science.gov (United States)

Tamborini, E; Faccini, S; Lidholm, J; Svensson, M; Brandazza, A; Longhi, R; Groenlund, H; Sidoli, A; Arosio, P

1997-11-01

Pollen from perennial rye grass (Lolium perenne), a major cause of type-I allergy worldwide, contains a complex mixture of allergenic proteins among which Lol p 1 is one of the most important. We describe the expression, purification and characterization of a recombinant Lol p 1 overproduced in Escherichia coli. The recombinant allergen, expressed in high yields and purified in milligram amounts, bound to specific IgE antibodies from human sera, induced histamine release from sensitized human basophils, and elicited rabbit antisera that recognize specifically recombinant Lol p 1 and natural Lol p 1 of pollen extract. Recombinant Lol p 1 was used to develop ImmunoCAP assays for analysis of 150 sera that were Radioallergosorbent test positive to L. perenne pollen. In 130 of them (87%) the assay detected a significant level of IgE antibodies to Lol p 1, reaching on average 37% of the level obtained with a test for IgE to the whole grass pollen extract. To map epitopes on Lol p 1, we produced three deletion mutants [des-(116-240)-Lol p 1, des-(1-88)-Lol p 1 and des-(133-189)-Lol p 1], which were efficiently expressed in bacteria. These all showed a strong reactivity with the specific rabbit IgG antibodies, but lacked most or all the allergenic properties of recombinant Lol p 1. A study of the antigenic structure of Lol p 1 was performed using the three deletion mutants and a set of 17-18-residue overlapping synthetic peptides covering the whole allergen sequence. The results indicate that human IgE and rabbit IgG antibodies bind to distinct regions of Lol p 1, and that at least some important IgE epitopes are mainly conformational. The findings suggest that recombinant allergens constitute useful reagents for further development of serological diagnosis of allergy, and that it should be possible to produce immunogenic fragments of allergenic proteins without allergenic properties.

Nanoparticle–allergen complexes for allergen immunotherapy

Directory of Open Access Journals (Sweden)

Di Felice G

2017-06-01

Full Text Available Gabriella Di Felice,1 Paolo Colombo2 1National Center for Drug Research and Evaluation, Istituto Superiore di Sanità, Rome, 2Institute of Biomedicine and Molecular Immunology, National Research Council, Palermo, Italy Abstract: Allergen-specific immunotherapy was introduced in clinical settings more than 100 years ago. It remains the only curative approach to treating allergic disorders that ameliorates symptoms, reduces medication costs, and blocks the onset of new sensitizations. Despite this clinical evidence and knowledge of some immunological mechanisms, there remain some open questions regarding the safety and efficacy of this treatment. This suggests the need for novel therapeutic approaches that attempt to reduce the dose and frequency of treatment administration, improving patient compliance, and reducing costs. In this context, the use of novel adjuvants has been proposed and, in recent years, biomedical applications using nanoparticles have been exploited in the attempt to find formulations with improved stability, bioavailability, favorable biodistribution profiles, and the capability of targeting specific cell populations. In this article, we review some of the most relevant regulatory aspects and challenges concerning nanoparticle-based formulations with immunomodulatory potential, their related immunosafety issues, and the nature of the nanoparticles most widely employed in the allergy field. Furthermore, we report in vitro and in vivo data published using allergen/nanoparticle systems, discuss their impact on the immune system in terms of immunomodulatory activity and the reduction of side effects, and show that this strategy is a novel and promising tool for the development of allergy vaccines. Keywords: allergy, nanocarriers, immunotoxicity, immune modulation, immunotherapy, allergens

Current glimpse of airborne allergenic pollen in Indian subcontinent

Directory of Open Access Journals (Sweden)

Kavita Ghosal

2015-12-01

Full Text Available Respiratory allergic diseases such as bronchial asthma, rhinitis, urticaria, atopic dermatitis have been steadily increasing all over the world, including India. Owing to its alarming trend, several aerobiological surveys have been undertaken in different parts of India to delineate the variety of pollen and spore load. In this review, we have reported the current state of aerobiological knowladge in India with particular reference to allergenic airborn pollen occurence in 2001–2015. Pollen have been found to contribute a significant proportion in the air and caused allergy symptoms in the local inhabitants. Aerobiological records, a questionnaire survey and hospitalization records have been employed for the analysis. Holoptelea integrifolia, Amaranthus spinosus in northern region, Sorghum vulgare, Pennisetum, Gynandropsis gynandra, Parthenium hysterophorus, Dolichandrone platycalyx in southern regions, and Parthenium hysterophorus from the western region; Cynodon dactylon, Cenchrus ciliaris in the central area; Acacia auriculiformis, Cleome gynandra, Catharanthus roseus, Phoenix sylvestris, Areca catechu, and Lantana camara in the eastern regions as potential aeroallergens in India. The statistical approach confirmed the correlation between hospitalization rate associated with allergy-related health troubles and the prevalent allergenic pollen in the air. The Poaceae group has been found to be dominant throughout India. Immuno-biochemical studies identified various protein with allergenic potential found in the pollen recorded. Epitope identification and homology of the major allergenic protein Cat r1 of Catharanthus sp and Par j 1 of Parietaria judaica have been found. Identification of allergenic pollen grains and the modern approach concerning cross-reactivity and epitope revelation of dominant airborne pollen have important clinical implications for the prevention, diagnosis and treatments of allergic diseases in India.

A protocol for a systematic review to identify allergenic tree nuts and the molecules responsible for their allergenic properties.

Science.gov (United States)

Javed, Bushra; Padfield, Philip; Sperrin, Matthew; Simpson, Angela; Mills, E N Clare

2017-08-01

Food regulations require that tree nuts and derived ingredients are included on food labels in order to help individuals with IgE-mediated allergies to avoid them. However, there is no consensus regarding which tree nut species should be included in this definition and specified on food labels. Allergen detection methods used for monitoring foods target allergen molecules, but it not clear which are the most relevant molecules to choose. A modified population-exposure-comparators-outcome (PECO) approach has been developed to systematically review the evidence regarding (1) which allergenic tree nuts should be included in food allergen labelling lists and (2) which are the clinically relevant allergens which should be used as analytical targets. A search strategy and criteria against which the evidence will be evaluated have been developed. The resulting evidence will be used to rank tree nuts with regards their ability to cause IgE-mediated allergies, and allergen molecules regarding their capacity to elicit an allergic reaction. The results of the systematic review will enable risk assessors and managers to identify tree nut species that should be included in food allergen labelling lists and ensure analytical methods for determination of allergens in foods are targeting appropriate molecules. Copyright © 2017. Published by Elsevier Ltd.

Patch test study of 90 patients with tattoo reactions: negative outcome of allergy patch test to baseline batteries and culprit inks suggests allergen(s) are generated in the skin through haptenization.

Science.gov (United States)

Serup, Jørgen; Hutton Carlsen, Katrina

2014-11-01

Tattoo reactions, especially in red tattoos, are often suggested as allergic in nature, however, systematic evaluation by patch testing has not performed in the past. To report the results of patch testing in 90 patients with non-infectious chronic tattoo reactions. From 2009 to 2013 at the 'Tattoo Clinic', Department of Dermatology, Bispebjerg University Hospital, 90 patients were patch tested with batteries of baseline allergens, disperse dyes/textile allergens, and a selection of tattoo ink stock products, which, according to case observations, were problematic, supplemented with individual culprit inks when accessible. Patients with reactions to the tattoo colour red, the most predominant colour associated with skin reactions, showed negative patch test results with common allergens. Outcomes were also negative in patients who had experienced concomitant reactions in another hitherto tolerated tattoo of the same colour as the problematic tattoo. The allergen or allergens responsible for tattoo reactions are not present directly in tattoo ink stock products. This is despite the fact that clinical histories suggest that the vast majority of clinical reactions, especially reactions to red and red nuances, are likely to be allergic events caused by the injected inks. We suggest that the responsible allergen results from a complicated and slow process of haptenization, which may even include photochemical cleavage of red azo pigment. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Consumer preferences for food allergen labeling.

Science.gov (United States)

Marra, Carlo A; Harvard, Stephanie; Grubisic, Maja; Galo, Jessica; Clarke, Ann; Elliott, Susan; Lynd, Larry D

2017-01-01

Food allergen labeling is an important tool to reduce risk of exposure and prevent anaphylaxis for individuals with food allergies. Health Canada released a Canadian food allergen labeling regulation (2008) and subsequent update (2012) suggesting that research is needed to guide further iterations of the regulation to improve food allergen labeling and reduce risk of exposure. The primary objective of this study was to examine consumer preferences in food labeling for allergy avoidance and anaphylaxis prevention. A secondary objective was to identify whether different subgroups within the consumer population emerged. A discrete choice experiment using a fractional factorial design divided into ten different versions with 18 choice-sets per version was developed to examine consumer preferences for different attributes of food labeling. Three distinct subgroups of Canadian consumers with different allergen considerations and food allergen labeling needs were identified. Overall, preferences for standardized precautionary and safety symbols at little or no increased cost emerged. While three distinct groups with different preferences were identified, in general the results revealed that the current Canadian food allergen labeling regulation can be improved by enforcing the use of standardized precautionary and safety symbols and educating the public on the use of these symbols.

Characterization of Cannabis sativa allergens.

Science.gov (United States)

Nayak, Ajay P; Green, Brett J; Sussman, Gordon; Berlin, Noam; Lata, Hemant; Chandra, Suman; ElSohly, Mahmoud A; Hettick, Justin M; Beezhold, Donald H

2013-07-01

Allergic sensitization to Cannabis sativa is rarely reported, but the increasing consumption of marijuana has resulted in an increase in the number of individuals who become sensitized. To date, little is known about the causal allergens associated with C sativa. To characterize marijuana allergens in different components of the C sativa plant using serum IgE from marijuana sensitized patients. Serum samples from 23 patients with a positive skin prick test result to a crude C sativa extract were evaluated. IgE reactivity was variable between patients and C sativa extracts. IgE reactivity to C sativa proteins in Western blots was heterogeneous and ranged from 10 to 70 kDa. Putative allergens derived from 2-dimensional gels were identified. Prominent IgE reactive bands included a 23-kDa oxygen-evolving enhancer protein 2 and a 50-kDa protein identified to be the photosynthetic enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase. Additional proteins were identified in the proteomic analysis, including those from adenosine triphosphate synthase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and luminal binding protein (heat shock protein 70), suggesting these proteins are potential allergens. Deglycosylation studies helped refine protein allergen identification and demonstrated significant IgE antibodies against plant oligosaccharides that could help explain cross-reactivity. Identification and characterization of allergens from C sativa may be helpful in further understanding allergic sensitization to this plant species. Copyright © 2013 American College of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Uptake of various trace elements during germination of wheat, buckwheat and quinoa.

Science.gov (United States)

Lintschinger, J; Fuchs, N; Moser, H; Jäger, R; Hlebeina, T; Markolin, G; Gössler, W

1997-01-01

The practice of sprouting is widely used to improve the nutritional value of grain seeds. Several nutritive factors such as vitamin concentrations and bioavailability of trace elements and minerals increase during germination. The objective of this work was to study the enrichment of various essential trace elements during germination of wheat (Triticum aestivum), buckwheat (Fagopyrum esculentum), and quinoa (Chenopodium quinoa) seeds in order to improve their nutritional role as a source of bioavailable trace elements. Seeds were sprouted either in distilled- or tap-water and in five different electrolyte solutions to investigate the concentration-dependent uptake. The time-dependence was investigated by analyzing aliquots of the sprouts after certain germination periods. Samples were analyzed after freeze drying for their Li, V, Cr, Fe, Mn, Co, Cu, Zn, Sr, Mo, As and Se concentrations with inductively-coupled plasma mass-spectrometry (ICP-MS). As a control for possible changes in the biochemical metabolism of the sprouts, the biosynthesis of vitamin C was also determined by using reversed-phase ion-pair HPLC. It was shown that quinoa was the most resistant to the applied electrolyte solutions and had the highest uptake rates for almost all elements, followed by buckwheat and wheat. Greatest increases were observed for Co, Sr, and Li. No significant changes in vitamin C biosynthesis were observed between sprouts grown in different electrolyte solutions. The time-dependent uptake for most elements was characterized by a significant absorption during soaking of the seeds, followed by a lag phase during the first day of germination and an increased uptake during the second and third day. Se and As showed distinctly different uptake behaviors.

Chemical and Biological Properties of Food Allergens

NARCIS (Netherlands)

Jedrychowski, L.; Wichers, H.J.

2009-01-01

This book provides epidemiological data on food allergens and information on the incidence of food allergies. It discusses the link between hypersensitivity and immune system health and covers methods used for assays on allergenic components, animal models for allergen analysis, and clinical methods

Avoidance Behavior against Positive Allergens Detected with a Multiple Allergen Simultaneous Test Immunoblot Assay in Patients with Urticaria: Factors Associated with Avoidance Success/Failure.

Science.gov (United States)

Lee, Min Kyung; Kwon, In Ho; Kim, Han Su; Kim, Heung Yeol; Cho, Eun Byul; Bae, Youin; Park, Gyeong Hun; Park, Eun Joo; Kim, Kwang Ho; Kim, Kwang Joong

2016-02-01

Avoidance behavior against positive allergens detected by using multiple allergen simultaneous test (MAST)-immunoblot assay in patients with urticaria has been rarely reported. We aimed to assess the avoidance behavior of patients with urticaria against positive allergens detected with a MAST. One hundred and one urticaria patients who showed positivity to at least one allergen on a MAST completed a questionnaire regarding their test results. The avoidance behavior of the patients was evaluated, and relevant determining factors of avoidance success/failure were statistically assessed. We detected 144 different data (n=51, food allergens; n=17, pollen allergens; and n=76, aeroallergens) from 101 patients with urticaria. The avoidance failure rates were 33.3% for food allergens, 70.6% for pollen allergens, and 30.3% for aeroallergens. The pollen group showed a significantly higher avoidance failure rate than the food and aeroallergen groups (psuccessfully avoid allergens (psuccess or failure against allergens in patients with urticaria when clinicians conduct allergen-specific immunoglobulin E tests.

Bioethanol production from waste bread samples made from mixtures of wheat and buckwheat flours

OpenAIRE

Ačanski, Marijana; Pastor, Kristian; Razmovski, Radojka; Vučurović, Vesna; Psodorov, Đorđe

2014-01-01

In this paper yields of bioethanol from seven samples of bread were compared. Samples of bread were produced and prepared in a laboratory by mixing wheat and buckwheat flour in amounts of 0, 20, 40, 50, 60, 80 and 100%. At first, the analysis of all seven samples of bread was done (dry matter, starch content and pH value of bread sample suspensions). Then the waste bread suspensions were hydrolyzed by applying commercial hydrolytic enzymes, Termamyl® SC and SAN Extra® L. The fermentation proc...

Allergen Sensitization Pattern by Sex: A Cluster Analysis in Korea.

Science.gov (United States)

Ohn, Jungyoon; Paik, Seung Hwan; Doh, Eun Jin; Park, Hyun-Sun; Yoon, Hyun-Sun; Cho, Soyun

2017-12-01

Allergens tend to sensitize simultaneously. Etiology of this phenomenon has been suggested to be allergen cross-reactivity or concurrent exposure. However, little is known about specific allergen sensitization patterns. To investigate the allergen sensitization characteristics according to gender. Multiple allergen simultaneous test (MAST) is widely used as a screening tool for detecting allergen sensitization in dermatologic clinics. We retrospectively reviewed the medical records of patients with MAST results between 2008 and 2014 in our Department of Dermatology. A cluster analysis was performed to elucidate the allergen-specific immunoglobulin (Ig)E cluster pattern. The results of MAST (39 allergen-specific IgEs) from 4,360 cases were analyzed. By cluster analysis, 39items were grouped into 8 clusters. Each cluster had characteristic features. When compared with female, the male group tended to be sensitized more frequently to all tested allergens, except for fungus allergens cluster. The cluster and comparative analysis results demonstrate that the allergen sensitization is clustered, manifesting allergen similarity or co-exposure. Only the fungus cluster allergens tend to sensitize female group more frequently than male group.

[Sampling of allergens in dust deposited in the workplace].

Science.gov (United States)

Perfetti, L; Galdi, E; Pozzi, V; Moscato, G

2001-01-01

Some workplaces share with domestic dwellings many characteristics favouring house dust mite growth. Moreover it has recently been shown that pets owners can bring allergens to public places with their clothes. So it is possible that significant exposure to indoor allergens can occur outside homes, at the workplace. The recent availability of immunoassays with monoclonal antibodies for indoor allergens has enabled many investigators to quantify exposure to such allergens in epidemiological studies. Analysis of allergens in settled dust is a simple method of quantification exposure to indoor allergens. The concentrations of indoor allergens in public places have already been investigated and high levels of indoor allergens have been reported. A study performed by our group in offices (banks and media) in different regions of Italy has also shown significant levels of indoor allergens. Thus, evaluating exposure to indoor allergens at the workplace is critical to evaluate risk factors for sensitization and elicitation of symptoms in sensitized subjects and such data help in addressing correctly the problem of reducing exposure levels.

Nasal mucosal blood flow after intranasal allergen challenge

International Nuclear Information System (INIS)

Holmberg, K.; Bake, B.; Pipkorn, U.

1988-01-01

The nasal mucosal blood flow in patients with allergic rhinitis was determined at nasal allergen challenges with the 133 Xenon washout method. Determinations were made in 12 subjects before and 15 minutes after challenge with diluent and increasing doses of allergen. The time course was followed in eight subjects by means of repeated measurements during 1 hour after a single allergen dose. Finally, the blood flow was measured after unilateral allergen challenge in the contralateral nasal cavity. A dose-dependent decrease in blood flow was found after nasal challenge with increasing doses of allergens, whereas challenge with diluent alone did not induce any changes. The highest allergen dose, which also induced pronounced nasal symptoms, resulted in a decrease in blood flow of 25% (p less than 0.001). The time-course study demonstrated a maximum decrease in blood flow 10 to 20 minutes after challenge and then a gradual return to baseline. Unilateral allergen challenge resulted in a decrease in blood flow in the contralateral, unchallenged nasal cavity, suggesting that part of the allergen-induced changes in blood flow were reflex mediated

Measurement of IgE antibodies against purified grass pollen allergens (Lol p 1, 2, 3 and 5) during immunotherapy.

Science.gov (United States)

Van Ree, R; Van Leeuwen, W A; Dieges, P H; Van Wijk, R G; De Jong, N; Brewczyski, P Z; Kroon, A M; Schilte, P P; Tan, K Y; Simon-Licht, I F; Roberts, A M; Stapel, S O; Aalberse, R C

1997-01-01

IgE titres tend to rise early after the start of immunotherapy, followed by a decline to pre-immunotherapy levels or lower. We were interested to know whether the early increase in IgE antibodies includes new specificities of IgE, and whether these responses persist. Sera of 64 patients undergoing grass pollen immunotherapy were tested for IgE against four purified grass pollen allergens: Lol p 1, 2, 3, and 5. At least two serum samples were taken, one before the start of therapy and one between 5 and 18 months after the first immunization (mean: 10 months). The mean IgE responses to Lol p 1, 2 and 3 showed a moderate but not significant increase. In contrast, the mean IgE response to Lol p 5 showed a significant decrease of > 30%. IgE against total Lohum perenne pollen extract moderately increased (> 20%), showing that a RAST for total pollen is not always indicative for the development of IgE against its major allergens. For > 40% of the patients it was found that IgE against one or more of the four allergens increased, while IgE against the remaining allergen(s) decreased. For 10 sera the ratio of IgE titres against at least two allergens changed by at least a factor of 5. The changes in specific IgE also included conversions from negative (< 0.1 RU) to positive (0.6 to 5.0 RU) for five patients. For two patients, the induction of these 'new' IgE antibodies against major allergens was shown to result in a response that was persistent over several years. Although active induction of new IgE specificities by immunotherapy was not really proven, the observations in this study indicate that monitoring of IgE against purified (major) allergens is necessary to evaluate changes in specific IgE in a reliable way.

Identification of enolases and aldolases as important fish allergens in cod, salmon and tuna: component resolved diagnosis using parvalbumin and the new allergens.

Science.gov (United States)

Kuehn, A; Hilger, C; Lehners-Weber, C; Codreanu-Morel, F; Morisset, M; Metz-Favre, C; Pauli, G; de Blay, F; Revets, D; Muller, C P; Vogel, L; Vieths, S; Hentges, F

2013-07-01

The majority of fish-allergic patients are sensitized to parvalbumin, known to be the cause of important IgE cross-reactivity among fish species. Little is known about the importance of fish allergens other than parvalbumin. The aim of this study was to characterize hitherto undefined fish allergens in three commonly consumed fish species, cod, salmon and tuna, and to evaluate their importance for in vitro IgE-diagnosis in addition to parvalbumin and fish gelatin. Sixty-two patients were diagnosed by clinical history, skin prick tests and specific IgE to fish extracts. Two new fish allergens from cod, salmon and tuna were identified by microsequencing. These proteins were characterized by immunoblot, ELISA and mediator release assay. Purified parvalbumin, enolase, aldolase and fish gelatin were used for quantification of specific IgE in ELISA. Parvalbumin and two other allergens of 50 and 40 kDa were detected in IgE-immunoblots of cod, salmon and tuna extracts by most patient sera. The 50 and 40 kDa proteins were identified as beta-enolase and fructose-bisphosphate aldolase A respectively. Both purified enzymes showed allergenic activity in the mediator release assay. Indeed, 72.6% of the patients were sensitized to parvalbumin, 20% of these had specific IgE to salmon parvalbumin only. IgE to enolases were found in 62.9% (0.5-95.0 kUA /L), to aldolases in 50.0% (0.4-26.0 kUA /L) and to fish gelatin in 19.3% (0.4-20.0 kUA /L) of the patients. Inter-species cross-reactivity, even though limited, was found for enolases and aldolases by IgE-inhibition ELISA. Fish enolase and aldolase have been identified as important new fish allergens. In fish allergy diagnosis, IgE to enolase and aldolase are especially relevant when IgE to parvalbumin are absent. © 2013 John Wiley & Sons Ltd.

Allergen-specific immunotherapy

Directory of Open Access Journals (Sweden)

Moote William

2011-11-01

Full Text Available Abstract Allergen-specific immunotherapy is a potentially disease-modifying therapy that is effective for the treatment of allergic rhinitis/conjunctivitis, allergic asthma and stinging insect hypersensitivity. However, despite its proven efficacy in these conditions, it is frequently underutilized in Canada. The decision to proceed with allergen-specific immunotherapy should be made on a case-by-case basis, taking into account individual patient factors such as the degree to which symptoms can be reduced by avoidance measures and pharmacological therapy, the amount and type of medication required to control symptoms, the adverse effects of pharmacological treatment, and patient preferences. Since this form of therapy carries the risk of anaphylactic reactions, it should only be prescribed by physicians who are adequately trained in the treatment of allergy. Furthermore, injections must be given under medical supervision in clinics that are equipped to manage anaphylaxis. In this article, the authors review the indications and contraindications, patient selection criteria, and the administration, safety and efficacy of allergen-specific immunotherapy.

Food, novel foods, and allergenicity

NARCIS (Netherlands)

Loveren H van; LPI

2002-01-01

Certain foods lead may to allergic responses in certain individuals. Main allergenic foods are Crustacea (shrimp, lobster, crab), egg, fish, milk, peanuts, soybeans, tree nuts, and wheat, and allergens are always proteins. A wide array of symptoms can result from food allergy (gastrointestinal,

Dominating IgE-binding epitope of Bet v 1, the major allergen of birch pollen, characterized by X-ray crystallography and site-directed mutagenesis

DEFF Research Database (Denmark)

Spangfort, Michael D; Mirza, Osman; Ipsen, Henrik

2003-01-01

Specific allergy vaccination is an efficient treatment for allergic disease; however, the development of safer vaccines would enable a more general use of the treatment. Determination of molecular structures of allergens and allergen-Ab complexes facilitates epitope mapping and enables a rational...

The human allergens of mesquite (Prosopis juliflora).

Science.gov (United States)

Killian, Sue; McMichael, John

2004-07-05

BACKGROUND: A computerized statistical analysis of allergy skin test results correlating patient reactivities initiated our interest in the cross-reactive allergens of mesquite tree pollen. In-vitro testing with mesquite-sensitized rabbits and a variety of deciduous tree pollens revealed so many cross-reactivities that it became apparent there could be more allergens in mesquite than previously described in the world literature. Our purpose was to examine the allergens of mesquite tree pollen (Prosopis juliflora) which elicit an IgE response in allergic humans so that future research could determine if these human allergens cross-react with various tree pollens in the same manner as did the mesquite antiserum from sensitized rabbits. METHODS: Proteins from commercial mesquite tree pollen were separated by polyacrylamide gel electrophoresis in the presence of sodium-dodecyl-sulphate. These mesquite proteins were subjected to Western blotting using pooled sera from ten mesquite-sensitive patients and goat anti-human IgE. The allergens were detected using an Amplified Opti-4-CN kit, scanned, and then interpreted by Gel-Pro software. RESULTS: Thirteen human allergens of mesquite pollen were detected in this study. CONCLUSION: The number of allergens in this study of mesquite exceeded the number identified previously in the literature. With the increased exposure to mesquite through its use in "greening the desert", increased travel to desert areas and exposure to mesquite in cooking smoke, the possible clinical significance of these allergens and their suggested cross-reactivity with other tree pollens merit further study.

Elaboration, Physicochemical and Sensory Analysis of a Sweet Bread Made with Buckwheat and Fruit Flours

OpenAIRE

C.S. Möller; V.R. De Oliveira; T.P. Amorim; R.U. Thoen; A. de O. Rios

2013-01-01

The aim of this study was to evaluate the viability of new gluten free sweet bread, as well as their physicochemical features and acceptance. As more and more people become aware of coeliac disease, it is important to increase the options in terms of gluten free bakery products. The adaptation of bakery product-sweet bread-to gluten free versions can be justified for its role in preventing coeliac problems. Wheat flour was substituted by common buckwheat flour mixed with fruit flours and thei...

21 CFR 680.1 - Allergenic Products.

Science.gov (United States)

2010-04-01

... ADDITIONAL STANDARDS FOR MISCELLANEOUS PRODUCTS § 680.1 Allergenic Products. (a) Definition. Allergenic Products are products that are administered to man for the diagnosis, prevention or treatment of allergies...

Incorporation of exogenous L-phenylalanine into C-glycosylflavones in buckwheat cotyledons

International Nuclear Information System (INIS)

Margna, U.; Margna, E.

1978-01-01

By tracer experiments it was demonstrated that in excised buckwheat cotyledons L-phenylalanine fed exogeneously was incorporated predominantly into the luteolinic C-glycosylflavones orientin and iso-orientin, but not into their simpler apigeninic analogues vitexin and isovitexin, as it could be expected theoretically. Evidence is presented that L-phenylalanine supplied exogenously does not mix with the endogenous pool of that precursor, and that ratios of L-phenylalanine distribution between pathways of apigeninic and luteolinic C-glycosylflavones are different depending on wether the flavonoids are synthesized from endogenous or exogenous material. This phenomenon is suggested to be caused by the differences between enzyme complexes responsible for the biosynthesis of separate C-glycosylflavones in their capability of consuming common endogenous precursors. (author)

Comparison of international food allergen labeling regulations.

Science.gov (United States)

Gendel, Steven M

2012-07-01

Food allergy is a significant public health issue worldwide. Regulatory risk management strategies for allergic consumers have focused on providing information about the presence of food allergens through label declarations. A number of countries and regulatory bodies have recognized the importance of providing this information by enacting laws, regulations or standards for food allergen labeling of "priority allergens". However, different governments and organizations have taken different approaches to identifying these "priority allergens" and to designing labeling declaration regulatory frameworks. The increasing volume of the international food trade suggests that there would be value in supporting sensitive consumers by harmonizing (to the extent possible) these regulatory frameworks. As a first step toward this goal, an inventory of allergen labeling regulations was assembled and analyzed to identify commonalities, differences, and future needs. Published by Elsevier Inc.

Triethylene glycol bis(2-ethylhexanoate) - a new contact allergen identified in a spectacle frame

DEFF Research Database (Denmark)

Andersen, Klaus Ejner; Vestergaard, M. E.; Christensen, Lars Porskjær

2014-01-01

Background. Allergic reactions to spectacle frames are not unusual. A patient had a reproducible strong allergic patch test reaction to scrapings from the plastic material, and negative patch test results with available spectacle frame chemicals. Objectives. To identify the culprit allergen...... bis(2-ethylhexanoate) was the causative allergen in the spectacle frame. Ten consecutive eczema patients tested as controls were negative. Conclusion. Triethylene glycol bis(2-ethylhexanoate) is a new, hitherto unreported contact allergen....... in this patient's spectacle frame. Materials and methods. An extract from the temple arms was analysed by gas chromatography mass spectrometry (GC-MS), and a major low molecular weight compound was detected. This compound was isolated by semi-preparative high-performance liquid chromatography and identified by GC...

Associations between baseline allergens and polysensitization

DEFF Research Database (Denmark)

Carlsen, B.C.; Menne, T.; Johansen, J.D.

2008-01-01

Background: Identification of patients at risk of developing polysensitization is not possible at present. An association between weak sensitizers and polysensitization has been hypothesized. Objectives: To examine associations of 21 allergens in the European baseline series to polysensitization....... Patients/Methods: From a database-based study with 14 998 patients patch tested with the European baseline series between 1985 and 2005, a group of 759 (5.1%) patients were polysensitized. Odds ratios were calculated to determine the relative contribution of each allergen to polysensitization. Results...... denominator for the association between the allergens and the polysensitization was apparent, and any association, whether positive or negative, was relatively low. Based on these results, sensitization to specific baseline allergens cannot be used as risk indicators for polysensitization Udgivelsesdato: 2008...

Associations between baseline allergens and polysensitization

DEFF Research Database (Denmark)

Carlsen, Berit Christina; Menné, Torkil; Johansen, Jeanne Duus

2008-01-01

BACKGROUND: Identification of patients at risk of developing polysensitization is not possible at present. An association between weak sensitizers and polysensitization has been hypothesized. OBJECTIVES: To examine associations of 21 allergens in the European baseline series to polysensitization....... PATIENTS/METHODS: From a database-based study with 14 998 patients patch tested with the European baseline series between 1985 and 2005, a group of 759 (5.1%) patients were polysensitized. Odds ratios were calculated to determine the relative contribution of each allergen to polysensitization. RESULTS...... denominator for the association between the allergens and the polysensitization was apparent, and any association, whether positive or negative, was relatively low. Based on these results, sensitization to specific baseline allergens cannot be used as risk indicators for polysensitization....

Food allergens and mucosal immune systems with special reference to recognition of food allergens by gut-associated lymphoid tissue

Directory of Open Access Journals (Sweden)

Shuichi Kaminogawa

1999-01-01

Full Text Available Food allergy, triggered by an aberrant immune response elicited by orally ingested food allergens, is generated through a complicated mechanism because the allergen interacts with the mucosal immune system (the gut- associated lymphoid tissue, GALT and the resulting immune response affects the generation of allergy. This review will describe the process by which antigens or allergens are recognized by the GALT and the characteristic immune responses induced thereafter. Orally administered antigens induce distinct immune responses in the Peyer's patches, lamina propria and the intestinal epithelium. In addition to these local immune responses in the gut, ingested antigens are known to affect systemic immunity. These may induce a suppressed state of systemic immune responsiveness, which is called oral tolerance, or in some cases they may elicit a systemic IgE antibody response which may lead to allergic reactions. Information on the regions on food allergens recognized by T cells and IgE antibodies is important in understanding the fates of food allergens after being recognized by the GALT. The structure of T and B cell epitopes on food allergens and the possibility of modulation of allergic reactions by amino-acid substituted analogs of allergen- derived peptides will also be discussed.

High correlation of specific IgE sensitization between birch pollen, soy and apple allergens indicates pollen-food allergy syndrome among birch pollen allergic patients in northern China.

Science.gov (United States)

Hao, Guo-Dong; Zheng, Yi-Wu; Wang, Zhi-Xiang; Kong, Xing-Ai; Song, Zhi-Jing; Lai, Xu-Xin; Spangfort, Michael D

2016-05-01

Birch pollen sensitization and associated pollen-food syndrome among Chinese allergic patients have not been investigated. Sera from 203 allergic patients from the northern part of China and collected during February to July 2014 were investigated. Specific immunoglobulin E (IgE) against birch pollen extract Bet v and major birch pollen allergen Bet v 1 were measured using the ADVIA Centaur. The presence of major apple allergen Mal d 1 and soy bean allergen Gly m 4 specific IgE was measured by ImmunoCAP 100. Among the 203 sera, 34 sera (16.7%) had specific IgE to Bet v and of these, 28 sera (82.4%) contained Bet v 1-specific IgE. Among the 28 sera with Bet v 1-specific IgE, 27 sera (96.4%) contained Mal d 1-specific IgE and 22 sera (78.6%) contained Gly m 4-specific IgE. Of the 34 Bet v-positive sera, 6 sera (17.6%) contained no specific IgE for Bet v 1, Mal d 1, or Gly m 4. Almost all Bet v-positive sera were donated during the birch pollen season. The prevalence of birch allergy among patients visiting health care during pollen season can be as high as 16.7% in Tangshan City. The majority of Chinese birch allergic patients are IgE-sensitized to the major birch pollen allergen Bet v 1 as well as to the major apple allergen Mal d 1 and soy bean allergen Gly m 4. A relatively high number of patients (17.6%) are IgE-sensitized to birch pollen allergen(s) other than Bet v 1. The high prevalence of specific IgE to Mal d 1 and Gly m 4 among Bet v 1-sensitized patients indicates that pollen-food allergy syndrome could be of clinical relevance in China.

Der p 11 Is a Major Allergen for House Dust Mite-Allergic Patients Suffering from Atopic Dermatitis

Czech Academy of Sciences Publication Activity Database

Banerjee, S.; Resch, Y.; Chen, K. W.; Swoboda, I.; Focke-Tejkl, M.; Blatt, K.; Novak, N.; Wickman, M.; van Hage, M.; Ferrara, R.; Mari, A.; Purohit, A.; Pauli, G.; Sibanda, E. N.; Ndlovu, P.; Thomas, W. R.; Krzyžánek, Vladislav; Tacke, S.; Malkus, U.; Valent, P.; Valenta, R.; Vrtala, S.

2015-01-01

Roč. 135, č. 1 (2015), s. 102-109 ISSN 0022-202X R&D Projects: GA MŠk(CZ) LO1212 Institutional support: RVO:68081731 Keywords : allergen * house dust mites * immunogold electron microscopy Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 6.915, year: 2015

Identification of autoclave-resistant Anisakis simplex allergens.

Science.gov (United States)

Carballeda-Sangiao, Noelia; Olivares, Fabiola; Rodriguez-Mahillo, Ana I; Careche, Mercedes; Tejada, Margarita; Moneo, Ignacio; González-Muñoz, Miguel

2014-04-01

Anisakis simplex is a fish parasite able to induce allergic reactions in humans infected when eating raw or undercooked fish parasitized with viable third-stage larvae. Some authors claim that exposure to nonviable Anisakis material can result in allergic symptoms in previously sensitized patients, indicating that parasite allergens are resistant to the thermal treatments of usual cooking procedures. Furthermore, some patients report symptoms after eating canned fish. The aim of this work was the analysis of parasite allergen stability in heating to 121 °C in an autoclave to simulate the thermal process applied to canned fish. Third-stage larvae were subjected to autoclaving for 20, 40, and 80 min, and parasite crude extracts were analyzed by electrophoresis, immunoblotting, and a flow-cytometric basophil activation test. Allergens resistant to autoclaving were separated by reversed-phase high-performance liquid chromatography and identified by ion trap mass spectrometry. Protein analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that autoclaving considerably reduced the number and intensity of identifiable protein bands in a time-dependent manner. Several allergens were detected by immunoblotting with a pool of A. simplex allergic patients' sera after autoclaving. Allergens of 9 and 14 kDa resistant to autoclaving were identified as Ani s 4 and Ani s 1 allergens, respectively. Functional analysis showed that allergens retain their capacity to activate basophils even after autoclaving for 80 min. In conclusion, some relevant A. simplex allergens retain their capacity to bind immunoglobulin E and activate basophils after being subjected to autoclaving, which is a method equivalent to that used in industrial canning processes.

The human allergens of mesquite (Prosopis juliflora

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Killian Sue

2004-07-01

Full Text Available Abstract Background A computerized statistical analysis of allergy skin test results correlating patient reactivities initiated our interest in the cross-reactive allergens of mesquite tree pollen. In-vitro testing with mesquite-sensitized rabbits and a variety of deciduous tree pollens revealed so many cross-reactivities that it became apparent there could be more allergens in mesquite than previously described in the world literature. Our purpose was to examine the allergens of mesquite tree pollen (Prosopis juliflora which elicit an IgE response in allergic humans so that future research could determine if these human allergens cross-react with various tree pollens in the same manner as did the mesquite antiserum from sensitized rabbits. Methods Proteins from commercial mesquite tree pollen were separated by polyacrylamide gel electrophoresis in the presence of sodium-dodecyl-sulphate. These mesquite proteins were subjected to Western blotting using pooled sera from ten mesquite-sensitive patients and goat anti-human IgE. The allergens were detected using an Amplified Opti-4-CN kit, scanned, and then interpreted by Gel-Pro software. Results Thirteen human allergens of mesquite pollen were detected in this study. Conclusion The number of allergens in this study of mesquite exceeded the number identified previously in the literature. With the increased exposure to mesquite through its use in "greening the desert", increased travel to desert areas and exposure to mesquite in cooking smoke, the possible clinical significance of these allergens and their suggested cross-reactivity with other tree pollens merit further study.

Influence of Thermal Treatment on the Characteristics of Major Oyster Allergen Cra g 1 (Tropomyosin).

Science.gov (United States)

Fang, Lei; Li, Guoming; Gu, Ruizeng; Cai, Muyi; Lu, Jun

2018-04-15

Shellfish, including oysters, often cause allergic reactions in adults. Thermal treatment is one of the most common technologies to deal with seafood, which may affect biological properties. The aim of this work was to evaluate the impact of heating on conformation and potential allergenicity of oyster-derived tropomyosin (Cra g 1). SDS-PAGE showed that there was an apparent band at 35KD of raw tropomyosin after purification and more significant polymers appeared in heated protein. Interestingly, the obviously changes in the intensity of CD signal and ANS-binding fluorescence were observed especially in the case of roasted form, which was associated with an increase in antibody reactivity. The degree of IgE binding of this treatment was demonstrated in the order roasted>boiled > raw. Furthermore, sequence alignment and amino acid composition revealed that Cra g 1 shared relatively high homology to tropomyosins from other shellfish and was abundant in lysine that was apt to be modified by reducing sugars during heating. Heated Cra g 1 produces higher IgE reactivity than raw one, owing to denaturation and formation of polymers. These findings will benefit more diagnosis and management of potential allergenicity due to shellfish. This article is protected by copyright. All rights reserved.

Allergens labeling on French processed foods - an Oqali study.

Science.gov (United States)

Battisti, Charlène; Chambefort, Amélie; Digaud, Olivier; Duplessis, Barbara; Perrin, Cécile; Volatier, Jean-Luc; Gauvreau-Béziat, Julie; Menard, Céline

2017-07-01

The French Observatory of Food Quality (Oqali) aims at collecting all nutritional data provided on labels of processed foods (nutritional information and composition), at branded products level, in order to follow nutritional labeling changes over time. This study carries out an overview of allergens labeling frequencies by distinguishing allergens used in recipes from those listed on precautionary statements, for the fourteen allergen categories for which labeling is mandatory according to European legislation. 17,309 products were collected, between 2008 and 2012, from 26 food categories. Products were classified per family and type of brand (national brands, retailer brands, entry-level retailer brands, hard discount, and specialized retailer brands). Allergenic ingredients were identified from ingredients lists and precautionary statements. 73% of the 17,309 products studied contained at least one allergen in their ingredients list and 39% had a precautionary statement for one or more allergens. Milk (53%), gluten (41%), and egg (22%) were the most commonly used allergens in ingredients lists. For precautionary statement, nuts (20%), egg (14%), peanut (13%), soybean (12%), and milk (11%) were the most common allergens listed. Precautionary statement was most frequently found among first-price products (hard discount and entry-level retailer brands). National brands seemed to use it less frequently. For all these results, differences depended both on food categories and allergen categories. This study will enable to follow allergens labeling and their use as ingredients over time, particularly by assessing an hypothetical increase in allergens presence in processed food.

Rubber elongation factor (REF, a major allergen component in Hevea brasiliensis latex has amyloid properties.

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Karine Berthelot

Full Text Available REF (Hevb1 and SRPP (Hevb3 are two major components of Hevea brasiliensis latex, well known for their allergenic properties. They are obviously taking part in the biosynthesis of natural rubber, but their exact function is still unclear. They could be involved in defense/stress mechanisms after tapping or directly acting on the isoprenoid biosynthetic pathway. The structure of these two proteins is still not described. In this work, it was discovered that REF has amyloid properties, contrary to SRPP. We investigated their structure by CD, TEM, ATR-FTIR and WAXS and neatly showed the presence of β-sheet organized aggregates for REF, whereas SRPP mainly fold as a helical protein. Both proteins are highly hydrophobic but differ in their interaction with lipid monolayers used to mimic the monomembrane surrounding the rubber particles. Ellipsometry experiments showed that REF seems to penetrate deeply into the monolayer and SRPP only binds to the lipid surface. These results could therefore clarify the role of these two paralogous proteins in latex production, either in the coagulation of natural rubber or in stress-related responses. To our knowledge, this is the first report of an amyloid formed from a plant protein. This suggests also the presence of functional amyloid in the plant kingdom.

Levels of house dust mite allergen in cars.

Science.gov (United States)

Mason, Howard J; Smith, Ian; Anua, Siti Marwanis; Tagiyeva, Nargiz; Semple, Sean; Devereux, Graham

2015-09-01

This small study investigated house dust mite (HDM) allergen levels in cars and their owners' homes in north-east Scotland. Dust samples from twelve households and cars were collected in a standardised manner. The dust samples were extracted and measured for the Dermatophagoides group 2 allergens (Der p 2 and Der f 2) and total soluble protein. Allergen levels at homes tended to be higher than in the cars, but not significantly. However, they significantly correlated with paired car dust samples expressed either per unit weight of dust or soluble protein (rho=0.657; p=0.02 and 0.769; p=0.003, respectively). This points to house-to-car allergen transfer, with the car allergen levels largely reflecting levels in the owner's home. Car HDM allergen levels were lower than those reported in Brazil and the USA. Twenty-five percent of the houses and none of the cars had allergen levels in dust greater than 2000 ng g(-1). This value is often quoted as a threshold for the risk of sensitisation, although a number of studies report increased risk of sensitisation at lower levels. This small study does not allow for characterisation of the distribution of HDM allergen in vehicles in this geographic area, or of the likely levels in other warmer and more humid areas of the UK. Cars and other vehicles are an under-investigated micro-environment for exposure to allergenic material.

Reduction and alkylation of peanut allergen isoforms Ara h 2 and Ara h 6; Characterization of intermediate- and end products

NARCIS (Netherlands)

Apostolovic, D.; Luykx, D.; Warmenhoven, H.; Verbart, D.; Stanic-Vucinic, D.; Jong, G.A.H. de; Velickovic, T.C.; Koppelman, S.J.

2013-01-01

Conglutins, the major peanut allergens, Ara h 2 and Ara h 6, are highly structured proteins stabilized by multiple disulfide bridges and are stable towards heat-denaturation and digestion. We sought a way to reduce their potent allergenicity in view of the development of immunotherapy for peanut

EAACI: A European Declaration on Immunotherapy. Designing the future of allergen specific immunotherapy

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Calderon Moises A

2012-10-01

Full Text Available Abstract Allergy today is a public health concern of pandemic proportions, affecting more than 150 million people in Europe alone. In view of epidemiological trends, the European Academy of Allergy and Clinical Immunology (EAACI predicts that within the next few decades, more than half of the European population may at some point in their lives experience some type of allergy. Not only do allergic patients suffer from a debilitating disease, with the potential for major impact on their quality of life, career progression, personal development and lifestyle choices, but they also constitute a significant burden on health economics and macroeconomics due to the days of lost productivity and underperformance. Given that allergy triggers, including urbanization, industrialization, pollution and climate change, are not expected to change in the foreseeable future, it is imperative that steps are taken to develop, strengthen and optimize preventive and treatment strategies. Allergen specific immunotherapy is the only currently available medical intervention that has the potential to affect the natural course of the disease. Years of basic science research, clinical trials, and systematic reviews and meta-analyses have convincingly shown that allergen specific immunotherapy can achieve substantial results for patients, improving the allergic individuals’ quality of life, reducing the long-term costs and burden of allergies, and changing the course of the disease. Allergen specific immunotherapy not only effectively alleviates allergy symptoms, but it has a long-term effect after conclusion of the treatment and can prevent the progression of allergic diseases. Unfortunately, allergen specific immunotherapy has not yet received adequate attention from European institutions, including research funding bodies, even though this could be a most rewarding field in terms of return on investments, translational value and European integration and, a field in

Identification and Characterization of a New Pecan [Carya illinoinensis (Wangenh.) K. Koch] Allergen, Car i 2.

Science.gov (United States)

Zhang, Yuzhu; Lee, BoRam; Du, Wen-Xian; Lyu, Shu-Chen; Nadeau, Kari C; Grauke, Larry J; Zhang, Yan; Wang, Shuo; Fan, Yuting; Yi, Jiang; McHugh, Tara H

2016-05-25

The 7S vicilin and 11S legumin seed storage globulins belong to the cupin protein superfamily and are major food allergens in many foods from the "big eight" food allergen groups. Here, for the first time, pecan vicilin was found to be a food allergen. Western blot experiments revealed that 30% of 27 sera used in this study and 24% of the sera from 25 patients with double-blind, placebo controlled clinical pecan allergy contained IgE antibodies specific to pecan vicilin. This allergen consists of a low-complexity region at its N-terminal and a structured domain at the C-terminal that contains two cupin motifs and forms homotrimers. The crystal structure of recombinant pecan vicilin was determined. The refined structure gave R/Rfree values of 0.218/0.262 for all data to 2.65 Å. There were two trimeric biological units in the crystallographic asymmetric unit. Pecan vicilin is also a copper protein. These data may facilitate the understanding of the nutritional value and the allergenicity relevance of the copper binding property of seed storage proteins in tree nuts.

Allergenicity of bony and cartilaginous fish - molecular and immunological properties.

Science.gov (United States)

Stephen, J N; Sharp, M F; Ruethers, T; Taki, A; Campbell, D E; Lopata, A L

2017-03-01

Allergy to bony fish is common and probably increasing world-wide. The major heat-stable pan-fish allergen, parvalbumin (PV), has been identified and characterized for numerous fish species. In contrast, there are very few reports of allergic reactions to cartilaginous fish despite widespread consumption. The molecular basis for this seemingly low clinical cross-reactivity between these two fish groups has not been elucidated. PV consists of two distinct protein lineages, α and β. The α-lineage of this protein is predominant in muscle tissue of cartilaginous fish (Chondrichthyes), while β-PV is abundant in muscle tissue of bony fish (Osteichthyes). The low incidence of allergic reactions to ingested rays and sharks is likely due to the lack of molecular similarity, resulting in reduced immunological cross-reactivity between the two PV lineages. Structurally and physiologically, both protein lineages are very similar; however, the amino acid homology is very low with 47-54%. Furthermore, PV from ancient fish species such as the coelacanth demonstrates 62% sequence homology to leopard shark α-PV and 70% to carp β-PV. This indicates the extent of conservation of the PV isoforms lineages across millennia. This review highlights prevalence data on fish allergy and sensitization to fish, and details the molecular diversity of the two protein lineages of the major fish allergen PV among different fish groups, emphasizing the immunological and clinical differences in allergenicity. © 2017 John Wiley & Sons Ltd.

Evolutionary distance from human homologs reflects allergenicity of animal food proteins.

Science.gov (United States)

Jenkins, John A; Breiteneder, Heimo; Mills, E N Clare

2007-12-01

In silico analysis of allergens can identify putative relationships among protein sequence, structure, and allergenic properties. Such systematic analysis reveals that most plant food allergens belong to a restricted number of protein superfamilies, with pollen allergens behaving similarly. We have investigated the structural relationships of animal food allergens and their evolutionary relatedness to human homologs to define how closely a protein must resemble a human counterpart to lose its allergenic potential. Profile-based sequence homology methods were used to classify animal food allergens into Pfam families, and in silico analyses of their evolutionary and structural relationships were performed. Animal food allergens could be classified into 3 main families--tropomyosins, EF-hand proteins, and caseins--along with 14 minor families each composed of 1 to 3 allergens. The evolutionary relationships of each of these allergen superfamilies showed that in general, proteins with a sequence identity to a human homolog above approximately 62% were rarely allergenic. Single substitutions in otherwise highly conserved regions containing IgE epitopes in EF-hand parvalbumins may modulate allergenicity. These data support the premise that certain protein structures are more allergenic than others. Contrasting with plant food allergens, animal allergens, such as the highly conserved tropomyosins, challenge the capability of the human immune system to discriminate between foreign and self-proteins. Such immune responses run close to becoming autoimmune responses. Exploiting the closeness between animal allergens and their human homologs in the development of recombinant allergens for immunotherapy will need to consider the potential for developing unanticipated autoimmune responses.

Lyral: a fragrance allergen.

Science.gov (United States)

Militello, Giuseppe; James, William

2005-03-01

Fragrances are a common cause of contact dermatitis and account for a large percentage of reactions to cosmetic products. Novel fragrance compounds that may not be detected by the common fragrance screening agents (including balsam of Peru and fragrance mix) are continually being produced. Lyral is one of those allergens found in many cosmetic and household products. This review will discuss the recent literature and the significance of this allergen to allergic contact dermatitis.

Allergen analysis of sea urchin roe using sera from five patients.

Science.gov (United States)

Tanaka, Kenichi; Kondo, Yasuto; Inuo, Chisato; Nakajima, Yoichi; Tsuge, Ikuya; Doi, Satoru; Yanagihara, Shigeto; Yoshikawa, Tetsushi; Urisu, Atsuo

2014-01-01

Sea urchin roe can cause anaphylactic reactions the first time they are consumed; therefore, careful clinical attention should be paid to their effects. However, no previous study has examined the allergens in sea urchin roe using sera from more than one patient. We attempted to identify sea urchin allergens using sera from 5 patients with sea urchin allergies. We enrolled 5 patients with relevant medical histories, positive results on a skin prick test and/or a food challenge test, and high levels of sea urchin-specific IgE in an enzyme-linked immunosorbent assay. We performed SDS-PAGE, immunoblotting, immunoblot inhibition, and N-terminal amino acid sequence detection. Ten protein bands ranging from 18 to 170 kDa were detected in more than 2 patients' sera. In immunoblotting, the protein band for the 170-kDa major yolk protein was recognized by 4 of the 5 sera. Furthermore, the reaction between IgE and the protein band for egg cortical vesicle protein (18 kDa) was inhibited by the addition of salmon roe extract. Major yolk protein was confirmed to be one of the main allergens in sea urchin roe. In addition, egg cortical vesicle protein (18 kDa) was demonstrated to be an important protein for cross-reactivity with salmon roe. © 2014 S. Karger AG, Basel.

Allergen and Epitope Targets of Mouse-Specific T Cell Responses in Allergy and Asthma

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Véronique Schulten

2018-02-01

Full Text Available Mouse allergy has become increasingly common, mainly affecting laboratory workers and inner-city households. To date, only one major allergen, namely Mus m 1, has been described. We sought to identify T cell targets in mouse allergic patients. PBMC from allergic donors were expanded with either murine urine or epithelial extract and subsequently screened for cytokine production (IL-5 and IFNγ in response to overlapping peptides spanning the entire Mus m 1 sequence, peptides from various Mus m 1 isoforms [major urinary proteins (MUPs], peptides from mouse orthologs of known allergens from other mammalian species and peptides from proteins identified by immunoproteomic analysis of IgE/IgG immunoblots of mouse urine and epithelial extracts. This approach let to the identification of 106 non-redundant T cell epitopes derived from 35 antigens. Three major T cell-activating regions were defined in Mus m 1 alone. Moreover, our data show that immunodominant epitopes were largely shared between Mus m 1 and other MUPs even from different species, suggesting that sequence conservation in different allergens is a determinant for immunodominance. We further identified several novel mouse T cell antigens based on their homology to known mammalian allergens. Analysis of cohort-specific T cell responses revealed that rhinitis and asthmatic patients recognized different epitope repertoires. Epitopes defined herein can be formulated into an epitope “megapool” used to diagnose mouse allergy and study mouse-specific T cell responses directly ex vivo. This analysis of T cell epitopes provides a good basis for future studies to increase our understanding of the immunopathology associated with MO-allergy and asthma.

Recombinant major urinary proteins of the mouse in specific IgE and IgG testing

NARCIS (Netherlands)

Krop, Esmeralda J. M.; Matsui, Elizabeth C.; Sharrow, Scott D.; Stone, Martin J.; Gerber, Peter; van der Zee, Jaring S.; Chapman, Martin D.; Aalberse, Rob C.

2007-01-01

BACKGROUND: Recombinant allergens are preferred over natural allergen extracts in measuring antibodies. We tested the use of recombinant variants of the major mouse allergen Mus m 1 in detection of mouse-specific antibodies in sera of laboratory animal workers and children. METHODS: Six recombinant

Tartary buckwheat improves cognition and memory function in an in vivo amyloid-β-induced Alzheimer model.

Science.gov (United States)

Choi, Ji Yeon; Cho, Eun Ju; Lee, Hae Song; Lee, Jeong Min; Yoon, Young-Ho; Lee, Sanghyun

2013-03-01

Protective effects of Tartary buckwheat (TB) and common buckwheat (CB) on amyloid beta (Aβ)-induced impairment of cognition and memory function were investigated in vivo in order to identify potential therapeutic agents against Alzheimer's disease (AD) and its associated progressive memory deficits, cognitive impairment, and personality changes. An in vivo mouse model of AD was created by injecting the brains of ICR mice with Aβ(25-35), a fragment of the full-length Aβ protein. Damage of mice recognition ability through following Aβ(25-35) brain injections was confirmed using the T-maze test, the object recognition test, and the Morris water maze test. Results of behavior tests in AD model showed that oral administration of the methanol (MeOH) extracts of TB and CB improved cognition and memory function following Aβ(25-35) injections. Furthermore, in groups receiving the MeOH extracts of TB and CB, lipid peroxidation was significantly inhibited, and nitric oxide levels in tissue, which are elevated by injection of Aβ(25-35), were also decrease. In particular, the MeOH extract of TB exerted a stronger protective activity than CB against Aβ(25-35)-induced memory and cognition impairment. The results indicate that TB may play a promising role in preventing or reversing memory and cognition loss associated with Aβ(25-35)-induced AD. Copyright © 2012 Elsevier Ltd. All rights reserved.

Authentication of food allergen quality by physicochemical and immunological methods

DEFF Research Database (Denmark)

Sancho, A I; Hoffmann-Sommergruber, K; Alessandri, S

2010-01-01

Purified allergens are required to detect cross-contamination with other allergenic foods and to understand allergen interaction with other components of the food matrix. Pure allergens are also used for the diagnosis and treatment of food allergies. For example, serological methods are being...... developed to improve the quality of diagnosis, and to reduce the need for food challenge tests. In addition, recombinant allergens are being evaluated as candidate vaccines for safe and efficacious specific immunotherapy. Pure allergens are indispensable as reference materials for the calibration...... and standardization of methods between different laboratories and operators for risk assessment in the food industry. Therefore, there is a need for well-defined purified food allergens. In this context, a panel of 46 food allergens from plant and animal sources has been purified, from either the food sources...

Effect of irradiation on biochemistry properties of shrimp allergen

International Nuclear Information System (INIS)

Gu Kefei; Gao Meixu; Li Chunhong; Li Shurong; Pan Jiarong

2007-01-01

Study on the effects of 60 Co γ-rays irradiation at the dose of 0,3,5,7,10 kGy on shrimp allergen biochemistry properties was conducted. The results indicated that the allergen protein molecule can be broken down to smaller molecules or coagulated to larger molecules by irradiation. The hydrophobicity and turbidity of irradiated allergen increased with the increase of absorbed dose. The results also show that allergen solution is more sensitive to irradiation than allergen in solid state or in the whole shrimp. (authors)

Dust Allergens within Rural Northern Rocky Mountain Residences.

Science.gov (United States)

Weiler, Emily; Semmens, Erin; Noonan, Curtis; Cady, Carol; Ward, Tony

2015-01-23

To date, few studies have characterized allergens within residences located in rural areas of the northern Rocky Mountain region. In this study, we collected dust samples from 57 homes located throughout western Montana and northern Idaho. Dust samples were collected and later analyzed for dust mite allergens Der f 1 and Der p 1 , Group 2 mite allergens ( Der p 2 and Der f 2 ), domestic feline ( Fel d 1 ), and canine ( Can f 1 ). Indoor temperature and humidity levels were also measured during the sampling program, as were basic characteristics of each home. Dog (96%) and cat (82%) allergens were the most prevalent allergens found in these homes (even when a feline or canine did not reside in the home). Results also revealed the presence of dust mites. Seven percent (7%) of homes tested positive for Der p 1 , 19% of homes were positive for Der f 1 , and 5% of homes were positive for the Group 2 mite allergens. Indoor relative humidity averaged 27.0 ± 7.6% within the homes. Overall, humidity was not significantly associated with dust mite presence, nor was any of the other measured home characteristics. This study provides a descriptive assessment of indoor allergen presence (including dust mites) in rural areas of the northern Rocky Mountains, and provides new information to assist regional patients with reducing allergen exposure using in-home intervention strategies.

Allergenic Proteins in Foods and Beverages

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Fernanda Cosme

2013-01-01

Full Text Available Food allergies can be defined as immunologically mediated hypersensitivity reactions; therefore, a food allergy is also known as food hypersensitivity. The reactions are caused by the immune system response to some food proteins. The eight most common food allergens are proteins from milk, eggs, peanuts, tree nuts, soya, wheat, fish and shellfish. However, many other foods have been identified as allergens for some people, such as certain fruits or vegetables and seeds. It is now recognized that food allergens are an important food safety issue. A food allergy occurs when the body’s immune system reacts to otherwise harmless substances in certain foods. For these reasons, one of the requirements from the European Union is that allergenic food ingredients should be labelled in order to protect allergic consumers. According to the European Federation of Allergy and Airways Diseases Patients’ Associations, about 8 % of children and 4 % of adults suffer from some type of food allergy. Food allergies often develop during infant or early childhood ages, affecting mainly the gastrointestinal tract (stomach and intestines. In some cases, the allergy may persist in adult age, for example, coeliac disease, which is an abnormal immune response to certain proteins present in gluten, a type of protein composite found in wheat and barley. Almost all allergens are proteins, and highly sensitive analytical methods have been developed to detect traces of these compounds in food, such as electrophoretic and immunological methods, enzyme-linked immunosorbent assay (ELISA and polyacrylamide gel electrophoresis. The purpose of this review is to describe the allergenic components of the most common causes of food allergies, followed by a brief discussion regarding their importance in the food industry and for consumer safety. The most important methods used to detect allergenicity in food will also be discussed.

Common indoor and outdoor aero-allergens in South Africa

African Journals Online (AJOL)

Aero-allergens in South Africa that are also encountered around the world are listed in Table I. In addition to this wide range of common aero-allergens, South Africans are also exposed to a full range of food allergens, some of which, e.g. perlemoen (Haliotis midae) and other seafood allergens, are unique to this region.

Detection of allergen composition and in vivo immunogenicity of depigmented allergoids of Betula alba.

Science.gov (United States)

Carnés, J; Himly, M; Gallego, M; Iraola, V; Robinson, D S; Fernández-Caldas, E; Briza, P

2009-03-01

Chemical modification of allergen vaccines to reduce IgE binding improves safety while maintaining clinical efficacy. However, this also complicates the characterization of allergoids using techniques as for native allergen extracts. The objective of this study was to analyse the molecular size of Betula alba depigmented allergoids, conservation of major allergens in the allergoids and in vivo antibody response to immunization. The molecular size of depigmented allergoids was evaluated by high performance-size exclusion chromatography and light scattering techniques. Protein composition was compared with native extracts by capillary liquid chromatography-tandem mass spectrometry based peptide mapping. Rabbits were immunized with depigmented allergoid of Betula pollen adsorbed onto aluminium hydroxide (Depigoid). IgG antibodies against individual allergens were determined by ELISA and immunoblot. Depigmented allergoids contained a range of high molecular weight particles, approximately 60% of which had a molecular weight of 1-3 MDa. Peptide sequencing confirmed the preservation of five isoforms of Bet v 1, as well as Bet v 2, Bet v 6 and Bet v 7. Sera from immunized rabbits showed high levels of specific IgG to rBet v 1.0101 and rBet v 2. The mean protein content was 544+/-106 microg per mg of freeze-dried material for depigmented allergoids and 434+/-71 for native extracts. They retain the capacity to induce specific IgG antibodies against individual allergens present in the native extract. These findings confirm the immunogenicity of depigmented allergoids and may explain why patients treated with these vaccines are protected against the native allergens. Analysis of molecular size and allergen content may be useful techniques for characterization and standardization of allergoid products.

Hierarchy and molecular properties of house dust mite allergens

Directory of Open Access Journals (Sweden)

Wayne R. Thomas

2015-10-01

Full Text Available The allergenic load of house dust mite allergy is largely constituted by a few proteins with a hierarchical pattern of allergenicity. The serodominant specificities are the group 1&2 and the group 23 faecal allergens. The collective IgE binding to the group 1&2 allergens can measure unequivocal HDM sensitisation better than HDM extracts although discrepancies have been found in regions with complex acarofauna suggesting a need to investigate the specificity with allergen components. The group 4, 5, 7&21 allergens that each induce responses in about 40% of subjects are mid-tier allergens accounting for most of the remaining IgE binding. Their titres are proportional to the concomitant responses to Der p1&2. Group 2 allergen variants have different antibody binding. Body proteins only occasionally induce sensitisation although a higher prevalence of binding by atopic dermatitis patients provides a new avenue of research. A broad spectrum of IgE binding has been associated with diverse symptoms but not with the severity of asthma which is associated with low IgG antibody. Some allergens such as the group 14 large lipid binding proteins and the recently described proteins Der f 24–33, need further investigation but with the cognoscence that other denominated allergens have been found to be minor sensitisers by comparative quantitative analyses. Scabies is a confounder for diagnosis with extracts, inducing cross-reactive antibodies with Der p 4&20 as is seafood allergy with cross reactivity to Der p 10 a minor HDM allergen. The HDM genome sequence can now be used to verify allelic and paralogous variations.

Acaroid mite allergens from the filters of air-conditioning system in China.

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Li, Chao-Pin; Guo, Wei; Zhan, Xiao-Dong; Zhao, Bei-Bei; Diao, Ji-Dong; Li, Na; He, Lian-Ping

2014-01-01

Accumulation of acaroid mites in the filters of air-conditioners is harmful to human health. It is important to clarify the allergen components of mites from the filters of local air-conditioning system. The present study was to detect the allergen types in the filters of air-conditioners and assesse their allergenicity by asthmatic models. Sixty aliquots of dust samples were collected from air conditioning filters in civil houses in Wuhu area. Total protein was extracted from the dust samples using PBS and quantified by Bradford method. Allergens I and II were also detected by Western blot using primary antibody (anti-Der f1/2, Der p1/Der f2/Der p2, respectively). Ten aliquots of the positive samples were randomly selected for homogenization and sensitized the mice for developing asthmatic animal models. Total serum IgE level and IFN-γ, IL-4 and IL-5 in the bronchoalveolar lavage fluid (BALF). The allergenicity of the extraction was assessed using pathological sections developed from the mouse pulmonary tissues. The concentration of extract from the 60 samples was ranged from 4.37 μg/ml to 30.76 μg/ml. After analyzing with Western blot, 31 of 60 samples were positive for 4 allergens of acaroid mites, and yet 16 were negative. The levels of total IgE from serum IL-4 and IL-5 from the BALF in the experimental group were apparently higher than that of negative control and PBS group (P 0.05). However,the IFN-γ level in BALF was lower compared with the negative control and PBS group (P 0.05). The pathological changes were evidently emerged in pulmonary tissues, which were similar to those of OVA group, compared with the PBS ground and negative controls. The air-conditioner filters in human dwellings of Wuhu area potentially contain the major group allergen 1 and 2 from D. farinae and D. pteronyssinus, which may be associated with seasonal prevalence of allergic disorders in this area.

Common ragweed (Ambrosia artemisiifolia L.): allergenicity and molecular characterization of pollen after plant exposure to elevated NO2.

Science.gov (United States)

Zhao, Feng; Elkelish, Amr; Durner, Jörg; Lindermayr, Christian; Winkler, J Barbro; Ruёff, Franziska; Behrendt, Heidrun; Traidl-Hoffmann, Claudia; Holzinger, Andreas; Kofler, Werner; Braun, Paula; von Toerne, Christine; Hauck, Stefanie M; Ernst, Dieter; Frank, Ulrike

2016-01-01

Ragweed pollen is the main cause of allergenic diseases in Northern America, and the weed has become a spreading neophyte in Europe. Climate change and air pollution are speculated to affect the allergenic potential of pollen. The objective of this study was to investigate the effects of NO2 , a major air pollutant, under controlled conditions, on the allergenicity of ragweed pollen. Ragweed was exposed to different levels of NO2 throughout the entire growing season, and its pollen further analysed. Spectroscopic analysis showed increased outer cell wall polymers and decreased amounts of pectin. Proteome studies using two-dimensional difference gel electrophoresis and liquid chromatography-tandem mass spectrometry indicated increased amounts of several Amb a 1 isoforms and of another allergen with great homology to enolase Hev b 9 from rubber tree. Analysis of protein S-nitrosylation identified nitrosylated proteins in pollen from both conditions, including Amb a 1 isoforms. However, elevated NO2 significantly enhanced the overall nitrosylation. Finally, we demonstrated increased overall pollen allergenicity by immunoblotting using ragweed antisera, showing a significantly higher allergenicity for Amb a 1. The data highlight a direct influence of elevated NO2 on the increased allergenicity of ragweed pollen and a direct correlation with an increased risk for human health. © 2015 John Wiley & Sons Ltd.

Airborne exposure to laboratory animal allergens

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Howard J Mason

2017-09-01

Full Text Available Exposure to laboratory animal allergens remains a significant cause of IgE-mediated occupational allergy and asthma. Since 2005, we have measured the major mouse and rat allergens (mus m 1 and rat n 1 collected on filters from air sampling in a range of UK and non-UK animal facilities. Supplied core data allowed us to construct an anonymized database of atmospheric results in ng m–3 containing 3080 mouse and 1392 rat analyses. Roughly twice as many static samples compared to personal samples had been sent for analysis. The medians (90th percentiles for the mouse and rat allergens employing personal atmospheric sampling were 2.6 (60.6 and 0.4 (12.4 ng m–3 respectively; for static samples the equivalent values were 0.2 (3.7 and 0.1 (1.4 ng m–3. Where unequivocal sample descriptors were provided with samples, results were categorised to activities/areas. Medians and 90th percentiles in these categories suggest that staff undertaking cleaning out, dumping of soiled bedding and cleaning cages can still have very substantial potential exposures in some facilities. The move to filtered cages appears to reduce general exposure, but filter changing and/or cleaning can lead to high exposures. In some facilities, animal receipt can cause significant exposures, as well as activities such as bleeding, culling and dosing; all activities involving the handling of animals outside of cages. We believe that the data presented may help those using air measurements in such facilities to improve their control of exposure to such aeroallergens, and thus reduce the risk of both sensitisation and subsequent allergic health problems, including the development of allergic asthma.

Reduction of allergenicity of irradiated ovalbumin in ovalbumin-allergic mice

International Nuclear Information System (INIS)

Seo, Ji-Hyun; Lee, Ju-Woon; Kim, Jae-Hun; Byun, Eui-Baek; Lee, Soo-Young; Kang, Il-Jun; Byun, Myung-Woo

2007-01-01

Egg allergy is one of the most serious of the immediate hypersensitivity reactions to foods. Such an allergic disorder is mediated by IgE antibodies stimulated by T-helper type 2 (Th2) lymphocytes. This study was undertaken to evaluate changes of allergenicity and cytokine profiles by exposure of irradiated ovalbumin (OVA), a major allergen of egg white, in the OVA-allergic mice model. OVA solutions (2 mg/ml in 0.01 M phosphate buffered saline (PBS) were gamma-irradiated to 50 and 100 kGy. The allergenicity in the OVA-allergy-induced mice model was remarkably reduced when challenged with irradiated OVA. Cultures of spleen cells harvested from OVA-sensitized mice showed a significant decrease in Th2 cytokine levels of ILs-4 and -5 with a concomitant increase in Th1 cytokine levels of IL-12 when co-cultured with irradiated OVA. However, IFN-γ level decreased dependant on the radiation dose of co-cultured OVA. The levels of IgEs and Th2-cytokine were reduced dependant on the radiation dose. These data show that the irradiated OVA could downregulate the activity of Th2 lymphocytes in OVA-sensitized mice

Authentication of food allergen quality by physicochemical and immunological methods

DEFF Research Database (Denmark)

Sancho, A I; Hoffmann-Sommergruber, K; Alessandri, S

2010-01-01

Purified allergens are required to detect cross-contamination with other allergenic foods and to understand allergen interaction with other components of the food matrix. Pure allergens are also used for the diagnosis and treatment of food allergies. For example, serological methods are being dev...

IgE sensitization to food allergens and airborne allergens in relation to biomarkers of type 2 inflammation in asthma.

Science.gov (United States)

Patelis, A; Alving, K; Middelveld, R; James, A; Ono, J; Ohta, S; Izuhara, K; Borres, M P; Forsberg, B; Janson, C; Malinovschi, A

2018-05-10

We have recently reported that sensitization to food allergens and sensitization to airborne allergens had independent associations with increased fraction of exhaled nitric oxide (FeNO) and blood eosinophils in middle-aged adults and in young subjects with asthma. To investigate the relation between IgE sensitization and several type 2 inflammation biomarkers in adult asthmatics. FeNO, urinary eosinophil-derived neurotoxin (U-EDN), serum eosinophil cationic protein (S-ECP) and periostin were measured in 396 asthmatics, aged 17-76 years, from the Swedish GA2LEN study. Sensitization to airborne allergens was examined with skin prick tests (≥3 mm wheal) and sensitization to food allergens with measurement of specific IgE (≥0.35 kU/L). Asthmatics sensitized to food allergens had higher FeNO, 22.3 ppb (18.6, 26.7) vs 16.1 ppb (14.2, 18.2) (P = .005), S-ECP, 17.7 mg/L (14.8, 21.1) vs 12.8 mg/L (10.9, 14.9) (P = .01), and periostin, 73.7 (67.5, 80.3) ng/mL vs 59.9 (55.8, 64.2) ng/mL (P = .003), than non-sensitized subjects. Periostin levels in this group were also significantly higher than in the group sensitized only to airborne allergens (P = .01). Sensitization to food allergens related independently to FeNO (P = .02), S-ECP (P = .006) and periostin (P = .004), whereas sensitization only to airborne allergens related only to FeNO (P = .02) after adjustments for age, sex, height, weight and smoking history. FeNO correlated weakly with S-ECP (r = .17, P < .001), periostin (r = .19, P < .001) and U-EDN (0.16, P < .001). S-ECP also correlated weakly with U-EDN (r = .12, P = .02). None of the correlations between the remaining pairs of markers of type 2 inflammation were significant. Sensitization to food allergens related to several local and systemic type 2 inflammation markers, such as FeNO, S-ECP and periostin. Assessing the profile of allergic sensitization, including to food allergens, might improve the understanding and

Digestion of peanut allergens Ara h 1, Ara h 2, Ara h 3, and Ara h 6: A comparative in vitro study and partial characterization of digestion-resistant peptides

NARCIS (Netherlands)

Koppelman, S.J.; Hefle, S.L.; Taylor, S.L.; Jong, G.A.H.de

2010-01-01

Scope: There are differences in stability to pepsin between the major allergens in peanut; however, data are from different reports using different digestion models. This study provides a comprehensive comparison of the digestibility of the major peanut allergens.Methods and results: Peanut

One-step multiplex PCR method for the determination of pecan and Brazil nut allergens in food products.

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Hubalkova, Zora; Rencova, Eva

2011-10-01

A one-step polymerase chain reaction (PCR) method for the simultaneous detection of the major allergens of pecan and Brazil nuts was developed. Primer pairs for the amplification of partial sequences of genes encoding the allergens were designed and tested for their specificity on a range of food components. The targeted amplicon size was 173 bp of Ber e 1 gene of Brazil nuts and 72 bp of vicilin-like seed storage protein gene in pecan nuts. The primer pair detecting the noncoding region of the chloroplast DNA was used as the internal control of amplification. The intrinsic detection limit of the PCR method was 100 pg mL(-1) pecan or Brazil nuts DNA. The practical detection limit was 0.1% w/w (1 g kg(-1)). The method was applied for the investigation of 63 samples with the declaration of pecans, Brazil nuts, other different nut species or nuts generally. In 15 food samples pecans and Brazil nuts allergens were identified in the conformity with the food declaration. The presented multiplex PCR method is specific enough and can be used as a fast approach for the detection of major allergens of pecan or Brazil nuts in food. Copyright © 2011 Society of Chemical Industry.

Survey of peanut levels in selected Irish food products bearing peanut allergen advisory labels.

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Robertson, Orla N; Hourihane, Jonathan O'B; Remington, Benjamin C; Baumert, Joseph L; Taylor, Steve L

2013-01-01

Peanut allergy affects up to 2% of consumers and is responsible for the majority of fatalities caused by food-induced anaphylaxis. Peanut-containing products must be clearly labelled. Manufacturers are not legally required to label peanut if its inclusion resulted from unintentional cross contact with foods manufactured in the same facility. However, the use of allergen advisory statements alerting consumers of the potential presence of peanut allergen has increased in recent years. In previous studies, the vast majority of foods with precautionary allergen statements did not contain detectable levels of peanut, but no data are available on Irish food products. Thirty-eight food products bearing peanut/nut allergen-related statements were purchased from multiple locations in the Republic of Ireland and analysed for the presence of peanut. Peanut was detected in at least one lot in 5.3% (2 of 38) of the products tested. The doses of peanut detected ranged from 0.14 mg to 0.52 mg per suggested serving size (0.035-0.13 mg peanut protein). No detectable levels of peanut were found in the products that indicated peanut/nuts as a minor ingredient. Quantitative risk assessment, based on the known distribution of individual threshold doses for peanut, indicates that only a very small percentage of the peanut-allergic population would be likely to experience an allergic reaction to those products while the majority of products with advisory labels appear safe for the peanut-allergic population. Food manufacturers should be encouraged to analyse products manufactured in shared facilities and even on shared equipment with peanuts for peanut residues to determine whether sufficient risk exists to warrant the use of advisory labelling. Although it appears that the majority of food products bearing advisory nut statements are in fact free of peanut contamination, advice to peanut allergy sufferers to avoid said foods should continue in Ireland and therefore in the wider European

Allergen immunotherapy for allergic respiratory diseases

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Cappella, Antonio; Durham, Stephen R.

2012-01-01

Allergen specific immunotherapy involves the repeated administration of allergen products in order to induce clinical and immunologic tolerance to the offending allergen. Immunotherapy is the only etiology-based treatment that has the potential for disease modification, as reflected by longterm remission following its discontinuation and possibly prevention of disease progression and onset of new allergic sensitizations. Whereas subcutaneous immunotherapy is of proven value in allergic rhinitis and asthma there is a risk of untoward side effects including rarely anaphylaxis. Recently the sublingual route has emerged as an effective and safer alternative. Whereas the efficacy of SLIT in seasonal allergy is now well-documented in adults and children, the available data for perennial allergies and asthma is less reliable and particularly lacking in children. This review evaluates the efficacy, safety and longterm benefits of SCIT and SLIT and highlights new findings regarding mechanisms, potential biomarkers and recent novel approaches for allergen immunotherapy. PMID:23095870

Comparison of immunoglobulin E measurements on IMMULITE and ImmunoCAP in samples consisting of allergen-specific mouse-human chimeric monoclonal antibodies towards allergen extracts and four recombinant allergens

DEFF Research Database (Denmark)

Szecsi, Pal B; Stender, Steen

2013-01-01

Specific immunoglobulin E (IgE) antibody in vitro tests are performed on enzyme immunoassay systems. Poor agreement among systems has been reported and comparisons have been made exclusively with allergen extracts - not with recombinant allergens. Here we compare the ImmunoCAP and the IMMULITE sy...

Evaluation of the potential allergenicity of the enzyme microbial transglutaminase using the 2001 FAO/WHO Decision Tree

DEFF Research Database (Denmark)

Pedersen, Mona H.; Hansen, Tine K.; Sten, Eva

2004-01-01

All novel proteins must be assessed for their potential allergenicity before they are introduced into the food market. One method to achieve this is the 2001 FAO/WHO Decision Tree recommended for evaluation of proteins from genetically modified organisms (GMOs). It was the aim of this study...... to investigate the allergenicity of microbial transglutaminase (m-TG) from Streptoverticillium mobaraense. Amino acid sequence similarity to known allergens, pepsin resistance, and detection of protein binding to specific serum immunoglobulin E (IgE) (RAST) have been evaluated as recommended by the decision tree...... meets the requirements of the decision tree. However, there is a match at the five contiguous amino acid level to the major codfish allergen Gad c1. The potential cross reactivity between m-TG and Gad c1 was investigated in RAST using sera from 25 documented cod-allergic patients and an extract of raw...

Effect of anti-IgE therapy on food allergen specific T cell responses in eosinophil associated gastrointestinal disorders

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Prussin Calman

2011-04-01

Full Text Available Abstract Background Anti-IgE therapy inhibits mast cell and basophil activation, blocks IgE binding to both FcεRI and CD23 and down regulates FcεRI expression by antigen (Ag presenting cells (APCs. In addition to its classical role in immediate hypersensitivity, IgE has been shown in vitro to facilitate Ag presentation of allergens, whereby APC bound IgE preferentially takes up allergens for subsequent processing and presentation. The purpose of this study was to determine whether anti-IgE therapy, by blocking facilitated Ag presentation in vivo, attenuates allergen specific Th2 cell responses. Methods To test this hypothesis, food allergen specific T cell responses were examined during a 16-week clinical trial of omalizumab in nine subjects with eosinophilic gastroenteritis and food sensitization. Allergen specific T cell responses were measured using carboxyfluorescein succinimidyl ester dye dilution coupled with intracellular cytokine staining and polychromatic flow cytometry. Four independent indices of allergen specific T cell response (proliferation, Ag dose response, precursor frequency, and the ratio of Th2:Th1 cytokine expression were determined. Results Eight of the 9 subjects had measurable food allergen specific responses, with a median proliferation index of 112-fold. Allergen specific T cell proliferation was limited to CD4 T cells, whereas CD8 T cell did not proliferate. Food allergen specific responses were Th2 skewed relative to tetanus specific responses in the same subjects. In contradistinction to the original hypothesis, anti-IgE treatment did not diminish any of the four measured indices of allergen specific T cell response. Conclusions In sum, using multiple indices of T cell function, this study failed to demonstrate that anti-IgE therapy broadly or potently inhibits allergen specific T cell responses. As such, these data do not support a major role for IgE facilitated Ag presentation augmenting allergen specific T cell

Down-regulation of the strawberry Bet v 1-homologous allergen in concert with the flavonoid biosynthesis pathway in colorless strawberry mutant

DEFF Research Database (Denmark)

Hjernø, Karin; Alm, Rikard; Canbäck, Björn

2006-01-01

Proteomic screening of strawberry (Fragaria ananassa) yielded a 58% success rate in protein identification in spite of the fact that no genomic sequence is available for this species. This was achieved by a combination of MALDI-MS/MS de novo sequencing of double-derivatized peptides and indel......-tolerant searching against local protein databases built on both EST and full-length nucleotide sequences. The amino acid sequence of a strawberry allergen, homologous to the well-known major birch pollen allergen Bet v 1, was partially determined. This strawberry allergen, named Fra a 1 according...... to the nomenclature for allergen proteins, showed sequence identity of 54 and 77%, respectively, with corresponding allergens from birch and apple. Differential expression, as evaluated by 2-D DIGE, occurred in 10% of protein spots when red strawberries were compared to a colorless (white) strawberry mutant. White...

The hazard communication of fragrance allergens must be improved.

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Klaschka, Ursula

2013-07-01

Contact allergy is a global health problem that could be alleviated considerably if the general public could reduce contact to sensitizers. Efficient hazard communication would be a valuable instrument to achieve this. What do current regulations concerning fragrance sensitizers in cosmetic products in Europe contribute? For example, there are bans and restrictions according to the Cosmetic Regulation, there is the "26 allergens rule" that requires that the names of some allergenic fragrance ingredients are listed on the containers, there is labeling and classification of hazardous products according to Regulation 1272/2008, and there is the regulation concerning the registration, evaluation, authorization and restriction of chemicals (REACH). Do these regulations increase consumer protection by suitable hazard communication instruments? Four main problems were identified. First, according to the 26 allergens rule, consumers carry a very large part of the responsibility for risk reduction management. They need to be capable and motivated to recognize the names of strong allergens listed in the ingredient list and decide for themselves whether they want to run the risk or not, provided that they are aware of their responsibility. Second, cosmetic products do not need to be classified and labeled like other consumer goods, according to the European Commission Regulation 1272/2008, if they contain hazardous substances. Third, some pictograms for hazardous substances, for example, the exclamation mark for sensitizers, are not well understood by the majority of the general public. Fourth, very often, the design of cosmetic containers implies health and well being, even if the respective products contain sensitizers or other hazardous substances. Against this background, the following improvements are proposed: 1) the 26 allergens rule needs revision, 2) the exception for cosmetic products from labeling and classification should be abolished, 3) a new self

Preparation of patient-related allergens for hyposensitization. Qualitative aspects

DEFF Research Database (Denmark)

Poulsen, L K; Søndergaard, I; Weeke, B

1988-01-01

An affinity chromatography method for preparation of patient-related antigens from commercially available allergen extracts has been investigated. IgG1,2,4 from a patient previously hyposensitized with dog hair and dandruff allergen was bound to protein A-sepharose. Secondly, commercial allergen ...

Overview of the most commonly used methods in allergen characterization

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TANJA CIRKOVIC VELICKOVIC

2005-03-01

Full Text Available The characterization of an allergen is a troublesome and difficult process, as it requires both the precise biochemical characterization of a (glycoprotein molecule and the establishment of its susceptibility to IgE antibodies, as they are themain link to histamine release in some hypersensitivity states (type I allergies. As the characterization of an allergen includes molecular weight determination of the allergenic molecule, its structure determination, physicochemical properties, IgE binding properties of the allergen molecule, and its allergenicity, an overal review of which biochemical and immunochemical methods are used in achieving this goal are presented in this paper. The information on the molecular level on the stuctures of allergens indicates that allergens are considerably heterogeneous protein structures, and that there is no particular aminoacid sequence which is responsible for the allergenicity. Therefore, information gained from detailed structural, functional and immunochemical studies of these intriguing molecules, which nowadaysmodulate a variety of pathophysiological conditions, would greatly improve our understanding of the underlying disease mechanisms, and the way to handle them.

Occupational allergens

NARCIS (Netherlands)

Terwoert, J.

2014-01-01

Allergens are substances that may cause a hypersensitivity (allergy) of the immune system. After acquiring this hypersensitivity, further exposure to the same substance may result in allergic skin disease such as allergic contact dermatitis, or allergic airway disease such as allergic rhinitis or

Oxidative Stress: Promoter of Allergic Sensitization to Protease Allergens?

NARCIS (Netherlands)

van Rijt, Leonie S.; Utsch, Lara; Lutter, René; van Ree, Ronald

2017-01-01

Allergies arise from aberrant T helper type 2 responses to allergens. Several respiratory allergens possess proteolytic activity, which has been recognized to act as an adjuvant for the development of a Th2 response. Allergen source-derived proteases can activate the protease-activated receptor-2,

Controlling allergens in animal rooms by using curtains

DEFF Research Database (Denmark)

Krohn, Thomas Cæcius; Itter, Gabi; Fosse, Richard

2006-01-01

The reduction and control of allergens in the animal facility is important for staff working with laboratory animals. This study was designed to evaluate the efficiency of perforated Makrolon curtains in front of racks as a method to reduce the amount of allergen in the animal room. The experimen......The reduction and control of allergens in the animal facility is important for staff working with laboratory animals. This study was designed to evaluate the efficiency of perforated Makrolon curtains in front of racks as a method to reduce the amount of allergen in the animal room....... The experimental situation we studied provides some information regarding allergen disposition in animal rooms but is clearly artificial and does not reflect a typical, ‘real-world’ environment in terms of preventing exposure of workers to allergens. Plastic curtains with holes were placed in front of racks......, and a corridor between the racks and a curtain was present. The room was ventilated with air, which was blown into the room through the middle of the corridor, flowing downstream and passing through the holes in the curtain. This set-up resulted in air flow from the corridor through the curtain. Air samples were...

Allergen-induced Increases in Sputum Levels of Group 2 Innate Lymphoid Cells in Subjects with Asthma.

Science.gov (United States)

Chen, Ruchong; Smith, Steven G; Salter, Brittany; El-Gammal, Amani; Oliveria, John Paul; Obminski, Caitlin; Watson, Rick; O'Byrne, Paul M; Gauvreau, Gail M; Sehmi, Roma

2017-09-15

Group 2 innate lymphoid cells (ILC2), a major source of type 2 cytokines, initiate eosinophilic inflammatory responses in murine models of asthma. To investigate the role of ILC2 in allergen-induced airway eosinophilic responses in subjects with atopy and asthma. Using a diluent-controlled allergen challenge crossover study, where all subjects (n = 10) developed allergen-induced early and late responses, airway eosinophilia, and increased methacholine airway responsiveness, bone marrow, blood, and sputum samples were collected before and after inhalation challenge. ILC2 (lin - FcεRI - CD45 + CD127 + ST2 + ) and CD4 + T lymphocytes were enumerated by flow cytometry, as well as intracellular IL-5 and IL-13 expression. Steroid sensitivity of ILC2 and CD4 + T cells was investigated in vitro. A significant increase in total, IL-5 + , IL-13 + , and CRTH2 + ILC2 was found in sputum, 24 hours after allergen, coincident with a significant decrease in blood ILC2. Total, IL-5 + , and IL-13 + , but not CRTH2 + , CD4 + T cells significantly increased at 24 and 48 hours after allergen in sputum. In blood and bone marrow, only CD4 + cells demonstrated increased activation after allergen. Airway eosinophilia correlated with IL-5 + ILC2 at all time points and allergen-induced changes in IL-5 + CD4 + cells at 48 hours after allergen. Dexamethasone significantly attenuated IL-2- and IL-33-stimulated IL-5 and IL-13 production by both cell types. Innate and adaptive immune cells are increased in the airways associated with allergic asthmatic responses. Total and type 2 cytokine-positive ILC2 are increased only within the airways, whereas CD4 + T lymphocytes demonstrated local and systemic increases. Steroid sensitivity of both cells may explain effectiveness of this therapy in those with mild asthma.

Crystallization and preliminary crystallographic study of the yeast Malassezia sympodialis allergen Mala s 1

International Nuclear Information System (INIS)

Vilhelmsson, Monica; Hallberg, B. Martin; Rasool, Omid; Zargari, Arezou; Scheynius, Annika; Achour, Adnane

2006-01-01

Crystals of the M. sympodialis allergen Mala s 1 have been obtained using the hanging-drop vapour-diffusion method. A diffraction data set has been collected from native crystals to 1.35 Å resolution. The opportunistic yeast Malassezia sympodialis can act as an allergen and elicit specific IgE- and T-cell reactivity in patients with atopic eczema. The first identified major allergen from M. sympodialis, Mala s 1, is present on the cell surface of the yeast. Recombinant Mala s 1 was expressed in Escherichia coli, purified and refolded in a soluble form. Crystals of Mala s 1 were obtained in 25% PEG 8K, 0.2 M (NH 4 ) 2 SO 4 . Crystals belong to space group P2 1 2 1 2, with unit-cell parameters a = 44.4, b = 163.7, c = 50.6 Å, and diffract to 1.35 Å resolution

Crystallization and preliminary crystallographic study of the yeast Malassezia sympodialis allergen Mala s 1

Energy Technology Data Exchange (ETDEWEB)

Vilhelmsson, Monica, E-mail: monica.vilhelmsson@medks.ki.se [Department of Medicine, Clinical Allergy Research Unit, Karolinska Institutet and Karolinska University Hospital, Stockholm (Sweden); Center for Infectious Medicine, Department of Medicine, Karolinska University Hospital, Huddinge, Stockholm (Sweden); Hallberg, B. Martin [Department of Biochemistry and Biophysics, Stockholm University, Stockholm (Sweden); Rasool, Omid; Zargari, Arezou; Scheynius, Annika [Department of Medicine, Clinical Allergy Research Unit, Karolinska Institutet and Karolinska University Hospital, Stockholm (Sweden); Achour, Adnane [Center for Infectious Medicine, Department of Medicine, Karolinska University Hospital, Huddinge, Stockholm (Sweden); Department of Medicine, Clinical Allergy Research Unit, Karolinska Institutet and Karolinska University Hospital, Stockholm (Sweden)

2006-02-01

Crystals of the M. sympodialis allergen Mala s 1 have been obtained using the hanging-drop vapour-diffusion method. A diffraction data set has been collected from native crystals to 1.35 Å resolution. The opportunistic yeast Malassezia sympodialis can act as an allergen and elicit specific IgE- and T-cell reactivity in patients with atopic eczema. The first identified major allergen from M. sympodialis, Mala s 1, is present on the cell surface of the yeast. Recombinant Mala s 1 was expressed in Escherichia coli, purified and refolded in a soluble form. Crystals of Mala s 1 were obtained in 25% PEG 8K, 0.2 M (NH{sub 4}){sub 2}SO{sub 4}. Crystals belong to space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 44.4, b = 163.7, c = 50.6 Å, and diffract to 1.35 Å resolution.

New Trends in Food Allergens Detection: Toward Biosensing Strategies.

Science.gov (United States)

Alves, Rita C; Barroso, M Fátima; González-García, María Begoña; Oliveira, M Beatriz P P; Delerue-Matos, Cristina

2016-10-25

Food allergens are a real threat to sensitized individuals. Although food labeling is crucial to provide information to consumers with food allergies, accidental exposure to allergenic proteins may result from undeclared allergenic substances by means of food adulteration, fraud or uncontrolled cross-contamination. Allergens detection in foodstuffs can be a very hard task, due to their presence usually in trace amounts, together with the natural interference of the matrix. Methods for allergens analysis can be mainly divided in two large groups: the immunological assays and the DNA-based ones. Mass spectrometry has also been used as a confirmatory tool. Recently, biosensors appeared as innovative, sensitive, selective, environmentally friendly, cheaper and fast techniques (especially when automated and/or miniaturized), able to effectively replace the classical methodologies. In this review, we present the advances in the field of food allergens detection toward the biosensing strategies and discuss the challenges and future perspectives of this technology.

Impact of allergen immunotherapy after two years of suspension in patients with asthma

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Jorge Sánchez

2016-05-01

Conclusion: The allergen immunotherapy improves asthma control and reduces the required doses of pharmacotherapy. These effects have an important impact on quality of life and perhaps economically for patients with asthma. The beginning at an early age seems to have a major impact.

Fish β-parvalbumin acquires allergenic properties by amyloid assembly.

Science.gov (United States)

Martínez, Javier; Sánchez, Rosa; Castellanos, Milagros; Fernández-Escamilla, Ana M; Vázquez-Cortés, Sonia; Fernández-Rivas, Montserrat; Gasset, María

2015-01-01

Amyloids are highly cross-β-sheet-rich aggregated states that confer protease resistance, membrane activity and multivalence properties to proteins, all essential features for the undesired preservation of food proteins transiting the gastrointestinal tract and causing type I allergy. Amyloid propensity of β-parvalbumin, the major fish allergen, was theoretically analysed and assayed under gastrointestinal-relevant conditions using the binding of thioflavin T, the formation of sodium dodecyl sulphate- (SDS-) resistant aggregates, circular dichroism spectroscopy and atomic force microscopy fibril imaging. Impact of amyloid aggregates on allergenicity was assessed with dot blot. Sequences of β-parvalbumin from species with commercial value contain several adhesive hexapeptides capable of driving amyloid formation. Using Atlantic cod β-parvalbumin (rGad m 1) displaying high IgE cross-reactivity, we found that formation of amyloid fibres under simulated gastrointestinal conditions accounts for the resistance to acid and neutral proteases, for the presence of membrane active species under gastrointestinal relevant conditions and for the IgE-recognition in the sera of allergic patients. Incorporation of the anti-amyloid compound epigallocatechin gallate prevents rGad m 1 fibrillation, facilitates its protease digestion and impairs its recognition by IgE. the formation of amyloid by rGad m 1 explains its degradation resistance, its facilitated passage across the intestinal epithelial barrier and its epitope architecture as allergen.

Modifications of allergenicity linked to food technologies.

Science.gov (United States)

Moneret-Vautrin, D A

1998-01-01

The prevalence of food allergies (FA) has increased over the past fifteen years. The reasons suggested are changes in dietary behaviour and the evolution of food technologies. New cases of FA have been described with chayote, rambutan, arguta, pumpkin seeds, custard apple, and with mycoproteins from Fusarium.... Additives using food proteins are at high risk: caseinates, lysozyme, cochineal red, papaïn, alpha-amylase, lactase etc. Heating can reduce allergenicity or create neo-allergens, as well as storage, inducing the synthesis of allergenic stress or PR proteins. Aeroallergens (miles, moulds) contaminate foods and can induce allergic reactions. Involuntary contamination by peanut proteins on production lines is a problem which is not yet solved. Genetically modified plants are at risk of allergenicity, requiring methodological steps of investigations: the comparison of the amino-acid sequence of the transferred protein with the sequence of known allergens, the evaluation of thermo degradability and of the denaturation by pepsin and trypsin are required, as well as the study with sera from patients allergic to the plant producing the gene. The combination of enzymatic hydrolysis, heating, or the development of genetically modified plants may offer new alternatives towards hypoallergenic foods (57 references).

Recombinant allergy vaccines based on allergen-derived B cell epitopes.

Science.gov (United States)

Valenta, Rudolf; Campana, Raffaela; Niederberger, Verena

2017-09-01

Immunoglobulin E (IgE)-associated allergy is the most common immunologically-mediated hypersensitivity disease. It affects more than 25% of the population. In IgE-sensitized subjects, allergen encounter can causes a variety of symptoms ranging from hayfever (allergic rhinoconjunctivitis) to asthma, skin inflammation, food allergy and severe life-threatening anaphylactic shock. Allergen-specific immunotherapy (AIT) is based on vaccination with the disease-causing allergens. AIT is an extremely effective, causative and disease-modifying treatment. However, administration of natural allergens can cause severe side effects and the quality of natural allergen extracts limits its application. Research in the field of molecular allergen characterization has allowed deciphering the molecular structures of the disease-causing allergens and it has become possible to engineer novel molecular allergy vaccines which precisely target the mechanisms of the allergic immune response and even appear suitable for prophylactic allergy vaccination. Here we discuss recombinant allergy vaccines which are based on allergen-derived B cell epitopes regarding their molecular and immunological properties and review the results obtained in clinical studies with this new type of allergy vaccines. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

Bla g 1 allergen levels in Zagreb area household dust.

Science.gov (United States)

Prester, Ljerka; Macan, Jelena

2011-03-01

Cockroach allergy is a health problem in many parts of the world. In urban environments, indoor exposure to cockroach allergens involves a risk of asthma. The aim of this study was to measure the mass fraction of Bla g 1, a major allergen of the German cockroach (Blatella germanica) in 30 house samples, collected at random from Zagreb area households, Croatia. Dust samples were collected on cellulose filters by vacuuming living rooms floors. After extraction, Bla g 1 was detected using the commercial enzyme-linked immunosorbent assay (ELISA). Only four of the thirty households had detectable Bla g 1 levels, and only in one was its concentration higher than 2.0 U g(-1), the threshold associated with sensitisation. The Bla g 1 ELISA proved highly sensitive, with the detection limit of 0.12 U g(-1). The within- and between-assay imprecision was 8.9 % and 14.4 %, respectively, and accuracy 85 % to 120 %. Low Bla g 1 levels in the household dust support previously reported low prevalence of skin sensitisation to B. germanica among Zagreb residents. Further monitoring should reveal if there are differences in cockroach allergen exposure and sensitisation between households from other geographic areas in Croatia.

Sensitising capacity of peptides from food allergens

DEFF Research Database (Denmark)

Bøgh, Katrine Lindholm

potential exist. Resistance to digestion is for this reason a test parameter included in the safety assessment of the allergenic potential of novel proteins in genetically modified foods. The association between resistance to digestion and allergenic potential has though been challenged in recent years...... and structures may contribute. In conclusion, the experimental data presented in this PhD thesis contribute to the understanding of induction of allergy by investigating the sensitising potential of peptides derived from a food allergen. It add knowledge to our understanding of the mechanisms underlying...

Common Allergens Identified Based on Patch Test Results in Patients with Suspected Contact Dermatitis of the Scalp.

Science.gov (United States)

Aleid, Nouf M; Fertig, Raymond; Maddy, Austin; Tosti, Antonella

2017-03-01

Contact dermatitis of the scalp is common and might be caused by many chemicals including metals, ingredients of shampoos and conditioners, dyes, or other hair treatments. Eliciting a careful history and patch tests are necessary to identify the responsible allergen and prevent relapses. To identify allergens that may cause contact dermatitis of the scalp by reviewing patch test results. We reviewed the records of 1,015 patients referred for patch testing at the Dermatology Department of the University of Miami. A total of 226 patients (205 females and 21 males) with suspected scalp contact dermatitis were identified, and the patch test results and clinical data for those patients were analyzed. Most patients were referred for patch testing from a specialized hair clinic at our institution. The most common allergens in our study population were nickel (23.8%), cobalt (21.0%), balsam of Peru (18.2%), fragrance mix (14.4%), carba mix (11.6%), and propylene glycol (PG) (8.8%). The majority of patients were females aged 40-59 years, and scalp itching or burning were reported as the most common symptom. Frequent sources of allergens for metals include hair clasps, pins, and brushes, while frequent sources of allergens for preservatives, fragrance mix, and balsam of Peru include shampoos, conditioners, and hair gels. Frequent sources of allergens for PG include topical medications.

Common Allergens Identified Based on Patch Test Results in Patients with Suspected Contact Dermatitis of the Scalp

Science.gov (United States)

Aleid, Nouf M.; Fertig, Raymond; Maddy, Austin; Tosti, Antonella

2017-01-01

Background Contact dermatitis of the scalp is common and might be caused by many chemicals including metals, ingredients of shampoos and conditioners, dyes, or other hair treatments. Eliciting a careful history and patch tests are necessary to identify the responsible allergen and prevent relapses. Objectives To identify allergens that may cause contact dermatitis of the scalp by reviewing patch test results. Methods We reviewed the records of 1,015 patients referred for patch testing at the Dermatology Department of the University of Miami. A total of 226 patients (205 females and 21 males) with suspected scalp contact dermatitis were identified, and the patch test results and clinical data for those patients were analyzed. Most patients were referred for patch testing from a specialized hair clinic at our institution. Results The most common allergens in our study population were nickel (23.8%), cobalt (21.0%), balsam of Peru (18.2%), fragrance mix (14.4%), carba mix (11.6%), and propylene glycol (PG) (8.8%). The majority of patients were females aged 40–59 years, and scalp itching or burning were reported as the most common symptom. Conclusion Frequent sources of allergens for metals include hair clasps, pins, and brushes, while frequent sources of allergens for preservatives, fragrance mix, and balsam of Peru include shampoos, conditioners, and hair gels. Frequent sources of allergens for PG include topical medications. PMID:28611994

Measurement of endogenous allergens in genetically modified soybeans--short communication.

Science.gov (United States)

Ladics, Gregory S; Budziszewski, Gregory J; Herman, Rod A; Herouet-Guicheney, Corinne; Joshi, Saurabh; Lipscomb, Elizabeth A; McClain, Scott; Ward, Jason M

2014-10-01

The measurement of endogenous allergens is required by the European Commission (EC) as part of the compositional analysis for GM products from host plants that are common causes of food allergy, such as soybean (EC Implementing Regulation No. 503/2013). In each case, the EC Implementing Regulation indicates that analysis be conducted on identified allergens as specified in the Organization of Economic Cooperation and Development (OECD) consensus documents on compositional considerations for new plant varieties. This communication discusses the methods available to measure endogenous allergens as well as the endogenous soybean allergens that should be analyzed. It is suggested herein that in conjunction with the 2012 OECD consensus document on soybean, any list of soybean allergens should be based on clinically relevant data among publicly available allergen databases and peer-reviewed scientific publications, and the ability to measure the identified allergen. Based on a detailed analysis of the scientific literature, the following key points are recommended: (1) the acceptance of serum-free, quantitative analytical method data as an alternative to traditional IgE reactivity qualitative or semi-quantitative data for evaluation of endogenous soybean allergen content; (2) eight of the 15 potential allergens listed in the OECD soybean consensus document (Gly m 3, Gly m 4, Gly m Bd28K, Gly m Bd30K, Gly m 5, Gly m 6, Gly m 8, and Kunitz trypsin inhibitor) have both appropriate supporting clinical data and sufficient sequence information to be evaluated in comparative endogenous soybean allergen studies; and (3) the remaining seven proteins (Gly m 1, Gly m 2, unknown 50kDa protein, unknown 39kDa protein, P-22-25, lipoxygenase and lectin) lack sufficient data for clear classification as confirmed allergens and/or available sequence information and should not be currently included in the measurement of endogenous soybean allergens in the compositional analysis for the EU

Cloning, expression, and immunological characterization of recombinant Lolium perenne allergen Lol p II.

Science.gov (United States)

Sidoli, A; Tamborini, E; Giuntini, I; Levi, S; Volonté, G; Paini, C; De Lalla, C; Siccardi, A G; Baralle, F E; Galliani, S

1993-10-15

The molecular cloning of the cDNA encoding for an isoallergenic form of Lol p II, a major rye grass (Lolium perenne) pollen allergen, was performed by polymerase chain reaction amplification on mRNA extracted from pollen. The amino acid sequence derived from the cDNA was truncated by 4 and 5 residues at the NH2- and COOH-terminal ends, respectively, and differed only in one position from that previously reported. This cDNA was expressed in Escherichia coli by fusion to the carboxyl terminus of the human ferritin H-chain. The molecule was produced in high yields as a soluble protein and was easily purified. The protein retains the multimeric quaternary structure of ferritin, and it exposes on the surface the allergenic moiety, which can be recognized in Western blotting and in enzyme-linked immunosorbent assay experiments by specific IgE from allergic patients. The recombinant allergen was used to analyze the sera of 26 patients allergic to L. perenne compared with control sera. The results were in good agreement with the values obtained with the radioallergosorbent test assay. In addition, histamine release experiments in whole blood from an allergic patient and skin prick tests showed that the recombinant allergen retains some of the biological properties of the natural compound. These findings indicate that the availability of homogeneous recombinant allergens may be useful for the development of more specific diagnostic and therapeutic procedures. Moreover, this expression system may be of more general interest for producing large amounts of soluble protein domains in E. coli.

IgE Sensitization Patterns to Commonly Consumed Foods Determined by Skin Prick Test in Korean Adults.

Science.gov (United States)

Kim, Sung Ryeol; Park, Hye Jung; Park, Kyung Hee; Lee, Jae-Hyun; Park, Jung-Won

2016-08-01

Offending food allergens can vary with regional preferences in food consumption. In this study, we analysed sensitization rates to commonly consumed foods in Korean adults suspected of having food allergy. One hundred and thirty four subjects underwent a skin prick test (SPT) with 55 food allergens, of which 13 were made by our laboratory and the rest were commercially purchased. Of the 134 patients, 73 (54.5%) were sensitized to one or more food allergens. Sensitization to chrysalis was detected most frequently, at a rate of 25.4%. Sensitization rates to other food allergens were as follows: maize grain (13.4%), shrimp (11.9%), almond (11.1%), wheat flour (8.2%), lobster (8.2%), buckwheat (8.2%), mackerel (5.2%), pollack (5.2%), halibut (4.5%), peanut (4.5%), anchovy (4.4%), squid (3.7%), saury (3.0%), common eel (3.0%), yellow corvina (3.0%), hairtail (2.2%), octopus (2.2%), and others. In addition to well-known food allergens, sensitivity to mackerel, chrysalis, pollack, and halibut, which are popular foods in Korea, was observed at high rates in Korean adults. We suggest that the SPT panel for food allergy in Korea should include these allergens.

IgE Sensitization Patterns to Commonly Consumed Foods Determined by Skin Prick Test in Korean Adults

Science.gov (United States)

2016-01-01

Offending food allergens can vary with regional preferences in food consumption. In this study, we analysed sensitization rates to commonly consumed foods in Korean adults suspected of having food allergy. One hundred and thirty four subjects underwent a skin prick test (SPT) with 55 food allergens, of which 13 were made by our laboratory and the rest were commercially purchased. Of the 134 patients, 73 (54.5%) were sensitized to one or more food allergens. Sensitization to chrysalis was detected most frequently, at a rate of 25.4%. Sensitization rates to other food allergens were as follows: maize grain (13.4%), shrimp (11.9%), almond (11.1%), wheat flour (8.2%), lobster (8.2%), buckwheat (8.2%), mackerel (5.2%), pollack (5.2%), halibut (4.5%), peanut (4.5%), anchovy (4.4%), squid (3.7%), saury (3.0%), common eel (3.0%), yellow corvina (3.0%), hairtail (2.2%), octopus (2.2%), and others. In addition to well-known food allergens, sensitivity to mackerel, chrysalis, pollack, and halibut, which are popular foods in Korea, was observed at high rates in Korean adults. We suggest that the SPT panel for food allergy in Korea should include these allergens. PMID:27478328

Structural analysis of linear and conformational epitopes of allergens

Science.gov (United States)

Ivanciuc, Ovidiu; Schein, Catherine H.; Garcia, Tzintzuni; Oezguen, Numan; Negi, Surendra S.; Braun, Werner

2009-01-01

In many countries regulatory agencies have adopted safety guidelines, based on bioinformatics rules from the WHO/FAO and EFSA recommendations, to prevent potentially allergenic novel foods or agricultural products from reaching consumers. We created the Structural Database of Allergenic Proteins (SDAP, http://fermi.utmb.edu/SDAP/) to combine data that had previously been available only as flat files on Web pages or in the literature. SDAP was designed to be user friendly, to be of maximum use to regulatory agencies, clinicians, as well as to scientists interested in assessing the potential allergenic risk of a protein. We developed methods, unique to SDAP, to compare the physicochemical properties of discrete areas of allergenic proteins to known IgE epitopes. We developed a new similarity measure, the property distance (PD) value that can be used to detect related segments in allergens with clinical observed crossreactivity. We have now expanded this work to obtain experimental validation of the PD index as a quantitative predictor of IgE cross-reactivity, by designing peptide variants with predetermined PD scores relative to known IgE epitopes. In complementary work we show how sequence motifs characteristic of allergenic proteins in protein families can be used as fingerprints for allergenicity. PMID:19121639

Purification and characterization of allergens from Xanthium strumarium pollen.

Science.gov (United States)

Jaggi, K S; Gangal, S V

1987-12-01

The allergenic components present in whole pollen extract of Xanthium strumarium were isolated by sequential ammonium sulphate precipitation, DEAE Sephadex A50 chromatography and gel filtration. The techniques of RAST inhibition and skin test were utilized to check the allergenicity of fractionated proteins revealing the presence of Xan Ib and Xan VIa as the important allergenic components. Xan Ib was found to be devoid of carbohydrate and had a molecular weight of 103,000 daltons. Xan VIa was a glycoprotein of molecular weight 17,000 daltons. The carbohydrate moiety of Xan VIa was found to be associated with allergenicity. The characteristic pattern of whole pollen extract on CIE and TLIEF showed 36 and 21 protein bands, respectively. The use of FPLC in isolation of partially purified allergens from Xanthium is discussed.

Cloning and characterization of 2S albumin, Car i 1, a major allergen in pecan.

Science.gov (United States)

Sharma, Girdhari M; Irsigler, Andre; Dhanarajan, Pushparani; Ayuso, Rosalia; Bardina, Luda; Sampson, Hugh A; Roux, Kenneth H; Sathe, Shridhar K

2011-04-27

Although pecans are associated with IgE-mediated food allergies, the allergens responsible remain to be identified and characterized. The 2S albumin gene was amplified from the pecan cDNA library. Dot-blots were used to screen the recombinant protein with pecan allergic patients' serum. The affinity purified native protein was analyzed by Edman sequencing and mass spectrometry/mass spectrometry (MS/MS) analysis. Cross-reactivity with walnut was determined by inhibition enzyme-linked immunosorbent assay (ELISA). Sequential epitopes were determined by probing the overlapping peptides with three different patients' serum pool. The 3-dimensional homology model was generated, and the locations of the pecan epitopes were compared with those of known sequential epitopes on other allergenic tree nut homologues. Of 28 patients tested by dot-blot, 22 (79%) bound to 2S albumin, designated as Car i 1. Edman sequencing and the MS/MS sequencing of native 2S albumin confirmed the identity of recombinant (r) Car i 1. Both pecan and walnut protein extracts inhibited the IgE-binding to rCar i 1. Sequential epitope mapping indicated weak, moderate, and strong reactivity against 12, 7, and 5 peptides, respectively. Of the 11 peptides recognized by all serum pools, 5 peptides were strongly reactive and located in 3 discrete regions of the Car i 1 (amino acids 43-57, 67-78, and 106-120). Three-dimensional modeling revealed IgE-reactive epitopes to be solvent accessible and share significant homology with other tree nuts providing a possible basis for previously observed cross-reactivity.

Molecular features of grass allergens and development of biotechnological approaches for allergy prevention.

Science.gov (United States)

Devis, Deborah L; Davies, Janet M; Zhang, Dabing

2017-09-01

Allergic diseases are characterized by elevated allergen-specific IgE and excessive inflammatory cell responses. Among the reported plant allergens, grass pollen and grain allergens, derived from agriculturally important members of the Poaceae family such as rice, wheat and barley, are the most dominant and difficult to prevent. Although many allergen homologs have been predicted from species such as wheat and timothy grass, fundamental aspects such as the evolution and function of plant pollen allergens remain largely unclear. With the development of genetic engineering and genomics, more primary sequences, functions and structures of plant allergens have been uncovered, and molecular component-based allergen-specific immunotherapies are being developed. In this review, we aim to provide an update on (i) the distribution and importance of pollen and grain allergens of the Poaceae family, (ii) the origin and evolution, and functional aspects of plant pollen allergens, (iii) developments of allergen-specific immunotherapy for pollen allergy using biotechnology and (iv) development of less allergenic plants using gene engineering techniques. We also discuss future trends in revealing fundamental aspects of grass pollen allergens and possible biotechnological approaches to reduce the amount of pollen allergens in grasses. Copyright © 2017. Published by Elsevier Inc.

Spreading of occupational allergens: laboratory animal allergens on hair-covering caps and in mattress dust of laboratory animal workers

NARCIS (Netherlands)

Krop, Esmeralda J. M.; Doekes, Gert; Stone, Martin J.; Aalberse, Rob C.; van der Zee, Jaring S.

2007-01-01

BACKGROUND: Family members of laboratory animal workers are at risk of developing allergy to laboratory animals. Little is known about the spreading of laboratory animal allergens outside the animal facilities. OBJECTIVE: To assess the presence of laboratory animal allergens in dust collected from

Predicting allergenicity of proteins using Physical–Chemical Property (PCP) motifs

Science.gov (United States)

Motivation: Quantitative guidelines to distinguish allergenic proteins from related, but non-allergenic ones are urgently needed for regulatory agencies, biotech companies and physicians. Cataloguing the SDAP database has indicated that allergenic proteins populate a relatively small number of prote...

Cosmetic Contact Allergens

Directory of Open Access Journals (Sweden)

An Goossens

2016-02-01

Full Text Available This article presents trends in the frequency of cosmetics as causal factors of allergic contact dermatitis during a 26-year period in 14,911 patients patch-tested between 1990 and 2014, and discusses the cosmetic allergens identified during the last six years (2010–2015 in 603 patients out of 3105 tested. The data were retrieved from, and evaluated with, a patient database developed in-house. The results show the increasing importance of cosmetic allergies, up to 25% of the patients tested during the last five-year period. As expected, fragrance materials, preservatives, and hair dyes were the most frequent culprits, but a great variety of other allergenic ingredients were involved as well. This underlines the need of additional and extensive patch testing with the patient’s products used and their ingredients.

Gal d 6 is the second allergen characterized from egg yolk.

Science.gov (United States)

Amo, Alvaro; Rodríguez-Pérez, Rosa; Blanco, Juan; Villota, Julian; Juste, Sonsoles; Moneo, Ignacio; Caballero, María Luisa

2010-06-23

Only one allergen from the egg yolk, alpha-livetin (Gal d 5) has been described thus far. A new egg yolk allergen was detected studying 27 egg allergic patients. The study was performed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and IgE-immunoblotting and IgE-immunoblotting-inhibition assays. An egg yolk extract was fractioned by reverse-phase high-performance liquid chromatography (RP-HPLC), and the new allergen detected was characterized by N-terminal amino acid analysis. A total of 5 of the 27 patients (18%) detected a yolk allergen of an apparent molecular weight of 35 kDa by SDS-PAGE. Heating and reduction treatments did not affect its allergenicity, although digestion with simulated gastric fluid diminished the IgE-binding capacity of the allergen. The N-terminal amino acid sequence corresponded with the YGP42 protein, a fragment of the vitellogenin-1 precursor. Thus, a second egg yolk allergen has been described and designated Gal d 6 by the World Health Organization (WHO)/International Union of Immunological Societies (IUIS) Allergen Nomenclature Subcommittee.

Food Production and Processing Considerations of Allergenic Food Ingredients: A Review

Science.gov (United States)

Alvarez, Pedro A.; Boye, Joyce I.

2012-01-01

Although most consumers show no adverse symptoms to food allergens, health consequences for sensitized individuals can be very serious. As a result, the Codex General Standard for the Labelling of Prepackaged Foods has specified a series of allergenic ingredients/substances requiring mandatory declaration when present in processed prepackaged food products. Countries adhering to international standards are required to observe this minimum of eight substances, but additional priority allergens are included in the list in some countries. Enforcement agencies have traditionally focused their effort on surveillance of prepackaged goods, but there is a growing need to apply a bottom-up approach to allergen risk management in food manufacturing starting from primary food processing operations in order to minimize the possibility of allergen contamination in finished products. The present paper aims to review food production considerations that impact allergen risk management, and it is directed mainly to food manufacturers and policy makers. Furthermore, a series of food ingredients and the allergenic fractions identified from them, as well as the current methodology used for detection of these allergenic foods, is provided. PMID:22187573

Modulation of the allergen-induced human IgE response in Hu-SCID mice: inhibitory effect of human recombinant IFN-gamma and allergen-derived lipopeptide.

Science.gov (United States)

Duez, C; Gras-Masse, H; Hammad, H; Akoum, H; Didierlaurent, A; André, C; Tonnel, A B; Pestel, J

2001-01-01

We have previously established a model to study the in vivo human IgE response using humanized SCID mice. Allergic SCID mice were obtained following intraperitoneal injection with mononuclear cells from Dermatophagoides pteronyssinus (Dpt)-sensitive patients, and sensitization by Dpt allergen intraperitoneal injection (immunization) or Dpt aerosol (inhalation). Human serum IgE was measured in allergic SCID mice after administration of human recombinant IFN-gamma or the lipopeptide LP 52-71 (derived from peptide p52-71 from Der p 1, Dpt major allergen, coupled to a lipophilic moiety), during the immunization or the inhalation phase. IFN-gamma inhibited human IgE production when given at the time of immunization, but not during inhalation. This effect was long-lasting as Dpt aerosol, given one month after immunization and IFN-gamma administration, failed to increase IgE levels. Unlike Dpt or p52-71, LP 52-71 failed to induce human IgE production at day 14 and 21 after its injection, but did inhibit the development of the IgE response after a secondary Dpt-challenge. Moreover, LP 52-71 administration 14 days after Dpt inhalation decreased IgE levels, in contrast to peptide 52-71, which increased IgE levels. Thus, taken together these results indicate that the development of the human IgE response in allergic SCID mice can be modulated by modified allergen and a Th1 cytokine.

Proficiency test for allergens in food 2014

NARCIS (Netherlands)

Bremer, M.G.E.G.; Alamenou, P.; Elbers, I.J.W.

2015-01-01

In the autumn of 2014 a proficiency test for allergens in baby cereal was organized by RIKILT, Wageningen UR. This PT-test enabled laboratories to evaluate their competence for the analysis of allergens in baby cereal. Both quantitative and qualitative methods were accepted. The proficiency test was

[Specific immunotherapy. Hyposensitization with allergens].

Science.gov (United States)

Wedi, B; Kapp, A

2004-04-01

Successful allergen-specific immunotherapy (SIT) induces complex immunologic chan-ges resulting in reduced allergic inflammatory reactions. SIT has long-term effects in mild forms of inhalant allergies and is effective even when standard pharmacotherapy fails. Moreover, the risk to develop additional allergic sensitizations and the development of asthma is significantly reduced in children with allergic rhinitis. SIT is the treatment of choice in patients with systemic reactions to hymenoptera venoms. Although the exact effector mechanisms of SIT still have to be clarified, the most probable effect is a modulation of regulatory T cells associated with a switch of allergen-specific B-cells towards IgG4 production. The critical point to insure efficacy and safety is the selection of patients and allergens, task best performed by a specialist trained in allergology. Further details are available in the position papers of the German allergy societies - DGAI(Deutsche Gesellschaft fiir Allergologie und Klinische Immunologie) and ADA (Arzte-verband Deutscher Allergologen) - which can be found at www.dgaki.de.

Degradation and removal of soybean allergen in Japanese soy sauce.

Science.gov (United States)

Magishi, Norihiro; Yuikawa, Naoya; Kobayashi, Makio; Taniuchi, Shoichiro

2017-08-01

Soy sauce is a traditional fermented seasoning of Japan and is available throughout the world. The two main raw ingredients of soy sauce are soybean and wheat, both of which are established food allergens. The present study examined the degradation and removal of soybean allergens in soy sauce by immunoblotting with anti‑soybean protein antibody from rabbit and sera from two children with soybean allergy. It was demonstrated that soybean allergens were gradually degraded during the fermentation process, but were not completely degraded in raw soy sauce. During the processes of heat‑treatment and filtration, the soluble soybean allergens in raw soy sauce were denatured to insoluble allergens by heat‑treatment and subsequently completely removed from soy sauce by filtration. Therefore, to reduce the allergenicity of soy sauce, heat‑treatment and filtration are very important processes in addition to the enzymatic degradation during the fermentation of soy sauce.

Nonadverse effects on allergenicity of isopentenyltransferase-transformed broccoli.

Science.gov (United States)

Liao, E C; Chen, J T; Chao, M L; Yu, S C; Chang, C Y; Chu, W S; Tsai, J J

2013-01-01

Genetically modified organisms (GMOs) provide modern agriculture with improvements in efficiency and the benefits of enhanced food production; however, the potential impact of GMOs on human health has not yet been clarified. To investigate the allergenicity of isopentenyltransferase (ipt)-transformed broccoli compared with non-GM broccoli. Sera from allergic individuals were used to identify the allergenicity of GM and non-GM broccoli. Immunoglobulin (Ig) binding of different lines of GM and non-GM broccoli was identified using immunoblotting, enzyme-linked immunosorbent assay, and the histamin release assay. Positive reactions to broccoli (Brassica Oleracea) were observed in 7.02% of individuals. Specific IgE to broccoli and total IgE fro allergic individuals were well correlated. The different tests performed showed no significant differences in the allergenicity of conventionally raised and GM broccoli, indicating the absence of unexpected effects on allergenicity in ipt-transformed plants. Using Western blot analysis we detected heterogeneous IgE-reactive allergenic components in broccoli-allergic sera, but no significant differences between GM an non-GM broccoli were observed in serum from the same patients. Our study demonstrates that there are no differences between GM (ipt-transformed) broccoli and non-GM broccoli, as determined by specific IgE in sera from broccoli-allergic patients. This indicates that there were no unexpected effects on allergenicity in this GM broccoli.

Purification and characterization of the glycoprotein allergen from Prosopis juliflora pollen.

Science.gov (United States)

Thakur, I S

1991-02-01

Highly active glycoprotein allergens have been isolated from pollen of Prosopis juliflora by a combination of Sephadex G-100 gel filtration and Sodium dodecyl sulphate-Poly-acrylamide gel electrophoresis. The glycoprotein fraction was homogeneous, and had molecular weight 20,000. The purified glycoprotein allergen contained 20% carbohydrate, mainly arabinose and galactose. Enzymatic digestion of glycoprotein with protease released glycopeptides of molecular weight ranging from less than 1,000 to more than 5,000 on Sephadex G-25 gel filtration. Antigenicity or allergenicity testing of these glycopeptides by immunodiffusion, immunoelectrophoresis, and radioallergosorbent test indicated complete loss of allergenic activity after digestion with protease whereas incubation with beta-D-galactosidase and periodate oxidation had little affect on the allergenic activity of the glycoprotein fraction. But incubation with alpha-D-glucosidase did not affect the allergenic activity significantly. All these tests indicated that protein played significant role in allergenicity of P. juliflora pollen.

ALLERGEN-SPECIFIC IMMUNOTHERAPY: VACCINES FOR ALLERGIC DISEASES

Directory of Open Access Journals (Sweden)

A. S. Fedorov

2015-01-01

Full Text Available Allergen-specific immunotherapy (ASIT is the most effective method of allergy treatment which consists of exposure to small doses of antigen responsible for development of allergic condition in the particular patient. Therefore, one may achieve desensitization to this antigen. The history of ASIT application lasts for more than 100 years, and, over this time, huge clinical evidence for the usage of the method has been accumulated. Use of ASIT causes reduction of allergy symptoms and treatment needs and, moreover, it has the potential for long-term clinical benefit, by preventing the development of allergy and its symptoms. The treatment affects basic immunological mechanisms responsible for the development of clinical symptoms. ASIT is an antiinflammatory, pathogenetic and prophylactic treatment of allergic airway disease. The review considers the results of major clinical trials of the ASIT applications for treatment of allergic diseases of the respiratory system (allergic rhinitis and bronchial asthma. Various schemes of ASIT are discussed including its different variants (injectable and sublingual ASIT, the issues of preparation choice for ASIT from those currently available on the pharmaceutical market, patient selection criteria, and the issues of modern molecular allergodiagnostic (allergic sensitization mapping of the patient at molecular level, in order to optimize them. Immunological mechanisms of ASIT are also considered, since appropriate views are rather contraversial. The ASIT effect is mediated through the following basic immunological mechanisms: the suppressed increase of the eosinophil concentrations, reduced duration of the delayed hypersensitivity phase, as well as initiation and maintenance of the Th2-to-Th1-like immune response transition. Regulatory T-cells play a major role in implementation of the immunological mechanism in ASIT, they have a significant impact on the Th2 response suppression. Such suppression may proceed

Level of Catechin, Myricetin, Quercetin and Isoquercitrin in Buckwheat (Fagopyrum esculentum Moench), Changes of Their Levels during Vegetation and Their Effect on The Growth of Selected Weeds

Czech Academy of Sciences Publication Activity Database

Kalinová, J.; Vrchotová, Naděžda

2009-01-01

Roč. 57, č. 7 (2009), s. 2719-2725 ISSN 0021-8561 Institutional research plan: CEZ:AV0Z60870520 Keywords : Allelopathy * buckwheat * catechin * quercetin * myricetin * weed Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 2.469, year: 2009

Immunochemical characterization of prosopis juliflora pollen allergens and evaluation of cross-reactivity pattern with the most allergenic pollens in tropical areas.

Science.gov (United States)

Assarehzadegan, Mohammad-Ali; Khodadadi, Ali; Amini, Akram; Shakurnia, Abdol-Hosein; Marashi, Seyed Saeid; Ali-Sadeghi, Hosein; Zarinhadideh, Farnoosh; Sepahi, Najmeh

2015-02-01

Allergy to Prosopis juliflora (mesquite) pollen is one of the common causes of respiratory allergy in tropical countries. Mesquite is widely used as street trees in towns and ornamental shade trees in parks and gardens throughout arid and semiarid regions of Iran. The inhalation of mesquite pollen and several species of Amaranthus/Chenopodiaceae family is the most important cause of allergic respiratory symptoms in Khuzestan province. This study was designed to evaluate IgE banding proteins of mesquite pollen extract and its IgE cross-reactivity with other allergenic plants. Twenty patients with allergic symptoms and positive skin prick tests (SPT) for mesquite pollen extract participated in the study. Crude pollen extract was prepared from local mesquite trees and used for the evaluation of allergenic profiles of P. juliflora pollen extract by Sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and IgE-immunoblotting. There were several protein bands in mesquite pollen extract using SDS-PAGE with the approximate range of molecular weight of 10-85 kDa. The most frequent IgE reactive bands among the patients' sera were approximately 20 and 66 kDa. However, there were other IgE reactive protein bands among the patients' sera with molecular weights of 10, 15, 35, 45, 55 and 85 kDa. Inhibition experiments revealed high IgE cross-reactivity between mesquite and acacia. There are several IgE-binding proteins in P. juliflora pollen extract. Results of this study indicate that proteins with a molecular weight of 10 to 85 kDa are the major allergens in P. juliflora pollen extract.

Managing Food Allergens in the U.K. Retail Supply Chain.

Science.gov (United States)

Walker, Michael J; Gowland, M Hazel; Points, John

2018-01-01

The U.K. food and grocery market is highly significant financially and dominated by 10 retailers within a regulated and extremely economically competitive environment. We summarize the approach of U.K. retailers to allergen risk assessment (RA) and risk management (RM) within the U.K. legal framework and explore public visibility of retailers' allergen policies. RA and RM of allergens appear effective in curtailing retail-triggered severe food allergy reactions. However, allergen recalls remain high, precautionary allergen labeling (PAL) remains an area of confusion, and there is no consistent Web-based provision of information for consumers who have allergies. Resolution of PAL awaits an agreed-on threshold framework, but a key challenge is to engage with patients and gain their trust rather than thrust education at them. It would be helpful for retailers to publish their allergen RA and RM policies. A target should be agreed on between government and retailers for a reduction in the proliferation of PAL wording variants by a given date within the next 3 years. A further hurdle is potentially flawed allergen analysis-development of reference methods and reference materials are acknowledged needs. Laboratories should report allergen results in an informative manner, communicating uncertainty and caveats. Ideally a laboratory representative would be included on any incident control team. Efforts must continue to standardize preparedness for protecting and defending food and drink from deliberate attack.

Identification of two distinct allergenic sites on ryegrass-pollen allergen, Lol p IV.

Science.gov (United States)

Jaggi, K S; Ekramoddoullah, A K; Kisil, F T; Dzuba-Fischer, J M; Rector, E S; Sehon, A H

1989-04-01

Lol p IV is an important allergen of ryegrass pollen. For the immunochemical identification of antigenic and/or allergenic site(s), murine monoclonal antibodies (MAbs) were prepared against Lol p IV. The hybridoma cell-culture supernatants were screened for anti-Lol p IV antibodies by a combination of ELISA and Western immunoblot analyses. The MAbs were finally purified from ascites on a Mono Q ion-exchange column. In a competitive radioimmunoassay with Lol p IV as the solid phase and 125I-labeled MAbs, it was established that MAbs 90, 91, 92, 93, and 94, although they differed in their relative affinities, recognized in common with one another an epitope designated as antigenic site A, whereas MAb 12 recognized a different epitope referred to as site B. Sites A and B were also demonstrated to constitute allergenic determinants of Lol p IV. Differences in the repertoire of specificities of the human IgE antibodies directed to Lol p IV were also demonstrated. Interestingly, it was found that sera from both allergic as well as from nonatopic individuals had IgG antibodies to sites A and/or B.

cDNA Cloning, expression and characterization of an allergenic 60s ribosomal protein of almond (prunus dulcis).

Science.gov (United States)

Abolhassani, Mohsen; Roux, Kenneth H

2009-06-01

Tree nuts, including almond (prunus dulcis) are a source of food allergens often associated with life-threatening allergic reactions in susceptible individuals. Although the proteins in almonds have been biochemically characterized, relatively little has been reported regarding the identity of the allergens involved in almond sensitivity. The present study was undertaken to identify the allergens of the almond by cDNA library approach. cDNA library of almond seeds was constructed in Uni-Zap XR lamda vector and expressed in E. coli XL-1 blue. Plaques were immunoscreened with pooled sera of allergic patients. The cDNA clone reacting significantly with specific IgE antibodies was selected and subcloned and subsequently expressed in E. coli. The amino acids deducted from PCR product of clone showed homology to 60s acidic ribosomal protein of almond. The expressed protein was 11,450 Dalton without leader sequence. Immunoreactivity of the recombinant 60s ribosomal protein (r60sRP) was evaluated with dot blot analysis using pooled and individual sera of allergic patients. The data showed that r60sRP and almond extract (as positive control) possess the ability to bind the IgE antibodies. The results showed that expressed protein is an almond allergen.Whether this r60sRP represents a major allergen of almond needs to be further studied which requires a large number of sera from the almond atopic patients and also need to determine the IgE-reactive frequencies of each individual allergen.

Allergenic food introduction and risk of childhood atopic diseases.

Directory of Open Access Journals (Sweden)

Niels J Elbert

Full Text Available The role of timing and diversity of allergenic food introduction in the development of childhood allergic sensitization and atopic diseases is controversial.To examine whether timing and diversity of allergenic food introduction are associated with allergic sensitization, allergy and eczema in children until age 10 years.This study among 5,202 children was performed in a population-based prospective cohort. Timing (age ≤6 months vs. >6 months and diversity (0, 1, 2 and ≥3 foods of allergenic food (cow's milk, hen's egg, peanut, tree nuts, soy and gluten introduction were assessed by questionnaires at ages 6 and 12 months. At age 10 years, inhalant and food allergic sensitization were measured by skin prick tests, and physician-diagnosed inhalant and food allergy by questionnaire. Data on parental-reported physician-diagnosed eczema were obtained from birth until age 10 years.Children introduced to gluten at age ≤6 months had a decreased risk of eczema (aOR (95% CI: 0.84 (0.72, 0.99, compared with children introduced to gluten at age >6 months. However, timing of allergenic food introduction was not associated with allergic sensitization or physician-diagnosed allergy. Children introduced to ≥3 allergenic foods at age ≤6 months had a decreased risk of physician-diagnosed inhalant allergy (0.64 (0.42, 0.98, compared with children not introduced to any allergenic food at age ≤6 months. However, diversity of allergenic food introduction was not associated with allergic sensitization, physician-diagnosed food allergy or eczema.Neither timing nor diversity of allergenic food introduction was consistently associated with childhood allergic sensitization, allergy or eczema.

Risk management of allergenic food ingredients in hospitality

Directory of Open Access Journals (Sweden)

Popov-Raljić Jovanka

2017-01-01

Full Text Available Food allergens have appeared in the last two decades as a concealed form of threat which significantly endangers public health, and their labelling on food products, drinks, and non pre-packed gastro-products is clearly defined with legal regulations. In practice, the chemical risk management is faced with several unexpected problems. Some of them are declarations or statements about allergenic ingredients, where a nutritional allergen that the food contains is labelled with an unusual name, or similar products from different manufacturers where one is safe and the other contains allergens. A hospitality facility which deals with production and distribution of unpackaged foods should, in addition to a developed HACCP concept and standardized recipes for food preparation, prepare a detailed, precise, and clearly defined plan for management of chemical risks.

Mass spectrometric analysis of electrophoretically separated allergens and proteases in grass pollen diffusates

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Geczy Carolyn L

2003-09-01

Full Text Available Abstract Background Pollens are important triggers for allergic asthma and seasonal rhinitis, and proteases released by major allergenic pollens can injure airway epithelial cells in vitro. Disruption of mucosal epithelial integrity by proteases released by inhaled pollens could promote allergic sensitisation. Methods Pollen diffusates from Kentucky blue grass (Poa pratensis, rye grass (Lolium perenne and Bermuda grass (Cynodon dactylon were assessed for peptidase activity using a fluorogenic substrate, as well as by gelatin zymography. Following one- or two-dimensional gel electrophoresis, Coomassie-stained individual bands/spots were excised, subjected to tryptic digestion and analysed by mass spectrometry, either MALDI reflectron TOF or microcapillary liquid chromatography MS-MS. Database searches were used to identify allergens and other plant proteins in pollen diffusates. Results All pollen diffusates tested exhibited peptidase activity. Gelatin zymography revealed high Mr proteolytic activity at ~ 95,000 in all diffusates and additional proteolytic bands in rye and Bermuda grass diffusates, which appeared to be serine proteases on the basis of inhibition studies. A proteolytic band at Mr ~ 35,000 in Bermuda grass diffusate, which corresponded to an intense band detected by Western blotting using a monoclonal antibody to the timothy grass (Phleum pratense group 1 allergen Phl p 1, was identified by mass spectrometric analysis as the group 1 allergen Cyn d 1. Two-dimensional analysis similarly demonstrated proteolytic activity corresponding to protein spots identified as Cyn d 1. Conclusion One- and two-dimensional electrophoretic separation, combined with analysis by mass spectrometry, is useful for rapid determination of the identities of pollen proteins. A component of the proteolytic activity in Bermuda grass diffusate is likely to be related to the allergen Cyn d 1.

Crystallization and preliminary X-ray analysis of birch-pollen allergen Bet v 1 in complex with a murine monoclonal IgG Fab' fragment

DEFF Research Database (Denmark)

Spangfort, M D; Mirza, Osman Asghar; Gajhede, M

1999-01-01

of the clinical symptoms of allergy. In order to study the structural basis of allergen-antibody interaction, a complex between the major birch-pollen allergen Bet v 1 and a Fab' fragment isolated from the murine monoclonal Bet v 1 antibody BV16 has been crystallized. Complex crystals belong to space group P1...

Adsorption of Cashew Allergens to Acid-Etched Zinc Metal Particles

Science.gov (United States)

Galvanized metal surfaces are approved by the FDA for use in many food processing steps. Food allergens can cause severe reactions even in very small amounts, and surfaces contaminated with allergens could pose a serious threat. The binding of cashew allergens to zinc particles was evaluated. Whi...

Allergen labelling in meat, dairy and cereal products from the Serbian market

Science.gov (United States)

Spirić, D.; Nikolić, D.; Ćirić, J.; Janković, S.; Stefanović, S.; Janković, V.; Teodorović, V.

2017-09-01

Allergens in food are a great health risk, because of the ratio of severity of problems compared to small amounts of ingested allergen. Since 2014, Serbian producers and importers of food have been obliged to declare allergens from the list of Codex Alimentarius on the product packaging. Surveillance of different meat, diary, and cereal product took place in 2016, with aim of checking if the Serbian regulatory requirements for labelling of allergens in food are being fulfilled. Out of 68 different meat products, 20 were not labelled for allergens. Thirty-six labels of various dairy products were examined revealing that allergen information was included on 27 of them. Only one of eight examined cereal products did not have allergen labelling.

Effect of Age on Allergen Responses of Allergic Patients in Southern Taiwan

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Wen Chiang

2004-07-01

Full Text Available To survey airborne and food allergen patterns in southern Taiwan and to analyze the effect of age on response to different allergens, we tested samples from 4,411 allergic patients at Kaohsiung Medical University Hospital using the MAST-CLA test (new Taiwan panel. A total of 2,212 (50.1% samples showed a positive response. We grouped allergic patients into five age groups. Milk and egg white were the main food allergens in the younger groups (< 3 years old and 3-6 years old. Shrimp, crab, and shellfish were the main allergens in the groups aged 7-12, 13-18, and more than 18 years. Among airborne allergens, house dust and mites Dermatophagoides farinae and D. pteronyssinus were the main allergens in all age groups, whereas the frequency of response to cockroach allergen was low in the group aged less than 3 years, but increased in the other age groups. There was a sharp increase in the frequency of response to airborne allergens after 3 years old and a sharp decrease in response to food allergens. Among subjects allergic to both airborne and food allergens, there was a positive MAST-CLA rate of 19.9% to 26% (all five age groups, no significant difference. When we compared our results with those from Taipei Veterans General Hospital in northern Taiwan, there were significant differences for yeast, peanut, feather mix, dog dander, cockroach, D. farinae and D. pteronyssinus allergens (p < 0.01. These differences were probably caused by differences in patient location, patient age, disease patterns and allergen panels.

Efficiency of Different Nitrogen Forms in Buckwheat (Fagopyrum esculentum Moench

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Burhan Kara

2016-06-01

Full Text Available The research was carried out with aim to determination the efficient of nitrogen forms (ammonium sulfate, ammonium nitrate and urea on nitrogen use efficient for buckwheat in Isparta during 2014 and 2015 years. All the examined characteristics were determined higher values in applied nitrogen forms according to non-nitrogen parcel. In compared to nitrogen forms, the highest grain yield (1456 and 1325 kg ha-1, biological yield (4873 and 4512 kg ha-1, 1000 grain weight (24.9 and 24.8 g, agronomic efficient (24.96% and 24.25%, recycling efficient (0.24% and 0.22% and utilization efficient (0.25% and 0.18% were obtained from ammonium sulfate, the highest protein content (11.37% and 12.44% and agro-physiological efficient (0.27% and 0.24% from ammonium nitrate in both years. Among the nitrogen forms weren’t significant differently in physiological efficient in both years, recycling and utilization efficient in the first year. The mineral nutrient content varied according to nitrogen forms. Generally, ammonium sulfate was positive effect to yield and some quality parameters.

Evolution of nutrient ingredients in tartary buckwheat seeds during germination.

Science.gov (United States)

Yiming, Zhou; Hong, Wang; Linlin, Cui; Xiaoli, Zhou; Wen, Tang; Xinli, Song

2015-11-01

Evolution of nutrient components and the antioxidative activity of seed sprouts of tartary buckwheat (Fagopyrum tataricum L. Gaertn) were investigated in the course of germination. Results showed that the contents of total flavonoids increased with germination time and leveled off after the third germination day with the changing trend of rutin and quercetin opposite to each other. The decrease of total protein and total sugar contents in the germinated seeds was accompanied respectively by an increase of amino acid and reducing sugar contents. The contents of vitamin C (Vc) and B1(V(B1)) exhibited a minimum with no appreciable changes found for vitamin B(2) (V(B2)) and B(6) (V(B6)). The contents of total chlorophyll, chlorophyll A and B all exhibited a maximum on the fifth germination day. The contents of fatty acids had no regular changing trend with germination time. The free radical-scavenging activities of the seeds increased with germination time and were caused by an increase in their antioxidative activity. Copyright © 2015 Elsevier Ltd. All rights reserved.

Modelling allergenic risk

DEFF Research Database (Denmark)

Birot, Sophie

combines second order Monte-Carlo simulations with Bayesian inferences [13]. An alternative method using second order Monte-Carlo simulations was proposed to take into account the uncertainty from the inputs. The uncertainty propagation from the inputs to the risk of allergic reaction was also evaluated...... countries is proposed. Thus, the allergen risk assessment can be performed cross-nationally and for the correct food group. Then the two probabilistic risk assessment methods usually used were reviewed and compared. First order Monte-Carlo simulations are used in one method [14], whereas the other one......Up to 20 million Europeans suffer from food allergies. Due to the lack of knowledge about why food allergies developed or how to protect allergic consumers from the offending food, food allergy management is mainly based on food allergens avoidance. The iFAAM project (Integrated approaches to Food...

Factors influencing the quality of Myrmecia pilosula (Jack Jumper) ant venom for use in in vitro and in vivo diagnoses of allergen sensitization and in allergen immunotherapy.

Science.gov (United States)

Wanandy, T; Dwyer, H E; McLean, L; Davies, N W; Nichols, D; Gueven, N; Brown, S G A; Wiese, M D

2017-11-01

Allergen immunotherapy uses pharmaceutical preparations derived from naturally occurring source materials, which contain water-soluble allergenic components responsible for allergic reactions. The success of in vivo and in vitro diagnoses in allergen sensitization and allergen immunotherapy largely depends on the quality, composition and uniformity of allergenic materials used to produce the active ingredients, and the formulation employed to prepare finished products. We aimed to examine the factors influencing batch-to-batch consistency of Jack Jumper (Myrmecia pilosula) ant venom (JJAV) in the form of active pharmaceutical ingredient (AI) and informed whether factors such as temperature, artificial light and container materials influence the quality of JJAV AIs. We also aimed to establish handling and storage requirements of JJAV AIs to ensure preservation of allergenic activities during usage in the diagnosis of allergen sensitization and in allergen immunotherapy. The quality and consistency of JJAV AIs were analysed using a combination of bicinchoninic acid assay for total protein quantification, HPLC-UV for JJAV allergen peptides quantification, ELISA inhibition for total allergenic potency, SDS-PAGE, AU-PAGE and immunoblot for qualitative assessment of JJAV components, and Limulus Amebocyte Lysate assay for the quantification of endotoxin concentration. API-ZYM and Zymogram assays were used to probe the presence of enzymatic activities in JJAV. Pharmaceutical-grade JJAV for allergen immunotherapy has good batch-to-batch consistency. Temporary storage at 4°C and light exposure do not affect the quality of JJAV. Exposure to temperature above 40°C degrades high MW allergens in JJAV. Vials containing JJAV must be stored frozen and in upright position during long-term storage. We have identified factors, which can influence the quality and consistency of JJAV AIs, and provided a framework for appropriate handling, transporting and storage of JJAV to be used