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Sample records for standardized allergenic extract

  1. Standardization of food allergen extracts for skin prick test

    DEFF Research Database (Denmark)

    Skamstrup Hansen, K; Bindslev-Jensen, C; Skov, P S

    2001-01-01

    The aim of the study was to standardize and evaluate technically optimized food allergen extracts for use in skin prick test (SPT). The standardization procedure comprised 36 allergic histories in 32 food allergic patients with 21 healthy, non-atopic individuals serving as controls. The patients......-blind, placebo-controlled food challenge, except for the hazelnut-allergic patients. The controls were subjected to an open food challenge with all the foods to ensure tolerance. The standardization was performed by means of titrated SPT in accordance with the guidelines on biological standardization from...

  2. Standardization of food allergen extracts for skin prick test

    DEFF Research Database (Denmark)

    Skamstrup Hansen, K; Bindslev-Jensen, C; Skov, P S

    2001-01-01

    The aim of the study was to standardize and evaluate technically optimized food allergen extracts for use in skin prick test (SPT). The standardization procedure comprised 36 allergic histories in 32 food allergic patients with 21 healthy, non-atopic individuals serving as controls. The patients...... had a history of allergic symptoms upon ingestion of either cow's milk (n=3), hen's egg (n=9), wheat (n=4), hazelnut (n=14) or cod (n=6). They also had specific IgE in serum to the food in question and a positive SPT with a fresh preparation of the food. The diagnosis had been confirmed by a double......-blind, placebo-controlled food challenge, except for the hazelnut-allergic patients. The controls were subjected to an open food challenge with all the foods to ensure tolerance. The standardization was performed by means of titrated SPT in accordance with the guidelines on biological standardization from...

  3. Safety evaluation of standardized allergen extract of Japanese cedar pollen for sublingual immunotherapy.

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    Mitobe, Yuko; Yokomoto, Yasuki; Ohashi-Doi, Katsuyo

    2015-04-01

    Japanese cedar (JC) pollinosis is caused by Japanese cedar pollen (JCP) and most common seasonal allergic disease in Japan. Subcutaneous immunotherapy (SCIT) with allergen extract of JCP (JCP-allergen extract) is well established for JC pollinosis treatment with improvement of symptoms. However, major drawbacks for SCIT are repeated painful injections, frequent hospital visits and anaphylactic risk. Currently, sublingual immunotherapy (SLIT) has received much attention as an advanced alternative application with lower incidence of systemic reactions because the liquid or tablet form of allergen is placed under the tongue. The aim of this study was safety evaluation of standardized JCP-allergen extract currently developed for SLIT in JC pollinosis. JCP-allergen extract showed no potential genotoxicity. No systemic effects were observed in rats administered JCP-allergen extract orally for 26 weeks followed by 4-week recovery period. Mild local reactions such as hyperplasia and increased globule leukocytes resulting from vehicle (glycerin)-induced irritation were observed in stomach. No-observed-adverse-effect level was greater than 10,000 JAU/kg/day for systemic toxicity, equivalent to 300-fold the human dose. No local irritation was found in rabbits oral mucosae by 7-day sublingual administration. These results demonstrate the safe profile of standardized JCP-allergen extract, suggesting it is suitable for SLIT in JC pollinosis. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Treatment of grass pollen allergy: focus on a standardized grass allergen extract - Grazax®.

    Science.gov (United States)

    Calderón, Moisés; Brandt, Tove

    2008-12-01

    Immunotherapy is the only treatment for allergy that has the potential to alter the natural course of the disease. Sublingual immunotherapy (SLIT) for grass pollen-induced rhino-conjunctivitis has been developed to make immunotherapy available to a broader group of allergic patients. In the largest clinical programme ever conducted with allergen-specific immunotherapy, over 1,700 adults and 260 children have been exposed to Grazax(®). Grazax is formulated as an oral lyophilisate (tablet) for sublingual administration, containing 75,000 SQ-T standardized allergen extract of grass pollen from Phleum pratense. Grazax is indicated for treatment of grass pollen-induced rhinitis and conjunctivitis in adult patients with clinically relevant symptoms and diagnosed with a positive skin prick test and/or specific IgE test to grass pollen. In phase I trials doses from 2,500 to 1,000,000 SQ-T were tested. All doses were well tolerated and 75,000 SQ-T, with approximately 15 mug major allergen protein, was chosen as the optimal dose. Three phase III trials are ongoing, one being a long-term trial. Results from GT-08 trial first and second treatment years showed a reduction of 30% and 36%, respectively, in daily rhino-conjunctivitis symptom scores and a reduction of 38% and 46% of daily rhino-conjunctivitis medication scores compared with placebo over the entire grass pollen season. Subjects treated with Grazax also had an increased number of well days and improved quality of life, and more subjects experienced excellent rhino-conjunctivitis control. The most common adverse events related to Grazax are local reactions, such as pruritus, edema mouth, ear pruritus, throat irritation, and sneezing. We conclude that Grazax is efficacious and safe for treatment of rhino-conjunctivitis due to grass pollen allergy.

  5. Treatment of grass pollen allergy: focus on a standardized grass allergen extract – Grazax®

    Directory of Open Access Journals (Sweden)

    Moisés Calderón

    2008-11-01

    Full Text Available Moisés Calderón1, Tove Brandt21Section of Allergy and Clinical Immunology, Royal Brompton Hospital, Imperial College, NHLI, London, UK; 2Group Clinical Development, ALK-Abelló A/S, Hørsholm, DenmarkAbstract: Immunotherapy is the only treatment for allergy that has the potential to alter the natural course of the disease. Sublingual immunotherapy (SLIT for grass pollen-induced rhino-conjunctivitis has been developed to make immunotherapy available to a broader group of allergic patients. In the largest clinical programme ever conducted with allergen-specific immunotherapy, over 1,700 adults and 260 children have been exposed to Grazax®. Grazax is formulated as an oral lyophilisate (tablet for sublingual administration, containing 75,000 SQ-T standardized allergen extract of grass pollen from Phleum pratense. Grazax is indicated for treatment of grass pollen-induced rhinitis and conjunctivitis in adult patients with clinically relevant symptoms and diagnosed with a positive skin prick test and/or specific IgE test to grass pollen. In phase I trials doses from 2,500 to 1,000,000 SQ-T were tested. All doses were well tolerated and 75,000 SQ-T, with approximately 15 µg major allergen protein, was chosen as the optimal dose. Three phase III trials are ongoing, one being a long-term trial. Results from GT-08 trial first and second treatment years showed a reduction of 30% and 36%, respectively, in daily rhino-conjunctivitis symptom scores and a reduction of 38% and 46% of daily rhinoconjunctivitis medication scores compared with placebo over the entire grass pollen season. Subjects treated with Grazax also had an increased number of well days and improved quality of life, and more subjects experienced excellent rhino-conjunctivitis control. The most common adverse events related to Grazax are local reactions, such as pruritus, edema mouth, ear pruritus, throat irritation, and sneezing. We conclude that Grazax is efficacious and safe for treatment

  6. Mite allergen extracts and clinical practice.

    Science.gov (United States)

    Carnés, Jerónimo; Iraola, Víctor; Cho, Seong H; Esch, Robert E

    2017-03-01

    To provide physicians, researchers, and other interested health care professionals with information about how mite source materials and allergen extracts are manufactured, including the critical process parameters that can affect the final composition of allergenic extracts available for clinical use. A PubMed search was performed using focused keywords combined with relevant regulatory documents and industry guidelines. The information obtained through literature and specialized books was evaluated and combined with the personal expertise and experience of the authors. Dermatophagoides farinae and Dermatophagoides pteronyssinus are the primary species responsible for allergen sensitizations and allergy symptoms in genetically predisposed individuals. Storage mites belonging to the families Glycyphagidae, Echimyopodidae, and Acaridae can also be relevant sources of indoor mite allergens. The cultivation and purification processes used to produce mite raw materials play a critical role in the final composition of mite allergen extracts. Mite extract standardization in the United States is based on total allergenic activity with respect to a single national standard, whereas in Europe consistency is ensured by in-house standards and international references. Because of the limitation of allergen avoidance and pharmacotherapy for patients with severe allergic rhinitis and asthma, house dust mite subcutaneous immunotherapy or sublingual immunotherapy can be an invaluable treatment option for them. Differences in manufacturing processes and extract standardization approaches may lead to differences in extract quality and potency. Physicians should be aware of these potential sources of mite extract variability. Use of well-standardized house dust mite extracts would be critical for success in the diagnosis and treatment of house dust mite allergy. Copyright © 2016 American College of Allergy, Asthma & Immunology. All rights reserved.

  7. Treatment of grass pollen allergy: focus on a standardized grass allergen extract ? Grazax ?

    OpenAIRE

    Calder?n, Mois?s; Brandt, Tove

    2008-01-01

    Immunotherapy is the only treatment for allergy that has the potential to alter the natural course of the disease. Sublingual immunotherapy (SLIT) for grass pollen-induced rhino-conjunctivitis has been developed to make immunotherapy available to a broader group of allergic patients. In the largest clinical programme ever conducted with allergen-specific immunotherapy, over 1,700 adults and 260 children have been exposed to Grazax?. Grazax is formulated as an oral lyophilisate (tablet) for su...

  8. Standardization of allergen products: 1. Detailed characterization of GMP-produced recombinant Bet v 1.0101 as biological reference preparation

    NARCIS (Netherlands)

    Himly, M.; Nony, E.; Chabre, H.; van Overtvelt, L.; Neubauer, A.; van Ree, R.; Buchheit, K.-H.; Vieths, S.; Moingeon, P.; Ferreira, F.

    2009-01-01

    Standardization of allergen extracts requires the availability of well-characterized recombinant allergens, which can be used as reference standards provided by the European regulatory authorities. The objective of this study was the detailed physicochemical and immunological characterization of

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

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    Blank, Simon; Etzold, Stefanie; Darsow, Ulf; Schiener, Maximilian; Eberlein, Bernadette; Russkamp, Dennis; Wolf, Sara; Graessel, Anke; Biedermann, Tilo; Ollert, Markus; Schmidt-Weber, Carsten B

    2017-10-03

    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 grade HBV extracts which are approved for immunotherapy in numerous countries. The extracts were analyzed for their content of the major allergens Api m 1, Api m 2, Api m 3, Api m 5 and Api m 10. Using allergen-specific antibodies we were able to demonstrate the underrepresentation of relevant major allergens such as Api m 3, Api m 5 and Api m 10 in particular therapeutic extracts. Taken together, standardization of therapeutic extracts by determination of the total allergenic potency might imply the intrinsic pitfall of losing information about particular major allergens. Moreover, the variable allergen composition of different therapeutic HBV extracts might have an impact on therapy outcome and the clinical management of HBV-allergic patients with specific IgE to particular allergens.

  10. Oral administration of allergen extracts from mugwort pollen desensitizes specific allergen-induced allergy in mice.

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    Ma, Wen-Jiang; Bao, Meng-Jing; Zhu, Jian-Ping; Yao, Hong-Yi; Xie, Yi-Cheng; Guan, Yan; Li, Fen-Fen; Dong, Xin-Wei; Zheng, Ying-Ming; Xie, Qiang-Min

    2012-02-14

    Clinically, sublingual immunotherapy (SLIT) using allergen extracts effectively alleviates the symptoms of allergic rhinitis and asthma. We hypothesized that oral administration of a high-dose of allergen extracts imitates SLIT, which may prevent IgE-related responses in allergic diseases. In the present study, we investigated the effects of oral administration of allergen extracts from mugwort pollen (MP) on allergen-induced inflammation and airway hyperresponsiveness (AHR) in an allergic mouse model. After administration of MPdrop containing Art v 1 and Art v 4 extracts derived from MP specifically in MP-sensitized mice, the effects of MPdrop on AHR, inflammatory cell accumulation, cytokine production in the bronchoalveolar lavage fluid and lung tissue, and serum IgE and IgG levels were investigated. The results indicated that MPdrop not only prevented the AHR in response to methacholine in a dose-dependent manner but also significantly reduced the total serum and allergen-specific IgE levels. All of the maximal effects were achieved at a dose of 100μg/(kgd) and were comparable to the effects of dexamethasone at a dose of 0.5mg/(kgd). Furthermore, oral administration of MPdrop dose-dependently elevated allergen-specific serum IgG2a levels, reduced total and allergen-specific IgE levels and normalized the imbalance between the Th1 cytokine IL-12 and Th2 cytokines IL-4 and IL-5. Finally, oral administration of MPdrop significantly reduced goblet cell hyperplasia and eosinophilia in the MP-sensitized allergic mouse model. These data suggest that MPdrop effectively improves specific allergen-induced inflammation and AHR in MP-sensitized and -challenged mice and provides the rationale for clinical use of MPdrop in the specific allergen-induced asthma. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. Ranking in importance of allergen extract characteristics for sublingual immunotherapy by Italian specialists.

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    Canonica, Giorgio Walter; Passalacqua, Giovanni; Incorvaia, Cristoforo; Cadario, Gianni; Fiocchi, Alessandro; Senna, Gianenrico; Rossi, Oliviero; Romano, Antonino; Scala, Enrico; Romano, Catello; Ingrassia, Antonino; Zambito, Marcello; Dell'Albani, Ilaria; Frati, Franco

    2014-01-01

    The efficacy of allergen immunotherapy (AIT) is well supported by evidence from trials and meta-analyses. However, its actual performance in daily practice may be diminished by several pitfalls, including inappropriate patient selection, and, especially, the use of allergen extracts of insufficient quality. We performed a survey, the Allergen Immunotherapy Decision Analysis, to evaluate which criteria specialists use to choose products for sublingual immunotherapy (SLIT) in adult patients suffering from allergic respiratory disease. We surveyed a total of 169 Italian allergists randomly chosen from a database belonging to a market research company (Lexis Ricerche, Milan, Italy). The survey was performed between October and November 2012 under the aegis of the European Center for Allergy Research Foundation and consisted of a questionnaire-based electronic survey prepared by a scientific board of 12 AIT experts. The questionnaire comprised two parts, the first of which contained 14 items to be ranked by each participant according to the importance assigned to each when choosing SLIT products. The physicians' rankings assigned major importance to the level of evidence-based validation of efficacy and safety, standardization of the product, efficacy based on personal experience, and defined content(s) of the major allergen(s) in micrograms. The results of this survey show that Italian allergists rank the quality-related characteristics of allergen extracts as highly important when choosing products for AIT. The allergists' preference for high-quality products should be addressed by regulatory agencies and by producers.

  12. Requirements for acquiring a high-quality house dust mite extract for allergen immunotherapy

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    Frati F

    2012-05-01

    Full Text Available Franco Frati,1 Cristoforo Incorvaia,2 Marie David,3 Silvia Scurati,3 Simona Seta,4 Guglielmo Padua,4 Eleonora Cattaneo,1 Carlo Cavaliere,5 Alessia Di Rienzo,6 Ilaria Dell'Albani,1 Paola Puccinelli11Medical and Scientific and Regulatory Department, Stallergenes, Milan, Italy; 2Allergy/Pulmonary Rehabilitation, ICP Hospital, Milan, Italy; 3Laboratoire Stallergenes, Antony, France; 4Marketing Department, Stallergenes, Milan, Italy; 5Ear, Nose and Throat Department, University Sapienza, Rome, Italy; 6Azienda Sanitaria Locale, Allergology Service, Frosinone, ItalyAbstract: The house dust mite is a major cause of respiratory allergy worldwide. The management of mite allergy is based on avoidance measures, drug treatment, and allergen immunotherapy, but only allergen immunotherapy is able to modify the natural history of the disease. Injectable subcutaneous immunotherapy was introduced a century ago, while sublingual immunotherapy was proposed in the 1980s and emerged in the ensuing years as an effective and safe option to subcutaneous immunotherapy. However, the quality of the extracts to be used in allergen immunotherapy is crucial for the success of treatment. The mite extract for sublingual immunotherapy known as Staloral 300 was developed to offer optimal characteristics concerning the mite culture medium, standardization, and allergen dose. Double-blind, placebo-controlled trials with Staloral 300 have provided a substantial part of the clinical evidence analyzed in a meta-analysis of the efficacy of allergen immunotherapy in mite-induced rhinitis and asthma. Safety and tolerability are very good, mild local reactions in the mouth being the most common side effect. This makes it feasible to carry out sublingual immunotherapy for the 3–5-year duration needed to achieve long-lasting tolerance to the specific allergen. The performance of Staloral 300 may provide optimal conditions for an effective and safe sublingual immunotherapy in patients with

  13. Optimized techniques for the extraction of grape allergens appropriate for in vivo and in vitro testing and diagnosis

    NARCIS (Netherlands)

    Vassilopoulou, Emilia V.; Zuidmeer, Laurian; Akkerdaas, Jaap; Rigby, Neil; Moren, F. Javier; Papadopoulos, Nikolaos G.; Saxoni-Papageorgiou, Photini; Milis, Clare; van Ree, Ronald

    2007-01-01

    Standardized allergen extracts are needed for diagnosis and therapy purposes. For grapes, standardization is hampered by low protein and high tannin and pectin concentrations. The aim of the current study was to develop an optimized method for the extraction of grape proteins and possibly extend

  14. Results of patch testing with a standard series of allergens at Manipal

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    Shenoi Shrutakirthi

    1994-01-01

    Full Text Available The study was designed to determine the common sensitizers in allergic contact dermatitis, to evolve a standard patch test tray for screening patients at our centre and to suggest allergens for multicentric trial in India. 212 patients (65 women, 147 men were patch tested with a standard series of allergens (23 allergens of European standard series extended with lanolin, cresol and gentamycin. The frequent sensitizers observed were gentamycin (14.2%, potassium dichromate (7.1%, colophony (6.6% and fragrance mix (6.1%. No positive reactions were observed for lanolin, quaternium - 15 and mercaptomix. Our standard tray will thus consist of all the allergens of European standard series except primin along with lanolin, cresol and gentamycin. Lanolin is included despite negativity as it was found to be the commonest sensitizer among topical medications in our previous study.

  15. 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...

  16. STUDYING IMMUNOBIOLOGICAL FEATURES OF PREPARED WATER_SALT EXTRACTS OF INSECT ALLERGENS

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    O.V. Radikova

    2008-01-01

    Full Text Available Through the indirect degranulation of mast cells, anaphylaxis test, solid phase immune enzyme analysis, the researchers evaluated the immuno biological features of serum of 20 patients suffering from atopic mild and moderate bronchial asthma with domestic sensitization. All received water salt extracts of insects (cockroach, black beetle, American cock roach, ash gray cockroach, cricket, fly, moth, mealworm, weevil, billbug demonstrated specific activity. An immune enzyme analysis has revealed more intensive allergic features of substances extracted from black beetle, cockroach, fly and moth.Key words: insect allergens, allergen specific immunotherapy, immuno biological features of allergic extracts, Ige anti bodies.

  17. Characterization of allergenic components in sap extract from the weeping fig (Ficus benjamina).

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    Axelsson, I G; Johansson, S G; Larsson, P H; Zetterström, O

    1990-01-01

    The allergen composition of crude extract from sap (latex) of the weeping fig (Ficus benjamina) was investigated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblotting. The allergenic components were detected by sera from 11 occupationally exposed plant keepers, of whom 7 were non-atopic, and from 9 non-occupationally exposed atopic patients with a positive radio-allergosorbent test to weeping fig. The allergen-antibody complexes were visualized by rabbit anti-IgE and beta-galactosidase-labelled sheep anti-rabbit IgG, using a chromogenic insoluble substrate. A total of 11 allergenic components were identified. Three of them were found to be major allergenic components, identified by more than 50% of the investigated sera. These 3 IgE-binding components had molecular weights of approximately 29,000, 28,000 and 25,000 daltons, respectively. The major allergenic components were denatured by heat in the temperature range of 60-90 degrees C.

  18. 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

    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....... 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...

  19. Standardization of Weed Pollen Extracts, Japanese Hop and Mugwort, in Korea.

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    Jeong, Kyoung Yong; Son, Mina; Choi, Soo Young; Park, Kyung Hee; Park, Hye Jung; Hong, Chein Soo; Lee, Jae Hyun; Park, Jung Won

    2016-03-01

    Japanese hop (Humulus spp.) and mugwort (Artemisia spp.) are notable causes of autumn pollinosis in East Asia. However, Japanese hop and mugwort pollen extracts, which are widely used for the diagnosis, have not been standardized. This study was performed to standardize Japanese hop and mugwort pollen extracts. Allergen extracts were prepared in a standardized way using locally collected Humulus japonicus and purchased Artemisia vulgaris pollens. The immunoglobulin E (IgE) reactivities of prepared extracts were compared with commercial extracts via IgE immunoblotting and inhibition analyses. Intradermal skin tests were performed to determine the bioequivalent allergy unit (BAU). The IgE reactive components of the extracts via IgE immunoblotting were similar to those of commercial extracts. A 11-kDa allergen showed the strongest IgE reactivity in Japanese hop, as did a 28-kDa allergen in mugwort pollen extracts. Allergenic potencies of the investigatory Japanese hop and mugwort extracts were essentially indistinguishable from the commercial ones. Sums of erythema of 50 mm by the intradermal skin test (ΣED50) were calculated to be 14.4th and 13.6th three-fold dilutions for Japanese hop and mugwort extracts, respectively. Therefore, the allergenic activity of the prepared extracts was 90827.4 BAU/mg for Japanese hop and 34412 BAU/mg for mugwort. We produced Japanese hop and mugwort pollen extracts using a standardized method. Standardized Japanese hop and mugwort pollen extracts will facilitate the production of improved diagnostic and immunotherapeutic reagents.

  20. Inter-laboratory optimization of protein extraction, separation, and fluorescent detection of endogenous rice allergens.

    Science.gov (United States)

    Satoh, Rie; Teshima, Reiko; Kitta, Kazumi; Lang, Gang-Hua; Schegg, Kathleen; Blumenthal, Kenneth; Hicks, Leslie; Labory-Carcenac, Bénédicte; Rouquié, David; Herman, Rod A; Herouet-Guicheney, Corinne; Ladics, Gregory S; McClain, Scott; Poulsen, Lars K; Privalle, Laura; Ward, Jason M; Doerrer, Nancy; Rascle, Jean-Baptiste

    2016-07-11

    In rice, several allergens have been identified such as the non-specific lipid transfer protein-1, the α-amylase/trypsin-inhibitors, the α-globulin, the 33 kDa glyoxalase I (Gly I), the 52-63 kDa globulin, and the granule-bound starch synthetase. The goal of the present study was to define optimal rice extraction and detection methods that would allow a sensitive and reproducible measure of several classes of known rice allergens. In a three-laboratory ring-trial experiment, several protein extraction methods were first compared and analyzed by 1D multiplexed SDS-PAGE. In a second phase, an inter-laboratory validation of 2D-DIGE analysis was conducted in five independent laboratories, focusing on three rice allergens (52 kDa globulin, 33 kDa glyoxalase I, and 14-16 kDa α-amylase/trypsin inhibitor family members). The results of the present study indicate that a combination of 1D multiplexed SDS-PAGE and 2D-DIGE methods would be recommended to quantify the various rice allergens.

  1. An alternative inhibition method for determining cross-reactive allergens

    NARCIS (Netherlands)

    Schmidt-Hieltjes, Yvonne; Teodorowicz, Gosia; Jansen, Ad; Hartog, Den Gerco; Elfvering-Berendsen, Lisette; Jong, De Nicolette W.; Savelkoul, Huub F.J.; Ruinemans-Koerts, Janneke

    2017-01-01

    Inhibition assays are an useful tool to identify the allergen of primary sensitization of cross-reactive allergens. Classical ELISA-based inhibition assays are limited by both the availability of commercial standardized allergen extracts and the experience and knowledge needed for making home-made

  2. Comparison of buffers for extraction of mite allergen der p 1 from dust.

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    Prester, Ljerka; Kovačić, Jelena; Macan, Jelena

    2012-09-01

    Der p 1 is the main allergen of house dust mite Dermatophagoides pteronyssinus, which has routinely been detected in residential dust. However, the procedure for extracting Der p 1 from reservoir dust has not been well defined. The aim of this study was to compare Der p 1 mass fractions in dust extracts prepared using the following extraction buffers: phosphate (pH 7.4), borate (pH 8.0), and ammonium bicarbonate (pH 8.0), all with 0.05 % Tween 20. Twenty-eight dust samples were divided into three aliquots and each portion was extracted with one of the three buffers at room temperature. Der p 1 mass fractions were measured in a total of 84 dust extracts using the enzyme immunoassay (range: 0.1 μg g-1 to 7.53 μg g-1). Statistical methods including intraclass correlation showed a high agreement between Der p 1 mass fractions irrespective of the extracting medium. Our results suggest that all three buffers are suitable for the extraction of mite allergens and routine Der p 1 analysis in dust.

  3. Occupational and food allergy: focus on allergen extracts

    NARCIS (Netherlands)

    N.W. de Jong (Nicolette)

    2004-01-01

    textabstractThe aim of this thesis is, first, the diagnostic work-up of occupational and food allergies in the absence of well-validated commercially available standardised extracts for Skin Prick Test. Second, to investigate cross-reactivity in occupational and food allergic patients. Third, the

  4. Pressurized liquid extraction-gas chromatography-mass spectrometry analysis of fragrance allergens, musks, phthalates and preservatives in baby wipes.

    Science.gov (United States)

    Celeiro, Maria; Lamas, J Pablo; Garcia-Jares, Carmen; Llompart, Maria

    2015-03-06

    Baby wipes and wet toilet paper are specific hygiene care daily products used on newborn and children skin. These products may contain complexes mixtures of harmful chemicals. A method based on pressurized liquid extraction (PLE) followed by gas chromatography-mass spectrometry (GC-MS) has been developed for the simultaneous determination of sixty-five chemical compounds (fragrance allergens, preservatives, musks, and phthalates) in wipes and wet toilet paper for children. These compounds are legislated in Europe according Regulation EC No 1223/2009, being twelve of them banned for their use in cosmetics, and one of them, 3-iodo-2-propynyl butylcarbamate (IPBC), is banned in products intended for children under 3 years. Also, propyl-, and butylparaben will be prohibited in leave-on cosmetic products designed for application on the nappy area of children under 3 years from April 2015. PLE is a fast, simple, easily automated technique, which permits to integrate a clean-up step during the extraction process reducing analysis time and stages. The proposed PLE-based procedure was optimized on real non-spiked baby wipe samples by means of experimental design to study the influence on extraction of parameters such as extraction solvent, temperature, extraction time, and sorbent type. Under the selected conditions, the method was validated showing satisfactory linearity, and intra-day, and inter-day precision. Recoveries were between 80-115% for most of the compounds with relative standard deviations (RSD) lower than 15%. Finally, twenty real samples were analyzed. Thirty-six of the target analytes were detected, highlighting the presence of phenoxyethanol in all analyzed samples at high concentration levels (up to 0.8%, 800μgg(-1)). Methyl paraben (MeP), and ethyl paraben (EtP) were found in 40-50% of the samples, and the recently banned isobutyl paraben (iBuP) and isopropyl paraben (iPrP), were detected in one and seven samples, respectively, at concentrations between

  5. 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.

  6. Evaluation of skin sensitivity in dogs bearing allergic dermatitis to standardized allergenic extract of house dust and storage mites Avaliação da sensibilidade de cães com dermatite alérgica a extratos padronizados de ácaros da poeira domiciliar

    Directory of Open Access Journals (Sweden)

    Victor E.S. Cunha

    2007-08-01

    Full Text Available The objective of the study was to evaluate whether allergenic extracts of five house dust and storage mite species standardized for humans might be used for the diagnosis of canine atopic dermatitis (CAD. Extracts of Dermatophagoides pteronyssinus (Pyroglyphidae, D. farinae (Pyroglyphidae, Blomia tropicalis (Glycyphagidae, Lepidoglyphus destructor (Glycyphagidae and Tyrophagus putrescentiae (Acaridae were evaluated by intradermal testing in 20 healthy dogs (control and 25 dogs with allergic dermatitis. A significant difference in the response was observed between the two groups (pO presente trabalho teve como objetivo avaliar se extratos alergênicos de cinco espécies de ácaros da poeira domiciliar e produtos armazenados, padronizados para humanos, podem ser utilizados no diagnóstico da dermatite atópica canina. Extratos de Dermatophagoides pteronyssinus (Pyroglyphidae, D. farinae (Pyroglyphidae, Blomia tropicalis (Glycyphagidae, Lepidoglyphus destructor (Glycyphagidae e Tyrophagus putrescentiae (Acaridae foram avaliados através de testes intradérmicos em 45 cães, dos quais 20 normais e 25 com dermatite alérgica. Uma diferença significativa foi observada no padrão de respostas obtidas dos dois grupos (p<0.05. Apenas um animal (5% do grupo controle reagiu ao teste cutâneo, enquanto que no grupo dos alérgicos 14 cães (56% apresentaram pelo menos uma reação positiva (odds ratio = 24.2. As maiores freqüências de reações positivas observadas no grupo dos alérgicos foram aos extratos de T. putres-centiae ou L. destructor, cada um induzindo reações em 10(40% cães. Os extratos de D. farinae, D. pteronyssinus e B. tropicalis foram responsáveis por reações positivas em 7(28%, 3(12% e 3(12% cães, respectivamente. Os extratos padronizados para humanos avaliados no presente estudo podem ser utilizados como complemento no diagnóstico da doença, assim como na seleção de alérgenos para a imunoterapia alérgeno-específica.

  7. Evaluation of reduced allergenicity of irradiated peanut extract using splenocytes from peanut-sensitized mice

    Energy Technology Data Exchange (ETDEWEB)

    Oh, Sejo; Jang, Da-In [Department of Pediatrics, Ajou University School of Medicine, Suwon 442-749 (Korea, Republic of); Lee, Ju-Woon; Kim, Jae-Hun; Byun, Myung-Woo [Team for Radiation Food Science and Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185 (Korea, Republic of); Lee, Soo-Young [Department of Pediatrics, Ajou University School of Medicine, Suwon 442-749 (Korea, Republic of)], E-mail: jsjs87@ajou.ac.kr

    2009-07-15

    Peanut (PN) allergy is one of the most serious forms of IgE-mediated food hypersensitivity. Gamma irradiation has been widely used for the preservation of food. The results of our previous studies showed that the IgE-binding capacity to several antigens were profoundly reduced after gamma irradiation. In this study, we evaluated the changes of allergenecity and cytokine production profiles after exposure of irradiated PN extract in a PN-allergy mouse model. Mice were sensitized to PN extract by intragastric administration on days 0, 1, 2, and 7, and then challenged on day 21. Four weeks later, we evaluated the cytokine production patterns and proliferation responses of splenocytes that were stimulated with intact PN extract, compared to 10 and 50 kGy irradiated PN extract. When the cells were stimulated with 10 kGy of irradiated PN extract, a higher level of production of IFN-{gamma} and IL-10 cytokines was observed. However, stimulation with 50 kGy of irradiated PN extract resulted in a higher level of production of only IFN-{gamma} cytokines. In addition, the Th1/Th2 ratio increased in response to treatment with gamma-irradiated PNs. The results of this study show that the allergenicity of PN extracts could be reduced by gamma irradiation which caused downregulation of Th2 lymphocyte activity in the PN-sensitized mice.

  8. Evaluation of reduced allergenicity of irradiated peanut extract using splenocytes from peanut-sensitized mice

    International Nuclear Information System (INIS)

    Oh, Sejo; Jang, Da-In; Lee, Ju-Woon; Kim, Jae-Hun; Byun, Myung-Woo; Lee, Soo-Young

    2009-01-01

    Peanut (PN) allergy is one of the most serious forms of IgE-mediated food hypersensitivity. Gamma irradiation has been widely used for the preservation of food. The results of our previous studies showed that the IgE-binding capacity to several antigens were profoundly reduced after gamma irradiation. In this study, we evaluated the changes of allergenecity and cytokine production profiles after exposure of irradiated PN extract in a PN-allergy mouse model. Mice were sensitized to PN extract by intragastric administration on days 0, 1, 2, and 7, and then challenged on day 21. Four weeks later, we evaluated the cytokine production patterns and proliferation responses of splenocytes that were stimulated with intact PN extract, compared to 10 and 50 kGy irradiated PN extract. When the cells were stimulated with 10 kGy of irradiated PN extract, a higher level of production of IFN-γ and IL-10 cytokines was observed. However, stimulation with 50 kGy of irradiated PN extract resulted in a higher level of production of only IFN-γ cytokines. In addition, the Th1/Th2 ratio increased in response to treatment with gamma-irradiated PNs. The results of this study show that the allergenicity of PN extracts could be reduced by gamma irradiation which caused downregulation of Th2 lymphocyte activity in the PN-sensitized mice.

  9. Standard skin prick testing and sensitization to inhalant allergens across Europe--a survey from the GALEN network

    NARCIS (Netherlands)

    Heinzerling, L.; Frew, A. J.; Bindslev-Jensen, C.; Bonini, S.; Bousquet, J.; Bresciani, M.; Carlsen, K.-H.; van Cauwenberge, P.; Darsow, U.; Fokkens, W. J.; Haahtela, T.; van Hoecke, H.; Jessberger, B.; Kowalski, M. L.; Kopp, T.; Lahoz, C. N.; Lodrup Carlsen, K. C.; Papadopoulos, N. G.; Ring, J.; Schmid-Grendelmeier, P.; Vignola, A. M.; Wöhrl, S.; Zuberbier, T.

    2005-01-01

    Skin prick testing (SPT) is the standard method for diagnosing allergic sensitization but is to some extent performed differently in clinical centres across Europe. There would be advantages in harmonizing the standard panels of allergens used in different European countries, both for clinical

  10. Allergen-Removed Rhus verniciflua Extract Induces Ovarian Cancer Cell Death via JNK Activation.

    Science.gov (United States)

    Kang, Se-Hui; Hwang, In-Hu; Son, Eunju; Cho, Chong-Kwan; Choi, Jong-Soon; Park, Soo-Jung; Jang, Byeong-Churl; Lee, Kyung-Bok; Lee, Zee-Won; Lee, Jong Hoon; Yoo, Hwa-Seung; Jang, Ik-Soon

    2016-01-01

    Nuclear factor-[Formula: see text]B (NF-[Formula: see text]B)/Rel transcription factors are best known for their central roles in promoting cell survival in cancer. NF-[Formula: see text]B antagonizes tumor necrosis factor (TNF)-[Formula: see text]-induced apoptosis through a process involving attenuation of the c-Jun-N-terminal kinase (JNK). However, the role of JNK activation in apoptosis induced by negative regulation of NF-[Formula: see text]B is not completely understood. We found that allergen-removed Rhus verniciflua Stokes (aRVS) extract-mediated NF-[Formula: see text]B inhibition induces apoptosis in SKOV-3 ovarian cancer cells via the serial activation of caspases and SKOV-3 cells are most specifically suppressed by aRVS. Here, we show that in addition to activating caspases, aRVS extract negatively modulates the TNF-[Formula: see text]-mediated I[Formula: see text]B/NF-[Formula: see text]B pathway to promote JNK activation, which results in apoptosis. When the cytokine TNF-[Formula: see text] binds to the TNF receptor, I[Formula: see text]B dissociates from NF-[Formula: see text]B. As a result, the active NF-[Formula: see text]B translocates to the nucleus. aRVS extract (0.5[Formula: see text]mg/ml) clearly prevented NF-[Formula: see text]B from mobilizing to the nucleus, resulting in the upregulation of JNK phosphorylation. This subsequently increased Bax activation, leading to marked aRVS-induced apoptosis, whereas the JNK inhibitor SP600125 in aRVS extract treated SKOV-3 cells strongly inhibited Bax. Bax subfamily proteins induced apoptosis through caspase-3. Thus, these results indicate that aRVS extract contains components that inhibit NF-[Formula: see text]B signaling to upregulate JNK activation in ovarian cancer cells and support the potential of aRVS as a therapeutic agent for ovarian cancer.

  11. Allergens in veterinary medicine.

    Science.gov (United States)

    Mueller, R S; Janda, J; Jensen-Jarolim, E; Rhyner, C; Marti, E

    2016-01-01

    Allergic diseases in animals are increasingly gaining importance in veterinary practice and as research models. For intradermal testing and allergen immunotherapy, a good knowledge of relevant allergens for the individual species is of great importance. Currently, the knowledge about relevant veterinary allergens is based on sensitization rates identified by intradermal testing or serum testing for allergen-specific IgE; crude extracts are the basis for most evaluations. Only a few studies provide evidence about the molecular structure of (particularly) dust mite, insect and mould allergens in dogs and horses, respectively. In those species, some major allergens differ from those in humans. This position paper summarizes the current knowledge about relevant allergens in dogs, cats and horses. © 2015 The Authors Allergy Published by John Wiley & Sons Ltd.

  12. [Study on safty of standardized specific mite-allergen immunotherapy to children with allergic rhinitis and/or asthma].

    Science.gov (United States)

    Wu, Yabin; Long, Zhen; Huang, Yang; Huang, Xuanzhao

    2011-07-01

    To evaluate the adverse reaction of standardized specific mite-allergen immunotherapy. One hundred and fifty-two patients diagnosed by the pediatric immunotherapy center of our hospital were treated with increasing doses of standardized specific mite-allergen injection. Before and 30 minutes after treatment, the peak expiratory flow (PEF) and pulmonary function for the maximum lung ventilation function were checked, and the adverse reactions were recorded. Six hundred and eighty-one injections were recorded. 84 injections (12.3%) caused immediate side effects, including 64 mild local adverse reactions (9.4%), 2 moderate local adverse reactions (0.3%), 18 systemic adverse reactions (2.6%) which were mild asthma, and no fatal anaphylactic shock and other serious adverse reactions were found. 50 injections (7.3%) cased delayed adverse reactions, all of which were mild local adverse reactions. The rate of immediate local adverse reactions and systemic adverse reactions in the maintenance treatment period was significantly higher than that in the initial treatment period (chi2 = 4.59, 19.82 respectively; P 0.05). The PEF change rate (-0.000 2 +/- 0.085 9) of the children at 681 injections and the MMEF change rate of the children at 109 injections (0.275 +/- 0.206) were not statistically different (t = -0.047, 1.39; P = 0.963, 0.166). Standardized specific mite-allergen immunotherapy is safe for children with allergic rhinitis and/or asthma.

  13. EFFECTIVENESS OF SUBCUTANEOUS ALLERGEN-SPECIFIC IMMUNOTHERAPY WITH TREE POLLEN ALLERGEN EXTRACT ADSORBED ON CALCIUM PHOSPHATE IN CHILDREN WITH ALLERGIC RHINOCONJUNCTIVITIS: RESULTS OF A 2 YEAR-LONG OBSERVATION

    Directory of Open Access Journals (Sweden)

    N. V. Shakhova

    2014-01-01

    Full Text Available The study was aimed at evaluating effectiveness and safety of subcutaneous allergen-specific immunotherapy (scASIT with tree pollen allergen extract adsorbed on calcium phosphate at allergic rhinoconjunctivitis in children. Patients and methods: open-label prospective study of 50 children and adolescents (5-17 years of age with allergic rhinoconjunctivitis caused by high sensitivity to tree pollen allergens. The first group involved the patients undergoing scASIT for 2 years 6 months (n = 23, the control group – the patients (n = 27 not undergoing any specific immunotherapy. Results: scASIT was accompanied by a statistically significant (in comparison with the control group reduction in intensity of rhinoconjunctivitis symptoms (6.1 ± 3.1; 11.8 ± 4.5; p = 0.00002, reduction in the use of symptomatic drugs (1.0 ± 0.4; 1.8 ± 0.3; p = 0.000004 and improvement of quality of all spheres of children’s life – physical (p = 0.001, social (p = 0.04, emotional (p = 0.001 and role functioning (p = 0.03. Systemic side reactions were not observed in the patients. Local reactions were observed in 23% of all allergen injections. Conclusions: the authors established high effectiveness and safety of scASIT with tree pollen allergen extract adsorbed on calcium phosphate suspension at allergic rhinoconjunctivitis in children. 

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Api m 10, a genuine A. mellifera venom allergen, is clinically relevant but underrepresented in therapeutic extracts.

    Science.gov (United States)

    Blank, S; Seismann, H; Michel, Y; McIntyre, M; Cifuentes, L; Braren, I; Grunwald, T; Darsow, U; Ring, J; Bredehorst, R; Ollert, M; Spillner, E

    2011-10-01

    Generalized systemic reactions to stinging hymenoptera venom constitute a potentially fatal condition in venom-allergic individuals. Hence, the identification and characterization of all allergens is imperative for improvement of diagnosis and design of effective immunotherapeutic approaches. Our aim was the immunochemical characterization of the carbohydrate-rich protein Api m 10, an Apis mellifera venom component and putative allergen, with focus on the relevance of glycosylation. Furthermore, the presence of Api m 10 in honeybee venom (HBV) and licensed venom immunotherapy preparations was addressed. Api m 10 was produced as soluble, aglycosylated protein in Escherichia coli and as differentially glycosylated protein providing a varying degree of fucosylation in insect cells. IgE reactivity and basophil activation of allergic patients were analyzed. For detection of Api m 10 in different venom preparations, a monoclonal human IgE antibody was generated. Both, the aglycosylated and the glycosylated variant of Api m 10 devoid of cross-reactive carbohydrate determinants (CCD), exhibited IgE reactivity with approximately 50% of HBV-sensitized patients. A corresponding reactivity could be documented for the activation of basophils. Although the detection of the native protein in crude HBV suggested content comparable to other relevant allergens, three therapeutical HBV extracts lacked detectable amounts of this component. Api m 10 is a genuine allergen of A. mellifera venom with IgE sensitizing potential in a significant fraction of allergic patients independent of CCD reactivity. Thus, Api m 10 could become a key element for component-resolved diagnostic tests and improved immunotherapeutic approaches in hymenoptera venom allergy. © 2011 John Wiley & Sons A/S.

  16. Immunological and physical properties of allergen solutions. Effects of nebulization

    DEFF Research Database (Denmark)

    Frølund, L; Poulsen, L K; Heinig, J H

    1991-01-01

    Lyophilised birch pollen allergen extracts, reconstituted with different diluents (H2O, saline, Albumin diluent (AD] were investigated to determine whether the allergen activity and quality of the extracts deteriorated by nebulization with different nebulizers (Pari, Wright, and Sandoz). Allergen...

  17. Inter-laboratory optimization of protein extraction, separation, and fluorescent detection of endogenous rice allergens

    DEFF Research Database (Denmark)

    Satoh, Rie; Teshima, Reiko; Kitta, Kazumi

    2016-01-01

    In rice, several allergens have been identified such as the non-specific lipid transfer protein-1, the α-amylase/trypsin-inhibitors, the α-globulin, the 33 kDa glyoxalase I (Gly I), the 52-63 kDa globulin, and the granule-bound starch synthetase. The goal of the present study was to define optimal...

  18. Safety of accelerated schedules of subcutaneous allergen immunotherapy with house dust mite extract in patients with atopic dermatitis.

    Science.gov (United States)

    Kim, Myoung-Eun; Kim, Jeong-Eun; Sung, Joon-Mo; Lee, Jin-Woo; Choi, Gil-Soon; Nahm, Dong-Ho

    2011-09-01

    The safety of accelerated schedules of allergen immunotherapy (ASAI) in patients with bronchial asthma (BA) has been reported but there are little data on the safety of ASAI for patients with atopic dermatitis (AD). In this study, we investigated the safety of ASAI in patients with AD. Sixty patients with AD and 18 patients with BA sensitized to house dust mites (HDM) were studied. A maximum maintenance dose of HDM extract, adsorbed to aluminum hydroxide, was administered to patients by subcutaneous injection with either a 3-day protocol (rush immunotherapy) or 1-day protocol (ultra-rush immunotherapy). Systemic reactions were observed 4 of 15 patients (26.7%) with AD during rush immunotherapy, 13 of 45 patients (28.9%) with AD during ultra-rush immunotherapy, and 4 of 18 patients (22.2%) with BA during rush immunotherapy (P > 0.05). No severe or near fatal systemic reactions occurred in 78 subjects of this study. Systemic reactions developed within 4 hr after administration of the maximum allergen dose in 20 of 21 patients (95.2%) with AD and BA who showed systemic reactions during rush or ultra-rush immunotherapy. In conclusion, ASAI was safe and well tolerated in patients with AD. ASAI can be a useful therapeutic option for AD.

  19. Efficacy and safety of subcutaneous allergen-specific immunotherapy with depigmented polymerized mite extract in atopic dermatitis.

    Science.gov (United States)

    Novak, Natalija; Bieber, Thomas; Hoffmann, Matthias; Fölster-Holst, Regina; Homey, Bernhard; Werfel, Thomas; Sager, Angelika; Zuberbier, Torsten

    2012-10-01

    Exposure to house dust mites (HDMs) aggravates the course of atopic dermatitis (AD) in patients sensitized to HDMs. This study investigated the efficacy and safety of subcutaneous allergen-specific immunotherapy with the use of depigmented polymerized mite extract as an add-on therapy to basic (ie, topical and, as necessary, systemic) medication. Patients (n = 168) were recruited in a randomized, double-blind, placebo-controlled parallel group phase III study conducted in Germany (21 sites), in adult patients with AD aggravated by HDMs. The primary end points of the study were the assessments of the area under the curves of the total Severity Scoring Atopic Dermatitis (SCORAD) score and of the use of basic medication during the 18-month treatment period. Post hoc subgroup analyses were also performed. Overall efficacy analysis of the intention-to-treat and per-protocol study populations showed no statistically significant differences between the active treatment and placebo groups. However, the subgroup of patients with severe AD (SCORAD > 50) showed a statistically significant reduction of the median total SCORAD by 18% (P = .02) compared with placebo. The frequency of adverse reactions was similar in both groups, suggesting the safety of the active treatment. Although subcutaneous allergen-specific immunotherapy showed no statistically significant difference in the overall population of patients with AD, statistically significant reduction of the total SCORAD could be achieved in a subgroup of patients with severe AD. Copyright © 2012 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

  20. The Consolidated Standards of Reporting Trials (CONSORT) Statement applied to allergen-specific immunotherapy with inhalant allergens: a Global Allergy and Asthma European Network (GA(2)LEN) article.

    Science.gov (United States)

    Bousquet, Philippe J; Calderón, Moisés A; Demoly, Pascal; Larenas, Désirée; Passalacqua, Giovanni; Bachert, Claus; Brozek, Jan; Canonica, G Walter; Casale, Thomas; Fonseca, Joao; Dahl, Ronald; Durham, Stephen R; Merk, Hans; Worm, Margitta; Wahn, Ulrich; Zuberbier, Torsten; Schünemann, Holger J; Bousquet, Jean

    2011-01-01

    Randomized trials provide evidence to inform treatment decisions. The Consolidated Standards of Reporting Trials (CONSORT) Statement is a set of recommendations for the reporting of trials. We sought to assess the quality of reporting allergen-specific immunotherapy trials according to CONSORT criteria. The reporting of the procedure, randomization, dropouts, strict conduct of intention-to-treat (ITT) analysis, and sample size calculation according to CONSORT were assessed in the 46 subcutaneous and 48 sublingual immunotherapy (SLIT) blind, placebo-controlled randomized trials published between 1996 and 2009 in English. One subcutaneous immunotherapy (2.2%) and 3 SLIT (6.6%) trials met CONSORT Statement criteria. These were used for the registration of sublingual tablets to the European Medicines Agency. In subcutaneous immunotherapy, 16 (35%) studies reported a CONSORT flow chart, and 12 (26%) provided a description of dropouts. Adequate randomization was reported in 9 (35%) studies, and incomplete randomization was reported in 15 (33%). Power analysis was reported in 15 (33%) studies. In SLIT, 20 (42%) studies reported a CONSORT flow chart, and 16 (32%) a description of dropouts. ITT analysis was carried out in 1 (2.2%) SLIT study, and a modified ITT analysis was used in 1 (2.2%) subcutaneous immunotherapy study and 2 (4.4%) SLIT studies. Adequate randomization was reported in 6 (12%) studies, and incomplete randomization was reported in 16 (32%). Power analysis was reported in 15 (27%) studies. As in other areas of medicine, the quality of reporting of most immunotherapy trials is low, and only 4.2% of SLIT randomized controlled trials met all of the criteria of the CONSORT Statement. Use of the CONSORT criteria should be encouraged. Copyright © 2010 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

  1. An alternative inhibition method for determining cross-reactive allergens.

    Science.gov (United States)

    Schmidt-Hieltjes, Yvonne; Teodorowicz, Malgorzata; Jansen, Ad; den Hartog, Gerco; Elfvering-Berendsen, Lisette; de Jong, Nicolette W; Savelkoul, Huub F J; Ruinemans-Koerts, Janneke

    2017-02-01

    Inhibition assays are an useful tool to identify the allergen of primary sensitization of cross-reactive allergens. Classical ELISA-based inhibition assays are limited by both the availability of commercial standardized allergen extracts and the experience and knowledge needed for making home-made extracts. Moreover the direct comparison of the inhibition ELISAs outcomes between different laboratories is difficult because of different sources of used allergen extracts and a number of methodological variations. Therefore, we propose a novel ImmunoCap (Phadia, Thermofisher Scientific) based immunoinhibition method with the use of commercially available Caps as the allergen source. The novel ImmunoCap based immunoinhibition method was developed and tested with sera from patients with a well-known cross-reactive sensitization for fig (Ficus carica) and ficus (Ficus benjamina). Results were compared with a classically applied inhibition method, i.e. addition of homemade allergen extract to patient serum. The amount of allergens (fig and ficus extracts) needed to reach a similar degree of inhibition was comparable for both inhibition methods. The ImmunoCap based inhibition assay, in addition to classical inhibition methods, is a valuable tool as the ImmunoCap analyzer and commercial allergens (Caps) are more widely available which makes the outcomes of inhibition tests comparable between different laboratories. Furthermore, in the ImmunoCap inhibition method the same protein source is used for both the inhibition of sIgE and sIgE measurement, which might be even more relevant when multiple cross-reactive allergens are tested.

  2. Comparative study of IQ-ultra and Finn Chambers test methodologies in detecting 10 common standard allergens that cause allergic contact dermatitis.

    Science.gov (United States)

    Doumit, Joseph; Pratt, Melanie

    2012-01-01

    Patch testing is routinely used in contact dermatitis clinics because it is the gold standard for the evaluation of potential allergic contact dermatitis. The study was undertaken to evaluate possible differences in reactivity between the Finn Chamber and IQ-Ultra patch-testing methodologies. Patients were patch-tested simultaneously with the Finn Chamber and IQ-Ultra patch tests. Ten standard allergens set by the North American Contact Dermatitis Group were used for both techniques. Both patch tests had a significant agreement in detecting all of the allergens. An "almost perfect agreement" was noted for ethylenediamine dihydrochloride, quaternium-15, mercapto mix, black rubber mix, balsam of Peru, and nickel sulfate; "substantial agreement" for formaldehyde, bisphenol A epoxy resin, and 4-tert-butylphenol formaldehyde resin; and "moderate agreement" for potassium dichromate. The Finn Chamber and IQ-Ultra patch tests had a good agreement in the detection of the 10 standard allergens that were tested.

  3. 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 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.

  4. [Hypersensitivity to allergens from the panels standard I and II in automobile tire manufacturers at Stomil plant].

    Science.gov (United States)

    Rubisz-Brzezińska, J; Bendkowski, W; Krauze, E; Suwała-Jurczyk, B

    1990-01-01

    Dermatological examination and patch tests were done in 114 manufacturers of tires, 78 women and 36 men, mean age 29 years, working in Stomil Plant. Patch tests included 15 substances in panels Standard I and II (excluding rubber components). The study demonstrated hypersensitivity to nickel and formalin (4.3% each), chromium and cobalt (3.5% each), and turpentine (2.6%), while hypersensitivity to other allergens was observed sporadically. A comparative analysis of the results of patch tests with antigens "Standard I and II" in confectioners and healthy and allergic persons showed that the per cent of positive tests in rubber industry workers was higher than in healthy subjects, and much lower than in patients with contact dermatitis.

  5. Inhaled allergen bronchoprovocation tests.

    Science.gov (United States)

    Diamant, Zuzana; Gauvreau, Gail M; Cockcroft, Don W; Boulet, Louis-Philippe; Sterk, Peter J; de Jongh, Frans H C; Dahlén, Barbro; O'Byrne, Paul M

    2013-11-01

    The allergen bronchoprovocation test is a long-standing exacerbation model of allergic asthma that can induce several clinical and pathophysiologic features of asthma in sensitized subjects. Standardized allergen challenge is primarily a research tool, and when properly conducted by qualified and experienced investigators, it is safe and highly reproducible. In combination with validated airway sampling and sensitive detection techniques, allergen challenge allows the study of several features of the physiology of mainly TH2 cell-driven asthma in relation to the kinetics of the underlying airway pathology occurring during the allergen-induced late response. Furthermore, given the small within-subject variability in allergen-induced airway responses, allergen challenge offers an adequate disease model for the evaluation of new (targeted) controller therapies for asthma in a limited number of subjects. In proof-of-efficacy studies thus far, allergen challenge showed a fair positive predicted value and an excellent negative predictive value for the actual clinical efficacy of new antiasthma therapies, underscoring its important role in early drug development. In this review we provide recommendations on challenge methods, response measurements, sample size, safety, and harmonization for future applications. Copyright © 2013 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

  6. The relative allergenicity of Stachybotrys chartarum compared to house dust mite extracts in a mouse model

    Science.gov (United States)

    A report by the Institute of Medicine suggested that more research is needed to better understand mold effects on allergic disease, particularly asthma development. The authors compared the ability of the fungus Stachybotrys chartarum (SCE) and house dust mite (HDM) extracts to i...

  7. Influence of enzymatic hydrolysis on the allergenicity of roasted peanut protein extract.

    Science.gov (United States)

    Cabanillas, Beatriz; Pedrosa, Mercedes M; Rodríguez, Julia; Muzquiz, Mercedes; Maleki, Soheila J; Cuadrado, Carmen; Burbano, Carmen; Crespo, Jesús F

    2012-01-01

    Peanut allergy is recognized as one of the most severe food allergies. Some studies have investigated the effects of enzymatic treatments on the in vitro immunological reactivity of members of the Leguminosae family, such as the soybean, chickpea and lentil. Nevertheless, there are only a few studies carried out with sera from patients with a well-documented allergy. Roasted peanut protein extract was hydrolyzed by the sequential and individual action of 2 food-grade enzymes, an endoprotease (Alcalase) and an exoprotease (Flavourzyme). Immunoreactivity to roasted peanut extract and hydrolyzed samples was evaluated by means of IgE immunoblot, ELISA and 2-dimensional electrophoresis using sera from 5 patients with a clinical allergy to peanuts and anti-Ara h 1, anti-Ara h 2 and anti-Ara h 3 immunoblots. Immunoblot and ELISA assays showed an important decrease of IgE reactivity and Ara h 1, Ara h 2 and Ara h 3 levels in the first 30 min of hydrolyzation with Alcalase. In contrast, individual treatment with Flavourzyme caused an increase in IgE reactivity detected by ELISA at 30 min and led to a 65% inhibition of IgE reactivity at the end of the assay (300 min). Ara h 1 and the basic subunit of Ara h 3 were still recognized after treatment with Flavourzyme for 300 min. Hydrolysis with the endoprotease Alcalase decreases IgE reactivity in the soluble protein fraction of roasted peanut better than hydrolysis with the exoprotease Flavourzyme. Copyright © 2011 S. Karger AG, Basel.

  8. Cigarette smoke facilitates allergen penetration across respiratory epithelium.

    Science.gov (United States)

    Gangl, K; Reininger, R; Bernhard, D; Campana, R; Pree, I; Reisinger, J; Kneidinger, M; Kundi, M; Dolznig, H; Thurnher, D; Valent, P; Chen, K-W; Vrtala, S; Spitzauer, S; Valenta, R; Niederberger, V

    2009-03-01

    The association between cigarette smoke exposure and allergic airway disease is a matter for debate. We sought to investigate in an in vitro system whether active smoking reduces the integrity and barrier function of the respiratory epithelium and thus facilitates allergen penetration. We cultured the human bronchial epithelial cell line 16HBE14o- in a transwell culture system as a surrogate for the intact respiratory epithelium. The cell monolayer was exposed to standardized cigarette smoke extract (CSE). The extent and effects of trans-epithelial allergen penetration were measured using 125I-labelled purified major respiratory allergens (rBet v 1, rPhl p 5 and rDer p 2) and histamine release experiments. Exposure of cells to concentrations of CSE similar to those found in smokers induced the development of para-cellular gaps and a decrease in trans-epithelial resistance. CSE exposure induced a more than threefold increase in allergen penetration. Increased subepithelial allergen concentrations provoked a substantial augmentation of histamine release from sensitized basophils. Our results indicate that cigarette smoke is a potent factor capable of reducing the barrier function of the respiratory epithelium for allergens and may contribute to increased allergic inflammation, exacerbation of allergic disease and boosting of IgE memory.

  9. Archive of Census Related Products (ACRP): 1990 Standard Extract Files

    Data.gov (United States)

    National Aeronautics and Space Administration — The 1990 Standard Extract Files portion of the Archive of Census Related Products (ACRP) contains population and housing data derived from the U.S. Census Bureau's...

  10. 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.

  11. Treatment of grass pollen allergy: focus on a standardized grass allergen extract – Grazax®

    OpenAIRE

    Calderón, Moisés

    2008-01-01

    Moisés Calderón1, Tove Brandt21Section of Allergy and Clinical Immunology, Royal Brompton Hospital, Imperial College, NHLI, London, UK; 2Group Clinical Development, ALK-Abelló A/S, Hørsholm, DenmarkAbstract: Immunotherapy is the only treatment for allergy that has the potential to alter the natural course of the disease. Sublingual immunotherapy (SLIT) for grass pollen-induced rhino-conjunctivitis has been developed to make immunotherapy available t...

  12. Impact of the vulcanization process on the structural characteristics and IgE recognition of two allergens, Hev b 2 and Hev b 6.02, extracted from latex surgical gloves.

    Science.gov (United States)

    Galicia, Christian; Mendoza-Hernández, Guillermo; Rodríguez-Romero, Adela

    2015-06-01

    Latex allergy is a health problem that mainly affects medical environments, causing anaphylactic shocks in extreme cases. Sensitization and reactions to this material is closely linked to the use of latex gloves. The objective of this study was to purify two of the major allergens from latex surgical gloves to study the biochemical and structural changes that could be generated during the product manufacture and to compare their IgE recognition with the non-processed allergens. Glycosylated allergen Hev b 2 (β-1,3-glucanase) and Hev b 6.02 (hevein) were purified from glove extracts using affinity (Concanavalin A) and reversed-phase chromatographies, respectively. ELISA experiments were performed with both proteins and sera from allergic patients to assess the IgE recognition, which was heterogeneous. Crystallographic methods were used to obtain the 3D structure of Hev b 6.02 from surgical gloves, which did not show evident modification when compared with the protein from the natural non-processed form. Despite having the same crystallographic structure, the IgE from some patients showed different recognition when the glove and the natural allergen were used in ELISA. Furthermore, using electrophoretic techniques, we identified three forms of Hev b 2: one corresponding to the complete polypeptide chain with posttranslational modifications, and two glycosylated fragments. The mixture of these three forms showed stronger recognition by IgE from latex-allergic patients than the pure non-processed allergen. In conclusion, IgE from subjects sensitized to latex products showed different recognition between the allergens obtained from a natural source and the processed material, even when the structure was maintained. This demonstrates the importance of using processed allergens in further investigations of diagnosis, prevalence, product allergenicity, and therapies. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Improving the extraction of Ara h 6 (a peanut allergen) from a chocolate-based matrix for immunosensing detection: Influence of time, temperature and additives.

    Science.gov (United States)

    Alves, Rita C; Pimentel, Filipa B; Nouws, Henri P A; Silva, Túlio H B; Oliveira, M Beatriz P P; Delerue-Matos, Cristina

    2017-03-01

    The extraction of Ara h 6 (a peanut allergen) from a complex chocolate-based food matrix was optimized by testing different temperatures, extraction times, and the influence of additives (NaCl and skimmed milk powder) in a total of 36 different conditions. Analyses were carried out using an electrochemical immunosensor. Three conditions were selected since they allowed the extraction of the highest levels of Ara h 6. These extractions were performed using 2g of sample and 20ml of Tris-HNO 3 (pH=8) containing: a) 0.1M NaCl and 2g of skimmed milk powder at 21°C for 60min; b) 1M NaCl and 1g of skimmed milk powder at 21°C for 60min; and c) 2g of skimmed milk powder at 60°C for 60min. Recoveries were similar or higher than 94.7%. This work highlights the importance to adjust extraction procedures regarding the target analyte and food matrix components. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Growth inhibitory and apoptosis-inducing effects of allergen-free Rhus verniciflua Stokes extract on A549 human lung cancer cells.

    Science.gov (United States)

    Jang, Ik-Soon; Park, Jae-Woo; Jo, Eun-Bi; Cho, Chong-Kwan; Lee, Yeon-Weol; Yoo, Hwa-Seung; Park, Junsoo; Kim, Jihye; Jang, Byeong-Churl; Choi, Jong-Soon

    2016-11-01

    Evidence suggests that Rhus verniciflua Stokes (RVS) or its extract has the potential to be used for the treatment of inflammatory and neoplastic diseases. However, direct use of RVS or its extract as a herbal medicine has been limited due to the presence of urushiol, an allergenic toxin. In the present study, we prepared an extract of the allergen‑removed RVS (aRVS) based on a traditional method and investigated its inhibitory effect on the growth of various types of human cancer cells, including lung (A549), breast (MCF-7) and prostate (DU-145) cancer cell lines. Notably, among the cell lines tested, treatment with the aRVS extract strongly inhibited proliferation of the A549 cells at a 0.5 mg/ml concentration for 24 h that was not cytotoxic to normal human dermal fibroblasts. Furthermore, aRVS extract treatment largely reduced the survival and induced apoptosis of the A549 cells. At the mechanistic levels, treatment with the aRVS extract led to the downregulation of Bcl-2 and Mcl-1 proteins, the activation of caspase-9/-3 proteins, an increase in cytosolic cytochrome c levels, the upregulation of Bax protein, an increase in phosphorylated p53 protein but a decrease in phosphorylated S6 protein in the A549 cells. Importantly, treatment with z-VAD‑fmk, a pan-caspase inhibitor attenuated aRVS extract-induced apoptosis in the A549 cells. These results demonstrate firstly that aRVS extract has growth inhibitory and apoptosis-inducing effects on A549 human lung cancer cells through modulation of the expression levels and/or activities of caspases, Bcl-2, Mcl-1, Bax, p53 and S6.

  15. Are cutaneous hypersensitivity tests to inhalant allergens a severity marker for vernal keratoconjunctivitis?

    Science.gov (United States)

    Augusto de Oliveira, Lauro; Mallozi, Marcia Carvalho; Sole, Dirceu; Freitas, Denise de; Sousa, Luciene Barbosa de; Mannis, Mark J

    2007-01-01

    The purpose of this study was to analyze the cutaneous sensitivity to a variety of allergens in patients with vernal keratoconjunctivitis (VKC) and to demonstrate the relation between skin response and clinical aspects of the disease. Twenty patients with vernal keratoconjunctivitis were randomly chosen from the External Disease and Cornea Sector. They were clinically evaluated, and a score ranging from 0 to 20 was applied based on signs and symptoms on ophthalmic examination. All subjects underwent a skin prick test against standardized allergens, such as house dust mites D. pteronyssinus, D. farinae, and Blomia tropicalis, as well as allergens from cat, dog, fungi and feather. Seventy-five per cent of patients were positive for at least one of the allergens tested. House dust mites were responsible for the majority of the cases (75%). There was a poor correlation between the clinical score and sensitivity to allergens (r= - 0.119 for fungi; r= - 0.174 for dog; r= - 0.243 for house dust mites; r= - 0.090 for feather). A significant correlation was found only for cat allergen extract (r = - 0.510; p=0.024). Our study demonstrated poor correlation between cutaneous hypersensitivity tests and clinical findings in patients with vernal keratoconjuntivitis. We concluded that skin response to inhalant allergens is not a useful test to identify clinical severity and chronicity of inflammatory process in this disease.

  16. Standard Setting for the Extractive Industries: A Critical Examination

    Directory of Open Access Journals (Sweden)

    Corinne Cortese

    2007-09-01

    Full Text Available This study examines the players involved in the setting of an international accountingstandard for the extractive industries. Publicly available data is used to exposeconnections between key constituents involved in the process, to enhance understandingof how the international accounting standard setting process occurred, and to identifyfuture research possibilities.

  17. The CREATE project: development of certified reference materials for allergenic products and validation of methods for their quantification

    NARCIS (Netherlands)

    van Ree, R.; Chapman, M. D.; Ferreira, F.; Vieths, S.; Bryan, D.; Cromwell, O.; Villalba, M.; Durham, S. R.; Becker, W. M.; Aalbers, M.; André, C.; Barber, D.; Cistero Bahima, A.; Custovic, A.; Didierlaurent, A.; Dolman, C.; Dorpema, J. W.; Di Felice, G.; Eberhardt, F.; Fernandez Caldas, E.; Fernandez Rivas, M.; Fiebig, H.; Focke, M.; Fötisch, K.; Gadermaier, G.; Das, R. G.; Gonzalez Mancebo, E.; Himly, M.; Kinaciyan, T.; Knulst, A. C.; Kroon, A. M.; Lepp, U.; Marco, F. M.; Mari, A.; Moingeon, P.; Monsalve, R.; Neubauer, A.; Notten, S.; Ooievaar-de Heer, P.; Pauli, G.; Pini, C.; Purohit, A.; Quiralte, J.; Rak, S.; Raulf-Heimsoth, M.; San Miguel Moncin, M. M.; Simpson, B.; Tsay, A.; Vailes, L.; Wallner, M.; Weber, B.

    2008-01-01

    Allergen extracts have been used for diagnosis and treatment of allergy for around 100 years. During the second half of 20th century, the notion increasingly gained foothold that accurate standardization of such extracts is of great importance for improvement of their quality. As a consequence,

  18. Single-allergen sublingual immunotherapy versus multi-allergen subcutaneous immunotherapy for children with allergic rhinitis.

    Science.gov (United States)

    Wang, Zhong-Xi; Shi, Han

    2017-06-01

    It has always been controversial whether a single allergen performs better than multiple allergens in polysensitized patients during the allergen-specific immunotherapy. This study aimed to examine the clinical efficacy of single-allergen sublingual immunotherapy (SLIT) versus multi-allergen subcutaneous immunotherapy (SCIT) and to discover the change of the biomarker IL-4 after 1-year immunotherapy in polysensitized children aged 6-13 years with allergic rhinitis (AR) induced by house dust mites (HDMs). The AR polysensitized children (n=78) were randomly divided into two groups: SLIT group and SCIT group. Patients in the SLIT group sublingually received a single HDM extract and those in the SCIT group were subcutaneously given multiple-allergen extracts (HDM in combination with other clinically relevant allergen extracts). Before and 1 year after the allergen-specific immunotherapy (ASIT), the total nasal symptom scores (TNSS), total medication scores (TMS) and IL-4 levels in peripheral blood mononuclear cells (PBMCs) were compared respectively between the two groups. The results showed that the TNSS were greatly improved, and the TMS and IL-4 levels were significantly decreased after 1-year ASIT in both groups (SLIT group: P0.05; for TMS: P>0.05; for IL-4 levels: P>0.05). It was concluded that the clinical efficacy of single-allergen SLIT is comparable with that of multi-allergen SCIT in 6-13-year-old children with HDM-induced AR.

  19. 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...

  20. Immunochemical and biological quantification of peanut extract

    DEFF Research Database (Denmark)

    Poulsen, Lars K.; Pedersen, Mona H; Platzer, Michael

    2003-01-01

    Biological standardization of allergen extracts is one of the steps in the characterization of an extract. The gold standard for determination of biological potency is the skin prick test, but histamine release (HR) has been used as a convenient ex vivo method for analyzing a large number...

  1. Determination of fragrance allergens in indoor air by active sampling followed by ultrasound-assisted solvent extraction and gas chromatography-mass spectrometry.

    Science.gov (United States)

    Lamas, J Pablo; Sanchez-Prado, Lucia; Garcia-Jares, Carmen; Llompart, Maria

    2010-03-19

    Fragrances are ubiquitous pollutants in the environment, present in the most of household products, air fresheners, insecticides and cosmetics. Commercial perfumes may contain hundreds of individual fragrance chemicals. In addition to the widespread use and exposure to fragranced products, many of the raw fragrance materials have limited available health and safety data. Because of their nature as artificial fragrances, inhalation should be considered as an important exposure pathway, especially in indoor environments. In this work, a very simple, fast, and sensitive methodology for the analysis of 24 fragrance allergens in indoor air is presented. Considered compounds include those regulated by the EU Directive, excluding limonene; methyl eugenol was also included due to its toxicity. The proposed methodology is based on the use of a very low amount of adsorbent to retain the target compounds, and the rapid ultrasound-assisted solvent extraction (UAE) using a very low volume of solvent which avoids further extract concentration. Quantification was performed by gas chromatography coupled to mass spectrometry (GC-MS). The influence of main factors involved in the UAE step (type of adsorbent and solvent, solvent volume and extraction time) was studied using an experimental design approach to account for possible factor interactions. Using the optimized procedure, 0.2 m(-3) air are sampled, analytes are retained on 25 mg Florisil, from which they are extracted by UAE (5 min) with 2 mL ethyl acetate. Linearity was demonstrated in a wide concentration range. Efficiency of the total sampling-extraction process was studied at several concentration levels (1, 5 and 125 microg m(-3)), obtaining quantitative recoveries, and good precision (RSD<10%). Method detection limits were < or =0.6 microg m(-3). Finally, the proposed method was applied to real samples collected in indoor environments in which several of the target compounds were determined. Copyright 2010 Elsevier B

  2. 78 FR 78368 - Allergenic Products Advisory Committee; Notice of Meeting

    Science.gov (United States)

    2013-12-26

    ... efficacy of RAGWITEK, a short ragweed pollen allergen extract tablet for sublingual use, manufactured by Merck, indicated for immunotherapy for diagnosed ragweed pollen induced allergic rhinitis, with or...

  3. Alternaria alternata and its allergens: a comprehensive review.

    Science.gov (United States)

    Kustrzeba-Wójcicka, Irena; Siwak, Emilia; Terlecki, Grzegorz; Wolańczyk-Mędrala, Anna; Mędrala, Wojciech

    2014-12-01

    Alternaria alternata is mainly an outdoor fungus whose spores disseminate in warm, dry air, so in temperate climates, their count peaks in the summers. Alternaria may also be found in damp, insufficiently ventilated houses, where its allergenic properties cocreate the sick building syndrome. Mold-induced respiratory allergies and research on Alternaria both have a lengthy history: the first was described as early as 1698 and the second dates back to 1817. However, the two were only linked in 1930 when Alternaria spores were found to cause allergic asthma. The allergenic extracts from Alternaria hyphae and spores still remain in use but are variable and insufficiently standardized as they are often a random mixture of allergenic ingredients and coincidental impurities. In contrast, contemporary biochemistry and molecular biology make it possible to obtain pure allergen molecules. To date, 16 allergens of A. alternata have been isolated, many of which are enzymes: Alt a 4 (disulfide isomerase), Alt a 6 (enolase), Alt a 8 (mannitol dehydrogenase), Alt a 10 (alcohol dehydrogenase), Alt a 13 (glutathione-S-transferase), and Alt a MnSOD (Mn superoxide dismutase). Others have structural and regulatory functions: Alt a 5 and Alt a 12 comprise the structure of large ribosomal subunits and mediate translation, Alt a 3 is a molecular chaperone, Alt a 7 regulates transcription, Alt a NTF2 facilitates protein import into the nucleus, and Alt a TCTP acts like a cytokine. The function of four allergenic proteins, Alt a 1, Alt a 2, Alt a 9, and Alt a 70 kDa, remains unknown.

  4. Mammalian airborne allergens

    NARCIS (Netherlands)

    Aalberse, Rob C.

    2014-01-01

    Historically, horse dandruff was a favorite allergen source material. Today, however, allergic symptoms due to airborne mammalian allergens are mostly a result of indoor exposure, be it at home, at work or even at school. The relevance of mammalian allergens in relation to the allergenic activity of

  5. [Quality standard study on Mori Cortex liquid extract].

    Science.gov (United States)

    Liu, Mao-feng; Xie, Mei-ping; Li, Lan; Lu, An-qi; Shi, Jian-gong; Wang, Su-juan

    2015-05-01

    A reasonable and practicable quality standard was developed for mori liquid extract from different sources by TLC, HPLC and fingerprint technology. In TLC method, the compounds were separated on polyamide film using glacial acetic acid-water (1: 3) as mobile phase at a UV wavelength of 365 nm. All qualified samples had the spots of the same color as the control herb and substance. The RP-HPLC method was used to determine the content of mulberroside A with mobile phase of methanol-water (25: 75) at a wave-length of 326 nm. The mulberroside A was in good linear with a regression equation of Y = 46.965X (r = 0.999 6) in the range of 4.6 - 228 mg x L(-1). In 14 batches of samples, the mulberroside A in 4 batches of them was less than 0.5 g x L(-1), and was more than 2.0 g x L(-1) in the other batches. It was suggested that the content limit of mulberroside A should be no less than 1.5 g x L(-1). The HPLC fingerprints were evaluated by the similarities. It has found that the similarities of different mori liquid extracts were very low and the chemical diversity of mori cortex was the major factor of similarity. Moreover, the process impact was minimal. Thus the fingerprint was not included in this quality standard.

  6. 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

    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......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...

  7. Efficacy and safety of sublingual tablets of house dust mite allergen extracts in adults with allergic rhinitis

    NARCIS (Netherlands)

    Bergmann, Karl-Christian; Demoly, Pascal; Worm, Margitta; Fokkens, Wytske J.; Carrillo, Teresa; Tabar, Ana I.; Nguyen, Hélène; Montagut, Armelle; Zeldin, Robert K.

    2014-01-01

    Preliminary studies have suggested the efficacy of sublingual tablets of house dust mite (HDM) extracts in adults with allergic rhinitis. We sought to assess the efficacy and safety of 2 doses of HDM sublingual tablets over 1 treatment year and the subsequent immunotherapy-free year. Adults with

  8. Grass Pollen Allergens

    Science.gov (United States)

    Augustin, Rosa

    1959-01-01

    Heat and pH stability studies and experiments with organic solvents show that the A-antigens discussed in the preceding paper (Augustin, 1959c) are much more labile than the I- (`inner ring') antigens. Breakdown products and/or aggregates are produced which no longer precipitate with antisera to the original extracts, but act as inhibitors. Solutions of pollen allergens, on the other hand, are found to withstand even autoclaving for 15 min. at 20 atm. and vigorous boiling over the naked flame of a bunsen burner. None of the carbohydrates tested has a demonstrable effect on skin reactivity which is, however, destroyed by crystalline pepsin, crystalline trypsin, a crystalline mould protease and a tissue protease (a partially purified extract from rabbit spleen). It follows that the bulk of the allergens—if not all—are proteins. The relation of skin reactivity, immuno-electrophoretic patterns, carbohydrate and protein reactions to the selective destruction of the pollen antigens is investigated. Pollen components prove to have a somewhat wider range of electrophoretic mobilities than serum proteins and are probably as complicated a mixture. The most and least highly negatively charged components are without skin reactivity in allergic subjects. The skin reactive allergens appear to have the mobilities of α- and β-globulins. Not all the hay fever subjects react equally to all the components, and Cocksfoot and Timothy activity patterns vary in different subjects. ImagesFIG. 5 PMID:13795119

  9. Plasticizer, di(2-ethylhexyl)phthalate (DEHP) enhances cockroach allergen extract-driven airway inflammation by enhancing pulmonary Th2 as well as Th17 immune responses in mice.

    Science.gov (United States)

    Alfardan, Ali S; Nadeem, Ahmed; Ahmad, Sheikh F; Al-Harbi, Naif O; Al-Harbi, Mohammad M; AlSharari, Shakir D

    2018-03-19

    In recent decades, there has been a gradual increase in the prevalence of asthma. Various factors including environmental pollutants have contributed to this phenomenon. Plasticizer, di(2-ethylhexyl)phthalate (DEHP) is one of the commonest environmental pollutants due to its association with plastic products. DEHP gets released from plastic products easily leading to respiratory exposure in humans. As a consequence, DEHP is associated with allergic asthma in humans and animals. DEHP is reported to act as an adjuvant in ovalbumin-induced mouse models of asthma at high doses. However, these studies mostly looked into the role of DEHP on Th2 cytokines/eosinophilic inflammation without investigating the role of airway epithelial cells (AECs)/dendritic cells (DCs)/Th17 cells. Its adjuvant activity with natural allergens such as cockroach allergens at tolerable daily intake needs to be explored. Cockroach allergens and DEHP may be inhaled together due to their coexistence in work place as well as household environments. Therefore, effect of DEHP was assessed in cockroach allergens extract (CE)-induced mouse model of asthma. Airway inflammation, histopathology, mucus secretion, and immune responses related to Th2/Th17/DCs and AECs were assessed in mice with DEHP exposure alone and in combination with CE. Our study shows that DEHP converts CE-induced eosinophilic inflammation into mixed granulocytic inflammation by promoting Th2 as well as Th17 immune responses. This was probably due to downregulation of E-cadherin in AECs, and enhancement of costimulatory molecules (MHCII/CD86/CD40)/pro-inflammatory cytokines (IL-6/MCP-1) in DCs by DEHP. This suggests that DEHP facilitates development of mixed granulocytic airway inflammation in the presence of a natural allergen. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Food allergen digestibility: The influence on allergenicity

    DEFF Research Database (Denmark)

    Bøgh, Katrine Lindholm; Madsen, Charlotte Bernhard

    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...... 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...... 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...

  11. An investigation of the possible immunological relationship between allergen extracts from birch pollen, hazelnut, potato and apple

    DEFF Research Database (Denmark)

    Andersen, Klaus Ejner; Løwenstein, H

    1978-01-01

    In a retrospective study on selected group of patients, the coincidence of birch pollen allergy and a clinically relevant positive prick test reaction to apples and potatoes was confirmed. Immunochemical comparison using the crossed line immunoelectrophoresis technique (CLIE) confirmed partial...... identity between birch pollen and hazelnut. By the same method no partial immunological identity between birch pollen and extracts and fresh peel from apples and potatoes was found. However, both apples and potatoes gave rise to non-immunological affinity precipitates. On this basis it is discussed...

  12. Evaluation of stability of allergen extracts for sublingual immunotherapy during transport under unfavourable temperature conditions with an innovative thermal insulating packaging.

    Science.gov (United States)

    Puccinelli, P; Natoli, V; Dell'albani, I; Scurati, S; Incorvaia, C; Barbieri, S; Masieri, S; Frati, F

    2013-10-01

    Many pharmaceutical and biotechnological products are temperature-sensitive and should normally be kept at a controlled temperature, particularly during transport, in order to prevent the loss of their stability and activity. Therefore, stability studies should be performed for temperature-sensitive products, considering product characteristics, typical environmental conditions, and anticipating environmental extremes that may occur during product transport in a specific country. Staloral products for sublingual immunotherapy are temperature sensitive and are labelled for maintenance under refrigerated conditions (2-8°C). Given the peculiar climatic context of Italy and the great temperature fluctuations that may occur during transport, this study was aimed at evaluating the impact of a new engineered thermal insulating packaging for Staloral. In particular, the purpose was to assess whether the new packaging could create a container condition able to preserve the stability and immunological activity of the product during the transport phase throughout Italy. The results showed that the range of temperatures that can affect the product, in the area surrounding the product packaging, may reach a peak of 63°C during transport under the most unfavourable climatic conditions, i.e. in a non-refrigerated van during the summer season, from the site of production in France to the patient's house in Catania, the city with the highest temperatures in Italy. However, the highest temperature reached inside the vaccine did not exceed 45°C over a period of about 2 h. The ELISA inhibition test on samples subjected to the extreme temperature conditions previously defined (45°C) showed an immunological activity higher than 75% of that initially measured and was comparable to those obtained with samples stored at controlled temperature (5°C). This means that, even in the worst case scenario, the structure of the allergen extracts is not influenced and the vaccine potency is

  13. Taxonomy of Allergenic Fungi.

    Science.gov (United States)

    Levetin, Estelle; Horner, W Elliott; Scott, James A

    2016-01-01

    The Kingdom Fungi contains diverse eukaryotic organisms including yeasts, molds, mushrooms, bracket fungi, plant rusts, smuts, and puffballs. Fungi have a complex metabolism that differs from animals and plants. They secrete enzymes into their surroundings and absorb the breakdown products of enzyme action. Some of these enzymes are well-known allergens. The phylogenetic relationships among fungi were unclear until recently because classification was based on the sexual state morphology. Fungi lacking an obvious sexual stage were assigned to the artificial, now-obsolete category, "Deuteromycetes" or "Fungi Imperfecti." During the last 20 years, DNA sequencing has resolved 8 fungal phyla, 3 of which contain most genera associated with important aeroallergens: Zygomycota, Ascomycota, and Basidiomycota. Advances in fungal classification have required name changes for some familiar taxa. Because of regulatory constraints, many fungal allergen extracts retain obsolete names. A major benefit from this reorganization is that specific immunoglobulin E (IgE) levels in individuals sensitized to fungi appear to closely match fungal phylogenetic relationships. This close relationship between molecular fungal systematics and IgE sensitization provides an opportunity to systematically look at cross-reactivity and permits representatives from each taxon to serve as a proxy for IgE to the group. Copyright © 2016 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

  14. How far can we simplify in vitro diagnostics for grass pollen allergy?: A study with 17 whole pollen extracts and purified natural and recombinant major allergens

    NARCIS (Netherlands)

    van Ree, R.; van Leeuwen, W. A.; Aalberse, R. C.

    1998-01-01

    BACKGROUND: Current diagnostics for grass pollen allergy are composed of mixtures of pollen of different grass species. Their complex composition hampers accurate standardization. OBJECTIVE: The aim of the study was to investigate whether mixtures of grass pollen extracts can be replaced by a single

  15. 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.

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

    Directory of Open Access Journals (Sweden)

    S.M. Nedelska

    2017-03-01

    Full Text Available Background. Allergen-specific immunotherapy as elimination procedures is the only method of treatment and prevention of allergic disorders formation and exacerbation of clinical symptoms. One of the approaches to molecular diagnosis is choice of allergen for allergen-specific immunotherapy (ASIT. The purpose of our study was to identify possible reasons of failure of ASIT with pollen allergens (predominantly weeds in children with seasonal allergic rhinitis (hay fever and/or bronchial asthma based on studying hypersensitivity to the allergens, analysis of anamnestic data. Materials and methods. Allergy skin prick tests were conducted to 192 children (middle age 9.8 ± 2.7 years with the seasonal symptoms of rhinitis and/or bronchial asthma according to standard methodology with pollen allergens of Immunolog Ltd (Vinnytsia. We evaluated positive results as 5 mm and higher diameter of papula/hyperemia. The anamnestic survey was carried out in 52 patients by means of the questionnaire that contained questions about cross allergy (pollen-food, oral allergy syndrome, concomitant pathology of upper respiratory tract and effectiveness of АSІТ, which is elaborated by us. Results. The skin prick tests show that in 192 patients, who have hay fever, ragweed, sunflower sensibilization predominates (56 and 58 %, correspondingly. About 10–20 % of children are sensitive to cereals (ryegrass, fescue. To the allergens of poplar, acacia, couch-grass, oak, mint, nettle, walnut, the positive reactions of prick tests were observed in 3–7 children, that is 1.5–3.6 %. According to our results, 23 % of patients had sensibilization to 5 and more pollen allergens. 52 % of children had concomitant food allergy, 15 % of patients have reactions of the lips, oral cavity when using certain products, mostly tomatoes, nuts, seeds (they were diagnosed oral allergy syndrome. Also, one third of children have varying degrees of adenoid hypertrophy, tonsils hypertrophy

  17. 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.

  18. Allergen specificity is relevant for immunotherapy prescription in polysensitised children

    Directory of Open Access Journals (Sweden)

    Ciprandi Giorgio

    2012-09-01

    Full Text Available Abstract The sensitization to more allergens, such as polysenitization, is becoming a frequent characteristic of allergic patients, since the childhood. However, this phenomenon is considered an obstacle to prescribe immunotherapy by many doctors. This study investigated the relevance of polysensitization in a cohort of allergic children and evaluated the number of allergen extracts prescribed for these children. There are allergens that are frequent, but not prescribed. This issue should be matter of adequate debate for Italian paediatricians.

  19. Allergenicity and crossreactivity of buffalo grass ( Stenotaphrum ...

    African Journals Online (AJOL)

    Background. In the subtropical climate of South Africa, grasses of the subfamily Panicoideae are predominant. Bermuda grass has previously been shown to be an important local allergen, and immunoglobulin E (IgE) epitopes of Bermuda grass extracts are known to be distinct from those of the Pooid pollen extracts.

  20. Hypersensitivity to latex and Ficus benjamina allergens.

    Science.gov (United States)

    Delbourg, M F; Moneret-Vautrin, D A; Guilloux, L; Ville, G

    1995-12-01

    Association between allergy to Ficus benjamina and Hevea brasiliensis, two botanically unrelated plants, was suspected in consequence of two clinical observations. Symptoms were rhinitis and asthma. This study was undertaken to assess the in vivo and in vitro cross-reactivity between Ficus benjamina and Hevea brasiliensis allergens. The two patients were asked about use and contact with latex devices and relationship between symptoms and Ficus benjamina exposure. Skin prick tests were performed with Ficus benjamina, Hevea brasiliensis extracts and common allergens. Double-blind nasal and bronchial challenge tests were done using the rinse fluid from a brand of latex gloves. Total and specific IgE antibodies to Ficus benjamina and Hevea brasiliensis were determined. In vitro cross-reactivity was investigated by means of CAP RAST and immunodot inhibition experiments. We observed that for the first patient the primary phenomenon is probably allergy to latex followed by allergy to Ficus benjamina. For the second patient, allergy to Ficus benjamina was diagnosed (improvement related to the avoidance of exposure to Ficus benjamina allergens) and positivity to latex skin prick tests may be due to the cross-reacting allergens. In vitro assays showed specific IgE antibodies to both allergens and cross-reactivity was confirmed in the two cases by reciprocal inhibition of the two extracts. The increasing risk of sensitization to widely used latex devices and extensive exposure to Ficus species in households and offices indicates increased allergenic risk from this newly recognized cross-reactivity.

  1. Contamination of nebulizers with environmental allergens.

    Science.gov (United States)

    Bollinger, Mary E; Butz, Arlene; Mudd, Kim; Hamilton, Robert G

    2005-11-01

    A previous article described cockroach allergen in the nebulizer reservoir of an asthmatic patient who experienced a life-threatening exacerbation after nebulizer use. To determine whether indoor allergens can be measured in home nebulizers. As part of a large study examining nebulizer use in underserved asthmatic children, visiting nurses replaced nebulizer sets in patients' homes. Twenty used sets were randomly selected for analysis, without linkage to clinical or home environmental data. Nebulizer reservoirs and negative controls (buffer and albuterol) were extracted overnight with 2 mL of buffer. For positive controls, nebulizer sets were placed in homes with cats and dogs, and other reservoirs were intentionally contaminated with cat (Fel d 1), dog (Can f 1), cockroach (Bla g 1 and Bla g 2), and mouse (Mus m 1) skin test solutions. Extracts were tested for allergens in a masked manner using enzyme-linked immunosorbent assay. Of 17 reservoirs with adequate specimens for allergen detection, 5 (29%) had measurable levels for at least 1 of 5 allergens tested. One reservoir had measurable Can f 1, 2 had Bla g, 3 had Mus m 1, and none had Fel d 1 allergen. Two of 3 homes with cats where nebulizer setups were placed had measurable Fel d 1 in the reservoir, and 1 of 2 homes with dogs had measurable Can f 1. Reservoirs kept in sealed plastic bags had no detectable allergen. Indoor allergens can be found in the nebulizer equipment of children with asthma, with the potential for adverse consequences. Storing nebulizer sets in sealed plastic bags may prevent contamination.

  2. Standardization of DNA extraction from invasive alien weed ...

    African Journals Online (AJOL)

    Tuoyo Aghomotsegin

    DNA isolation from the weed, Parthenium hysterophorus is complicated due to the presence of high amount of allelochemicals in the form of secondary metabolites that causes hindrance in extraction and enzymatic reactions. A modified and efficient DNA extraction from P. hysterophorus leaf has been developed.

  3. 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...... found in immune complexes with a molecular weight of greater than 300 kdalton equivalent to 2 or more molecules of IgG. The possibility of employing a similar method with IgE instead of IgG for preparation of patient-related allergens instead of antigens, is discussed....

  4. Modelling allergenic risk

    DEFF Research Database (Denmark)

    Birot, Sophie

    of allergic reaction in the population. In allergen risk assessment, the emphasis was on the threshold data, and no effort was made on consumption data. Moreover, no pan-European consumption data suitable for allergen risk assessment are available. A procedure for grouping food products automatically across...... 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...... Allergen and Allergy Management) aims at developing strategies for food allergies based on evidences. Especially, food allergen risk assessment helps food producers or authorities to make decisions on withdrawing a food product from the market or adding more information on the label when allergen presence...

  5. 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...... in allergic patients. Sublingual allergen immunotherapy (SLIT) has been approved, notably in the European Union, as an effective alternative to subcutaneous allergen immunotherapy (SCIT) for allergic rhinitis patients. Compared with SCIT, SLIT has a better safety profile. This is possibly because oral antigen...

  6. Specific IgE and IgG measured by the MeDALL allergen-chip depend on allergen and route of exposure: The EGEA study.

    Science.gov (United States)

    Siroux, Valérie; Lupinek, Christian; Resch, Yvonne; Curin, Mirela; Just, Jocelyne; Keil, Thomas; Kiss, Renata; Lødrup Carlsen, Karin; Melén, Erik; Nadif, Rachel; Pin, Isabelle; Skrindo, Ingebjørg; Vrtala, Susanne; Wickman, Magnus; Anto, Josep Maria; Valenta, Rudolf; Bousquet, Jean

    2017-02-01

    The nature of allergens and route and dose of exposure may affect the natural development of IgE and IgG responses. We sought to investigate the natural IgE and IgG responses toward a large panel of respiratory and food allergens in subjects exposed to different respiratory allergen loads. A cross-sectional analysis was conducted in 340 adults of the EGEA (Epidemiological study of the Genetics and Environment of Asthma, bronchial hyperresponsiveness and atopy) (170 with and 170 without asthma) cohort. IgE and IgG responses to 47 inhalant and food allergen components were analyzed in sera using allergen microarray and compared between 5 French regions according to the route of allergen exposure (inhaled vs food allergens). Overall 48.8% of the population had allergen-specific IgE levels of 0.3 ISAC standardized units (ISU) or more to at least 1 of the 47 allergens with no significant differences across the regions. For ubiquitous respiratory allergens (ie, grass, olive/ash pollen, house dust mites), specific IgE did not show marked differences between regions and specific IgG (≥0.5 ISU) was present in most subjects everywhere. For regionally occurring pollen allergens (ragweed, birch, cypress), IgE sensitization was significantly associated with regional pollen exposure. For airborne allergens cross-reacting with food allergens, frequent IgG recognition was observed even in regions with low allergen prevalence (Bet v 1) or for allergens less frequently recognized by IgE (profilins). The variability in allergen-specific IgE and IgG frequencies depends on exposure, route of exposure, and overall immunogenicity of the allergen. Allergen contact by the oral route might preferentially induce IgG responses. Copyright © 2016 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

  7. GA(2)LEN skin test study I: GA(2)LEN harmonization of skin prick testing: novel sensitization patterns for inhalant allergens in Europe

    DEFF Research Database (Denmark)

    Heinzerling, L M; Burbach, G J; Edenharter, G

    2009-01-01

    BACKGROUND: Skin prick testing is the standard for diagnosing IgE-mediated allergies. However, different allergen extracts and different testing procedures have been applied by European allergy centres. Thus, it has been difficult to compare results from different centres or studies across Europe...

  8. Monoclonal antibodies against the major allergen of Plantago lanceolata pollen, Pla l 1: affinity chromatography purification of the allergen and development of an ELISA method for Pla l 1 measurement.

    Science.gov (United States)

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

    2001-05-01

    Plantago lanceolata (English plantain) pollen is a relevant cause of pollinosis in temperate regions. The major allergen of this pollen, Pla l 1, is recognized by the specific IgE from more than 80% of plantain-sensitive patients. It displays significant sequence homology with the major olive-pollen allergen Ole e 1. The objective was to develop a monoclonal antibody-based ELISA to quantify Pla l 1, and to assess the correlation of Pla l 1 content with the biologic activity of plantain pollen extracts. We also aimed to establish the specificity of the monoclonal antibodies against the potentially cross-reactive allergen Ole e 1, and to investigate the presence of Pla l 1-like proteins in psyllium and melon that have been reported to cross-react with P. lanceolata pollen. After fusion of myeloma cells with spleen cells from a BALB/c mouse, two Pla l 1-specific monoclonal antibodies secreting hybridomas were selected, and the antibodies characterized. One of them (2A10) was used as the capture antibody in an ELISA for Pla l 1 quantitation. An anti-P. lanceolata rabbit serum was used as the second antibody. Pla l 1 was purified by immunoaffinity chromatography and used as the standard in the assay. The ELISA developed was highly reproducible and sensitive, with a detection limit of 0.1 ng/ml, and a practical working range of 0.4-12 ng/ml. The specificity was demonstrated against a large battery of allergens, including Ole e 1. The concentration of Pla l 1 was measured in 19 extracts of P. lanceolata pollen, and a good correlation was observed between the Pla l 1 content and the allergenic activity of the extracts. Pla l 1 was not detected in psyllium or melon extracts. The results prove the usefulness of the Pla l 1-ELISA for the standardization of extracts of P. lanceolata pollen intended for clinical use.

  9. Allergens of mites

    Directory of Open Access Journals (Sweden)

    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.

  10. Standard Operation Procedure of Gaharu Oil Extraction Process

    International Nuclear Information System (INIS)

    Peng, C.S.; Mohd Fajri Osman; Norella Bahari; Rusli Zakaria

    2015-01-01

    Gaharu oil extraction system developed by Nuclear Malaysia is a hydro-distillation system innovated from the tradition method and improvised with science and technology to enhance the process efficiency and safety. The main component of gaharu oil extraction system included the extraction vessel, condenser and collection decanter. Apart from the extraction unit, the gaharu oil extraction process also incorporated two major supporting systems. These systems are the liquefied petroleum gas (LPG) system and the water cooling system. The LPG system is solely used for heating the vessel, which contains a mixture of treated and grind gaharu bark and fluid, to produce steam. Meanwhile the water cooling system is used to supply the necessary cool water into the condenser unit and facilitate in the transformation of the mentioned steam into gaharu oil and distilled water. The dissipate water from this process will be channeled back into the water cooling ponds to let excess heat to be removed and allow the cool water to reuse again by the process. (author)

  11. Watermelon and ragweed share allergens.

    Science.gov (United States)

    Enberg, R N; Leickly, F E; McCullough, J; Bailey, J; Ownby, D R

    1987-06-01

    A biotin-avidin amplified ELISA was used to measure antigen-specific IgE for ragweed, representative members of the gourd family (watermelon, cantaloupe, honeydew melon, zucchini, and cucumber), and banana in the sera of 192 allergic patients, each with an IgE greater than or equal to 180 microns/ml. Sixty-three percent (120/192) of the sera contained antiragweed IgE, and of these patients, 28% to 50% contained IgE specific for any single gourd family member. In contrast, no greater than 11% of the sera positive for a given gourd or banana were negative for ragweed. Correlations between ragweed and gourd-specific IgE levels were significant (p less than 0.001), and correlation coefficients between any two gourds exceeded 0.79. In an ELISA system, the extracts of watermelon and ragweed inhibited each other in a dose-dependent manner; the resulting nonparallel inhibition curves indicate that some, but not all, of the allergens in the two extracts are cross-reactive. Isoelectric focusing of watermelon and ragweed extracts in narrow range gel (pH 4 to 6) followed by immunoblotting demonstrated six watermelon allergen bands with isoelectric points identical to those of ragweed allergens. Several remaining bands in the two extracts had differing isoelectric points, however. Six of 26 patients interviewed with watermelon-specific IgE reported developing oropharyngeal symptoms (itching and/or swelling of the lips, tongue, or throat) after ingesting at least one of the study foods, whereas only one of 25 patients interviewed without detectable watermelon-specific IgE reported similar symptoms (p = 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

  12. Allergens in indoor spaces; Allergene im Innenraumbereich

    Energy Technology Data Exchange (ETDEWEB)

    Wahn, U. [Universitaetskinderklinik, FU Berlin (Germany)

    1994-02-01

    Research into environmental factors and how they affect the health of children needs to address a number of questions: Does the environment play any part in this at all? What are the affecting environmental factors? What is the scope of specific environmental factors relative to an induction or an enhancement of a disease? The search for and identification of allergens has priority relative to allergic developments in children. The author describes major allergen sources, the pathogenesis of allergies, the analysis of allergens and a study conducted by several pediatric clinics on the development of allergies in infants and young children, while focussing on risk factor of house-dust mites and options to reduce the exposure to mite-associated allergens. (Uhe) [Deutsch] Bei der Erforschung von Umweltfaktoren und ihrem Einfluss auf die Gesundheit von Kindern interessieren mehrere Fragen: spielt die Umwelt ueberhaupt ein Rolle? Welche Umwelteinfluesse sind es, wie gross ist die krankheitsindzierende oder krankheitsverstaerkte Rolle bestimmter Umwelteinfluesse? Fuef die Allergieentwicklung bei Kindern stehen die Allergene an erster Stelle. Neben der Beschreibung der wichtigsten Allergiequellen und der Entstehung von Allergien und Analytik von Allergenen wird ausfuehrlich eine Studie verschiedener Kinderkliniken zur Allergieentwicklung im Kindesalter beschrieben. Dabei wird vor allem auf den Risikofaktor Hausstaubmilbe und die Moeglichkeiten der Reduzierung der Milbenallergenbelastung eingegangen. (Uhe)

  13. [New aero-allergens].

    Science.gov (United States)

    De Blay, F; Bessot, J C; Pauli, G

    1996-01-01

    As the number of proteins recognized as causing allergic respiratory diseases increases, new aero allergens have appeared in the animal and vegetable realms, both in home and professional environments. Lepidoglyphus destructor and Blomia tropicalis, two mites found in storage areas, are particularly important in agricultural areas and in homes. Over the last ten years, the frequency of reactions to cockroaches has also increased in several countries. The allergenicity of non-biting insects is a frequent cause of allergy in certain countries including Japan. Chironomides cause respiratory diseases in professional and outdoor environments. The important role of Alternaria, a mold, in producing severe asthma has also been demonstrated. The pathophysiology of pollen-induced asthma has been shown to result from pollen allergens carried by particles less than 5 microns in diameter. Cyprus and ash tree pollen also cause an increasing number of pollinoses and flowers can cause rhinitis and asthma. Respiratory allergy to Ficus benjamina inaugurated a new type of allergies caused airborne allergens from non-pollinating plants. Allergy to latex raises a particular problem for health care workers. The immunochemical structures of the major and minor airborne allergens are now better known and the homologous structures of different allergens largely explains certain cross-reactions. In the future, recombinant allergens will probably be used to better understand the role of allergens in inducing and maintaining the allergic reaction and should help in our approach to diagnosis and therapy.

  14. Allergens: An Enhanced Focus.

    Science.gov (United States)

    Spotz, Kristen

    2018-01-01

    Food Allergy Awareness Week was created with the purpose of placing a spotlight on the seriousness of food allergies. Recognized in the United States in mid-May every year, Food Allergy Awareness Week serves as a reminder of the over 15 million Americans who suffer from food allergies. The importance of allergies and allergen labeling can be seen when looking at U.S. Food and Drug Administration recall data: of the 764 recalls in 2016, 305 (representing more than 40%) were due to undeclared allergens. However, recalls for undeclared allergens are a complex issue with numerous factors. The implementation of prevention-based systems with the necessary management components and further error-proofing the systems, along with allergen awareness embedded throughout a company's food safety culture, can likely help reduce the number of recalls for undeclared allergens. As a resource to manufacturers, the Grocery Manufacturers Association and the Food Allergy Research and Resource Program have developed several resources to assist with developing robust allergen management programs. By reducing the number of recalls for undeclared allergens, the food industry will likely increase and maintain consumer confidence and trust of the food-allergic community. This enhanced consumer confidence and trust could eventually open the door for further collaboration with the food-allergic community and, potentially, advance allergen-related policies.

  15. 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.

  16. 76 FR 14414 - Allergenic Products Advisory Committee; Notice of Meeting

    Science.gov (United States)

    2011-03-16

    ... pollen allergen extracts by the Laboratory; (3) statistical considerations for the design and... attendance of the public at its advisory committee meetings and will make every effort to accommodate persons...

  17. Comparison of supercritical fluid extraction and Soxhlet extraction for the determination of polychlorinated biphenyls in environmental matrix standard reference materials.

    Science.gov (United States)

    Schantz, M M; Bøwadt, S; Benner, B A; Wise, S A; Hawthorne, S B

    1998-08-14

    Supercritical fluid extraction (SFE) was compared to traditional Soxhlet extraction for the determination of polychlorinated biphenyl congeners in three standard reference materials: SRM 1941a (Organics in Marine Sediment), SRM 1944 (New York/New Jersey Waterway Sediment) and SRM 2974 [Organics in Mussel Tissue (Mytilus edulis) (Freeze-Dried)]. The concentrations determined using SFE compared well with the certified concentrations for the majority of the polychlorinated biphenyl congeners.

  18. Anti-inflammatory and antinociceptive activities of Phyllanthus niruri spray-dried standardized extract

    Directory of Open Access Journals (Sweden)

    Cínthia R. C. Porto

    2013-02-01

    Full Text Available Phyllanthus niruri L., Euphorbiaceae, spray-dried standardized extract was studied for its anti-inflammatory and antinociceptive activities in adult albino rats and mice. The anti-inflammatory effect of spray-dried standardized extract was observed in carrageenan-induced paw edema and thioglycolate-induced leukocyte migration, while antinociceptive effects were observed using Randall & Selitto, tail flick, and hot plate tests. This study showed that intraperitoneal spray-dried standardized extract at 100, 200, 800, or 1600 mg/kg reduced the vascular response in the inflammatory process of paw edema induced by 1% carrageenan. Oral spray-dried standardized extract at 100 or 200 mg/kg inhibited leukocyte migration to the site of inflammation induced by 3% thioglycolate. In rats, at 100 and 200 mg/kg intraperitoneally, the extract exhibited a marked peripheral analgesic effect in a Randall & Selitto assay and showed significant central analgesic activity in a hot plate and tail flick assay. In conclusion, this study suggested that Phyllanthus niruri spray-dried standardized extract has potent inflammatory and antinociceptive activities and that these activities are not modified by standard drying process, making it feasible to use the dry extract standardized to obtain a phytotherapic preparation and thus validating its use for the treatment of pain and inflammation disorders.

  19. Basidiomycete allergens: comparison of three Ganoderma species.

    Science.gov (United States)

    Horner, W E; Helbling, A; Lehrer, S B

    1993-02-01

    High atmospheric concentrations of basidiospores occur in various parts of the world. Ganoderma basidiospores are distinctive, easily identifiable in aeroallergen surveys, and widely abundant. Previous studies showed that Ganoderma basidiospores caused respiratory allergies. Thus, we investigated various extracts (spore, cap, and/or mycelial) of G. meredithae, G. lucidum, and G. applanatum for allergen components. Analyses included radioallergosorbent test (RAST) inhibition and IgE blots from isoelectric focusing (IEF) and SDS-PAGE. RAST inhibition with spores and caps of G. meredithae and G. lucidum showed that spores inhibited caps better than caps inhibited spores. Species differences were minor. Coomassie blue (CB) staining of IEF gels detected at least 23 protein bands (pI 3.6-6.6) in caps of G. meredithae and G. lucidum. G. meredithae spore extracts contained 17 of these (pI 3.6-5.0, 6.6). Spores and caps of G. meredithae contained 13 and 11 allergen bands, respectively, on IEF blots. SDS-PAGE of G. meredithae spore and cap showed one and four bands, respectively, by CB staining, but IgE blots showed 13 bands in cap and 17 in spore. Culture mycelia of G. lucidum and G. applanatum attained significant and essentially constant RAST activity by day 4. Activity was also present in culture supernatant by day 4. Blots of mycelium and supernatant detected a single allergen in day-8 mycelia and subsequently six allergen bands in day-16 mycelia and eight in day-16 supernatant (one appeared as a doublet). These data show that Ganoderma extracts contain a complex mixture of allergens. Differences among species were minor; spores and mycelia are apparently better sources of allergens than caps.

  20. free radical scavenging and cytotoxic activity of standardized extracts

    African Journals Online (AJOL)

    Ledy

    Cerecero et al., 2011; Hernández-Pérez and Herrera-. Arellano, 2011). These compounds have standardized which can use as quality markers, ensuring the composition of assets present. ACKNOWLEDGEMENTS. We thank Consejo Nacional de Ciencia y Tecnología. (CONACYT) for their support (204643). We also thank.

  1. Cardioprotective effect of melatonin-standardized ethanol extract of ...

    African Journals Online (AJOL)

    out using TLC densitometry and quantitated using the calibration curve of melatonin (0-2.0 mg/ml). Treatment of animals. Male albino Wistar rats aged 7 - 8 weeks were used in this study. The animals were housed in air conditioned room and were kept in standard laboratory condition, which included a 12-h light- dark cycle ...

  2. A four-step sandwich radioimmunoassay for direct selection of monoclonal antibodies to allergen molecules

    International Nuclear Information System (INIS)

    Ley, V.; Corbi, A.L.; Sanchez-Madrid, F.; Carreira, J.C.

    1985-01-01

    A 4-step radioimmunoassay has been devised for direct identification of monoclonal antibodies (MAb) directed to IgE-binding molecules. Polyvinyl chloride wells coated with purified anti-mouse kappa chain MAb (187-1) were successively incubated with: (1) MAb-containing hybridoma supernatants, (2) allergen extract, (3) allergic patients' serum pool, and (4) 125 I-labeled anti-human IgE antiserum, to detect MAb-allergen-IgE complexes. MAb to allergens from Parietaria judaica pollen and Dermatophagoides mites have been selected with this screening procedure. The affinity-purified allergen molecules competed the binding of IgE to allergen extracts coated to paper discs in a RAST inhibition assay, confirming the anti-allergen specificity of the selected MAb. This screening method is sensitive enough to allow detection of MAb directed to poorly represented allergens. (Auth.)

  3. The European Union CREATE project: a model for international standardization of allergy diagnostics and vaccines

    NARCIS (Netherlands)

    Chapman, Martin D.; Ferreira, Fatima; Villalba, Mayte; Cromwell, Oliver; Bryan, Donna; Becker, Wolf-Meinhard; Fernández-Rivas, Montserrat; Durham, Stephen; Vieths, Stefan; van Ree, Ronald; Aalbers, M.; Notten, S.; Ooievaar-de Heer, P.; Ferreira, F.; Gademaier, G.; Wallner, M.; Villalba, M.; Rodriguez, R.; Becker, W.-M.; Eberhardt, F.; Lepp, U.; Raulf-Heimsoth, M.; Valenta, R.; Focke, M.; Bryan, D.; Dolman, C.; Das, R. G.; Vieths, S.; Fötisch, K.; Di Felice, G.; Pini, C.; Cromwell, O.; Fiebig, H.; Weber, B.; van Schijndel, H.; Dorpema, J. W.; Marco, F. M.; Monsalve, R.; Barber, D.; Caldas, E. Fernandez; Moingeon, P.; Didierlaurent, A.; André, C.; Kroon, A.; Neubauer, A.; Chapman, M.; Vailes, L.; Tsay, A.; Durham, S.; Custovic, A.; Simpson, B.; Knulst, A.; Rivas, M. Fernández; Mancebo, E. Gonzalez; Bahima, A. Cistero; Moncin, M. M. San Miguel; Mari, A.; Kinaciyan, T.; Quiralte, J.; Pauli, G.; de Blay, F.; Purohit, A.; Rak, S.

    2008-01-01

    Allergen measurements are used extensively in the formulation of allergy diagnostics and vaccines, yet no purified international allergen standards are available for calibration purposes. The aims of the European Union CREATE project were to develop international standards with verifiable allergen

  4. Allergen analysis of mugwort pollen (Artemisia vulgaris) by means of crossed radioimmunoelectrophoresis (CRIE) and RAST inibition

    International Nuclear Information System (INIS)

    Diener, C.; Wiebicke, K.; Schlenvoigt, G.; Jaeger, L.; Fuster, R.

    1988-01-01

    43 antigens of mugwort pollen (Artemisia vulgaris), 31 migrating anodically and 12 cathodically, were identified by means of crossed immunoelectrophoresis (CIE). Eleven antigens bound human IgE antibodies by CRIE. Two major allergens were determined, the allergens 23 and 40. The allergenic activity of mugwort pollen extract was examined by means of RAST inhibition. Specificity of this reaction could be determined by a parallel experiment with grass pollen extract. (author)

  5. Use of patch testing for identifying allergen causing chronic urticaria

    Directory of Open Access Journals (Sweden)

    Sharma Ashimav

    2008-01-01

    Full Text Available Background and Aims: The aim of this present study was to evaluate the role of patch testing for the etiological diagnosis of chronic urticaria (CU by using the Indian standard battery of patch test allergens approved by Contact and Occupational Dermatitis Forum of India (CODFI. Methods: A total of 57 cases with chronic urticaria were tested with the Indian standard battery of allergens. All those cases that showed allergy to patch test allergens were advised to avoid contact with the allergen(s to whom they had allergy; they were also advised to avoid/restrict allergens in the diet. This avoidance/restriction was advised for a period of six weeks. During this period, clinical improvement of each patient was evaluated and recorded at weekly intervals. Results: Out of the 57 cases of CU, 11 patients showed positive reactions to one or more patch test allergens. Nine out of eleven showed complete disappearance of CU by 2-3 weeks on avoidance of the allergen and this improvement continued till the end of six weeks. The remaining two cases showed partial recovery from CU during the same period. Conclusion: Patch testing is a safe, simple and inexpensive alternative that can be used for the etiological diagnosis of chronic urticaria before undertaking expensive investigations.

  6. Direct contact between dendritic cells and bronchial epithelial cells inhibits T cell recall responses towards mite and pollen allergen extracts in vitro

    DEFF Research Database (Denmark)

    Papazian, Dick; Wagtmann, Valery R; Hansen, Soren

    2015-01-01

    Background: Airway epithelial cells (AECs) form a polarized barrier along the respiratory tract. They are the first point of contact with airborne antigens and are able to instruct resident immune cells to mount appropriate immune responses by either soluble or contact-dependent mechanisms....... Objective: We hypothesize that a healthy, polarized epithelial cell layer inhibits inflammatory responses toward allergens to uphold homeostasis. Methods: Using an in vitro co-culture model of the airway epithelium, where a polarized cell layer of bronchial epithelial cells can interact with dendritic cells...... cell recall responses towards Bet v 1, Phl p 5 and Der p 1 in vitro, suggesting that AECs-DC contact in vivo constitute a key element in mucosal homeostasis. This article is protected by copyright. All rights reserved....

  7. Quantification of Peach Fruit Allergen Lipid Transfer Protein by a Double Monoclonal Antibody-based Sandwich ELISA

    NARCIS (Netherlands)

    Gao, Zhong-Shan; Ma, Ying-tao; Zhou, Xiang; Yang, Zhao-Wei; Jia, Hui-juan; Gao, Ling; Wu, Shan-Dong; Han, Lu-yang; Yi, Xue-ying; Wang, Hui-Ying; Akkerdaas, Jaap H.; van Ree, Ronald

    2016-01-01

    Fruit is one of the most commonly reported food allergy sources in China, and peach lipid transfer protein (Pru p 3) has been identified as the major allergen inducing systematic symptoms. Crude allergen extracts and single component allergens have been used in food allergy diagnosis and

  8. 21 CFR 680.1 - Allergenic Products.

    Science.gov (United States)

    2010-04-01

    ... the source material. (2) Molds. (i) Molds (excluding rusts and smuts) used as source material in the... manufacturing step during and subsequent to harvest, as specified in the standard operating procedures. Before use of the mold as a source material for Allergenic Products, in accordance with 21 CFR 601.2, the...

  9. 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.

  10. 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.)

  11. Chloroatranol, an extremely potent allergen hidden in perfumes

    DEFF Research Database (Denmark)

    Johansen, J.D.; Andersen, Klaus Ejner; Svedman, C.

    2003-01-01

    Oak moss absolute is a long-known, popular natural extract widely used in perfumes. It is reported as the cause of allergic reactions in a significant number of those with perfume allergy. Oak moss absolute has been the target of recent research to identify its allergenic components. Recently....... The dose eliciting a reaction in 50% of the test subjects at patch testing was 0.2 p.p.m. In conclusion, the hidden exposure to a potent allergen widely used in perfumes has caused a highly sensitized cohort of individuals. Judged from the elicitation profile, chloroatranol is the most potent allergen...

  12. Antibacterial activity of Achillea tenuifolia Lam. extract against standard bacteria and isolated strains

    Directory of Open Access Journals (Sweden)

    Sahar Omidpanah

    2016-12-01

    Full Text Available Researchers have been trying to develop new broad-spectrum antibiotics against the infectious diseases caused by bacteria, fungi, viruses, and parasites for many decades. Prolonged usage of the antibiotics has led to the emergence of drug resistance among bacteria; therefore, there is a tremendous need for novel antimicrobial agents from different sources such as plants which are used in traditional medicine. The aim of this study was to evaluate antibacterial effect of Achillea tenuifolia. The plant material was extracted by maceration method using methanol three times at room temperature. The extract was concentrated after removing the solvent by rotary evaporator and then lyophilized using freeze dryer. Inhibitory effect of the extract was examined against four standard bacteria strains and two isolated strains from diseased hen using disk diffusion method and microdilution method to evaluate their inhibition zone diameter (IZD and minimum inhibitory concentration (MIC, respectively. The results showed that the extract of the plant was active against standard strains including Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis with IZDs of 10.3±0.5, 14±0.0, 12±0.0 and 11.6±0.5, respectively. However, growths of isolated strains were not inhibited in the presence of the extract. Although, the growths of isolated strains were not inhibited by the plant extract, the standard strains were moderately susceptible to the extract; among those P. aeroginosa was more sensible than other tested strains

  13. 466 Bee venom Immunotherapy with Standardized Extract, Two Case Comunication and Clinical Progress

    Science.gov (United States)

    Cardona, Aristoteles Alvarez; Nieto, Leticia Hernandez; Melendez, Alvaro Pedroza

    2012-01-01

    Background Bee venom immunotherapy is a safe and effective treatment, indicated in patients with previous history of severe systemic reactions to bee venom, demonstrating succesful desensitization in more than 90% of cases with standardized extract. Currently in Mexico there is no standardized extract commercially available for treatment, despite of having high activity of beekeeping and occupational exposure with at least 17,478 registered stings per year and an annually honey production of nearly 70 tons. Methods We present the clinical progress of 2 patients with history of severe systemic reactions to bee venom and occupational exposure, both with demonstrated sensitization by specific IgE and who underwent specific immunotherapy with standardized extract (Alk-US) reaching a maintenance weekly dose of 100 mcg (PLA2) for the last 4 years. Results Both patients sufered of accidental stings after reached the maintenance dose presenting mild local reactions to stings. Both patients had very different clinical course presenting a wide variety of adverse reactions during desensitization protocol; from mild local to generalized reactions all generally well tolerated allowed to reach the maintenance dose with succesful desensitization proved by accidental exposure without severe systemic reactions. Conclusions Bee venom specific immunotherapy with standardized extract is a well tolerated and efective treatment preventing the development of life threathening reactions in sensitized patients. It is important to promote the use and availability of standardized extract in developing countries with poor safety measures and high occupational exposure.

  14. A standardized method for sampling and extraction methods for quantifying microplastics in beach sand.

    Science.gov (United States)

    Besley, Aiken; Vijver, Martina G; Behrens, Paul; Bosker, Thijs

    2017-01-15

    Microplastics are ubiquitous in the environment, are frequently ingested by organisms, and may potentially cause harm. A range of studies have found significant levels of microplastics in beach sand. However, there is a considerable amount of methodological variability among these studies. Methodological variation currently limits comparisons as there is no standard procedure for sampling or extraction of microplastics. We identify key sampling and extraction procedures across the literature through a detailed review. We find that sampling depth, sampling location, number of repeat extractions, and settling times are the critical parameters of variation. Next, using a case-study we determine whether and to what extent these differences impact study outcomes. By investigating the common practices identified in the literature with the case-study, we provide a standard operating procedure for sampling and extracting microplastics from beach sand. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. 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

  16. Allergen Immunotherapy and Tolerance

    Directory of Open Access Journals (Sweden)

    Tomokazu Matsuoka

    2013-01-01

    Full Text Available Successful allergen-specific immunotherapy (AIT is associated with a marked decrease in symptoms on allergen exposure, a reduced requirement for 'rescue' anti-allergic drugs and improvement in patients' quality of life. These benefits persist for at least several years following discontinuation of immunotherapy - the hallmark of clinical and immunological tolerance. AIT has been shown to modulate both innate and adaptive immunological responses. Early suppression of innate effector cells of allergic inflammation (mast cells, basophils, regulation of pro-allergic T helper 2 type (Th 2 responses and IgE+ B cell responses have been shown to occur both in the tissue and in the peripheral blood during AIT. The allergen-tolerant state is associated with local and systemic induction of distinct populations of allergen-specific T regulatory cells including IL-10+ Tregs (Tr1 cells, TGF-P+ Tregs and FoxP3+ memory T regs. B cells are switched in favour of producing IgG (particularly IgG4 antibodies and associated blocking activity for IgE-dependent events, including basophil activation and IgE-facilitated allergen binding to B cells. An induction of IL-10+ B regulatory cells and alterations in dendritic cell subsets have also recently been described. These events are followed by the induction of T regulatory cells, suppression of allergen-specific T cell proliferation and immune deviation from Th2 in favour of Th1 responses. Alternative mechanisms of tolerance include apoptosis/deletion of antigen-specific memory Th2 cells and/or a failure of co-stimulation leading to T cell anergy.

  17. Human dander as a potential allergen source in atopic dogs : allergen characterization and IgE-profiling

    OpenAIRE

    Resk, Nicole

    2006-01-01

    Perennial indoor allergens, among them house dust mites and molds are known to be important inducers of IgE-mediated hypersensitivity reactions in dogs. Human dander (HD), an important constituent of house dust may also be a major cause of atopic disease in dogs. So far, HD has been poorly characterized as an allergenic source and the role HD plays in veterinary medicine is unclear. The aims of this study were to characterize two available HD extracts, thus completing a representativ...

  18. Content of Asthmagen Natural Rubber Latex Allergens in Commercial Disposable Gloves.

    Science.gov (United States)

    Bittner, C; Garrido, M V; Krach, L H; Harth, V

    The use of natural rubber latex (NRL) gloves in many occupations may lead to latex sensitization, allergic asthma, and skin reactions. Due to their good properties and environmental safety NRL gloves are still being used in the healthcare setting, but also in the food industry, by hairdressers, cleaners, etc. The aim of our study was to assess the protein and NRL allergen content in commercial gloves by different methods, including a new assay. Twenty commercially available NRL gloves were analyzed. Protein extraction was performed according to the international standard ASTM D-5712. Total protein content was measured with a modified Lowry method, NRL content with the CAP Inhibition Assay, the Beezhold ELISA Inhibition Assay, and an innovative ELISA with IgY-antibodies extracted from eggs of NRL-immunized hens (IgY Inhibition Assay). We found a high protein content in a range of 215.0-1304.7 μg/g in 8 out of the 20 NRL gloves. Seven of the 20 gloves were powdered, four of them with a high protein content. In gloves with high protein content, the immunological tests detected congruently high levels of NRL allergen. We conclude that a high percentage of commercially available NRL gloves still represent a risk for NRL allergy, including asthma. The modified Lowry Method allows to infer on the latex allergen content.

  19. Standardization of DNA extraction from sand flies: Application to genotyping by next generation sequencing.

    Science.gov (United States)

    Casaril, Aline Etelvina; de Oliveira, Liliane Prado; Alonso, Diego Peres; de Oliveira, Everton Falcão; Gomes Barrios, Suellem Petilim; de Oliveira Moura Infran, Jucelei; Fernandes, Wagner de Souza; Oshiro, Elisa Teruya; Ferreira, Alda Maria Teixeira; Ribolla, Paulo Eduardo Martins; de Oliveira, Alessandra Gutierrez

    2017-06-01

    Standardization of the methods for extraction of DNA from sand flies is essential for obtaining high efficiency during subsequent molecular analyses, such as the new sequencing methods. Information obtained using these methods may contribute substantially to taxonomic, evolutionary, and eco-epidemiological studies. The aim of the present study was to standardize and compare two methods for the extraction of genomic DNA from sand flies for obtaining DNA in sufficient quantities for next-generation sequencing. Sand flies were collected from the municipalities of Campo Grande, Camapuã, Corumbá and Miranda, state of Mato Grosso do Sul, Brazil. Three protocols using a silica column-based commercial kit (ReliaPrep™ Blood gDNA Miniprep System kit, Promega ® ), and three protocols based on the classical phenol-chloroform extraction method (Uliana et al., 1991), were compared with respect to the yield and quality of the extracted DNA. DNA was quantified using a Qubit 2.0 fluorometer. The presence of sand fly DNA was confirmed by PCR amplification of the IVS6 region (constitutive gene), followed by electrophoresis on a 1.5% agarose gel. A total of 144 male specimens were analyzed, 72 per method. Significant differences were observed between the two methods tested. Protocols 2 and 3 of phenol-chloroform extraction presented significantly better performance than all commercial kit extraction protocols tested. For phenol-chloroform extraction, protocol 3 presented significantly better performance than protocols 1 and 2. The IVS6 region was detected in 70 of 72 (97.22%) samples extracted with phenol, including all samples for protocols 2 and 3. This is the first study on the standardization of methods for the extraction of DNA from sand flies for application to next-generation sequencing, which is a promising tool for entomological and molecular studies of sand flies. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Study on the developmental toxicity of a standardized extract of Orthosiphon stamineus in rats

    Directory of Open Access Journals (Sweden)

    Hussin Muhammad

    2013-06-01

    Full Text Available Infusions of Orthosiphon stamineus Benth., Lamiaceae, leaves are widely used in Southeastern Asia to treat different illnesses. Nonetheless, no data is available on the safety of O. stamineus for pregnant women and their babies. This study was undertaken to evaluate the developmental toxicity of O. stamineus standardized aqueous extract in female Sprague Dawley rats (n=21 at 0, 250, 500, 1000 and 2000 mg/kg/day, by gavage on gestation days 6-20. Clinical signs of maternal toxicity, body weight gain, and food and water consumption were recorded. Caesarean sections were performed on gestation day 21; resorptions and living and dead fetuses were counted. Fetuses were weighed and examined for external abnormalities. Half of the fetuses from each litter were cleared and stained with Alizarin red S for skeleton evaluation. O. stamineus standardized aqueous extract did not alter pregnancy body weight gain and food and water consumption and caused no other sign of maternal toxicity. Embryolethality and prenatal growth retardation were not observed either. O. stamineus standardized aqueous extract increased a few skeleton variations and a skull bone malformation (hyoid bone absent in a non-dose dependent manner. Anogenital distance was increased in male and female fetuses exposed to the highest O. stamineus standardized aqueous extract dose, an indication that the extract could possibly contain androgenic compounds.

  1. Inhibition of cyclooxygenase activity by standardized hydroalcoholic extracts of four Asteraceae species from the Argentine Puna

    Directory of Open Access Journals (Sweden)

    M.R. Alberto

    2009-09-01

    Full Text Available We determined the anti-inflammatory activity of standardized extracts of four medicinal plant species (Baccharis incarum, B. boliviensis, Chuquiraga atacamensis, Parastrephia lucida that grow in the Argentine Puna (3800 m above sea level and that are used to reduce oxidative stress and alleviate gout and arthritic pain. The extracts of plant aerial parts were standardized in terms of total phenolic compounds and flavone/flavanone content and free radical scavenging activity. All extracts showed high phenolic compound concentration (0.5-1.6 mg/mL, mainly flavones and flavonols (0.1-0.8 mg/mL. The extracts showed hydrogen donating ability (DPPH and ABTS and reactive oxygen species scavenging activity (O2●-, OH-, H2O2. The ability of the extracts to inhibit cyclooxygenase enzymes (COX-1 and COX-2 was determined by calculating percent inhibition of PGE2 production measured by enzyme immunoassay. All extracts inhibited both enzymes with IC50 values of 2.0 to 16.7 µg/mL. The anti-inflammatory activity of B. incarum and C. atacamensis extracts was higher than that of B. boliviensis and P. lucida. The IC50 values obtained for indomethacin were 0.11 and 0.78 µM for COX-1 and COX-2, respectively. The present results are consistent with the anecdotal use of these species in phytotherapic preparations.

  2. [Establish quality evaluation method based on standard decoction of Danshen extract].

    Science.gov (United States)

    Dong, Qing; Yu, Hua-Tao; Dai, Yun-Tao; Chao, Jung; Fan, Zi-Quan; Wang, Dan-Dan; Zhu, Chao; Chen, Shi-Lin

    2017-03-01

    The quality of Danshen extract granules on market is largely different from each other mainly due to the heterogeneous quality of raw materials of Salvia miltiorrhiza, various producing procedures and lack of good quality evaluation method. Formula granule and "standard decoction" have the same quality. In this paper, a systematic evaluation method for the quality of Danshen decoction was established from the perspective of "standard decoction", in order to explore the main factors affecting the quality uniformity of Danshen extract granules. Danshen standard decoction was prepared; then the fingerprint method was developed to determine the content of salvianolic acid B; and the main peaks in the fingerprint were identified with UPLC-QTOF/MS to clarify the chemical compositions of Danshen decoction. Three indexes were calculated to evaluate the stability of whole process, including the extraction ratio; transfer rate of index components and pH value. The results showed that the main components of Danshen decoction were phenolic acids, while the extraction rate, the transfer rate of salvianolic acid B and pH value were in a relatively stable level, and the similarity in the fingerprint of standard decoction was high, indicating that the preparation procedure was stable. The level of salvianolic acid B in the standard decoction was in a large range, which was mainly due to the difference in the quality of Salviae Miltiorrhizae Radix et Rhizoma. Copyright© by the Chinese Pharmaceutical Association.

  3. Allergens from fish and egg

    DEFF Research Database (Denmark)

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

  4. Immunochemical and biological quantification of peanut extract

    DEFF Research Database (Denmark)

    Poulsen, Lars K.; Pedersen, Mona H; Platzer, Michael

    2003-01-01

    Biological standardization of allergen extracts is one of the steps in the characterization of an extract. The gold standard for determination of biological potency is the skin prick test, but histamine release (HR) has been used as a convenient ex vivo method for analyzing a large number...... of samples. We describe the use of rabbit basophils as a tool in biological standardization. Using peanut as a model allergen, it is described how rabbits immunized for production of antiserum may become sensitized and their basophils used for histamine release experiments. It is also possible to use rabbit...... antiserum to passively sensitize basophils derived from naive rabbits, but the sensitivity of this method is so far 100-1000 times lower than the direct histamine release. The rabbit histamine release results are compared to an ELISA developed by means of the same antisera and by passive sensitization...

  5. Strychnos nux-vomica seeds: Pharmacognostical standardization, extraction, and antidiabetic activity

    Directory of Open Access Journals (Sweden)

    Rajesh Bhati

    2012-01-01

    Full Text Available Background: Strychnos nux-vomica, commonly known as kuchla, contains strychnine and brucine as main constituents. Minor alkaloids present in the seeds are protostrychnine, vomicine, n-oxystrychnine, pseudostrychnine, isostrychnine, chlorogenic acid, and a glycoside. Seeds are used traditionally to treat diabetes, asthma, aphrodisiac and to improve appetite. Objective: The present study was aimed to evaluate the various pharmacognostical characters and antidiabetic activity of S. nux-vomica seed. Materials and Methods: Pharmacognostical characters were performed as per the WHO guideline. Extraction was carried out in petroleum ether, chloroform, alcohol, hydroalcoholic, aqueous, and phytochemical constituents present in extracts were detected by different chemical tests. Among these extracts hydroalcoholic, aqueous extracts were evaluated for antidiabetic activity on the basis of extractive yield and phytoconstituents, in alloxan-induced diabetic rats using gliclazide as standard. Results: Various analytical values of S. nux-vomica extract were established. Phytoconstituents present in S. nux-vomica extracts were detected. Conclusion: S. nux-vomica extracts show antihyperglycemic activity in experimental animals.

  6. The cashew allergens

    NARCIS (Netherlands)

    Reitsma, Marit

    2017-01-01

    Cashew nut allergy can be a severe food allergy of which the prevalence appears to be increasing. The aim of this thesis was a comprehensive molecular and serological characterisation of the cashew nut allergens Ana o 1, 2 and 3 for improved diagnosis and characterisation of patient populations.

  7. New Cosmetic Contact Allergens

    Directory of Open Access Journals (Sweden)

    An Goossens

    2015-02-01

    Full Text Available Allergic and photo-allergic contact dermatitis, and immunologic contact urticaria are potential immune-mediated adverse effects from cosmetics. Fragrance components and preservatives are certainly the most frequently observed allergens; however, all ingredients must be considered when investigating for contact allergy.

  8. Phytochemical analysis and standardization of Strychnos nux-vomica extract through HPTLC techniques

    Directory of Open Access Journals (Sweden)

    Dinesh Kumar Patel

    2012-05-01

    Full Text Available Objective: The objective is to develop a noval qualitative and quantitative method by which we can determine different phytoconstituents of Strychnos nux-vomica L. Methods: To profile the phyconstituents of Strychnos nux-vomica, in the present study hydroalcoholic extract of Strychnos nux-vomica was subjected to preliminary phytochemical analysis, antimicrobial activities against certain pathogenic microorganisms, solubility test, loss on drying and pH value. Extract was also subjected to the quantitative analysis including total phenol, flavonoid and heavy metal analysis. Quantitative analysis was performed through HPTLC methods using strychnine and brucine as a standard marker. Results: Phytochemical analysis revealed the presence of alkaloid, carbohydrate, tannin, steroid, triterpenoid and glycoside in the extract. Total flavonoid and phenol content of Strychnos nux-vomica L extract was found to be 0.40 % and 0.43%. Result showed that the level of heavy metal (lead, arsenic, mercury and cadmium complie the standard level. Total bacterial count, yeast and moulds contents were found to be under the limit whereas E. coli and salmonella was found to be absent in the extract. Content of strychnine and brucine were found to be 4.75% and 3.91%. Conclusions: These studies provide valluable information for correct identification and selection of the drug from various adulterations. In future this study will be helpful for the quantitative analysis as well as standardization of the Strychnos nux-vomica L.

  9. Advances in patent applications related to allergen immunotherapy.

    Science.gov (United States)

    Silva, Eduardo Santos da; Pinheiro, Carina Silva; Quintella, Cristina Maria; Ferreira, Fatima; C Pacheco, Luis Gustavo; Alcântara-Neves, Neuza Maria

    2016-06-01

    Allergies are among the most prevalent chronic diseases worldwide. Allergen-specific immunotherapy is used as an alternative treatment to pharmacotherapy. These immunotherapies are performed with crude extracts, which have disadvantages when compared to the new approaches, among them are recombinant proteins and hypoallergens. This review aims to assess immunotherapy for allergies through patent application analysis spanning recent decades. Patents referring to allergen immunotherapies used in allergy treatment. Data were obtained from the Espacenet® website, using the Cooperative Patent Classification (CPC) system. Two-hundred-and-one patent applications were analyzed, taking into consideration their classification by the type of technology and applicant. Allergen-specific immunotherapy represents the only potentially curative therapeutic intervention for the treatment of allergic diseases. The extract-based immunotherapy is being replaced by the use of recombinant allergens, highlighting the hypoallergenic forms, which have low IgE-binding while retaining T-cell reactivity. It is expected that the development of hypoallergens will expand the scope of allergen-specific immunotherapy, especially if associated with alternative systems for expression and delivery systems with future potential. Furthermore, these new developments will likely address the problem of long-term protocols in allergen-specific immunotherapy, thus allowing better patient adherence and compliance.

  10. Reducing peanut allergens by high pressure combined with polyphenol oxidase

    Science.gov (United States)

    Chung, Si-Yin; Houska, Milan; Reed, Shawndrika

    2013-12-01

    Polyphenol oxidase (PPO) has been shown to reduce major peanut allergens. Since high pressure (HP) can increase enzyme activity, we postulated that further reduction of peanut allergens can be achieved through HP combined with PPO. Peanut extracts containing caffeic acid were treated with each of the following: (1) HP; (2) HP+PPO; (3) PPO; and (4) none. HP was conducted at 300 and 500 MPa, each for 3 and 10 min, 37 °C. After treatment, SDS-PAGE was performed and allergenic capacity (IgE binding) was determined colorimetrically in inhibition enzyme-linked immunosorbent assay and Western blots, using a pooled plasma from peanut-allergic patients. Data showed that HP alone had no effect on major peanut allergens. However, HP at 500 MPa combined with PPO (HP500/PPO) induced a higher (approximately twofold) reduction of major peanut allergens and IgE binding than PPO alone or HP300/PPO. There was no difference between treatment times. We concluded that HP500/PPO at 3-min enhanced a twofold reduction of the allergenic capacity of peanut extracts, as compared to PPO itself.

  11. Vaccine development for allergen-specific immunotherapy based on recombinant allergens and synthetic allergen peptides: Lessons from the past and novel mechanisms of action for the future.

    Science.gov (United States)

    Valenta, Rudolf; Campana, Raffaela; Focke-Tejkl, Margit; Niederberger, Verena

    2016-02-01

    In the past, the development of more effective, safe, convenient, broadly applicable, and easy to manufacture vaccines for allergen-specific immunotherapy (AIT) has been limited by the poor quality of natural allergen extracts. Progress made in the field of molecular allergen characterization has now made it possible to produce defined vaccines for AIT and eventually for preventive allergy vaccination based on recombinant DNA technology and synthetic peptide chemistry. Here we review the characteristics of recombinant and synthetic allergy vaccines that have reached clinical evaluation and discuss how molecular vaccine approaches can make AIT more safe and effective and thus more convenient. Furthermore, we discuss how new technologies can facilitate the reproducible manufacturing of vaccines of pharmaceutical grade for inhalant, food, and venom allergens. Allergy vaccines in clinical trials based on recombinant allergens, recombinant allergen derivatives, and synthetic peptides allow us to target selectively different immune mechanisms, and certain of those show features that might make them applicable not only for therapeutic but also for prophylactic vaccination. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  12. Contact allergens for armpits--allergenic fragrances specified on deodorants.

    Science.gov (United States)

    Klaschka, Ursula

    2012-11-01

    According to the so-called "26 allergens rule" 26 supposedly allergenic fragrances must be specified on the containers of cosmetic products if they are present above 0.001% in leave-on products and, 0.01% in rinse-off products. This declaration is meant to inform the consumers of potential risks of skin sensitizers in the products. As many consumers of deodorants suffer from allergic or irritant contact dermatitis in the axillae, the presence of allergens in deodorants deserves special attention. The objective of this study was to find answers to the following questions: Does compulsory labeling lead to omission of strong allergenic fragrances in deodorants? Is there a difference in the use patterns of strong and weak allergens? What is the quantitative exposure to fragrances by deodorants? Is the situation in Germany different from other European countries? Is there a difference between deodorants for men and for women? I tested the implementation of the "26 allergens rule" and compiled which allergenic fragrances are specified on the containers of deodorants. Three market studies were conducted in Germany in 2008, 2010 and 2011. The labels of a total number of 374 deodorants were analyzed as to whether any of the "26 allergens" were listed. The frequency of each allergen in the deodorants was compared with results from previous studies by other authors. It was found that up to 83% of the deodorants contain at least one of the "26 allergens" and that up to 30% of all products contain strong allergens above the threshold for labeling (0.001% in the product). The most frequently listed allergens are medium or weak allergens. In comparison with other authors, the frequency of the "26 allergens" in products is slightly smaller in these recent studies for the German market. There is no significant difference between deodorants for men and women, as far as the labeling of the "26 allergens" is concerned. The results show that the mandatory labeling procedure as designed

  13. Identification and characterisation of heat-stable allergens from Sarcoptes scabiei

    Directory of Open Access Journals (Sweden)

    Simson Tarigan

    2006-03-01

    Full Text Available Animals or human recovered from Sarcoptes scabiei infestation acquired protective immunity against reinfestation. The protective immunity is considered to be associated with a type-1-hypersensitivity reaction against allergens instigated by the mites during infestation. It is assumed that these allergens have the potential to be used as the main component of an anti-scabies vaccine. The purpose of this study is to identify and characterise the sarcoptic allergens. For this purpose, 645 mg of mites, collected from mangy goats, were homogenised in PBS to prepare soluble mite proteins. Fractionation of proteins was initially performed on a Q-sepharose column but the results were unsatisfactory. Consequently, SDS PAGE was used as an alternative. Proteins from the gel were transferred onto a nitrocellulose membrane. The membrane was cut into strips so each strip contained proteins with molecular weights of ³ 90, 80-90, 70-80, 60-70, 50-60, 40-50, 30-40, 25-30, 20-25, 15-20 and 10-15 kDa, respectively. The heat stability of the allergens was determined by heating the suspension at 60ºC for 60 minutes, whereas their dialysability was evaluated using a 10-kDa-cut-off ultramembrane. The activity of the allergens was assayed by an intradermal test on sensitised goats. This study showed that mite protein extract was very potent allergens since mite extract containing as little as 1 ng mite proteins still caused an obvious hypersensitive reaction. The mite extract contained heat-stable, dialysable and non-dialysable allergens. All fractions recovered from a Q-sepharose column contained allergens with almost equal potency. Fractionation with the SDS-PAGE revealed that the allergens had molecular weights of 35 and <10 kDa. The former allergen is assumed to be a member of group 10 allergens, whereas the later belong to haptenic allergens.

  14. The influence of digestibility on the allergenicity of food allergens

    DEFF Research Database (Denmark)

    Bøgh, Katrine Lindholm; Madsen, Charlotte Bernhard

    2012-01-01

    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...... 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...... 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...

  15. Presenting a New Standard Drug Model for Turmeric and Its Prized Extract, Curcumin

    Directory of Open Access Journals (Sweden)

    Franco Cavaleri

    2018-01-01

    Full Text Available Various parts of the turmeric plant have been used as medicinal treatment for various conditions from ulcers and arthritis to cardiovascular disease and neuroinflammation. The rhizome’s curcumin extract is the most studied active constituent, which exhibits an expansive polypharmacology with influence on many key inflammatory markers. Despite the expansive reports of curcucmin’s therapeutic value, clinical reliability and research repeatability with curcumin treatment are still poor. The pharmacology must be better understood and reliably mapped if curcumin is to be accepted and used in modern medical applications. Although the polypharmacology of this extract has been considered, in mainstream medicine, to be a drawback, a perspective change reveals a comprehensive and even synergistic shaping of the NF-kB pathway, including transactivation. Much of the inconsistent research data and unreliable clinical outcomes may be due to a lack of standardization which also pervades research standard samples. The possibility of other well-known curcumin by-products contributing in the polypharmacology is also discussed. A new flowchart of crosstalk in transduction pathways that lead to shaping of nuclear NF-kB transactivation is generated and a new calibration or standardization protocol for the extract is proposed which could lead to more consistent data extraction and improved reliability in therapy.

  16. Extracting and standardizing medication information in clinical text - the MedEx-UIMA system.

    Science.gov (United States)

    Jiang, Min; Wu, Yonghui; Shah, Anushi; Priyanka, Priyanka; Denny, Joshua C; Xu, Hua

    2014-01-01

    Extraction of medication information embedded in clinical text is important for research using electronic health records (EHRs). However, most of current medication information extraction systems identify drug and signature entities without mapping them to standard representation. In this study, we introduced the open source Java implementation of MedEx, an existing high-performance medication information extraction system, based on the Unstructured Information Management Architecture (UIMA) framework. In addition, we developed new encoding modules in the MedEx-UIMA system, which mapped an extracted drug name/dose/form to both generalized and specific RxNorm concepts and translated drug frequency information to ISO standard. We processed 826 documents by both systems and verified that MedEx-UIMA and MedEx (the Python version) performed similarly by comparing both results. Using two manually annotated test sets that contained 300 drug entries from medication list and 300 drug entries from narrative reports, the MedEx-UIMA system achieved F-measures of 98.5% and 97.5% respectively for encoding drug names to corresponding RxNorm generic drug ingredients, and F-measures of 85.4% and 88.1% respectively for mapping drug names/dose/form to the most specific RxNorm concepts. It also achieved an F-measure of 90.4% for normalizing frequency information to ISO standard. The open source MedEx-UIMA system is freely available online at http://code.google.com/p/medex-uima/.

  17. Presenting a New Standard Drug Model for Turmeric and Its Prized Extract, Curcumin.

    Science.gov (United States)

    Cavaleri, Franco

    2018-01-01

    Various parts of the turmeric plant have been used as medicinal treatment for various conditions from ulcers and arthritis to cardiovascular disease and neuroinflammation. The rhizome's curcumin extract is the most studied active constituent, which exhibits an expansive polypharmacology with influence on many key inflammatory markers. Despite the expansive reports of curcucmin's therapeutic value, clinical reliability and research repeatability with curcumin treatment are still poor. The pharmacology must be better understood and reliably mapped if curcumin is to be accepted and used in modern medical applications. Although the polypharmacology of this extract has been considered, in mainstream medicine, to be a drawback, a perspective change reveals a comprehensive and even synergistic shaping of the NF-kB pathway, including transactivation. Much of the inconsistent research data and unreliable clinical outcomes may be due to a lack of standardization which also pervades research standard samples. The possibility of other well-known curcumin by-products contributing in the polypharmacology is also discussed. A new flowchart of crosstalk in transduction pathways that lead to shaping of nuclear NF-kB transactivation is generated and a new calibration or standardization protocol for the extract is proposed which could lead to more consistent data extraction and improved reliability in therapy.

  18. Introducing mapping standards in the quality assessment of buildings extracted from very high resolution satellite imagery

    Science.gov (United States)

    Freire, S.; Santos, T.; Navarro, A.; Soares, F.; Silva, J. D.; Afonso, N.; Fonseca, A.; Tenedório, J.

    2014-04-01

    Many municipal activities require updated large-scale maps that include both topographic and thematic information. For this purpose, the efficient use of very high spatial resolution (VHR) satellite imagery suggests the development of approaches that enable a timely discrimination, counting and delineation of urban elements according to legal technical specifications and quality standards. Therefore, the nature of this data source and expanding range of applications calls for objective methods and quantitative metrics to assess the quality of the extracted information which go beyond traditional thematic accuracy alone. The present work concerns the development and testing of a new approach for using technical mapping standards in the quality assessment of buildings automatically extracted from VHR satellite imagery. Feature extraction software was employed to map buildings present in a pansharpened QuickBird image of Lisbon. Quality assessment was exhaustive and involved comparisons of extracted features against a reference data set, introducing cartographic constraints from scales 1:1000, 1:5000, and 1:10,000. The spatial data quality elements subject to evaluation were: thematic (attribute) accuracy, completeness, and geometric quality assessed based on planimetric deviation from the reference map. Tests were developed and metrics analyzed considering thresholds and standards for the large mapping scales most frequently used by municipalities. Results show that values for completeness varied with mapping scales and were only slightly superior for scale 1:10,000. Concerning the geometric quality, a large percentage of extracted features met the strict topographic standards of planimetric deviation for scale 1:10,000, while no buildings were compliant with the specification for scale 1:1000.

  19. Application of porous foams for size-selective measurements of airborne wheat allergen

    NARCIS (Netherlands)

    Bogdanovic, J.; Pater, A.J. de; Doekes, G.; Wouters, I.M.; Heederik, D.J.J.

    2006-01-01

    Background: Exposure to airborne wheat allergen is a well-known cause of bakers' allergy and asthma. Airborne wheat allergen can be measured by enzyme immunoassays (EIAs) in extracts of inhalable dust samples, but only limited knowledge is available on the size distribution of wheat

  20. 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.

  1. 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.

  2. 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...

  3. 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.

  4. Determination of diagnostic standards on saturated soil extracts for cut roses grown in greenhouses.

    Science.gov (United States)

    Franco-Hermida, John Jairo; Quintero, María Fernanda; Cabrera, Raúl Iskander; Guzman, José Miguel

    2017-01-01

    This work comprises the theoretical determination and validation of diagnostic standards for the analysis of saturated soil extracts for cut rose flower crops (Rosa spp.) growing in the Bogota Plateau, Colombia. The data included 684 plant tissue analyses and 684 corresponding analyses of saturated soil extracts, all collected between January 2009 and June 2013. The tissue and soil samples were selected from 13 rose farms, and from cultivars grafted on the 'Natal Briar' rootstock. These concurrent samples of soil and plant tissues represented 251 production units (locations) of approximately 10,000 m2 distributed across the study area. The standards were conceived as a tool to improve the nutritional balance in the leaf tissue of rose plants and thereby define the norms for expressing optimum productive potential relative to nutritional conditions in the soil. To this end, previously determined diagnostic standard for rose leaf tissues were employed to obtain rates of foliar nutritional balance at each analyzed location and as criteria for determining the diagnostic norms for saturated soil extracts. Implementing this methodology to foliar analysis, showed a higher significant correlation for diagnostic indices. A similar behavior was observed in saturated soil extracts analysis, becoming a powerful tool for integrated nutritional diagnosis. Leaf analyses determine the most limiting nutrients for high yield and analyses of saturated soil extracts facilitate the possibility of correcting the fertigation formulations applied to soils or substrates. Recommendations are proposed to improve the balance in soil-plant system with which the possibility of yield increase becomes more probable. The main recommendations to increase and improve rose crop flower yields would be: continuously check pH values of SSE, reduce the amounts of P, Fe, Zn and Cu in fertigation solutions and carefully analyze the situation of Mn in the soil-plant system.

  5. Critical appraisal of Timothy grass pollen extract GRAZAX in the management of allergic rhinitis.

    Science.gov (United States)

    Scaparrotta, Alessandra; Attanasi, Marina; Petrosino, Marianna I; Di Filippo, Paola; Di Pillo, Sabrina; Chiarelli, Francesco

    2015-01-01

    Allergic rhinitis is one of the most common diseases of adult and pediatric age, associated with grass pollen (GP) allergy in >50% cases, with a consistent impact on quality of life of affected patients. A grass allergen tablet, containing standardized extract derived from Timothy grass (Phleum pratense) pollen and ~15 μg major allergen P. pratense (rPhl p 5), may be the future of allergen-specific immunotherapy (IT) for GP allergy. The aim of this review was to critically evaluate the role of Timothy GP extract IT for the management of allergic rhinitis. For this purpose, we have tried to analyze potential mechanisms of action at the basis of Timothy GP extract, we have reviewed efficacy studies to establish potential benefits and clinical response, and we have also evaluated safety and tolerability profiles and patient focus perspective, such as quality of life, satisfaction and acceptability, and compliance to this IT.

  6. Improving the efficacy of the standard DNA differential extraction method for sexual assault evidence.

    Science.gov (United States)

    Timken, Mark D; Klein, Sonja B; Buoncristiani, Martin R

    2018-05-01

    The efficacy of a DNA differential extraction procedure relies on reducing the amount of non-sperm female DNA carryover into the sperm fraction, while providing a sufficient recovery of male DNA from the sperm cell component. A standard approach to this extraction is to use a mild initial lysis step to digest the female (epithelial cell) component in the mixture, followed by a series of centrifugation and wash steps to further purify the resulting sperm-pellet fraction. This sperm fraction is then digested in the presence of a chemical reducing agent in preparation for DNA extraction. This method has been employed with relatively few changes since its introduction in the mid-1980s, despite numerous attempts to develop new or improved procedures. In this report, we demonstrate that it is possible to improve the efficacy of the standard differential extraction by applying simple modifications that can reduce the amount of female DNA carryover into the sperm fraction, with no adverse effects on the recovery of male DNA. In one modification, the addition of a second mild lysis step at the beginning of the differential extraction procedure improved the average male-to-female DNA ratio in the sperm fraction by 3- to 6-fold. In another modification, a "tube transfer" step was added to move the re-suspended sperm pellet to a new tube for the second mild lysis and subsequent wash steps. With this modification, the average male-to-female DNA ratio in the sperm fraction was improved by 4- to 90-fold, relative to results obtained for the non-modified differential extraction method. These modifications may be accomplished using tools and reagents that are already present in most forensic DNA laboratories, so that implementation should be relatively low-cost and practical. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. 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.

  8. Specific Antibodies for the Detection of Alternaria Allergens and the Identification of Cross-Reactive Antigens in Other Fungi

    Science.gov (United States)

    Twaroch, Teresa E.; Curin, Mirela; Sterflinger, Katja; Focke-Tejkl, Margit; Swoboda, Ines; Valenta, Rudolf

    2017-01-01

    Background The mould Alternaria alternata is an important source of respiratory allergens. A. alternata extracts show great variations regarding allergenic potency. The aim of this study was to generate antibody probes specific for important Alternaria allergens and to use them to study allergen expression, depending on different culture conditions, as well as to search for cross-reactive allergens in other mould species. Methods Synthetic peptides from antigenic regions of A. alternata allergens (Alt a 1, Alt a 2, Alt a 3, Alt a 6 and Alt a 8) were used to raise highly specific rabbit antibodies. These antibodies and IgE from allergic patients were used to detect allergens by immunoblotting in extracts of 4 A. alternata strains grown under varying culturing conditions, in commercial skin-prick extracts and in closely (Cladosporium herbarum and Aureobasidium pullulans) or distantly related (Aspergillus niger and Penicillium chrysogenum) mould species. Results There was a wide variation of expression of the individual A. Alternata allergens, depending on the strain and culture conditions, but the antibody probes allowed us to distinguish strains and culture conditions with low and high allergen expression. In the commercial skin-prick solutions, varying levels of Alt a 1 were found, but no other allergens were detectable. Alt a 1 was identified as species-specific A. Alternata allergen, whereas Alt a 3, 6- and Alt a 8-cross-reactive antigens were found in C. herbarum and/or A. pullulans. Conclusions and Clinical Relevance Peptide-specific antibodies are useful to analyze diagnostic and therapeutic mould extracts, to study the presence of A. Alternata allergens in biological samples and to search for cross-reactive allergens in other mould species. PMID:27780168

  9. Antidepressant effect and pharmacological evaluation of standardized extract of flavonoids from Byrsonima crassifolia.

    Science.gov (United States)

    Herrera-Ruiz, M; Zamilpa, A; González-Cortazar, M; Reyes-Chilpa, R; León, E; García, M P; Tortoriello, J; Huerta-Reyes, M

    2011-11-15

    Byrsonima crassifolia (Malpighiaceae) has been used in traditional medicine for the treatment of some mental-related diseases; however, its specific neuropharmacological activities remain to be defined. The present study evaluates the anxiolytic, anticonvulsant, antidepressant, sedative effects produced by the extracts of Byrsonima crassifolia, and their influence on motor activity in ICR mice. Additionally, we determine the acute toxicity profiles of the Byrsonima crassifolia extracts and the presence of neuroactive constituents. Our results show that the methanolic extract of Byrsonima crassifolia produces a significant (P0.05). Although the main compound of the methanolic extract was identified as quercetin 3-O-xyloside (12 mg/kg), our findings suggest that flavonoids, such as rutin (4.4 mg/kg), quercetin (1.4 mg/kg) and hesperidin (0.7 mg/kg), may be involved in the antidepressant effects. To the best of our knowledge, the present study constitutes the first report on the presence of the flavonoids with neuropharmacological activity rutin and hesperidin in Byrsonima crassifolia. In conclusion, the present results showed that the methanolic extract standardized on flavonoids content of Byrsonima crassifolia possesses potential antidepressant-like effects in the FST in mice, and could be considered as relatively safe toxicologically with no deaths of mice when orally administered at 2000 mg/kg. Copyright © 2011 Elsevier GmbH. All rights reserved.

  10. Extracting the properties of dark matter particles in minimal extensions of the standard model

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Maira Dutra Vasconcelos dos; Santos, Antonio Carlos Oliveira; Silva, Paulo Sergio Rodrigues da; Pires, Carlos Antonio de Sousa; Siqueira, Clarissa [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil); Queiroz, Farinaldo da Silva [University of California (United States)

    2013-07-01

    Full text: Nature has provided a striking evidence for physics beyond the Standard Model, namely dark matter. Observations coming from a variety of sources point to the existence of a non-baryonic matter that accounts for roughly 27% of the total abundance of the universe and is composed of neutral, massive, stable and weakly interacting particles. Once the Standard Model has no candidate that fulfills all these properties we must extend it. There are many interesting proposals in the literature that have a good dark matter candidate. Essentially, all of them invoke an extended scalar or gauge sector. Here we aim to extract information about the underlying beyond Standard Model theory able to address the dark matter and many other theoretical puzzles through minimal extensions of the standard model. The minimality perspective it is a worthwhile approach because we can focus on the dark side of many particle physics models. We will carry on our investigation in a pedagogic way Firstly, we will add a neutral fermion, which is our dark matter candidate, and one neutral scalar, both being singlet under the Standard Model gauge group. In this model we compute the abundance of our dark matter candidate and the scattering cross sections off nuclei in order to face our results with the current direct detection experiments data. Secondly, we add a charged scalar field, which is predicted in many standard model extensions, to the first model and investigate the role of this scalar in our results. Lastly, we add a Z' boson to the latter model, and study how our results are affected, with the purpose of, further on, exploring the complementarity between direct detection and collider physics regarding the search of this boson. Thus, we will be able to extract precise information about the beyond Standard Model theory and the properties of the dark matter particles. (author)

  11. Citral a fragrance allergen and irritant.

    Science.gov (United States)

    Heydorn, S; Menné, T; Andersen, K E; Bruze, M; Svedman, C; White, I R; Basketter, D A

    2003-07-01

    Citral is a well known contact allergen and a contact irritant. Routine patch testing in the past may have been restricted because of possible irritant (IR) patch test responses. 586 consecutive patients, with hand eczema, were patch tested with a selection of fragrances including citral 2% petrolatum and the European standard series. 28 of the patients showed a positive patch test reaction (+ to +++) to citral and 82 at least 1 IR patch test reaction and no positive patch test reaction to citral. A statistically significant association between a positive patch test reaction to citral and positive patch test reactions to other fragrances compared with IR reactions (n = 82) was established. The difference regarding fragrance history found between those with IR and positive reactions to citral was not significant. Citral could be an allergen and/or irritant, worthy of further more extensive studies.

  12. 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.

  13. 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,

  14. Allergenic Ingredients in Personal Hygiene Wet Wipes.

    Science.gov (United States)

    Aschenbeck, Kelly A; Warshaw, Erin M

    Wet wipes are a significant allergen source for anogenital allergic contact dermatitis. The aim of the study was to calculate the frequency of potentially allergenic ingredients in personal hygiene wet wipes. Ingredient lists from brand name and generic personal hygiene wet wipes from 4 large retailers were compiled. In the 54 personal hygiene wet wipes evaluated, a total of 132 ingredients were identified (average of 11.9 ingredients per wipe). The most common ingredients were Aloe barbadensis (77.8%), citric acid (77.8%), fragrance (72.2%), sorbic acid derivatives (63.0%), tocopherol derivatives (63.0%), glycerin (59.3%), phenoxyethanol (55.6%), disodium cocoamphodiacetate (53.7%), disodium ethylenediaminetetraacetic acid (EDTA) (42.6%), propylene glycol (42.6%), iodopropynyl butylcarbamate (40.7%), chamomile extracts (38.9%), sodium benzoate (35.2%), bronopol (22.2%), sodium citrate (22.2%), lanolin derivatives (20.4%), parabens (20.4%), polyethylene glycol derivatives (18.5%), disodium phosphate (16.7%), dimethylol dimethyl hydantoin (DMDM) (14.8%), and cocamidopropyl propylene glycol (PG)-dimonium chloride phosphate (11.1%). Of note, methylisothiazolinone (5.6%) was uncommon; methylchloroisothiazolinone was not identified in the personal hygiene wet wipes examined. There are many potential allergens in personal hygiene wet wipes, especially fragrance and preservatives.

  15. 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...

  16. Rule-based Approach on Extraction of Malay Compound Nouns in Standard Malay Document

    Science.gov (United States)

    Abu Bakar, Zamri; Kamal Ismail, Normaly; Rawi, Mohd Izani Mohamed

    2017-08-01

    Malay compound noun is defined as a form of words that exists when two or more words are combined into a single syntax and it gives a specific meaning. Compound noun acts as one unit and it is spelled separately unless an established compound noun is written closely from two words. The basic characteristics of compound noun can be seen in the Malay sentences which are the frequency of that word in the text itself. Thus, this extraction of compound nouns is significant for the following research which is text summarization, grammar checker, sentiments analysis, machine translation and word categorization. There are many research efforts that have been proposed in extracting Malay compound noun using linguistic approaches. Most of the existing methods were done on the extraction of bi-gram noun+noun compound. However, the result still produces some problems as to give a better result. This paper explores a linguistic method for extracting compound Noun from stand Malay corpus. A standard dataset are used to provide a common platform for evaluating research on the recognition of compound Nouns in Malay sentences. Therefore, an improvement for the effectiveness of the compound noun extraction is needed because the result can be compromised. Thus, this study proposed a modification of linguistic approach in order to enhance the extraction of compound nouns processing. Several pre-processing steps are involved including normalization, tokenization and tagging. The first step that uses the linguistic approach in this study is Part-of-Speech (POS) tagging. Finally, we describe several rules-based and modify the rules to get the most relevant relation between the first word and the second word in order to assist us in solving of the problems. The effectiveness of the relations used in our study can be measured using recall, precision and F1-score techniques. The comparison of the baseline values is very essential because it can provide whether there has been an improvement

  17. The role of contact allergens in chronic idiopathic urticaria.

    Science.gov (United States)

    Hession, Meghan T; Scheinman, Pamela L

    2012-01-01

    The objective of this study was to determine whether contact allergens play a role in chronic idiopathic urticaria (CIU). We conducted a longitudinal prospective study of 23 patients with CIU. Patients were patch tested to a modified North American Contact Dermatitis Group standard, fragrance, and cosmetic series; other series were tested as warranted by relevant history and physical examination. Readings were performed at 48 and 72 hours. Patients were counseled to avoid proven contact allergens and were followed up 2 to 9 months after testing. Twenty-one of 23 patients were female. The mean age was 46 years. The mean duration of urticaria was 32 months. Of the 23 patients, 8 (35%) experienced improvement of their symptoms with allergen avoidance. Four (17%) experienced a complete remission, and 4 (17%) experienced partial improvement. Two of the complete responders challenged themselves to proven contact allergens and developed urticaria, which resolved upon allergen avoidance. The most common allergens were potassium dichromate (n = 9), nickel sulfate (n = 7), Myroxylon pereirae (n = 6), cobalt chloride, neomycin, p-phenylenediamine (n = 5); fragrance mix I, fragrance mix II (n = 4); cinnamic aldehyde (n = 3); and formaldehyde (n = 2). Patch testing may be helpful in the evaluation of CIU patients for whom previous workup has failed to reveal an etiology.

  18. Prevalence of food and airborne allergens in allergic patients in Kerman

    Directory of Open Access Journals (Sweden)

    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.

  19. Expression levels of parvalbumins determine allergenicity of fish species.

    Science.gov (United States)

    Griesmeier, U; Vázquez-Cortés, S; Bublin, M; Radauer, C; Ma, Y; Briza, P; Fernández-Rivas, M; Breiteneder, H

    2010-02-01

    Parvalbumins are the most important fish allergens. Polysensitization to various fish species is frequently reported and linked to the cross-reactivity of their parvalbumins. Studies on cross-reactivity and its association to the allergenicity of purified natural parvalbumins from different fish species are still lacking. In addition, some studies indicate that dark muscled fish such as tuna are less allergenic. Total protein extracts and purified parvalbumins from cod, whiff, and swordfish, all eaten frequently in Spain, were tested for their IgE-binding properties with 16 fish allergic patients' sera from Madrid. The extent of cross-reactivity of these parvalbumins was investigated by IgE ELISA inhibition assays. Additionally, the cDNA sequences of whiff and swordfish parvalbumins were determined. Extractable amounts of parvalbumins from cod were 20 times and from whiff 30 times higher than from swordfish. Parvalbumins were recognized by 94% of the patients in extracts of cod and whiff, but only by 60% in swordfish extracts. Nevertheless, a high cross-reactivity was determined for all purified parvalbumins by IgE inhibition. The amino acid sequence identities of the three parvalbumins were in a range of 62-74%. The parvalbumins of cod, whiff and swordfish are highly cross-reactive. The high amino acid sequence identity among cod, whiff and swordfish parvalbumins results in the observed IgE cross-reactivity. The low allergenicity of swordfish is due to the low expression levels of its parvalbumin.

  20. The identification of allergen proteins in sugar beet (Beta vulgaris) pollen causing occupational allergy in greenhouses.

    Science.gov (United States)

    Luoto, Susanne; Lambert, Wietske; Blomqvist, Anna; Emanuelsson, Cecilia

    2008-08-11

    During production of sugar beet (Beta vulgaris) seeds in greenhouses, workers frequently develop allergic symptoms. The aim of this study was to identify and characterize possible allergens in sugar beet pollen. Sera from individuals at a local sugar beet seed producing company, having positive SPT and specific IgE to sugar beet pollen extract, were used for immunoblotting. Proteins in sugar beet pollen extracts were separated by 1- and 2-dimensional electrophoresis, and IgE-reactive proteins analyzed by liquid chromatography tandem mass spectrometry. A 14 kDa protein was identified as an allergen, since IgE-binding was inhibited by the well-characterized allergen Che a 2, profilin, from the related species Chenopodium album. The presence of 17 kDa and 14 kDa protein homologues to both the allergens Che a 1 and Che a 2 were detected in an extract from sugar beet pollen, and partial amino acid sequences were determined, using inclusion lists for tandem mass spectrometry based on homologous sequences. Two occupational allergens were identified in sugar beet pollen showing sequence similarity with Chenopodium allergens. Sequence data were obtained by mass spectrometry (70 and 25%, respectively for Beta v 1 and Beta v 2), and can be used for cloning and recombinant expression of the allergens. As for treatment of Chenopodium pollinosis, immunotherapy with sugar beet pollen extracts may be feasible.

  1. An experimental evaluation of adaptogenic potential of standardized epipremnum aureum leaf extract

    Directory of Open Access Journals (Sweden)

    Sreemoy Kanti Das

    2017-01-01

    Full Text Available Background: Stress is a normal part of everyday life but chronic stress can lead to a variety of stress-related illnesses including hypertension, anxiety, and depression. In the present investigation, standardized leaf extract of Epipremnumaureum was evaluated for its anti-stress potential. Materials and Methods: For the evaluation of anti-stress activity, groups of mice (n = 6 were subjected to forced swim stress and anoxic stress tolerance test in mice 1h after daily treatment of E.aureumextract. Diazepam (5 mg/kg was taken as a reference standard. Urinary vanillylmandelic acid (VMA and ascorbic acid were selected as noninvasive biomarkers to assess the anti-stress activity and plasma cortisol, blood ascorbic acid, and weight of adrenal were measured. The 24 h urinary excretion of VMA and ascorbic acid were determined by spectrophotometric methods in all groups under normal and stressed conditions. The hematological parameters (neutrophils, lymphocytes, and eosinophils were also determined. Results: Administration of E.aureumat doses of 400 and 600 mg/kg wasfound to be effective in inhibiting the stress induced urinary biochemical changes in a dose-dependent manner. Treatment with E. aureum extract prevents the rise in blood ascorbic acid and plasma cortisol. Moreover, the extract prevented the increase in weight of adrenal gland also significantly increased the anoxia stress tolerance time. Dose-dependent significant reduction in white blood cell count was observed in anoxic stress tolerance test as compared to stressed group. Conclusion: Hence, the present study provides scientific support for the positiveadaptogenic effect of E. aureum extract.

  2. Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae

    Directory of Open Access Journals (Sweden)

    Yubao Cui

    2012-08-01

    Full Text Available Crude extracts of house dust mites are used clinically for diagnosis and immunotherapy of allergic diseases, including bronchial asthma, perennial rhinitis, and atopic dermatitis. However, crude extracts are complexes with non-allergenic antigens and lack effective concentrations of important allergens, resulting in several side effects. Dermatophagoides farinae (Hughes; Acari: Pyroglyphidae is one of the predominant sources of dust mite allergens, which has more than 30 groups of allergen. The cDNA coding for the group 5 allergen of D. farinae from China was cloned, sequenced and expressed. According to alignment using the VECTOR NTI 9.0 software, there were eight mismatched nucleotides in five cDNA clones resulting in seven incompatible amino acid residues, suggesting that the Der f 5 allergen might have sequence polymorphism. Bioinformatics analysis revealed that the matured Der f 5 allergen has a molecular mass of 13604.03 Da, a theoretical pI of 5.43 and is probably hydrophobic and cytoplasmic. Similarities in amino acid sequences between Der f 5 and allergens of other domestic mite species, viz. Der p 5, Blo t 5, Sui m 5, and Lep d 5, were 79, 48, 53, and 37%, respectively. Phylogenetic analysis indicated that Der f 5 and Der p 5 clustered together. Blo t 5 and Ale o 5 also clustered together, although Blomia tropicalis and Aleuroglyphus ovatus belong to different mite families, viz. Echimyopodidae and Acaridae, respectively.

  3. Detection of allergen coding sequences of kiwi, peach, and apple in processed food by qPCR.

    Science.gov (United States)

    Graziano, Sara; Gullì, Mariolina; Marmiroli, Nelson

    2017-12-06

    Food traceability becomes lifesaving for persons suffering severe allergy or intolerance, and therefore need a complete avoidance of the immune-trigger food. This paper describes how to fingerprint the presence of some allergenic species (kiwi, peach, and apple) in foods by quantitative real-time PCR (qPCR). Five DNA extraction procedures were tested on fruits and foods. The results were statistically evaluated, and discussed. Analysis by qPCR with SYBR Green was developed to detect traces of these allergenic species in foods. Plasmids containing the target sequences of kiwi, peach and apple were employed as internal reference standard. Analysis of spiked food samples showed a limit of detection of 25 mg kg -1 for kiwi, 20 mg kg -1 for peach and 50 mg kg -1 for apple. The qPCR method here developed, combined with the use of internal plasmid reference standard, represents a specific system for the quick detection of allergenic species in complex food matrices, with a limit of detection comparable with those reported using more time-consuming methods. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  4. Contact allergens in oral antihistamines.

    Science.gov (United States)

    McEnery-Stonelake, Melissa; Silvestri, Dianne L

    2014-01-01

    Excipients in various formulations of active drugs occasionally include known contact allergens. Their ingestion may trigger dermatitis or cause it to become widespread or refractory to therapy. The aim of this study was to investigate the prevalence of common contact allergens among the excipients of oral antihistamines available in this country. We gathered the complete ingredient lists of 2119 different preparations of 12 oral antihistamines from the National Library of Medicine data bank and entered them into an electronic database for analysis. More than half the formulations (55.0%) contained at least 1 member of the 10 allergen families assessed. Most brompheniramine and doxepin preparations included potentially allergenic excipients, whereas fexofenadine was most often free of them. Sorbitan group members, azo dyes, and propylene glycol were the allergens found most frequently in the antihistamines, each present in over 25% of the products. Elixirs, liquids, solutions, suspensions, and syrups were more likely than nonchewable caplets, capsules, and tablets to contain the allergens tabulated (100% vs 39.3%, respectively). Chewable pills frequently contained azo dyes. Ingestion of antihistamines could precipitate a systemic contact dermatitis in a patient sensitized to an allergen present as an excipient in the medicine.

  5. Monoclonal antibody-based ELISA to quantify the major allergen of Artemisia vulgaris pollen, Art v 1.

    Science.gov (United States)

    Jimeno, L; Duffort, O; Serrano, C; Barber, D; Polo, F

    2004-09-01

    Pollen of Artemisia vulgaris (mugwort) is a relevant cause of pollinosis in temperate and humid regions. Recently, the major allergen of this pollen, Art v 1, has been characterized. To develop a monoclonal antibody (mAb)-based enzyme-linked immunosorbent assay (ELISA) to quantify Art v 1, and to assess the correlation of Art v 1 content with the biological activity of mugwort pollen extracts. Art v 1-specific mAbs were obtained from a BALB/c mouse immunized with high-performance liquid chromatography (HPLC)-purified Art v 1. One of these antibodies (Av 3.7), which recognizes the N-terminal defensin-like domain of Art v 1, was used as the capture antibody in an ELISA method for allergen quantitation. An anti-A. vulgaris rabbit serum was used as the second antibody. Art v 1 was purified by immunoaffinity chromatography and used as the standard in the assay. The purity and identity of the affinity-purified Art v 1 was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), mass spectrometry, amino acid composition, and N-terminal amino acid sequencing. The prevalence of specific IgE against Art v 1, determined by radioallergosorbent test (RAST) in a population of 44 mugwort-allergic patients, was 79%. The Art v 1-ELISA developed displays a detection limit of 0.1 ng/ml, and a practical working range of 0.2-10 ng/ml. The concentration of Art v 1 was measured in 10 A. vulgaris pollen extracts, and a good correlation was observed between the Art v 1 content and the allergenic activity of the extracts. The results prove the usefulness of the Art v 1-ELISA for the standardization of A. vulgaris pollen extracts intended for clinical use.

  6. The allergens causing contact sensitization in textile industry workers

    Directory of Open Access Journals (Sweden)

    Özlem Su

    2014-09-01

    Full Text Available Background and Design: Our aim was to determine the frequency of contact sensitization to textile materials and the most common textile allergens in patients who work in the textile industry and have been diagnosed with allergic contact dermatitis (ACD. Materials and Methods: Fifty textile industry workers, who attended our outpatient clinic with the diagnosis of ACD, between October 2005-December 2009, were enrolled in this study. While 50 patients were tested with the Thin layer-Rapid-Use-Epicutaneous (TRUE test, 36 patients were tested with the TRUE test and textile series allergen. The results were analyzed as percentage and statistically. Results: Of the 50 patients, 34 (68% were men, 16 (32% were women. The mean of the subjects was 37.4 years. In 38.8 of the patients in whom TRUE test and textile series allergens applied together, at least one allergic reaction was seen. Of the 36 patients applied textile series in addition to the TRUE test, 33.3% of patients had positive reaction to only textile allergens, 2.7% to only standard series allergens, and 2.7% of subjects had positive reaction to both textile series and standard series allergens. 76.9% of these reactions were to dyes, 15.3% to resins and 7.6% of them were to both of them. Disperse blue 106 (8.3%, acid red 359 (8.3% and disperse red 17 (5.5% were the most positive reaction seen dyes. 18% of 50 patients tested with TRUE test alone showed at least one positive reaction. The most common standard series allergens were nickel sulphate (6% and ethylenediamine dihydrocloride (6%. Both the relationship between atopy and contact sensitization and also the relationship with hand localization and contact sensitization to textile allergens were not statistically significant. Conclusion: In occupational textile dermatitis, contact sensitization is common and especially seen to disperse dyes. For the contact sensitization to textile materials, standard series allergens cannot be adequate in

  7. Standardization of Tragopogon graminifolius DC. Extract Based on Phenolic Compounds and Antioxidant Activity

    Directory of Open Access Journals (Sweden)

    Mohammad Hosein Farzaei

    2014-01-01

    Full Text Available Tragopogon graminifolius DC. (TG, Compositae family, is traditionally used for the treatment of various diseases like gastrointestinal and hepatic disorders. The aim of the present study is to standardize extracts from TG used for preparation of different dosage forms in traditional Iranian medicine (TIM based on phenolic compounds. For this purpose, total phenolic content and some phenolic compounds were determined in ethanolic extracts from aerial part and root of TG by HPLC method. Furthermore, antioxidant activity was evaluated using DPPH-HPLC methods. Caffeic acid, gallic acid, ρ-coumaric acid, ferulic acid, and catechin were detected in root and aerial part of TG. ρ-Coumaric acid (6.357 ± 0.014 mg·g−1 was dominant phenolic compound in aerial part followed by ferulic acid (1.24 ± 0.018 mg·g−1. Also, ρ-coumaric acid (2.685 ± 0.031 mg·g−1 was highly abundant in root, followed by catechin (2.067 ± 0.021 mg·g−1. Antioxidant activity of root extract (460.45 ± 0.78 µg Vit.E.E·mL−1 was better than that of aerial part. Generally, phenolic compounds are one of the major constituents of TG and could be used as markers for standardization of dosage forms prepared from this plant. Also, TG demonstrated significant antioxidant activity using DPPH-HPLC method. Phenolic compounds of TG may be responsible for its marked antioxidant properties.

  8. Hepatoprotective and antioxidant polyphenols from a standardized methanolic extract of the leaves of Liquidambar styraciflua L.

    Directory of Open Access Journals (Sweden)

    Hanaa H. Eid

    2015-12-01

    Full Text Available The methanolic extract of the leaves of Liquidambar styraciflua L. (Altingiaceae (LSE was evaluated for hepatoprotective and antioxidant activities in carbon tetrachloride liver-damaged rats. Hepatotoxicity was induced via intraperitoneal injection of CCl4 1:9 in olive oil, at a dose of 0.5 ml/kg b.wt. The animals received the extract, orally, at two dose levels (250 and 500 mg/kg b.wt. The administration regimen was twice a week, for six consecutive weeks. LSE exhibited a significant dose-dependent protective effect by lowering the serum levels of alanine aminotransferase (ALT, aspartate aminotransferase (AST, alkaline phosphatase (ALP, malondialdehyde (MDA and ameliorating the level of serum protein. In addition, LSE showed antioxidant activity through improving the levels of blood glutathione (GSH, vitamin C, vitamin E and hepatic total protein contents. The LSE revealed activity approached that of silymarin, a known hepatoprotective agent. These biochemical observations were supported by examination of the histopathological features of the liver. Chromatographic fractionation of LSE afforded seven phenolic compounds. These were identified on the basis of chromatographic, chemical and spectroscopic analyses as: gallic acid (1, isorugosin B (2, casuarictin (3, quercetin-3-O-β-d-4C1-glucopyranoside (4, myricetin-3-O-α-l-1C4-rhamnopyranoside (myricetrin (5, quercetin (6 and myricetin (7. The isolated phenolics probably account for the antioxidant and hepatoprotective effects exhibited by the parent extract. Furthermore, a validated RP-HPLC method was devised for standardization of LSE, in view to fulfill the requirements of efficient research methodology for evaluation of bioactive herbal drugs. The above findings suggested that LSE could be considered as a standardized herbal product with antifibrotic, hepatoprotective and antioxidant potential.

  9. Gastrointestinal digestion of food allergens: effect on their allergenicity.

    Science.gov (United States)

    Moreno, F Javier

    2007-01-01

    This paper reviews the in vitro digestion models developed to assess the stability digestion of food allergens, as well as the factors derived from the methodology and food structure that may affect the assay results. The adequacy of using the digestion stability of food allergens as a criterion for assessing potential allergenicity is also discussed. Data based on the traditional pepsin digestibility test in simulated gastric fluid are discussed in detail, with special attention to the influence of the pH and pepsin: allergen ratio in the pepsinolysis rate. This review points out the importance of using physiologically relevant in vitro digestion systems for evaluating digestibility of allergens. This would imply the sequential use of digestive enzymes in physiological concentrations, simulation of the stomach/small intestine environment (multi-phase models) with addition of surfactants such as phospholipids or bile salts, as well as the consideration of the gastrointestinal transit and the effect of the food matrices on the allergen digestion and subsequent absorption through the intestinal mucosa. In vitro gastrointestinal digestion protocols should be preferably combined with immunological assays in order to elucidate the role of large digestion-resistant fragments and the influence of the food matrix on the stimulation of the immune system.

  10. Does allergen-specific immunotherapy induce contact allergy to aluminium?

    Science.gov (United States)

    Netterlid, Eva; Hindsén, Monica; Siemund, Ingrid; Björk, Jonas; Werner, Sonja; Jacobsson, Helene; Güner, Nuray; Bruze, Magnus

    2013-01-01

    Persistent, itching nodules have been reported to appear at the injection site after allergen-specific immuno-therapy with aluminium-precipitated antigen extract, occasionally in conjunction with contact allergy to aluminium. This study aimed to quantify the development of contact allergy to aluminium during allergen-specific immunotherapy. A randomized, controlled, single-blind multicentre study of children and adults entering allergen-specific immunotherapy was performed using questionnaires and patch-testing. A total of 205 individuals completed the study. In the 3 study groups all subjects tested negative to aluminium before allergen-specific immunotherapy and 4 tested positive after therapy. In the control group 4 participants tested positive to aluminium. Six out of 8 who tested positive also had atopic dermatitis. Positive test results were found in 5/78 children and 3/127 adults. Allergen-specific immunotherapy was not shown to be a risk factor for contact allergy to aluminium. Among those who did develop aluminium allergy, children and those with atopic dermatitis were more highly represented.

  11. Matrix metalloproteinase, hyaluronidase and elastase inhibitory potential of standardized extract of Centella asiatica.

    Science.gov (United States)

    Nema, Neelesh Kumar; Maity, Niladri; Sarkar, Birendra Kumar; Mukherjee, Pulok Kumar

    2013-09-01

    Centella asiatica (L.) Urban (Apiaceae), a valuable herb described in Ayurveda, is used in the indigenous system of medicine as a tonic to treat skin diseases. Centella asiatica methanol extract and its ethyl acetate, n-butanol and aqueous fraction, were subjected for the evaluation of skin care potential through the in vitro hyaluronidase, elastase and matrix metalloproteinase-1 (MMP-1) inhibitory assay. The C. asiatica plant was extracted with methanol and fractionated with ethyl acetate, n-butanol and water. The enzymatic activities were evaluated using ursolic acid and oleanolic acid as standards. Isolate molecule asiaticoside was quantified in the crude extract and fractions through high-performance liquid chromatography (HPLC) and structural was characterized by liquid chromatography-mass spectroscopy (LC-MS) and ¹H nuclear magnetic resonance (NMR). Isolated compound was also evaluated for in vitro enzyme assays. Extract exhibited anti-hyaluronidase and anti-elastase activity with IC₅₀ of 19.27 ± 0.37 and 14.54 ± 0.39 µg/mL, respectively, as compared to ursolic acid. Centella asiatica n-butanol fraction (CAnB) and isolated compound showed significant hyaluronidase (IC₅₀ = 27.00 ± 0.43 and 18.63 ± 0.33 µg/mL) and elastase (IC₅₀ = 29.15 ± 0.31 and 19.45 ± 0.25 µg/mL) inhibitory activities, respectively, and also showed significant MMP-1 inhibition (p asiatica may be a prospective agent for skin care.

  12. The variability of standard artificial soils: Behaviour, extractability and bioavailability of organic pollutants

    Energy Technology Data Exchange (ETDEWEB)

    Hofman, Jakub, E-mail: hofman@recetox.muni.cz [Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Kamenice 753/5, Brno CZ-62500 (Czech Republic); Hovorková, Ivana [Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Kamenice 753/5, Brno CZ-62500 (Czech Republic); Semple, Kirk T. [Lancaster Environment Centre, Lancaster University, Lancaster LA1 4YQ (United Kingdom)

    2014-01-15

    Highlights: • Artificial soils from different laboratories revealed different fates, behaviour and bioavailability of lindane and phenanthrene. • Lindane behaviour was related to organic carbon. • Phenanthrene behaviour was significantly affected by degrading microorganisms from peat. • Sterilization of artificial soils might reduce unwanted variability. -- Abstract: Artificial soil is an important standard medium and reference material for soil ecotoxicity bioassays. Recent studies have documented the significant variability of their basic properties among different laboratories. Our study investigated (i) the variability of ten artificial soils from different laboratories by means of the fate, extractability and bioavailability of phenanthrene and lindane, and (ii) the relationships of these results to soil properties and ageing. Soils were spiked with {sup 14}C-phenanthrene and {sup 14}C-lindane, and the total residues, fractions extractable by hydroxypropyl-β-cyclodextrin, and the fractions of phenanthrene mineralizable by bacteria were determined after 1, 14, 28 and 56 days. Significant temporal changes in total residues and extractable and mineralizable fractions were observed for phenanthrene, resulting in large differences between soils after 56 days. Phenanthrene mineralization by indigenous peat microorganisms was suggested as the main driver of that, outweighing the effects of organic matter. Lindane total residues and extractability displayed much smaller changes over time and smaller differences between soils related to organic matter. Roughly estimated, the variability between the artificial soils was comparable to natural soils. The implications of such variability for the results of toxicity tests and risk assessment decisions should be identified. We also suggested that the sterilization of artificial soils might reduce unwanted variability.

  13. Optimization of two methods based on ultrasound energy as alternative to European standards for soluble salts extraction from building materials.

    Science.gov (United States)

    Prieto-Taboada, N; Gómez-Laserna, O; Martinez-Arkarazo, I; Olazabal, M A; Madariaga, J M

    2012-11-01

    The Italian recommendation NORMAL 13/83, later replaced by the UNI 11087/2003 norm, were used as standard for soluble salts extraction from construction materials. These standards are based on long-time stirring (72 and 2h, respectively) of the sample in deionized water. In this work two ultrasound based methods were optimized in order to reduce the extraction time while efficiency is improved. The instrumental variables involved in the extraction assisted by ultrasound bath and focused ultrasounds were optimized by experimental design. As long as it was possible, the same non-instrumental parameters values as those of standard methods were used in order to compare the results obtained on a mortar sample showing a black crust by the standards and the optimized methods. The optimal extraction time for the ultrasounds bath was found to be of two hours. Although the extraction time was equal to the standard UNI 11087/2003, the obtained extraction recovery was improved up to 119%. The focused ultrasound system achieved also better recoveries (up to 106%) depending on the analyte in 1h treatment time. The repeatabilities of the proposed ultrasound based methods were comparables to those of the standards. Therefore, the selection of one or the other of the ultrasound based methods will depend on topics such as laboratory facilities or number of samples, and not in aspects related with their quality parameters. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. 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.

  15. Allergen immunotherapy for allergic rhinoconjunctivitis

    DEFF Research Database (Denmark)

    Nurmatov, Ulugbek; Dhami, Sangeeta; Arasi, Stefania

    2017-01-01

    Background: The European Academy of Allergy and Clinical Immunology (EAACI) is developing Guidelines on Allergen Immunotherapy (AIT) for Allergic Rhinoconjunctivitis (ARC). To inform the development of recommendations, we sought to critically assess the systematic review evidence on the effective...

  16. Important variations in parvalbumin content in common fish species: a factor possibly contributing to variable allergenicity.

    Science.gov (United States)

    Kuehn, A; Scheuermann, T; Hilger, C; Hentges, F

    2010-01-01

    Although 95% of fish-allergic patients are sensitized to the major fish allergen parvalbumin, clinical reactions to different fish species vary considerably in symptoms, intensity and frequency in allergic subjects. This study aimed at the quantification of parvalbumin levels in salmon, trout, cod, carp, mackerel, herring, redfish and tuna. Fish muscle extracts were separated by SDS-PAGE and parvalbumin content was estimated by densitometric band quantification. Individual antisera were raised in BALB/c mice against parvalbumins purified from seven fish species. Parvalbumin content was quantified in fish (raw/processed) and skin prick test (SPT) solutions by ELISA using the corresponding anti-serum for detection and the purified parvalbumins as standards. Using SDS-PAGE scanning, parvalbumin contents were 2 mg for cod, carp, redfish and herring. Using ELISA, parvalbumin content ranged from 2.5 mg per gram raw muscle for carp, herring and redfish. The parvalbumin content of processed samples (cooked/commercial) was 20-60% lower. Allergen content in SPT samples ranged from 20 to 70 μg parvalbumin/ml of extract. No parvalbumin was found in tuna SPT solution. The parvalbumin content of most commonly consumed fish species varies considerably. Differences range from severalfold to one hundredfold. This has to be taken into account when designing food challenge tests and advising fish-allergic patients. Copyright © 2010 S. Karger AG, Basel.

  17. Allergen and allergy risk assessment, allergen management, and gaps in the European Food Information Regulation (FIR): Are allergic consumers adequately protected by current statutory food safety and labelling regulations?

    Science.gov (United States)

    Reese, Imke; Holzhauser, Thomas; Schnadt, Sabine; Dölle, Sabine; Kleine-Tebbe, Jörg; Raithel, Martin; Worm, Margitta; Zuberbier, Torsten; Vieths, Stefan

    Individuals suffering from IgE-mediated food allergies are obliged to systematically eliminate the culprit allergen from their diet. To support allergic consumers in avoiding food allergens to make informed and safe purchasing choices, the European Union (EU) Food Information Regulation (FIR) imposes a requirement to label the 13 most common allergenic foods (food groups) as ingredients in pre-packed and non-pre-packed foods. The as yet unregulated labelling of unintended presence of allergens has lead to a widespread use of precautionary allergen labelling (PAL) - despite established allergen management in many companies. This PAL significantly hampers making a safe food choice, or renders it largely unfeasible, since it is not possible to estimate the actual extent to which allergens are present in a product. Not only food-allergic consumers, but also the food industry, physicians, dieticians/nutritionists, as well as food regulators and law enforcement officers would benefit from a standardized labelling regulation for unintended presence of allergens. The following position paper highlights the chances of such a regulation on the basis of available data and the analytical methods for detecting allergenic components in the context of effective allergen management. On the basis of evidence-based allergen and allergy-risk assessment, the use of PAL should be restricted to only those allergens that are unavoidable and which represent an unacceptable risk for allergic consumers, e. g., when allergens are present at levels that exceed a clinically meaningful reference dose or in the case of unintended presence of particulate allergens (e. g., nuts or sesame seeds).

  18. Identification of the major allergens of Indian scad (Decapterus russelli).

    Science.gov (United States)

    Misnan, Rosmilah; Murad, Shahnaz; Jones, Meinir; Taylor, Graham; Rahman, Dinah; Arip, Masita; Abdullah, Noormalin; Mohamed, Jamaluddin

    2008-12-01

    The purpose of this study was to characterize major allergens of Indian scad (Decapterus russelli) which is among the most commonly consumed fish in Malaysia. Raw and cooked extracts of the fish were prepared. Protein profiles and IgE binding patterns were produced by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting using sera from subjects with fish allergy. The major allergens of the fish were then identified by two-dimensional electrophoresis (2-DE), followed by mass spectrometry of the peptide digests. The SDS-PAGE of the raw extract revealed 27 protein fractions over a wide molecular weight range, while the cooked extract demonstrated only six protein fractions. The 1-DE immunoblotting detected 14 IgE-binding proteins, with a molecular weight range from 90 to fish. The approximately 12 kDa band was a heat-resistant protein while the approximately 51 and 46 kDa proteins were sensitive to heat. The 2-DE gel profile of the raw extract demonstrated > 100 distinct protein spots and immunoblotting detected at least 10 different major IgE reactive spots with molecular masses as expected and isoelectric point (pI) values ranging from 4.0 to 7.0. A comparison of the major allergenic spot sequences of the 12 kDa proteins with known protein sequences in databases revealed extensive similarity with fish parvalbumin. In conclusion, this study demonstrated that a parvalbumin which is similar to Gad c 1 is the major allergen of Indian scad. Interestingly, we also detected heat-sensitive proteins as major allergenic components in our fish allergy patients.

  19. Allergenicity and cross-reactivity of booklice (Liposcelis bostrichophila): a common household insect pest in Japan.

    Science.gov (United States)

    Fukutomi, Yuma; Kawakami, Yuji; Taniguchi, Masami; Saito, Akemi; Fukuda, Azumi; Yasueda, Hiroshi; Nakazawa, Takuya; Hasegawa, Maki; Nakamura, Hiroyuki; Akiyama, Kazuo

    2012-01-01

    Booklice (Liposcelis bostrichophila) are a common household insect pest distributed worldwide. Particularly in Japan, they infest 'tatami' mats and are the most frequently detected insect among all detectable insects, present at a frequency of about 90% in dust samples. Although it has been hypothesized that they are an important indoor allergen, studies on their allergenicity have been limited. To clarify the allergenicity of booklice and the cross-reactivity of this insect allergen with allergens of other insects, patients sensitized to booklice were identified from 185 Japanese adults with allergic asthma using skin tests and IgE-ELISA. IgE-inhibition analysis, immunoblotting and immunoblotting-inhibition analysis were performed using sera from these patients. Allergenic proteins contributing to specific sensitization to booklice were identified by two-dimensional electrophoresis and two-dimensional immunoblotting. The booklouse-specific IgE antibody was detected in sera from 41 patients (22% of studied patients). IgE inhibition analysis revealed that IgE reactivity to the booklouse allergen in the sera from one third of booklouse-sensitized patients was not inhibited by preincubation with extracts from any other environmental insects in this study. Immunoblotting identified a 26-kD protein from booklouse extract as the allergenic protein contributing to specific sensitization to booklice. The amino acid sequence of peptide fragments of this protein showed no homology to those of previously described allergenic proteins, indicating that this protein is a new allergen. Sensitization to booklice was relatively common and specific sensitization to this insect not related to insect panallergy was indicated in this population. Copyright © 2011 S. Karger AG, Basel.

  20. Quantification in mass units of group 1 grass allergens by a monoclonal antibody-based sandwich ELISA.

    Science.gov (United States)

    Arilla, M C; Ibarrola, I; Eraso, E; Aguirre, M; Martínez, A; Asturias, J A

    2001-08-01

    Grass pollen extracts currently used for allergy diagnosis and immunotherapy are a complex mixture of proteins of which only a few have allergenic activity. Lol p 1 is one of the most important allergens in grass pollen extracts. To develop a two-site enzyme-linked immunosorbent assay for the quantification of Lol p 1 and other group 1 allergens from grass species, and to assess its suitability for quantifying this group of allergens. Balb/c mice immunized with recombinant Lol p 1 were used for the production of monoclonal antibodies. Screening of hybridomas was performed by direct ELISA, and selected monoclonal antibodies were immobilized on ELISA plates and incubated with samples containing group 1 allergens. Bound allergens were detected by a combination of biotinylated Lol p 1-specific monoclonal antibody and peroxidase-streptavidin conjugate. The assay is based on three Lol p 1-specific monoclonal antibodies with different epitope specificities. The optimized ELISA measured Lol p 1 concentrations ranging from 125 to 1000 ng/mL and could quantify group 1 allergen from grass species belonging to the Pooidea subfamily. The assay does not depend on anti-sera production or availability of human sera and thus reactives can be produced in unlimited amounts. This sensitive and specific Lol p 1 assay will be helpful both for quantifying the group 1 allergen content of Pooideae pollen extracts intended for clinical use and for studying cross-reactivities among pollen extracts.

  1. 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.

  2. Anticholinesterase activity of standardized extract of Illicium verum Hook. f. fruits.

    Science.gov (United States)

    Bhadra, Santanu; Mukherjee, Pulok K; Kumar, N Satheesh; Bandyopadhyay, A

    2011-04-01

    Illicium verum is a well known spice in traditional Indian system for its therapeutic potential. The present study was aimed to evaluate the acetylcholinesterase (AChE) and butyrylcholinesterase inhibitory (BChE) activity of standardized extracts of I. verum and its oil. Present study confirmed that anethole contributed to the anticholinesterase activity of I. verum, with more specificity towards AChE. IC(50) for AChE and BChE inhibitory activity of anethole was 39.89±0.32 μg/mL and 75.35±1.47 μg/mL, whereas for the oil, 36.00±0.44 μg/mL and 70.65±0.96 μg/mL respectively. Therefore I. verum can be a good lead as anti-cholinesterase agent from natural resources. Copyright © 2010 Elsevier B.V. All rights reserved.

  3. Inhibition of human platelet aggregation in vitro by standardized extract of Wendtia calycina

    Directory of Open Access Journals (Sweden)

    Milagros Garcia Mesa

    2011-10-01

    Full Text Available Wendtia calycina (Griseb. Griseb., Vivianiaceae, is a Paraguayan herbaceous plant commonly known as burrito. Our previous study indicated that burrito leaves are a very good source of phenylpropanoid glycosides, principally verbascoside. From W. calycina leaves, a standardized, water-soluble extract rich in phenylpropanoid glycosides (WSE has been developed on an industrial scale to be used as a food supplement, cosmetic, phytomedicine, and ingredient of different formulations. In this study, we investigated the effect of the WSE on human platelet aggregation in vitro induced by adenosine diphosphate (ADP, epinephrine (EPN, collagen (COL or arachidonic acid (AA. WSE, concentration-dependently, inhibited ADP and EP-induced human platelet aggregation (IC50 were 0.82±0.15 mg/mL and 0.41±0.02 mg/mL, respectively. It did not inhibit collagen-induced platelet aggregation, thus suggesting a selectivity for the ADP-induced platelet activation pathways.

  4. Inhibition of human platelet aggregation in vitro by standardized extract of Wendtia calycina

    Directory of Open Access Journals (Sweden)

    Milagros Garcia Mesa

    2011-06-01

    Full Text Available Wendtia calycina (Griseb. Griseb., Vivianiaceae, is a Paraguayan herbaceous plant commonly known as burrito. Our previous study indicated that burrito leaves are a very good source of phenylpropanoid glycosides, principally verbascoside. From W. calycina leaves, a standardized, water-soluble extract rich in phenylpropanoid glycosides (WSE has been developed on an industrial scale to be used as a food supplement, cosmetic, phytomedicine, and ingredient of different formulations. In this study, we investigated the effect of the WSE on human platelet aggregation in vitro induced by adenosine diphosphate (ADP, epinephrine (EPN, collagen (COL or arachidonic acid (AA. WSE, concentration-dependently, inhibited ADP and EP-induced human platelet aggregation (IC50 were 0.82±0.15 mg/mL and 0.41±0.02 mg/mL, respectively. It did not inhibit collagen-induced platelet aggregation, thus suggesting a selectivity for the ADP-induced platelet activation pathways.

  5. Fish allergens at a glance: Variable allergenicity of parvalbumins, the major fish allergens

    OpenAIRE

    Annette eKuehn; Ines eSwoboda; Karthik eArumugam; Christiane eHilger; François eHentges; François eHentges

    2014-01-01

    Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand,...

  6. Fish Allergens at a Glance: Variable Allergenicity of Parvalbumins, the Major Fish Allergens

    OpenAIRE

    Kuehn, Annette; Swoboda, Ines; Arumugam, Karthik; Hilger, Christiane; Hentges, François

    2014-01-01

    Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand,...

  7. Evaluation of antifungal activity of standardized extract of Salvia rhytidea Benth. (Lamiaceae) against various Candida isolates.

    Science.gov (United States)

    Salari, S; Bakhshi, T; Sharififar, F; Naseri, A; Ghasemi Nejad Almani, P

    2016-12-01

    Salvia species have long been described in traditional medicine for various indications. Owing to the widespread use of this genus by ethnic populations, especially for various infections ranging from skin disease to gastrointestinal disorders, we were encouraged to determine whether Salvia rhytidea could be effective against fungal infections. Given the increased incidence of candidiasis in the past decade, limits on the use of antifungal drugs, emergence of azole-resistant Candida species and increased incidence of treatment failures, it is necessary to identify a novel agent with antifungal properties. Aim of the study was to evaluate the antifungal properties of S. rhytidea against various Candida isolates. In this study, at first rosmarinic acid content of plant extract was determined. A total of 96 Candida isolates were tested, including the following species: Candida albicans (n=42), Candida glabrata (n=16), Candida tropicalis (n=11), Candida krusei (n=9), Candida parapsilosis (n=9), Candida lusitaniae (n=7) and Candida guilliermondii (n=2). The in vitro antifungal activity of methanolic extracts of S. rhytidea Benth. was evaluated against Candida isolates and compared with that of the standard antifungal drug nystatin by using a broth microdilution method, according to CLSI. Phytochemical screening results showed that the methanolic extract of S. rhytidea Benth. was rich in flavonoids and tannins. The minimal inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of S. rhytidea Benth. ranged from 3.125 to>100μg/ml and 6.25 to>100μg/ml respectively. The growth inhibition value displayed that C. tropicalis, C. krusei and C. albicans isolates were most susceptible to S. rhytidea. Findings show that S. rhytidea possesses an antifungal effect against Candida isolates. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  8. Antidepressant, antioxidant and neurotrophic properties of the standardized extract of Cocos nucifera husk fiber in mice.

    Science.gov (United States)

    Lima, Eliane Brito Cortez; de Sousa, Caren Nádia Soares; Vasconcelos, Germana Silva; Meneses, Lucas Nascimento; E Silva Pereira, Yuri Freitas; Ximenes, Naiara Coelho; Santos Júnior, Manuel Alves; Matos, Natália Castelo Branco; Brito, Rayanne; Miron, Diogo; Leal, Luzia Kalyne Almeida Moreira; Macêdo, Danielle; Vasconcelos, Silvânia Maria Mendes

    2016-07-01

    The plant Cocos nucifera and its derivatives have shown antidepressant-like effects, although its hydroalcoholic extract has not been studied with this end in mind. Therefore, we decided to determine the antidepressant-like effects of the standardized hydroalcoholic extract of Cocos nucifera husk fiber (HECN) as well as oxidative alterations in the prefrontal cortex (PFC), hippocampus (HC) and striatum (ST), and the levels of brain-derived neurotrophic factor (BDNF) in the HC of mice. The extract was characterized based on the content of total polyphenols as well as two phenol compounds-catechin and chlorogenic acid-by HPLC-PDA. Male animals were treated per os (p.o.) for 7 days with distilled water or HECN (50, 100 or 200 mg/kg), or intraperitoneally with vitamin E (Vit E 400 mg/kg). One hour after the last drug administration, the animals were submitted to the open field test, forced swimming test (FST), tail suspension test (TST) and, immediately after the behavioral tests, had their brain removed for neurochemical determinations. The results showed that HECN100 decreased the immobility time in the FST and TST presenting, thus demonstrating an antidepressant-like effect. The administration of HECN decreased malondialdehyde levels in all doses and brain areas studied with the exception of HECN50 in the HC. The administration of HECN also decreased nitrite levels in all doses and brain regions studied. HECN100 also increased the levels of BDNF in HC of mice. In conclusion, we demonstrated that HECN has antidepressant-like properties, probably based on its antioxidant and neurotrophic effects, and is thus relevant for the treatment of depression.

  9. Safety assessment of a standardized polyphenolic extract of clove buds: Subchronic toxicity and mutagenicity studies

    Directory of Open Access Journals (Sweden)

    Liju Vijayasteltar

    2016-01-01

    Full Text Available Despite the various reports on the toxicity of clove oil and its major component eugenol, systematic evaluations on the safety of polyphenolic extracts of clove buds have not been reported. Considering the health beneficial pharmacological effects and recent use of clove polyphenols as dietary supplements, the present study investigated the safety of a standardized polyphenolic extract of clove buds (Clovinol, as assessed by oral acute (5 g/kg b.wt. for 14 days and subchronic (0.25, 0.5 and 1 g/kg b.wt. for 90 days toxicity studies on Wistar rats and mutagenicity studies employing Salmonella typhimurium strains. Administration of Clovinol did not result in any toxicologically significant changes in clinical/behavioural observations, ophthalmic examinations, body weights, organ weights, feed consumption, urinalysis, hematology and clinical biochemistry parameters when compared to the untreated control group of animals, indicating the no observed-adverse-effect level (NOAEL as 1000 mg/kg b.wt./day; the highest dose tested. Terminal necropsy did not reveal any treatment-related histopathology changes. Clovinol did not show genotoxicity when tested on TA-98, TA-100 and TA-102 with or without metabolic activation; rather exhibited significant antimutagenic potential against the known mutagens, sodium azide, NPD and tobacco as well as against 2-acetamidoflourene, which needed metabolic activation for mutagenicity.

  10. Evaluation of genotoxicity and subchronic toxicity of standardized rose hip extract.

    Science.gov (United States)

    Nagatomo, A; Oguri, M; Nishida, N; Ogawa, M; Ichikawa, A; Tanaka-Azuma, Y

    2017-01-01

    Rose hip is the fruit of the rose plant, which is widely used in food, cosmetics and as a traditional medicine. Therefore, rose hip is considered safe and has a sufficient history of consumption as food. However, few studies have reported on the safety of rose hip extracts in toxicological analyses. Thus, to evaluate the safety of rosehip polyphenol MJ (RHPMJ), an aqueous ethanol extract standardized with the trans-tiliroside content, we performed genotoxicity and 90-day repeated oral dose toxicity studies in compliance with the Organisation for Economic Co-operation and Development-Good Laboratory Practice. RHPMJ did not induce gene mutations in reverse mutation tests of Salmonella typhimurium TA98, TA100, TA1535, TA1537 and Escherichia coli WP2 uvrA strains and did not induce chromosomal aberrations in cultured Chinese hamster lung (CHL/IU) cells. Moreover, micronucleus tests using rat bone marrow showed RHPMJ had no micronucleus-inducing potential. Finally, 90-day repeated oral dose toxicity studies (100-1000 mg/kg) in male and female rats showed no treatment-related toxicity in rats. These data indicate that the RHPMJ had no genotoxicity and a no-observed-adverse-effect level greater than 1000 mg/kg in rats.

  11. Pharmacokinetics of prenylated hop phenols in women following oral administration of a standardized extract of hops.

    Science.gov (United States)

    van Breemen, Richard B; Yuan, Yang; Banuvar, Suzanne; Shulman, Lee P; Qiu, Xi; Alvarenga, René F Ramos; Chen, Shao-Nong; Dietz, Birgit M; Bolton, Judy L; Pauli, Guido F; Krause, Elizabeth; Viana, Marlos; Nikolic, Dejan

    2014-10-01

    Women seeking alternatives to hormone-replacement therapy for menopausal symptoms often try botanical dietary supplements containing extracts of hops (Humulus lupulus L.). Hops contain 8-prenylnaringenin (8-PN), a potent phytoestrogen, the related flavanones 6-prenylnaringenin and isoxanthohumol (IX), and the prenylated chalcone xanthohumol (XN). After chemically and biologically standardizing an extract of spent hops to these marker compounds, an escalating dose study was carried out in menopausal women to evaluate safety and pharmacokinetics. 8-PN, 6-prenylnaringenin, IX, and XN, sex hormones, and prothrombin time were determined in blood samples and/or 24 h urine samples. There was no effect on sex hormones or blood clotting. The maximum serum concentrations of the prenylated phenols were dose-dependent and were reached from 2 to 7 h, indicating slow absorption. The marker compounds formed glucuronides that were found in serum and urine. Secondary peaks at 5 h in the serum concentration-time curves indicated enterohepatic recirculation. The serum concentration-time curves indicated demethylation of IX to form 8-PN and cyclization of XN to IX. Slow absorption and enterohepatic recirculation contributed to half-lives exceeding 20 h. This human study indicated long half-lives of the estrogenic and proestrogenic prenylated phenols in hops but no acute toxicity. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    Science.gov (United States)

    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.

  13. Serological identification of house dust mite allergens in dogs with atopic dermatitis

    Directory of Open Access Journals (Sweden)

    Victor E.S. Cunha

    2012-09-01

    Full Text Available House dust mite antigens have been used for decades to diagnose allergic diseases in humans and animals. The objective of this study was to identify allergens in commercial Dermatophagoides farinae and Blomia tropicalis extracts by immunoblotting using sera from allergic dogs and anti-dog IgE conjugate. The analysis of antigens present in the D. farinae extract (FDA Allergenic using sera from 10 dogs allergic to D. farinae showed that eight sera recognized a band of approximately 102 kDa, eight recognized two bands of 52 to 76 kDa, five recognized one band of approximately 76 kDa, four recognized one band of 31 to 38 kDa, and two recognized one band of 12 to 17 kDa. Immunoblot assays of the B. tropicalis extract (FDA Allergenic using sera from 10 animals allergic to B. tropicalis showed that five sera recognized two bands of 52 to 76 kDa. These results demonstrate the importance of the two house dust mite species for the pathogenesis of canine atopic dermatitis in Brazil. In addition, the results indicate which allergens should be present in allergenic extracts used for diagnosis and allergen-specific immunotherapy.

  14. Beer-induced anaphylaxis: identification of allergens.

    Science.gov (United States)

    Figueredo, E; Quirce, S; del Amo, A; Cuesta, J; Arrieta, I; Lahoz, C; Sastre, J

    1999-06-01

    We report on a 21-year-old atopic woman who developed urticaria, angioedema of the face, and wheezy dyspnea shortly after drinking beer and after eating a corn-made snack. Skin prick tests and specific IgE determinations to beer ingredients and cereal extracts were performed. Immunoblotting inhibition assays were carried out to investigate possible common allergens shared by barley and malt with corn. Skin prick tests and specific IgE measurements with beer, barley, malt, wheat, corn, rye, rice, and oat flour were positive. Ten pollen-allergic patients showed negative skin tests to beer. Double-blind, placebo-controlled, oral challenge tests with sodium metabisulfite and wheat flour were negative. Immunoblotting demonstrated several IgE-binding bands at 31-56 kDa in malt and barley extracts, and a major band at 38 kDa in the beer extract. Immunoblot inhibition assays showed that malt extract was able to inhibit most of the IgE-binding bands in wheat and corn extracts, whereas corn did not produce significant inhibition to barley and malt extracts. This patient developed type I hypersensitivity to barley/malt and corn. Although she also showed IgE reactivity to wheat and other cereals, no symptoms were elicited upon ingestion of these cereals, probably indicating latent sensitization to them.

  15. Standardization of Berberis aristata extract through conventional and modern HPTLC techniques

    Directory of Open Access Journals (Sweden)

    Dinesh K. Patel

    2012-05-01

    Full Text Available Objective: Berberis aristata (Berberidaceae is an important medicinal plant, found in the different region of the world. It has significant medicinal value in the traditional Indian and Chinese system of medicine. The aim of the present investigation includes qualitative and quantitative analysis of Berberis aristata extract. Methods: Present study includes determination of phytochemical analysis, solubility test, heavy metal analysis, antimicrobial study and quantitative analysis by HPTLC method. Results: Preliminary phytochemical analysis showed the presence of carbohydrate, glycoside, alkaloid, protein, amino acid, saponin, tannin and flavonoid. Solubility in water and alcohal were found to be 81.90% in water and 84.52% in 50% in alcohal. Loss on drying was found to be 5.32%. Total phenol and flavonoid content were found to be 0.11% and 2.8%. Level of lead, arsenic, mercury and cadmium complies the standard level. E. coli and salmonella was found to be absent whereas total bacterial count, yeast and moulds contents were found to be under the limit. Content of berberine was found to be 13.47% through HPTLC techniques. Conclusions: The results obtained from the present studies could be used as source of valuable information which can play an important role for the food scientists, researchers and even the consumers for its standards.

  16. Epilobi Hirsuti Herba Extracts Influence the In Vitro Activity of Common Antibiotics on Standard Bacteria

    Directory of Open Access Journals (Sweden)

    Pirvu Lucia

    2016-01-01

    Full Text Available Epilobium genus has been confirmed as an effective source of natural antimicrobials. However, the influence of Epilobi hirsuti herba derived products on usual antibiotics activity has not been studied. In this study, several standardized Epilobi hirsuti herba extracts (EHE were evaluated in order to asses their potential effects on usual antibiotics tested on standard Gram-positive and Gram-negative bacterial strains in vitro. The results emphasized that the bacterial strains ranged from sensitive (MIC values between 50–200 μg GAE mL-1 (S. epidermidis ATCC 12228 to very resistant (E. coli strains, E. faecalis ATCC 29212 being practically immune to EHE. In terms of synergistic interaction, Tetracycline and Ampicillin combinations lead to the most important stimulatory effects, the diameters of the inhibition zone being even 60% bigger compared to the antibiotic alone. Synergistic effects between myricetin(galloyl derivates and Tetracycline were also revealed on P. aeruginosa and E. coli strains. Together, it clearly demonstrated not only EHE’s own antimicrobial properties, but also their capacity to influence the antimicrobial potency of some common antibiotics. These results could be useful for the area of herbal medicines and as potential candidates in managing microbial resistance, but also for physicians and pharmacists using combined antibacterial therapy.

  17. Crystal structure of peanut (Arachis hypogaea) allergen Ara h 5

    Science.gov (United States)

    Profilins from numerous species are known to be allergens, including food allergens, such as peanut (Arachis hypogaea) allergen Ara h 5, and pollen allergens, such as birch allergen Bet v 2. Patients with pollen allergy can also cross-react to peanut. Structural characterization of allergens will al...

  18. A biodegradable device for the controlled release of Piper nigrum (Piperaceae) standardized extract to control Aedes aegypti (Diptera, Culicidae) larvae.

    Science.gov (United States)

    Custódio, Kauê Muller; Oliveira, Joice Guilherme de; Moterle, Diego; Zepon, Karine Modolon; Prophiro, Josiane Somariva; Kanis, Luiz Alberto

    2016-01-01

    The significant increase in dengue, Zika, and chikungunya and the resistance of the Aedes aegypti mosquito to major insecticides emphasize the importance of studying alternatives to control this vector. The aim of this study was to develop a controlled-release device containing Piper nigrum extract and to study its larvicidal activity against Aedes aegypti. Piper nigrum extract was produced by maceration, standardized in piperine, and incorporated into cotton threads, which were inserted into hydrogel cylinders manufactured by the extrusion of carrageenan and carob. The piperine content of the extract and thread reservoirs was quantified by chromatography. The release profile from the device was assessed in aqueous medium and the larvicidal and residual activities of the standardized extract as well as of the controlled-release device were examined in Aedes aegypti larvae. The standardized extract contained 580mg/g of piperine and an LC50 value of 5.35ppm (24h) and the 3 cm thread reservoirs contained 13.83 ± 1.81mg of piperine. The device showed zero-order release of piperine for 16 days. The P. nigrum extract (25ppm) showed maximum residual larvicidal activity for 10 days, decreasing progressively thereafter. The device had a residual larvicidal activity for up to 37 days. The device provided controlled release of Piper nigrum extract with residual activity for 37 days. The device is easy to manufacture and may represent an effective alternative for the control of Aedes aegypti larvae in small water containers.

  19. Navigating through the Jungle of Allergens: Features and Applications of Allergen Databases.

    Science.gov (United States)

    Radauer, Christian

    2017-01-01

    The increasing number of available data on allergenic proteins demanded the establishment of structured, freely accessible allergen databases. In this review article, features and applications of 6 of the most widely used allergen databases are discussed. The WHO/IUIS Allergen Nomenclature Database is the official resource of allergen designations. Allergome is the most comprehensive collection of data on allergens and allergen sources. AllergenOnline is aimed at providing a peer-reviewed database of allergen sequences for prediction of allergenicity of proteins, such as those planned to be inserted into genetically modified crops. The Structural Database of Allergenic Proteins (SDAP) provides a database of allergen sequences, structures, and epitopes linked to bioinformatics tools for sequence analysis and comparison. The Immune Epitope Database (IEDB) is the largest repository of T-cell, B-cell, and major histocompatibility complex protein epitopes including epitopes of allergens. AllFam classifies allergens into families of evolutionarily related proteins using definitions from the Pfam protein family database. These databases contain mostly overlapping data, but also show differences in terms of their targeted users, the criteria for including allergens, data shown for each allergen, and the availability of bioinformatics tools. © 2017 S. Karger AG, Basel.

  20. 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.

  1. Assessment of endogenous allergenicity of genetically modified plants exemplified by soybean - Where do we stand?

    Science.gov (United States)

    Selb, R; Wal, J M; Moreno, F J; Lovik, M; Mills, C; Hoffmann-Sommergruber, K; Fernandez, A

    2017-03-01

    According to EU regulation, genetically modified (GM) plants considered to be allergenic have to be assessed concerning their endogenous allergens before placement on the EU market, in line with the international standards described in Codex Alimentarius. Under such premises, a quantitative relevant increase in allergens might occur in GM plants as an unintended effect compared with conventionally produced crops, which could pose a risk to consumers. Currently, data showing a connection between dose and allergic sensitisation are scarce since the pathophysiological mechanisms of sensitisation are insufficiently understood. In contrast, data on population dose-distribution relationships acquired by oral food challenge are available showing a connection between quantity of allergenic protein consumed and the population of allergic individuals experiencing reactions. Soybean is currently the only recognised allergenic GM food by law for which EFSA has received applications and was therefore taken as an example for defining an assessment strategy. Identification of potential allergens, methodology for quantification as well as risk assessment considerations, are discussed. A strategy is proposed for the identification, assessment and evaluation of potential hazards/risks concerning endogenous allergenicity in food derived from plants developed by biotechnology. This approach could be expanded to other allergenic foods in the future, whenever required. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. 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

  3. Sensitising capacity of peptides from food allergens

    DEFF Research Database (Denmark)

    Bøgh, Katrine Lindholm

    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 food allergens. One of the features believed...... to be a general characteristic of food allergens is resistance to digestion. This is based on studies showing that allergenic dietary proteins in general are more resistant to digestion than dietary proteins with no proven allergenicity, concluding that a correlation between stability to digestion and allergenic...... 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...

  4. Associations between baseline allergens and polysensitization

    DEFF Research Database (Denmark)

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

    2008-01-01

    : Seven allergens - parabens mix, N-isopropyl-N-phenyl-p-phenylenediamine, sesquiterpene lactone mix, wool alcohols, potassium dichromate, Myroxylon pereirae, and cobalt chloride - showed statistically significant positive associations to polysensitization. Five allergens p-phenylenediamine, neomycin...

  5. Propolis standardized extract (EPP-AF®), an innovative chemically and biologically reproducible pharmaceutical compound for treating wounds.

    Science.gov (United States)

    Berretta, Andresa Aparecida; Nascimento, Andresa Piacezzi; Bueno, Paula Carolina Pires; Vaz, Mirela Mara de Oliveira Lima Leite; Marchetti, Juliana Maldonado

    2012-01-01

    The aim of this study was to develop a formulation, containing the propolis standardized extract (EPP-AF(®)), which can assist in the healing of skin lesions. To achieve this objective the antimicrobial activity and chemical composition of the propolis extract was determined. The final product was subjected to in vitro and in vivo pre-clinical evaluation. The broth macrodilution method was used to determine the antimicrobial activity of the extracts and formulations against the microorganisms most commonly found in burns, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Staphylococcus epidermidis. Wistar rats with puncture wounded skin were used to evaluate the wound healing properties of propolis. The results of chemical and biological characterization demonstrated the batch-to-batch reproducibility of the standardized extract which is an unprecedented result. The antimicrobial and wound healing activity of the pharmaceutical studied showed the best results when samples contain 3.6% propolis, suggesting that this is the most promising composition.

  6. The Allergen Bank: a source of extra contact allergens for the dermatologist in practice

    DEFF Research Database (Denmark)

    Andersen, Klaus Ejner; Rastogi, S C; Carlsen, L

    1996-01-01

    The Allergen Bank was established to give dermatologists easy access to special test materials in order to make early diagnoses of special cases of allergic contact dermatitis. The Allergen Bank comprises a computer system to register several hundred contact allergens in appropriate patch test...... concentrations available at the allergy laboratory and the patch test results. At the request of dermatologists in practice for Allergen Bank may supply special contact allergens for aimed patch testing of contact dermatitis patients. The organization of the Allergen Bank and the procedure of its use...... third of the positive reactions were caused by the 16 most frequently ordered allergens, which amounted to 340 allergen samples. The allergens included plant chemicals, acrylates, animal feed additives, fragrance chemicals and preservatives. Selected allergens were investigated for stability during...

  7. Pharmacokinetics of a Standardized Extract of Centella asiatica ECa 233 in Rats.

    Science.gov (United States)

    Anukunwithaya, Tosapol; Tantisira, Mayuree H; Tantisira, Boonyong; Khemawoot, Phisit

    2017-05-01

    ECa 233, a standardized extract of Centella asiatica , has been found to exhibit various positive neurological effects and to have a good safety profile. The present study aimed to explore the disposition kinetics of ECa 233, containing madecassoside (53.1 %) and asiaticoside (32.3 %), in rats. The extract was intravenously or orally administered at doses from 50 to 200 mg/kg. Plasma, tissues, urine, and feces were collected at time points from 0 to 48 h after dosing. The levels of madecassoside and asiaticoside, as well as their postulated triterpenic metabolites, madecassic acid and asiatic acid, in biological samples, were simultaneously measured by liquid chromatography-tandem mass spectrometry. The results showed that all animals had a good tolerability for ECa 233, whereas madecassic and asiatic acids were found in negligible amounts after pharmacokinetic assessment. Madecassoside and asiaticoside demonstrated rather similar absorption and tissue distribution profiles. They were rapidly absorbed, reaching maximum levels within 5-15 min after oral administration, but they had poor oral bioavailability, less than 1 %. Both triterpenoids were extensively distributed in the brain, stomach, and skin within 1 h and remained there for at least 4 h after dosing. Madecassoside and asiaticoside in ECa 233 were mainly excreted as an unchanged form after being injected, and exclusively as triterpenic acid metabolites in feces after oral administration. The pharmacokinetic results obtained could provide some guidance for an appropriate dosing regimen of ECa 233 in future studies. This study also provided the first evidence demonstrating the presence of madecassoside and asiaticoside in their target tissues. Georg Thieme Verlag KG Stuttgart · New York.

  8. An experimental evaluation of the antidiabetic and antilipidemic properties of a standardized Momordica charantia fruit extract

    Science.gov (United States)

    Fernandes, Nafisa PC; Lagishetty, Chakradhar V; Panda, Vandana S; Naik, Suresh R

    2007-01-01

    Background The MCE, Momordica charantia fruit extract Linn. (Cucurbitaceae) have been documented to elicit hypoglycemic activity on various occasions. However, due to lack of standardization of these extracts, their efficacy remains questionable. The present study was undertaken by selecting a well standardised MCE. This study reports hypoglycemic and antilipidemic activities of MCE employing relevant animal models and in vitro methods. Methods Diabetes was induced in Wistar rats by a s.c., subcutaneous injection of alloxan monohydrate (100 mg/kg) in acetate buffer (pH 4.5). MCE and glibenclamide were administered orally to alloxan diabetic rats at doses of 150 mg/kg, 300 mg/kg & 600 mg/kg, and 4 mg/kg respectively for 30 days, blood was withdrawn for glucose determination on 0, 7, 14, 21 and 30th days. On the 31st day, overnight fasted rats were sacrificed and blood was collected for various biochemical estimations including glycosylated haemoglobin, mean blood glucose, serum insulin, cholesterol, triglcerides, protein and glycogen content of liver. The hemidiaphragms and livers were also isolated, carefully excised and placed immediately in ice cooled perfusion solution and processed to study the glucose uptake/transfer processes. Hypolipidemic activity in old obese rats was evaluated by treating two groups with MCE (150 mg/kg & 300 mg/kg) orally for 30 days and determining total cholesterol, triglyceride and HDL-CH, LDL-CH and VLDL-CH levels from serum samples. Results Subchronic study of MCE in alloxan induced diabetic rats showed significant antihyperglycemic activity by lowering blood glucose and GHb%, percent glycosylated haemoglobin. Pattern of glucose tolerance curve was also altered significantly. MCE treatment enhanced uptake of glucose by hemidiaphragm and inhibited glycogenolysis in liver slices in vitro. A significant reduction in the serum cholesterol and glyceride levels of obese rats following MCE treatment was also observed. Conclusion Our

  9. An experimental evaluation of the antidiabetic and antilipidemic properties of a standardized Momordica charantia fruit extract.

    Science.gov (United States)

    Fernandes, Nafisa P C; Lagishetty, Chakradhar V; Panda, Vandana S; Naik, Suresh R

    2007-09-24

    The MCE, Momordica charantia fruit extract Linn. (Cucurbitaceae) have been documented to elicit hypoglycemic activity on various occasions. However, due to lack of standardization of these extracts, their efficacy remains questionable. The present study was undertaken by selecting a well standardised MCE. This study reports hypoglycemic and antilipidemic activities of MCE employing relevant animal models and in vitro methods. Diabetes was induced in Wistar rats by a s.c., subcutaneous injection of alloxan monohydrate (100 mg/kg) in acetate buffer (pH 4.5). MCE and glibenclamide were administered orally to alloxan diabetic rats at doses of 150 mg/kg, 300 mg/kg & 600 mg/kg, and 4 mg/kg respectively for 30 days, blood was withdrawn for glucose determination on 0, 7, 14, 21 and 30th days. On the 31st day, overnight fasted rats were sacrificed and blood was collected for various biochemical estimations including glycosylated haemoglobin, mean blood glucose, serum insulin, cholesterol, triglcerides, protein and glycogen content of liver. The hemidiaphragms and livers were also isolated, carefully excised and placed immediately in ice cooled perfusion solution and processed to study the glucose uptake/transfer processes. Hypolipidemic activity in old obese rats was evaluated by treating two groups with MCE (150 mg/kg & 300 mg/kg) orally for 30 days and determining total cholesterol, triglyceride and HDL-CH, LDL-CH and VLDL-CH levels from serum samples. Subchronic study of MCE in alloxan induced diabetic rats showed significant antihyperglycemic activity by lowering blood glucose and GHb%, percent glycosylated haemoglobin. Pattern of glucose tolerance curve was also altered significantly. MCE treatment enhanced uptake of glucose by hemidiaphragm and inhibited glycogenolysis in liver slices in vitro. A significant reduction in the serum cholesterol and glyceride levels of obese rats following MCE treatment was also observed. Our experimental findings with respect to the

  10. An experimental evaluation of the antidiabetic and antilipidemic properties of a standardized Momordica charantia fruit extract

    Directory of Open Access Journals (Sweden)

    Naik Suresh R

    2007-09-01

    Full Text Available Abstract Background The MCE, Momordica charantia fruit extract Linn. (Cucurbitaceae have been documented to elicit hypoglycemic activity on various occasions. However, due to lack of standardization of these extracts, their efficacy remains questionable. The present study was undertaken by selecting a well standardised MCE. This study reports hypoglycemic and antilipidemic activities of MCE employing relevant animal models and in vitro methods. Methods Diabetes was induced in Wistar rats by a s.c., subcutaneous injection of alloxan monohydrate (100 mg/kg in acetate buffer (pH 4.5. MCE and glibenclamide were administered orally to alloxan diabetic rats at doses of 150 mg/kg, 300 mg/kg & 600 mg/kg, and 4 mg/kg respectively for 30 days, blood was withdrawn for glucose determination on 0, 7, 14, 21 and 30th days. On the 31st day, overnight fasted rats were sacrificed and blood was collected for various biochemical estimations including glycosylated haemoglobin, mean blood glucose, serum insulin, cholesterol, triglcerides, protein and glycogen content of liver. The hemidiaphragms and livers were also isolated, carefully excised and placed immediately in ice cooled perfusion solution and processed to study the glucose uptake/transfer processes. Hypolipidemic activity in old obese rats was evaluated by treating two groups with MCE (150 mg/kg & 300 mg/kg orally for 30 days and determining total cholesterol, triglyceride and HDL-CH, LDL-CH and VLDL-CH levels from serum samples. Results Subchronic study of MCE in alloxan induced diabetic rats showed significant antihyperglycemic activity by lowering blood glucose and GHb%, percent glycosylated haemoglobin. Pattern of glucose tolerance curve was also altered significantly. MCE treatment enhanced uptake of glucose by hemidiaphragm and inhibited glycogenolysis in liver slices in vitro. A significant reduction in the serum cholesterol and glyceride levels of obese rats following MCE treatment was also

  11. Standardization and anti-inflammatory activity of aqueous extract of Psittacanthus plagiophyllus Eichl. (Loranthaceae).

    Science.gov (United States)

    Bezerra, Adrielle Nara Serra; Massing, Laís Tatiele; de Oliveira, Ricardo Bezerra; Mourão, Rosa Helena Veras

    2017-04-18

    The hemiparasitic species Psittacanthus plagiophyllus Eichl. (Loranthaceae), also known as erva de passarinho, is used in folk medicine in the Santarém region in the state of Pará, Brazil, to treat gastritis and a variety of inflammatory disorders. In view of the lack of pharmacological studies on this species in the literature and the fact that it is used constantly by the local population, this study sought to standardize the extract of the leaves of P. plagiophyllus (AEPp) and to assess its anti-inflammatory potential in in vivo tests. Quality control and standardization of AEPp were performed following the 5th edition of the Brazilian Pharmacopeia. To assess the in vivo anti-inflammatory activity of AEPp, the carrageenan-induced and dextran-induced rat paw edema models were initially used. To investigate the effect of AEPp on carrageenan-induced leukocyte recruitment and exudate production, the air pouch inflammation model was used. In addition, the antioxidant activity of AEPp was assessed in vitro by the DPPH radical scavenging method. The chromatographic profile of AEPp indicated the presence of flavonoids, coumarins and hydrolyzable and condensed tannins. Measurement of phenolics revealed the following percentages in the extract: 12.62±0.18% total phenolics, 5.39±0.01% total tannins, 12.54±0.24% hydrolyzable tannins, 8.37±0.32% condensed tannins and 1.23±0.02% total flavonoids. In 500 and 1000mg/kg doses (p.o.) AEPp had significant edema-inhibiting activity (ptested [250, 500 and 1000mg/kg (p.o.)], AEPp inhibited total leukocyte and neutrophil migration and reduced the amount of exudate in the air pouch in a dose-dependent manner. Both effects were statistically significant (pconcentration of 75µg/mL), indicating that it has antioxidant activity. AEPp, therefore, exhibited considerable in vivo anti-inflammatory activity and in vitro antioxidant activity. This may be due to its high phenolics content. These findings provide evidence to support the

  12. Polyphenol-rich pomegranate juice reduces IgE binding to cashew nut allergens.

    Science.gov (United States)

    Li, Yichen; Mattison, Christopher P

    2018-03-01

    Food allergy negatively impacts quality of life and can be life-threatening. Cashew nuts can cause severe reactions in very small amounts, and they are included in a group of foods most commonly responsible for causing food allergy. Polyphenols and polyphenol-rich juices have been demonstrated to complex with peanut allergens. Here, the interaction between cashew nut allergens and polyphenol-rich juices is evaluated biochemically and immunologically. Various juices, including pomegranate (POM), blueberry (BB), and concord grape (CG) juices, were evaluated for polyphenol content and formation of polyphenol-cashew allergen complexes. Among the various juices studied, POM juice showed a greater capacity to form complexes with cashew proteins. Dynamic light scattering (DLS) demonstrated a sharp increase in cashew protein extract particle size to around 3580 nm, and fewer cashew proteins were resolved by electrophoresis after treatment with POM juice. Immunoassays demonstrated reduced IgG and IgE binding to cashew allergens due to allergen precipitation by POM juice. These observations support the formation of complexes between polyphenol and cashew proteins that can prevent antibody recognition of cashew allergens through allergen precipitation. POM juice treatment of cashew extract effectively reduces antibody binding through allergen precipitation, and these findings could be applied to the development of less allergenic cashew nut products and oral immunotherapy. Published 2017. This article is a U.S. Government work and is in the public domain in the USA. Published 2017. This article is a U.S. Government work and is in the public domain in the USA.

  13. 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

  14. Fish allergens at a glance: variable allergenicity of parvalbumins, the major fish allergens.

    Science.gov (United States)

    Kuehn, Annette; Swoboda, Ines; Arumugam, Karthik; Hilger, Christiane; Hentges, François

    2014-01-01

    Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand, some individuals have IgE antibodies directed against unique, species-specific parvalbumin epitopes, and these patients show clinical symptoms only with certain fish species. Furthermore, different parvalbumin isoforms and isoallergens are present in the same fish and might display variable allergenicity. This was shown for salmon homologs, where only a single parvalbumin (beta-1) isoform was identified as allergen in specific patients. In addition to the parvalbumins, several other fish proteins, enolases, aldolases, and fish gelatin, seem to be important allergens. New clinical and molecular insights advanced the knowledge and understanding of fish allergy in the last years. These findings were useful for the advancement of the IgE-based diagnosis and also for the management of fish allergies consisting of advice and treatment of fish-allergic patients.

  15. Fish allergens at a glance: Variable allergenicity of parvalbumins, the major fish allergens

    Directory of Open Access Journals (Sweden)

    Annette eKuehn

    2014-04-01

    Full Text Available Fish is a common trigger of severe, food-allergic reactions. Only a limited number of proteins induce specific IgE-mediated immune reactions. The major fish allergens are the parvalbumins. They are members of the calcium-binding EF-hand protein family characterized by a conserved protein structure. They represent highly cross-reactive allergens for patients with specific IgE to conserved epitopes. These patients might experience clinical reactions with various fish species. On the other hand, some individuals have IgE antibodies directed against unique, species-specific parvalbumin epitopes, and these patients show clinical symptoms only with certain fish species. Furthermore, different parvalbumin isoforms and isoallergens are present in the same fish and might display variable allergenicity. This was shown for salmon homologs, where only a single parvalbumin (beta-1 isoform was identified as allergen in specific patients. In addition to the parvalbumins, several other fish proteins, enolases, aldolases and fish gelatin, seem to be important allergens.New clinical and molecular insights advanced the knowledge and understanding of fish allergy in the last years. These findings will be useful for the advancement of the IgE-based diagnosis but also for the management of fish allergies consisting of advice and treatment of fish-allergic patients.

  16. Standardization of HPLC Method of Scopoletin in Different Extracts of Convolvulus pluricaulis

    OpenAIRE

    Upadhyay Vipul; Sharma Neeru; Tiwari Kumar Amit; Joshi H. M; Malik Amreesh; Singh Brijpal; Kalakoti Singh Bahadur

    2013-01-01

    Different extract of Convolvulus pluricaulis choisy, (Methanol, hydro-alcohol (50%) and water) were prepared and tested of scopoletin. The maximum scopoletin content was observed in 50% ethanolic extract followed by methanol and water extracts. It was 0.1738%, 0.0932% and 0.0435% in hydro-alcoholic, Methanol and water extract respectively. A simple HPLC (High Performance Liquid Chromatography) was developed for the determination of Scopoletin in Convolvulus pluricaulis choisy. Shankhpushpi is...

  17. Neuritogenic effect of standardized extract of Centella asiatica ECa233 on human neuroblastoma cells

    Science.gov (United States)

    2013-01-01

    Background In order to gain insight into neuroprotective effects of ECa 233, a standardized extract of Centella asiatica, previously demonstrated in animal models of memory impairment induced by transient global ischemia or intracerebroventricular injection of β-amyloid, the effect of ECa 233 on neurite outgrowth of human IMR-32 neuroblastoma cell line was investigated. Methods Cells were seeded and incubated with various concentrations of ECa 233. Morphometric analysis was carried out by a measurement of the longest neurite growth of cells at 24 and 48 h. Contributing signaling pathways possibly involved were subsequently elucidated by western blot analysis. Results While ECa 233 had only limited effects on cell viability, it significantly enhanced neurite outgrowth of IMR-32 cells at the concentrations of 1–100 μg/ml. Western blot analysis revealed that ECa 233 significantly upregulated the level of activated ERK1/2 and Akt of the treated cells suggesting their involvement in the neuritogenic effect observed, which was subsequently verified by the finding that an addition of their respective inhibitors could reverse the effect of ECa 233 on these cells. Conclusions The present study clearly demonstrated neurite outgrowth promoting activity of ECa 233. ERK1/2 and Akt signaling pathways seemed to account for the neurotrophic effect observed. In conjunction with in vivo neuroprotective effect of ECa 233 previously reported, the results obtained support further development of ECa 233 for clinical use in neuronal injury or neurodegenerative diseases. PMID:23915016

  18. Anxiolytic effects of standardized extract of Centella asiatica (ECa 233) after chronic immobilization stress in mice.

    Science.gov (United States)

    Wanasuntronwong, Aree; Tantisira, Mayuree H; Tantisira, Boonyong; Watanabe, Hiroshi

    2012-09-28

    Centella asiatica has long been used for various neurological disturbances in Southeast Asian countries. The present study aims to demonstrate the anxiolytic effect of ECa 233, a standardized extract of C. asiatica containing triterpenoids not less than 80%, in comparison to diazepam. The test compound was given orally to non-stressed mice and mice subjected to chronic immobilization stress. Anxiolytic effect was assessed by an elevated plus maze (EPM), a dark-light box and an open-field tests. Anxiolytic effect of ECa 233 was clearly demonstrated in non-stressed mice subjected to acute stress in all behavioral tests employed. In the EPM test, chronically stressed mice showed significant decrease in the number of open arm entries, shortening the time spent in open arms and an increase of the latency to leave the central area, suggesting their release from the stress. In addition, ameliorating effect of ECa 233 was observed on the body weight and serum corticosterone which were adversely affected by immobilization stress. Madecassoside and asiaticoside, equal to their respective contents of the effective doses of ECa 233, exclusively presented anxiolytic effects in EPM, while no distinct effect was observed on the body weight and serum corticosterone. The present study demonstrated anxiolytic effect of ECa 233 in both acutely and chronically stressed animals. These effects could be mainly accounted by madecassoside and asiaticoside, suggesting a possible use of ECa 233 for the treatment of both acute and chronic anxiety in the pathological state. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  19. Standardized Kaempferia parviflora Extract Enhances Exercise Performance Through Activation of Mitochondrial Biogenesis.

    Science.gov (United States)

    Kim, Mi-Bo; Kim, Taeyoon; Kim, Changhee; Hwang, Jae-Kwan

    2018-01-01

    Exercise enhances mitochondrial biogenesis in skeletal muscle. Increased mitochondrial function and content can contribute to the improvement in skeletal muscle function and the benefits of exercise by increasing the response to energy demands. The effect of standardized Kaempferia parviflora extract (KPE) on exercise performance was accessed in L6 myotubes and C57BL/6J mice. KPE significantly activated peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and increased mitochondrial density in L6 myotubes. KPE also upregulated the expression of transcription factors for mitochondrial biogenesis (estrogen-related receptor-α [ERRα], nuclear respiratory factor-1 [NRF-1], and mitochondrial transcription factor A [Tfam]) through activation of PGC-1α in L6 myotubes. In vivo models including normal diet mice and high-fat diet obese mice showed that KPE effectively enhanced running endurance and increased the skeletal muscle weight/body weight ratio. Furthermore, these observations were associated with a significant upregulation of mitochondrial biogenesis regulatory genes in skeletal muscle tissue. KPE enhanced the protein expression of the sirtuin 1 (SIRT1)/adenosine monophosphate (AMP)-activated protein kinase (AMPK)/PGC-1α/peroxisome proliferator-activated receptor-δ (PPARδ) signaling pathway components in vitro and in vivo, acting as an exercise metabolism regulator. These results suggest that KPE has the potential to enhance exercise performance through mitochondrial biogenesis and the SIRT1/AMPK/PGC-1α/PPARδ signaling pathways.

  20. Physiological aspects of male libido enhanced by standardized Trigonella foenum-graecum extract and mineral formulation.

    Science.gov (United States)

    Steels, Elizabeth; Rao, Amanda; Vitetta, Luis

    2011-09-01

    The aim of the clinical study was to evaluate the effect of Testofen, a standardized Trigonella foenum-graecum (Fenugreek) extract and mineral formulation, on male libido (sexual drive, urge or desire) in a double blind randomized placebo controlled study. The study recruited 60 healthy males aged between 25 and 52, without erectile dysfunction and randomized to an oral dose (two tablets per day) of the active treatment (600 mg Testofen per day) or placebo for 6 weeks. The primary outcome measure was the DISF-SR (male) self-administered QOL total score and the four domain scores. The secondary outcome was specific quality of life parameters. Testofen had an overall positive effect on physiological aspects of libido. In particular, there was a significant increase in the subdomains of sexual arousal and orgasm. Testofen had a positive effect on QOL in self-reported satisfaction with muscle strength, energy and well-being but did not have an effect on mood or sleep. Serum prolactin and testosterone levels remained within the reference range. It was concluded that Testofen demonstrated a significant positive effect on physiological aspects of libido and may assist to maintain normal healthy testosterone levels. Copyright © 2011 John Wiley & Sons, Ltd.

  1. Sputum RNA signature in allergic asthmatics following allergen bronchoprovocation test

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    Rob G.J.A. Zuiker

    2016-07-01

    Full Text Available Background: Inhaled allergen challenge is a validated disease model of allergic asthma offering useful pharmacodynamic assessment of pharmacotherapeutic effects in a limited number of subjects. Objectives: To evaluate whether an RNA signature can be identified from induced sputum following an inhaled allergen challenge, whether a RNA signature could be modulated by limited doses of inhaled fluticasone, and whether these gene expression profiles would correlate with the clinical endpoints measured in this study. Methods: Thirteen non-smoking, allergic subjects with mild-to-moderate asthma participated in a randomised, placebo-controlled, 2-period cross-over study following a single-blind placebo run-in period. Each period consisted of three consecutive days, separated by a wash-out period of at least 3 weeks. Subjects randomly received inhaled fluticasone ((FP MDI; 500 mcg BID×5 doses in total or placebo. On day 2, house dust mite extract was inhaled and airway response was measured by FEV1 at predefined time points until 7 h post-allergen. Sputum was induced by NaCl 4.5%, processed and analysed at 24 h pre-allergen and 7 and 24 h post-allergen. RNA was isolated from eligible sputum cell pellets (<80% squamous of 500 cells, amplified according to NuGEN technology, and profiled on Affymetrix arrays. Gene expression changes from baseline and fluticasone treatment effects were evaluated using a mixed effects ANCOVA model at 7 and at 24 h post-allergen challenge. Results: Inhaled allergen-induced statistically significant gene expression changes in sputum, which were effectively blunted by fluticasone (adjusted p<0.025. Forty-seven RNA signatures were selected from these responses for correlation analyses and further validation. This included Th2 mRNA levels for cytokines, chemokines, high-affinity IgE receptor FCER1A, histamine receptor HRH4, and enzymes and receptors in the arachidonic pathway. Individual messengers from the 47 RNA signatures

  2. Allergenicity of Peanut Proteins is Retained Following Enzymatic Hydrolysis

    Science.gov (United States)

    Rationale: Hydrolysis of peanut proteins by food-grade enzymes may reduce allergenicity and could lead to safer forms of immunotherapy. Methods: Light roasted peanut flour extracts were digested with pepsin (37°C, pH 2), Alcalase (60°C pH 8), or Flavourzyme (50°C, pH 7) up to 1 hr, or sequentially w...

  3. Standardization of solvent extraction procedure for determination of uranium in seawater

    International Nuclear Information System (INIS)

    Sukanta Maity; Sahu, S.K.; Pandit, G.G.

    2015-01-01

    Solvent extraction procedure using ammonium pyrolidine dithiocarbamate complexing agent in methyl isobutyl ketone organic phase and acid exchange back-extraction is described for the simultaneous quantitative pre-concentration of uranium in seawater followed by its determination by differential pulse adsorptive stripping voltammetry. Solvent extraction time is optimized for extraction of uranium from seawater. Solvent extraction efficiency for uranium in seawater at different pH was carried out. The method gives a recovery of 98 ± 2 % for 400 mL sample at pH 3.0 ± 0.02, facilitating the rapid and interference free analysis of seawater samples. (author)

  4. Alpha-actinin is a new type of house dust mite allergen.

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    Su An

    Full Text Available Main indoor allergens for humans are from house dust mites. There are more than 30 allergens in Dermatophagoides farinae but only fourteen allergens have been identified from this mite including Der f 1-3, 6, 7, 10, 11, 13-18, and 22. A native allergen protein (Der f 24, 90 kDa was purified from D. farinae by gel filtration and anionic exchange liquid chromatography combined with IgE immunodetection. Its primary structure was determined by Edman degradation, mass spectrometry analysis and cDNA cloning. Enzyme-linked immunosorbent assay inhibition tests (ELISA-IT, immunoblots, basophil activation test (BAT and skin prick test (SPT were performed to evaluate the allergenicity. It was identified as an alpha (α-actinin containing a CaM-like domain with EF-hand motifs. Der f 24 reacted to sera from 85.4% (35/41 of patients on western blot analysis. It reduced ∼20% sera IgE reactivity to D. farinae extracts on a competitive ELISA. Eighty percent (8/10 of patients with D. farinae allergy showed positive reactions to Der f 24 in skin prick test. The expression of CD63 on basophils from patients was up-regulated by Der f 24 by ∼5.4-fold. Alpha-actinin was identified as a new type of house dust mite allergen. To the best of our knowledge, this is the first report of α-actinin as an allergen.

  5. [Immunoproteomics of non water-soluble allergens from 4 legumes flours: peanut, soybean, sesame and lentil].

    Science.gov (United States)

    Bouakkadia, Hayette; Boutebba, Aissa; Haddad, Iman; Vinh, Joëlle; Guilloux, Laurence; Sutra, Jean-Pierre; Sénéchal, Hélène; Poncet, Pascal

    2015-01-01

    Peanut, soybean, sesame and lentil are members of legumes worldwide consumed by human that can induce food allergy in genetically predisposed individuals. Several protein allergens, mainly water-soluble, have been described. We studied the non water-soluble fraction from these 4 food sources using immunoproteomics tools and techniques. Flour extracts were solubilized in detergent and chaotropes and analysed in 1 and 2 dimensional gel electrophoresis (2D). Results showed numerous proteins exhibiting wide ranges of isoelectric points and relative molecular masses. When IgE immunoreactivities of 18 food allergy patients were individually tested in 1 and 2D western-blots, a very diversified IgE repertoire was observed, reflecting extensive cross-reactivities but also co-sensitizations. Besides already well known and characterized allergens, mass spectrometry analysis allowed the identification of 22 allergens undescribed until now: 10 in peanut, 2 in soybean, 3 in sesame and 7 in lentil. Three allergens are legume storage proteins and the others belong to transport proteins, nucleotide binding proteins and proteins involved in the regulation of metabolism. Seven proteins are potentially similar to allergens described in plants and fungi and 11 are not related to any known allergen. Our results contribute to increase the repertoire of legume allergens that may improve the diagnosis, categorize patients and thus provide a better treatment of patients.

  6. A milestone in house dust-mite-allergen immunotherapy: the new sublingual tablet S-524101 (actair).

    Science.gov (United States)

    Bahceciler, Nerin N; Babayigit Hocaoglu, Arzu; Galip, Nilufer

    2014-12-01

    Subcutaneous allergen-specific immunotherapy has long been used in the treatment of allergic rhinitis and/or asthma and its efficacy has been confirmed. However, due to the discomfort of injections and the risk of severe adverse reactions, alternative routes of allergen administration have emerged. Delivery of allergens through the mucosal route had been proposed and investigated thoroughly, confirming the sublingual route to be the most efficacious. Later, the efficacy and safety of this route have been documented by numerous controlled trials both for house dust mite (HDM) and pollens. Recently, sublingual orodispersable grass pollen allergen tablets were in use followed by the newly developed HDM allergen tablets with satisfactory clinical results: Moreover, very recently 1 year of HDM tablet treatment was demonstrated to exert its clinical efficacy 1 year after discontinuation of tablet IT. The persistence of efficacy after only 1 year of treatment is a new and promising era. Currently, Sublingual Immunotherapy is the most easily administered and safe treatment option until more immunogenic, less allergenic and more efficient allergen extracts are developed.

  7. Association Between Allergic Diseases and Food Allergens Based on Skin Prick Test in Bushehr Province

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    Saman Keshvari

    2017-04-01

    Full Text Available Background: The Frequency of allergic diseases is growing in recent years. Identification of frequency of food allergens in different areas play an important role in diagnosis and treatment of these diseases. The aim of this study was to determine frequency and association of common food allergens in patients with allergic diseases based on Skin Prick Test in Bushehr province. Material and Methods: In this descriptive cross-sectional study, 1100 patients were enrolled with allergic diseases which had a sensitivity to at least one allergen.  This test was carried out with 21 common food allergens extract. Results: In all patients, association between the severity of the reaction prick allergy test and severity of allergic diseases with shrimp, cow's Milk and peanuts were (P= 0.01, (P= 0.02 and (P=0.04 respectively. In this study, the frequency of allergic rhinitis, asthma, chronic and acute urticaris and atopic eczema were 54.2%, 23%, 12.4%, 4.1% and 12%, respectively. While the the most common food allergens were peanuts (46.6%, egg yolk (43.1% and shrimp (42% respectively. Conclusion: This study indicated that food allergens such as shrimp, cow's Milk and peanuts have a greater role in severity of allergic diseases and this food allergens showed the highest frequency in patients.

  8. Extended boiling of peanut progressively reduces IgE allergenicity while retaining T cell reactivity.

    Science.gov (United States)

    Tao, B; Bernardo, K; Eldi, P; Chegeni, N; Wiese, M; Colella, A; Kral, A; Hayball, J; Smith, W; Forsyth, K; Chataway, T

    2016-07-01

    Current peanut oral immunotherapy is hampered by frequent adverse events. It has been shown that boiling can reduce peanut allergenicity. Hypoallergenic peanut products have the potential to reduce treatment-related reactions during desensitization. To show that extended boiling (for up to 12 h) can progressively reduce peanut allergenicity while retaining T cell reactivity. Raw peanuts were boiled for half, 1, 2, 4 and 12 h in deionized water. After dehydration, boiled and raw peanuts were ground, defatted and soluble proteins extracted in PBS and cooking water (leachate) retained. SDS-PAGE, Western blot, inhibition ELISA, mass spectrometry and skin prick test were used to characterize changes to peanut allergens and human IgE reactivity. T cell responses to raw and boiled peanut extracts were determined by proliferation of CD4+/CD25+/CD134+ T cells in peanut-allergic and non-allergic individuals. Extended boiling progressively reduced peanut allergenicity through a combination of leaching of allergens into cooking water, fragmentation of allergens and denaturation of conformational epitopes. Two-hour boiling led to an eightfold reduction in IgE binding capacity of boiled peanuts as determined by inhibition ELISA, while 12-h boiling led to a 19-fold reduction. Mass spectrometry revealed an increasing number of unique allergen peptides with longer boiling times. Raw, 2- and 12-h boiled peanut extracts were equivalent in their ability to stimulate T cell activation and proliferation. Progressive reduction in peanut allergenicity with extended boiling does not affect T cell reactivity. Boiled peanuts may be a candidate for oral immunotherapy. © 2016 John Wiley & Sons Ltd.

  9. 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

  10. EAACI Guidelines on Allergen Immunotherapy

    DEFF Research Database (Denmark)

    Roberts, G; Pfaar, O; Akdis, C A

    2018-01-01

    Allergic rhinoconjunctivitis (AR) is an allergic disorder of the nose and eyes affecting about a fifth of the general population. Symptoms of AR can be controlled with allergen avoidance measures and pharmacotherapy. However, many patients continue to have ongoing symptoms and an impaired quality...

  11. Viwithan, a StandardizedWithania somniferaRoot Extract Induces Apoptosis in Murine Melanoma Cells.

    Science.gov (United States)

    Sudeep, H V; Gouthamchandra, K; Venkatesh, B J; Prasad, K Shyam

    2018-01-01

    Withania somnifera is an Indian medicinal herb known for the multipotential ability to cure various therapeutic ailments as described in the ayurvedic system of medicine. In the present study, we have evaluated the antiproliferative activity of a standardized W. somnifera root extract (Viwithan) against different human and murine cancer cell lines. The cytotoxicity of Viwithan was determined using thiazolyl blue tetrazolium blue assay and crystal violet staining. The apoptotic changes in B16F1 cells following treatment with Viwithan were observed by acridine orange/ethidium bromide (AO/EB) staining and DNA fragmentation assay. The binding affinity of withanolides in Viwithan with antiapoptotic proteins B-cell lymphoma 2, B-cell lymphoma-extra large, and myeloid cell leukemia 1 (MCL-1) were studied using in silico approach. The half maximal inhibitory concentration (IC50) values of Viwithan against liver hepatocellular carcinoma, Henrietta Lacks cervical carcinoma cells, human colorectal carcinoma cell line, and Ehrlich ascites carcinoma cells were 1830, 968, 2715, and 633 μg/ml, respectively. Interestingly, Viwithan was highly effective against B16F1 cells with an IC50 value of 220 μg/ml after 24 h treatment. The morphological alterations of apoptotic cell death were clearly observed in the AO/EB-stained cells after treatment with Viwithan. Viwithan induced late apoptotic changes in treated B16F1 cells as evident by the ladder formation of fragmented DNA in a time-dependent manner. The findings of molecular docking showed that withanolides present in Viwithan have a more binding affinity with the antiapoptotic proteins, particularly MCL-1. We have reported for the first time that Viwithan with 5% withanolides has a potent cytotoxic effect, particularly against B16F1 murine melanoma cells among the different cancer cell lines tested. The present study reports for the first time that Viwithan, a standardized 5% Withania somnifera root extract, has potent

  12. A biodegradable device for the controlled release of Piper nigrum (Piperaceae standardized extract to control Aedes aegypti (Diptera, Culicidae larvae

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    Kauê Muller Custódio

    Full Text Available Abstract: INTRODUCTION: The significant increase in dengue, Zika, and chikungunya and the resistance of the Aedes aegypti mosquito to major insecticides emphasize the importance of studying alternatives to control this vector. The aim of this study was to develop a controlled-release device containing Piper nigrum extract and to study its larvicidal activity against Aedes aegypti. METHODS: Piper nigrum extract was produced by maceration, standardized in piperine, and incorporated into cotton threads, which were inserted into hydrogel cylinders manufactured by the extrusion of carrageenan and carob. The piperine content of the extract and thread reservoirs was quantified by chromatography. The release profile from the device was assessed in aqueous medium and the larvicidal and residual activities of the standardized extract as well as of the controlled-release device were examined in Aedes aegypti larvae. RESULTS The standardized extract contained 580mg/g of piperine and an LC50 value of 5.35ppm (24h and the 3 cm thread reservoirs contained 13.83 ± 1.81mg of piperine. The device showed zero-order release of piperine for 16 days. The P. nigrum extract (25ppm showed maximum residual larvicidal activity for 10 days, decreasing progressively thereafter. The device had a residual larvicidal activity for up to 37 days. CONCLUSIONS: The device provided controlled release of Piper nigrum extract with residual activity for 37 days. The device is easy to manufacture and may represent an effective alternative for the control of Aedes aegypti larvae in small water containers.

  13. Inhibition of colon carcinogenesis by a standardized Cannabis sativa extract with high content of cannabidiol.

    Science.gov (United States)

    Romano, Barbara; Borrelli, Francesca; Pagano, Ester; Cascio, Maria Grazia; Pertwee, Roger G; Izzo, Angelo A

    2014-04-15

    Colon cancer is a major public health problem. Cannabis-based medicines are useful adjunctive treatments in cancer patients. Here, we have investigated the effect of a standardized Cannabis sativa extract with high content of cannabidiol (CBD), here named CBD BDS, i.e. CBD botanical drug substance, on colorectal cancer cell proliferation and in experimental models of colon cancer in vivo. Proliferation was evaluated in colorectal carcinoma (DLD-1 and HCT116) as well as in healthy colonic cells using the MTT assay. CBD BDS binding was evaluated by its ability to displace [(3)H]CP55940 from human cannabinoid CB1 and CB2 receptors. In vivo, the effect of CBD BDS was examined on the preneoplastic lesions (aberrant crypt foci), polyps and tumours induced by the carcinogenic agent azoxymethane (AOM) as well as in a xenograft model of colon cancer in mice. CBD BDS and CBD reduced cell proliferation in tumoral, but not in healthy, cells. The effect of CBD BDS was counteracted by selective CB1 and CB2 receptor antagonists. Pure CBD reduced cell proliferation in a CB1-sensitive antagonist manner only. In binding assays, CBD BDS showed greater affinity than pure CBD for both CB1 and CB2 receptors, with pure CBD having very little affinity. In vivo, CBD BDS reduced AOM-induced preneoplastic lesions and polyps as well as tumour growth in the xenograft model of colon cancer. CBD BDS attenuates colon carcinogenesis and inhibits colorectal cancer cell proliferation via CB1 and CB2 receptor activation. The results may have some clinical relevance for the use of Cannabis-based medicines in cancer patients. Copyright © 2013 Elsevier GmbH. All rights reserved.

  14. Enhanced Cutaneous Wound Healing In Vivo by Standardized Crude Extract of Poincianella pluviosa.

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    Fernanda Giacomini Bueno

    Full Text Available Wound healing is a complex process that involves several biological events, and a delay in this process may cause economic and social problems for the patient. The search continues for new alternative treatments to aid healing, including the use of herbal medicines. Members of the genus Caesalpinia are used in traditional medicine to treat wounds. The related species Poincianella pluviosa (DC. L.P. Queiroz increases the cell viability of keratinocytes and fibroblasts and stimulates the proliferation of keratinocytes in vitro. The crude extract (CE from bark of P. pluviosa was evaluated in the wound-healing process in vivo, to validate the traditional use and the in vitro activity. Standardized CE was incorporated into a gel and applied on cutaneous wounds (TCEG and compared with the formulation without CE (Control for 4, 7, 10, or 14 days of treatment. The effects of the CE on wound re-epithelialization; cell proliferation; permeation, using photoacoustic spectroscopy (PAS; and proteins, including vascular endothelial growth factor (VEGF, superoxide dismutase 2 (SOD-2 and cyclooxygenase 2 (COX-2 were evaluated. The TCEG stimulated the migration of keratinocytes at day 4 and proliferation on the following days, with a high concentration of cells in metaphase at 7 days. Type I collagen formed more rapidly in the TCEG. PAS showed that the CE had permeated through the skin. TCEG stimulated VEGF at day 4 and SOD-2 and COX-2 at day 7. The results suggest that the CE promoted the regulation of proteins and helped to accelerate the processes involved in healing, promoting early angiogenesis. This led to an increase in the re-epithelialized surface, with significant mitotic activity. Maturation of collagen fibers was also enhanced, which may affect the resistance of the extracellular matrix. PAS indicated a correlation between the rate of diffusion and biological events during the healing process. The CE from P. pluviosa appears promising as an aid in

  15. Allergen Microarray Indicates Pooideae Sensitization in Brazilian Grass Pollen Allergic Patients.

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    Priscila Ferreira de Sousa Moreira

    Full Text Available Grass pollen, in particular from Lolium multiflorum is a major allergen source in temperate climate zones of Southern Brazil. The IgE sensitization profile of Brazilian grass pollen allergic patients to individual allergen molecules has not been analyzed yet.To analyze the IgE sensitization profile of a Brazilian grass pollen allergic population using individual allergen molecules.We analyzed sera from 78 grass pollen allergic patients for the presence of IgE antibodies specific for 103 purified micro-arrayed natural and recombinant allergens by chip technology. IgE-ELISA inhibition experiments with Lolium multiflorum, Phleum pratense extracts and a recombinant fusion protein consisting of Phl p 1, Phl p 2, Phl p 5 and Phl p 6 were performed to investigate cross-reactivities.Within the Brazilian grass pollen allergic patients, the most frequently recognized allergens were Phl p 1 (95%, Phl p 5 (82%, Phl p 2 (76% followed by Phl p 4 (64%, Phl p 6 (45%, Phl p 11 (18% and Phl p 12 (18%. Most patients were sensitized only to grass pollen allergens but not to allergens from other sources. A high degree of IgE cross-reactivity between Phleum pratense, Lolium multiflorum and the recombinant timothy grass fusion protein was found.Component-resolved analysis of sera from Brazilian grass pollen allergic patients reveals an IgE recognition profile compatible with a typical Pooideae sensitization. The high degree of cross-reactivity between Phleum pratense and Lolium multiflorum allergens suggests that diagnosis and immunotherapy can be achieved with timothy grass pollen allergens in the studied population.

  16. An autoclave treatment reduces the solubility and antigenicity of an allergenic protein found in buckwheat flour.

    Science.gov (United States)

    Tomotake, Hiroyuki; Yamazaki, Rikio; Yamato, Masayuki

    2012-06-01

    The effects of an autoclave treatment of buckwheat flour on a 24-kDa allergenic protein were investigated by measuring reduction in solubility and antibody binding. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that the intensity of the major bands, including that of the 24-kDa allergen, was reduced by the autoclave treatment. The protein solubility in buckwheat flour was variably decreased by the autoclave treatment. Enzyme-linked immunosorbent assay analysis using a monoclonal antibody specific for buckwheat 24-kDa protein showed that the reactivity of protein extracts (10 μg/ml) from buckwheat flour was lowered by the autoclave treatment. The autoclave treatment may reduce the major allergen content of buckwheat. Future studies will determine if autoclaving treatments affect the allergenicity of the 24-kDa buckwheat protein.

  17. Enzymatic hydrolysis: a method in alleviating legume allergenicity.

    Science.gov (United States)

    Kasera, Ramkrashan; Singh, A B; Lavasa, S; Prasad, Komarla Nagendra; Arora, Naveen

    2015-02-01

    Legumes are involved in IgE mediated food allergy in many countries. Avoidance of allergenic food is the only way to avoid symptomatic reaction. The present study investigated the effect of enzymatic hydrolysis on the allergenicity of three legumes - kidney bean (Phaseolus vulgaris), black gram (Vigna mungo) and peanut (Arachis hypogaea). Soluble protein extracts of the study legumes were sequentially treated by Alcalase(®) and Flavourzyme(®). Allergenicity of hydrolysates was then determined by ELISA, immunoblot, stripped basophil histamine release and skin prick test (SPT). Hydrolysis resulted in the loss of all IgE binding fractions determined by immunoblot in the three legumes. Specific IgE binding in ELISA was reduced by 62.2 ± 7.7%, 87.1 ± 9.6% and 91.8 ± 7.2% in the hydrolysates of kidney bean, black gram and peanut, respectively (p < 0.01). The release of histamine was decreased significantly when sensitized basophils were challenged with hydrolysates as compared to raw extracts. Significant reduction in the biopotency of hydrolysates was also observed in SPT where only 1/10 kidney bean-sensitive individuals, 2/6 black gram-sensitive individuals and 1/7 peanut-sensitive individuals were found positive to their respective hydrolysates. In conclusion, enzymatic hydrolysis is effective in attenuating allergenicity of legume proteins and may be employed for preparing hypoallergenic food extracts. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. NUTIRTION LABELLING OF FOOD AND ALLERGEN IN FOOD

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    Jozef Golian

    2012-10-01

    Full Text Available The new regulation introduced mandatory nutrition labelling and ordering food manufacturers provide information on energy and six nutrients: fat, saturated fatty acids, carbohydrates, sugars, protein and salt - in that order, and per 100 g or 100 ml. This information should be included in the nutritional table in one visual field (usually on the back cover, moreover, can also be expressed on per serving. It is important to realize that this regulation requires manufacturers indicate the nutritional value in one field of vision, usually on the "back cover" designation in the principal field (e.g. "on the front cover" remains voluntary. 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 labelling of ‘‘priority allergens. 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 labelling regulations was assembled and analyzed to identify commonalities, differences, and future needs.doi:10.5219/230

  19. Quantitation of house dust mites and house dust mite allergens in the microenvironment of dogs.

    Science.gov (United States)

    Randall, Amy; Hillier, Andrew; Cole, Lynette K; Kwochka, Kenneth W; Needham, Glen; Wassom, Donald L

    2003-10-01

    To quantitate the density of Dermatophagoides farinae and D pteronyssinus and concentrations of house dust mite (HDM) allergens (Der f 1, Der p 1, and Group 2 allergens) in the indoor microenvironment of dogs. Sample Population-50 homes in Columbus, Ohio. n each home, samples of dust were collected from 3 locations in which dogs spent most time. Whenever possible, the species of mites collected was identified. Mite density (mites/g of dust) was assessed, and allergen concentrations were assayed by standardized ELISAs. Relative humidity and temperature in each home were monitored during a 5-day period. Characteristics of homes and sample sources were evaluated. Dust samples from all 50 homes contained > or = 1 HDM allergen; Der f 1 and Der p 1 were detected in 100 and 74% of homes, respectively. Fifteen homes had HDMs; compared with D pteronyssinus, D farinae was found more commonly (14/15 homes) and at a higher density. Basements, homes without central air-conditioning, and dog beds that were > or = 1 year old had high HDM allergen concentrations. Homes with > or = 2 microg of Der f 1 or Group 2 allergens/g of dust or > or = 100 mites/g of dust were significantly more likely to have a maximum relative humidity > or = 75%. Results indicate the presence of HDMs and HDM allergens in the specific microenvironment of dogs in homes. Factors associated with high levels of exposure were identified, which may be associated with increased risk for sensitization and development of atopic diseases.

  20. Extensive protein hydrolyzation is indispensable to prevent IgE-mediated poultry allergen recognition in dogs and cats.

    Science.gov (United States)

    Olivry, Thierry; Bexley, Jennifer; Mougeot, Isabelle

    2017-08-17

    The central premise for the commercialization of diets with hydrolyzed ingredients is that the small-sized digested peptides would be unable to crosslink allergen-specific IgE at the surface of tissue mast cells and induce their degranulation. Evidence for the validity of this concept to diagnose food allergies in dogs and cats is limited, however. Our objectives were to study the recognition of standard and variably hydrolyzed poultry extracts by sera from dogs and cats with elevated chicken-specific serum IgE. Forty sera from dogs and 40 from cats with undetectable, low, medium or high serum levels of chicken-specific IgE were tested by ELISA on plates coated with the positive controls chicken, duck and turkey meat extracts and the negative controls beef meat (dogs) or wheat (cats). Plates were also coated with a non-hydrolyzed chicken meal, and mildly- or extensively-hydrolyzed poultry feather extracts. The frequencies of dogs with positive IgE against the various extracts were: chicken meat: 100%, duck and turkey meats: 97%, beef meat: 3%, non-hydrolyzed chicken meal: 73%, mildly-hydrolyzed poultry feathers: 37% and extensively-hydrolyzed poultry feathers: 0%. For cats, these respective percentages were (with wheat replacing beef as a negative control): 100, 84, 97, 7, 7, 0 and 0%. To detect any allergenic cross-reactivity between poultry meat-based and feather hydrolysate-derived extracts, an IgE ELISA inhibition was also done. Ten canine sera with the highest level of anti-poultry IgE in the previous experiment were incubated overnight with a previously optimized 50 μg amount of each of the extracts used above. We performed ELISA on plates coated with chicken, duck or turkey meats with or without inhibitors. The median inhibition percentages after incubation with the non-hydrolyzed chicken meal were ~22%, with the mildly-hydrolyzed poultry feathers: 14-22%, and those with the extensively-hydrolyzed poultry feathers: 5 to 10%; the last inhibition level was

  1. Skeletal muscle relaxant effect of a standardized extract of Valeriana officinalis L. after acute administration in mice

    Directory of Open Access Journals (Sweden)

    Dorian Caudal

    2018-04-01

    Full Text Available Valeriana officinalis L. root extracts are traditionally taken for their sedative and anxiolytic properties and are also used for muscle relaxation. Relaxant effects were clearly observed on smooth muscle whereas data on effects on skeletal muscle are scarce and inconsistent. The aim of this study was to assess whether a standardized extract (SE of V. officinalis had myorelaxant effects by decreasing skeletal muscle strength and/or neuromuscular tone in mice. Mice received an acute dose of V. officinalis SE (2 or 5 g/kg per os or tetrazepam (10 mg/kg ip, a standard myorelaxant drug. Thirty minutes later, the maximal muscle strength was measured using a grip test, while global skeletal muscle function (endurance and neuromuscular tone was assessed in a wire hanging test. Compared to tetrazepam, both doses of V. officinalis SE induced a pronounced decrease in skeletal muscle strength without any significant effects on endurance and neuromuscular tone. This study provides clear evidence that the extract of V. officinalis tested has a relaxant effect on skeletal muscle. By decreasing skeletal muscle strength without impacting endurance and neuromuscular tone, V. officinalis SE could induce less undesirable side effects than standard myorelaxant agents, and be particularly useful for avoiding falls in the elderly. Keywords: Valeriana officinalis, Skeletal muscle relaxant, Strength, Hydroethanolic root extract, Acute treatment, Mouse

  2. Eugenia uniflora fruit (red type) standardized extract: a potential pharmacological tool to diet-induced metabolic syndrome damage management.

    Science.gov (United States)

    Oliveira, Pathise Souto; Chaves, Vitor Clasen; Bona, Natália Pontes; Soares, Mayara Sandrielly Pereira; Cardoso, Juliane de Souza; Vasconcellos, Flávia Aleixo; Tavares, Rejane Giacomelli; Vizzotto, Marcia; Silva, Luísa Mariano Cerqueira da; Grecco, Fabiane Borelli; Gamaro, Giovana Duzzo; Spanevello, Roselia Maria; Lencina, Claiton Leoneti; Reginatto, Flávio Henrique; Stefanello, Francieli Moro

    2017-08-01

    The aim of this study was to investigate the effect of Eugenia uniflora fruit (red type) extract on metabolic status, as well as on neurochemical and behavioral parameters in an animal model of metabolic syndrome induced by a highly palatable diet (HPD). Rats were treated for 150days and divided into 4 experimental groups: standard chow (SC) and water orally, SC and E. uniflora extract (200mg/kg daily, p.o), HPD and water orally, HPD and extract. Our data showed that HPD caused glucose intolerance, increased visceral fat, weight gain, as well as serum glucose, triacylglycerol, total cholesterol and LDL cholesterol; however, E. uniflora prevented these alterations. The extract decreased lipid peroxidation and prevented the reduction of superoxide dismutase and catalase activities in the prefrontal cortex, hippocampus and striatum of animals submitted to HPD. We observed a HPD-induced reduction of thiol content in these cerebral structures. The extract prevented increased acetylcholinesterase activity in the prefrontal cortex caused by HPD and the increase in immobility time observed in the forced swim test. Regarding chemical composition, LC/MS analysis showed the presence of nine anthocyanins as the major compounds. In conclusion, E. uniflora extract showed benefits against metabolic alterations caused by HPD, as well as exhibited antioxidant and antidepressant-like effects. Copyright © 2017. Published by Elsevier Masson SAS.

  3. In vitro allergy investigation: Does a multiple allergen testing system give useful information?

    Science.gov (United States)

    Sinclair, David; Saas, Myk; Cook, L John

    2005-01-01

    The Hitachi CLA allergy test gives individual results on a wide range of common allergens. This study looked at the effect on patient management of the extra information gleaned from the use of this test compared to the same patient being investigated under a protocol of a total IgE and 4 allergen specific IgE tests using the Pharmacia UNICAP system. Fifty-four patients who presented to our Dermatology Department with possible Type I hypersensitivity: 19 males (10 were <16 years); 36 females (12 were <16 years). Our standard investigative protocol using the Pharmacia UNICAP was applied to all samples; each was also tested using the Hitachi CLA system and the two sets of data compared. The CLA system identified 22/54 patients in which one or more potentially significant allergens were found that would not have been identified by our current system of total IgE plus 4 allergen specific IgE results. Multiple positive allergens were detected by the CLA system in 13/54 cases. 12/54 patients had no allergen that was positive CLA and the UNICAP. 3/54 had positive results using our Pharmacia-based protocol that were not available by use of the extended panel. 4/54 patients had profiles in which both systems offered differing clinically useful information. These data support the use of a multiple allergen testing system to identify potentially significant individual allergens, patients in whom a Type I allergic reaction is unlikely or patients with multiple positive results for whom allergen avoidance is unlikely to be effective.

  4. A randomized, double-blinded, placebo-controlled oral challenge study to evaluate the allergenicity of commercial, food-grade fish gelatin

    DEFF Research Database (Denmark)

    Hansen, Tine K; Poulsen, Lars K.; Stahl Skov, Per

    2004-01-01

    Recent interest in the labeling of foods and food proteins derived from allergenic sources necessitates determination of the potential allergenicity of such food ingredients. Fish gelatin is extracted from the skin of fish species known to elicit allergic reactions in sensitized individuals....

  5. EAACI Guidelines on allergen immunotherapy

    DEFF Research Database (Denmark)

    Pajno, G. B.; Fernandez-Rivas, M.; Arasi, S.

    2017-01-01

    . This Guideline, prepared by the European Academy of Allergy and Clinical Immunology (EAACI) Task Force on Allergen Immunotherapy for IgE-mediated Food Allergy, aims to provide evidence-based recommendations for active treatment of IgE-mediated food allergy with FA-AIT. Immunotherapy relies on the delivery......Food allergy can result in considerable morbidity, impairment of quality of life, and healthcare expenditure. There is therefore interest in novel strategies for its treatment, particularly food allergen immunotherapy (FA-AIT) through the oral (OIT), sublingual (SLIT), or epicutaneous (EPIT) routes...... with an extensive experience in FA-AIT. Patients and their families should be provided with information about the use of FA-AIT for IgE-mediated food allergy to allow them to make an informed decision about the therapy....

  6. Fish allergy and fish allergens

    DEFF Research Database (Denmark)

    Kuehn, A; Hilger, Christiane; Ollert, Markus

    2016-01-01

    Fish is one of the main elicitors for food allergies. For a long time, the clinical picture of fish allergy was reduced to the following features. First, fish-allergic patients suffer from a high IgE cross-reactivity among fishes so that they have to avoid all species. Second, clinically relevant...... symptoms are linked to the presence of IgE-antibodies recognizing parvalbumin, the fish panallergen. This view was challenged by results from recent studies as follows. 1. Allergic reactions which are limited to single or several fish species (mono-or oligosensitisations) apply not only to single cases...... review gives an overview on the clinical characteristics of fish allergy and the molecular properties of relevant fish allergens. The advancement of the IgE-based diagnosis using a panel of well-defined fish allergens from different species is in the focus of the discussion. © 2016 Dustri-Verlag Dr. Karl...

  7. Evaluation of Cholesterol-lowering Activity of Standardized Extract ofMangifera indicain Albino Wistar Rats.

    Science.gov (United States)

    Gururaja, G M; Mundkinajeddu, Deepak; Kumar, A Senthil; Dethe, Shekhar Michael; Allan, J Joshua; Agarwal, Amit

    2017-01-01

    Cholesterol lowering activity of Mangifera indica L. has been determined by earlier researchers and kernel, leaf and bark have shown significant activity. However, the specific cholesterol lowering activity of leaf methanol extract has not been determined. The present study involved evaluation of cholesterol lowering potential of methanol extract of M. indica leaves using high cholesterol diet model in albino Wistar rats. The acute oral toxicity at a dose of 5000 mg/ kg body weight was also determined in female albino Wistar rats. Phytoconstituents Iriflophenone 3-C-β-D-glucoside and mangiferin were quantified in methanol extracts of different varieties of mango leaves using high performance liquid chromatography. Significant cholesterol lowering activity was observed with methanol extract of M. indica leaves, at dose of 90 mg/kg body weight in rats and it was also found to be safe at dose of 5000 mg/kg rat body. Iriflophenone 3-C-β-D-glucoside and mangiferin were found to be in the range of 1.2 to 2.8% w/w and 3.9 to 4.6% w/w, respectively which along with 3 β taraxerol and other sterols could be contributing to the cholesterol lowering activity of mango leaves extract. The phytosterols rich extract of Mangifera indica leaves is a good source of nutraceutical ingredient that have the potential to lower serum cholesterol levels. The Mangifera indica leaves methanolic extract showed significant cholesterol lowering activity in high cholesterol diet induced hypercholesterolaemia model in rats when evaluated at a dose of 90 mg/kg rat body weight. The extract was found to contain Iriflophenone 3-C-β-D-glucoside and mangiferin which along with 3 β taraxerol and other sterols could be contributing to the cholesterol lowering activity.

  8. Evaluation of Cholesterol-lowering Activity of Standardized Extract of Mangifera indica in Albino Wistar Rats

    Science.gov (United States)

    Gururaja, G. M.; Mundkinajeddu, Deepak; Kumar, A. Senthil; Dethe, Shekhar Michael; Allan, J. Joshua; Agarwal, Amit

    2017-01-01

    Introduction: Cholesterol lowering activity of Mangifera indica L. has been determined by earlier researchers and kernel, leaf and bark have shown significant activity. However, the specific cholesterol lowering activity of leaf methanol extract has not been determined. Materials and Methods: The present study involved evaluation of cholesterol lowering potential of methanol extract of M. indica leaves using high cholesterol diet model in albino Wistar rats. The acute oral toxicity at a dose of 5000 mg/ kg body weight was also determined in female albino Wistar rats. Phytoconstituents Iriflophenone 3-C-β-D-glucoside and mangiferin were quantified in methanol extracts of different varieties of mango leaves using high performance liquid chromatography. Results and Discussion: Significant cholesterol lowering activity was observed with methanol extract of M. indica leaves, at dose of 90 mg/kg body weight in rats and it was also found to be safe at dose of 5000 mg/kg rat body. Iriflophenone 3-C-β-D-glucoside and mangiferin were found to be in the range of 1.2 to 2.8% w/w and 3.9 to 4.6% w/w, respectively which along with 3 β taraxerol and other sterols could be contributing to the cholesterol lowering activity of mango leaves extract. Conclusions: The phytosterols rich extract of Mangifera indica leaves is a good source of nutraceutical ingredient that have the potential to lower serum cholesterol levels. SUMMARY The Mangifera indica leaves methanolic extract showed significant cholesterol lowering activity in high cholesterol diet induced hypercholesterolaemia model in rats when evaluated at a dose of 90 mg/kg rat body weight. The extract was found to contain Iriflophenone 3-C-β-D-glucoside and mangiferin which along with 3 β taraxerol and other sterols could be contributing to the cholesterol lowering activity. PMID:28250649

  9. A Targeted LC-MS/MS Method for the Simultaneous Detection and Quantitation of Egg, Milk, and Peanut Allergens in Sugar Cookies.

    Science.gov (United States)

    Boo, Chelsea C; Parker, Christine H; Jackson, Lauren S

    2018-01-01

    Food allergy is a growing public health concern, with many individuals reporting allergies to multiple food sources. Compliance with food labeling regulations and prevention of inadvertent cross-contact in manufacturing requires the use of reliable methods for the detection and quantitation of allergens in processed foods. In this work, a novel liquid chromatography-tandem mass spectrometry multiple-reaction monitoring method for multiallergen detection and quantitation of egg, milk, and peanut was developed and evaluated in an allergen-incurred baked sugar cookie matrix. A systematic evaluation of method parameters, including sample extraction, concentration, and digestion, were optimized for candidate allergen peptide markers. The optimized method enabled the reliable detection and quantitation of egg, milk, and peanut allergens in sugar cookies, with allergen concentrations as low as 5 ppm allergen-incurred ingredient.

  10. Allergen immunotherapy for allergic rhinoconjunctivitis

    DEFF Research Database (Denmark)

    Dhami, Sangeeta; Nurmatov, Ulugbek; Roberts, Graham

    2016-01-01

    BACKGROUND: The European Academy of Allergy and Clinical Immunology (EAACI) is in the process of developing the EAACI Guidelines for Allergen Immunotherapy (AIT) for the Management of Allergic Rhinoconjunctivitis. We seek to critically assess the effectiveness, cost-effectiveness and safety of AI...... appraised using established instruments. Data will be descriptively and, if possible and appropriate, quantitatively synthesised. CONCLUSION: The findings from this review will be used to inform the development of recommendations for EAACI's Guidelines on AIT....

  11. 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.

  12. Successful transdermal allergen delivery and allergen-specific immunotherapy using biodegradable microneedle patches.

    Science.gov (United States)

    Kim, Ji Hye; Shin, Jung U; Kim, Seo Hyeong; Noh, Ji Yeon; Kim, Hye Ran; Lee, Jungsoo; Chu, Howard; Jeong, Kyoung Yong; Park, Kyung Hee; Kim, Jung Dong; Kim, Hong Kee; Jeong, Do Hyeon; Yong, Tai-Soon; Park, Jung-Won; Lee, Kwang Hoon

    2018-01-01

    Allergen-specific immunotherapy (SIT) is an effective treatment modality for allergic diseases such as atopic dermatitis (AD). However, frequent visits over a 3-year period as well as looming adverse events tend to discourage patient compliance. Therefore, a more convenient, effective, and safe method of SIT is needed. For several decades, use of microneedles has been promoted as an efficient and precise transdermal drug delivery method. In this study, we developed Dermatophagoides farinae (D. farinae) extract (DfE)-loaded microneedle patches, and evaluated their safety and efficacy as a novel SIT method. After 4 weeks of patch application, efficient allergen delivery and successful induction of immune response to DfE were demonstrated in mice, with no apparent adverse events. AD-induced NC/Nga mice received microneedle immunotherapy (MNIT) (10 μg), subcutaneous immunotherapy (SCIT) (10 μg), SCIT (100 μg), or placebo. Both MNIT (10 μg) and SCIT (100 μg) treatments improved clinical and histologic manifestations of AD skin lesions, altered immunoglobulin production, dampened Th2 cellular response, and boosted Treg infiltrates, without significant side effects; whereas SCIT (10 μg) or placebo subsets failed to show any effects. Based on the favorable safety and efficacy profiles demonstrated in mice by MNIT in the current study, we believe that MNIT may serve as a new SIT modality. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Topically applied standardized aqueous extract of Curcuma longa Linn. suppresses endotoxin-induced uveal inflammation in rats.

    Science.gov (United States)

    Agarwal, Renu; Gupta, S K; Agarwal, Puneet; Srivastava, Sushma

    2013-10-01

    Aqueous extract of C. longa when administered 4 h after induction of E. coli lipopolysaccharide-induced uveitis in rats showed significantly suppressed inflammation with a significantly lower mean clinical grade, histopathological grade and aqueous humor (AH) protein level compared to vehicle treated group. Although, prednisolone group showed significantly lower clinical grade, histopathological grades and AH protein levels compared to C. longa group, TNF-alpha levels did not differ significantly. Moreover, when the aqueous extract was administered starting from 3 days before induction of uveitis, the mean clinical and histopathological grade as well as AH protein and TNF-alpha levels were comparable to C. longa group when treatment was administered 4 h after induction of uveitis. It is concluded that topically applied standardized aqueous extract of C. longa suppresses endotoxin-induced uveitis in rats by reducing TNF-alpha activity.

  14. Aeroallergen analyses and their clinical relevance. I. Immunochemical quantification of allergens by RAST-inhibition, Mab-ELISA, basophil histamine release, and counter current immuno electrophoresis

    DEFF Research Database (Denmark)

    Johnsen, C R; Abrahamsen, L; Stahl Skov, P

    1991-01-01

    of indoor aeroallergens, cat, dog, and Derm. pter. allergen extracts were selected for the experiments. To evaluate unspecific interference, these allergens were compared mutually and with Cladosporium herbarum. Allergen extracts in varying dilutions were mixed with crushed glass fibre filter materials......, eluted, recovered by centrifugation, and allergen concentration quantified by the assays. Equal sensitivity was found for both IgE- and IgG4-RI assaying cat allergen (in the range 5-50 SQ-U/ml) and dog allergen (in the range 10(2)-10(3) SQ-U/ml). The IgG4-RI assaying Derm. pter. was more sensitive (50 SQ......-U/ml) than IgE-RI (2*10(3) SQ-U/ml). The ranges of allergen detection limits for the Mab-ELISA were equal for cat and Derm. pter. (10-10(2) SQ-U/ml). The range of allergen detection limits for CCIE, assaying dog were 10(4)-10(5) SQ-U/ml. The ranges of allergen detection limits for HR were equal for cat...

  15. Associations between baseline allergens and polysensitization

    DEFF Research Database (Denmark)

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

    2008-01-01

    : Seven allergens - parabens mix, N-isopropyl-N-phenyl-p-phenylenediamine, sesquiterpene lactone mix, wool alcohols, potassium dichromate, Myroxylon pereirae, and cobalt chloride - showed statistically significant positive associations to polysensitization. Five allergens p-phenylenediamine, neomycin...... sulfate, epoxy resin, primin, and nickel sulfate showed statistically significant negative associations to polysensitization. For the allergens with the strongest associations, only every second individual with these particular allergies had two or more additional allergies. Conclusions: No common...

  16. ANALYSIS OF IFN- CONCENTRATION IN WISTAR RAT BLOOD AFTER ORAL ADMINISTRATION OF STANDARDIZED GREEN TEA WATER EXTRACT

    Directory of Open Access Journals (Sweden)

    Djoko Agus Purwanto

    2010-12-01

    Full Text Available Green tea and its polyphenols have been studied extensively as cancer chemopreventive agents in recent years. However, the mechanisms of action are still not clearly understood. Some researchers suggest that immune system plays important role to destroy cancer cells. Because of that reason, the present study was designed to analyse the effects of oral administration standardized green tea water extract on increasing of IFN-g blood concentration and to elucidate possible mechanisms involved in the inhibitory action of the cancer development. Two groups (male and female of 5 rats have given p.o. administration 1.25% of standardized green tea water extract and got 300 mg of (--epigallocatechin gallate (EGCG/kg body weight, while two groups others (male and female were used as control. We found that IFN-g blood concentration on male and female Wistar rat are significantly increase with 13.11% and 17.59%, respectively (p

  17. 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

  18. 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

  19. Differential responses to natural and recombinant allergens in a murine model of fish allergy.

    Science.gov (United States)

    van der Ventel, Michelle L; Nieuwenhuizen, Natalie E; Kirstein, Frank; Hikuam, Christoph; Jeebhay, Mohamed F; Swoboda, Ines; Brombacher, Frank; Lopata, Andreas L

    2011-01-01

    Aerosolized fish proteins are an important cause of allergic airway reactions in both the domestic and the occupational environment. The aim of this study was to investigate inhalant fish-induced allergy in a mouse model and compare immune responses generated by raw and heat-treated fish extracts as well as natural and recombinant forms of the major fish allergen parvalbumin. Mice were sensitized with raw or cooked pilchard extract and challenged intranasally with cooked pilchard extract, purified natural pilchard parvalbumin or recombinant carp parvalbumin (rCyp c1.01). Cooked pilchard extract predominantly sensitized mice to parvalbumin and induced specific IgG1 and IgE antibodies against both pilchard parvalbumin and rCyp c1.01, whereas additional allergens were recognized by mice sensitized with raw extract, including a 36 kDa allergen that was also recognized by fish processing workers and was identified as glyceraldehyde-3-phosphate dehydrogenase. Mice challenged with cooked extract and purified pilchard parvalbumin had increased Th2 cytokine production in mediastinal lymph node cells and splenocytes, whereas mice challenged with rCyp c1.01 did not. This study identifies a new IgE-binding protein that may be important in occupational allergy to fish and demonstrates the feasibility of testing recombinant allergens for immunotherapeutic potential in vivo. Copyright © 2010 Elsevier Ltd. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Differences in the presence of allergens among several types of indoor environments

    Directory of Open Access Journals (Sweden)

    Barbara Brunetto

    2009-12-01

    Full Text Available Exposure to indoor allergens can occur both at home and in public places such as schools and workplaces. To investigate and compare the presence of indoor allergens in different kind of environments (schools, offices and homes, dust samples were collected from furniture, desks, mattresses and floors with a standardized procedure. Samples were analyzed for Der p 1, Der f 1, Mite group 2 (mites and Fel d 1(cat by monoclonal antibody ELISA assay. Mite allergens were detected with low frequencies in schools and workplaces and with high frequency in homes. Fel d 1 was found with high frequency in every examined environment. Homes rather than public places can represent the environment where people can easier incur in mite allergy. All environments could be at risk for cat allergen exposure.

  2. Evaluation of the ISO Standard 11063 DNA Extraction Procedure for Assessing Soil Microbial Abundance and Community Structure

    Science.gov (United States)

    Griffiths, Robert; Dequiedt, Samuel; Lelievre, Mélanie; Regnier, Tiffanie; Nowak, Virginie; Bailey, Mark; Lemanceau, Philippe; Bispo, Antonio; Chabbi, Abad; Maron, Pierre-Alain; Mougel, Christophe; Ranjard, Lionel

    2012-01-01

    Soil DNA extraction has become a critical step in describing microbial biodiversity. Historically, ascertaining overarching microbial ecological theories has been hindered as independent studies have used numerous custom and commercial DNA extraction procedures. For that reason, a standardized soil DNA extraction method (ISO-11063) was previously published. However, although this ISO method is suited for molecular tools such as quantitative PCR and community fingerprinting techniques, it has only been optimized for examining soil bacteria. Therefore, the aim of this study was to assess an appropriate soil DNA extraction procedure for examining bacterial, archaeal and fungal diversity in soils of contrasting land-use and physico-chemical properties. Three different procedures were tested: the ISO-11063 standard; a custom procedure (GnS-GII); and a modified ISO procedure (ISOm) which includes a different mechanical lysis step (a FastPrep ®-24 lysis step instead of the recommended bead-beating). The efficacy of each method was first assessed by estimating microbial biomass through total DNA quantification. Then, the abundances and community structure of bacteria, archaea and fungi were determined using real-time PCR and terminal restriction fragment length polymorphism approaches. Results showed that DNA yield was improved with the GnS-GII and ISOm procedures, and fungal community patterns were found to be strongly dependent on the extraction method. The main methodological factor responsible for differences between extraction procedure efficiencies was found to be the soil homogenization step. For integrative studies which aim to examine bacteria, archaea and fungi simultaneously, the ISOm procedure results in higher DNA recovery and better represents microbial communities. PMID:22984486

  3. Evaluation of the ISO standard 11063 DNA extraction procedure for assessing soil microbial abundance and community structure.

    Directory of Open Access Journals (Sweden)

    Pierre Plassart

    Full Text Available Soil DNA extraction has become a critical step in describing microbial biodiversity. Historically, ascertaining overarching microbial ecological theories has been hindered as independent studies have used numerous custom and commercial DNA extraction procedures. For that reason, a standardized soil DNA extraction method (ISO-11063 was previously published. However, although this ISO method is suited for molecular tools such as quantitative PCR and community fingerprinting techniques, it has only been optimized for examining soil bacteria. Therefore, the aim of this study was to assess an appropriate soil DNA extraction procedure for examining bacterial, archaeal and fungal diversity in soils of contrasting land-use and physico-chemical properties. Three different procedures were tested: the ISO-11063 standard; a custom procedure (GnS-GII; and a modified ISO procedure (ISOm which includes a different mechanical lysis step (a FastPrep ®-24 lysis step instead of the recommended bead-beating. The efficacy of each method was first assessed by estimating microbial biomass through total DNA quantification. Then, the abundances and community structure of bacteria, archaea and fungi were determined using real-time PCR and terminal restriction fragment length polymorphism approaches. Results showed that DNA yield was improved with the GnS-GII and ISOm procedures, and fungal community patterns were found to be strongly dependent on the extraction method. The main methodological factor responsible for differences between extraction procedure efficiencies was found to be the soil homogenization step. For integrative studies which aim to examine bacteria, archaea and fungi simultaneously, the ISOm procedure results in higher DNA recovery and better represents microbial communities.

  4. A standardized extract of Ginkgo biloba prevents locomotion impairment induced by cassava juice in Wistar rats

    Science.gov (United States)

    Rivadeneyra-Domínguez, Eduardo; Vázquez-Luna, Alma; Rodríguez-Landa, Juan F.; Díaz-Sobac, Rafael

    2014-01-01

    The long-term consumption of cassava (Manihot esculenta Crantz) juice produce neurotoxic effects in the rat, characterized by an increased motor activity in the open field test and presence of uncoordinated swim (i.e., lateral swimming), in the swim test; which has been associated with damage in the hippocampus (CA1). On the other hand, flavonoids content in the Ginkgo biloba extract has been reported to produces neuroprotective effects at experimental level; therefore we hypothesized that G. biloba extract may prevents the motor alterations produced by cassava juice and reduce cellular damage in hippocampal neurons of the rat. In present study the effect of vehicle, cassava juice (linamarin, 0.30 mg/kg), G. biloba extract (dry extract, 160 mg/kg), and combination of treatment were evaluated in the open field and swim tests to identify locomotor and hippocampal alterations in adult male Wistar rats. All treatments were administered once per day, every 24 h, for 28 days, by oral rout. The effect was evaluated at 0, 7, 14, 21, and 28 days of treatment. The results show that cassava group from day 14 of treatment increase crossing and rearing in the open field test, as compared with the vehicle group; while in the swim test produces an uncoordinated swim characterized by the lateral swim. In this same group an increase in the number of damage neurons in the hippocampus (CA1) was identified. Interestingly, both behavioral and neuronal alterations produced by cassava juice administration were prevented by treatment with G. biloba extract. The results shown that G. biloba extract exert a protective effect against behavioral and neuronal damage associated with consumption of cassava juice in the rat. These effects are possibly related with flavonoid content in the G. biloba extract. PMID:25309441

  5. Chapter 1: an overview of allergens.

    Science.gov (United States)

    Shah, Rachna; Grammer, Leslie C

    2012-01-01

    Most allergens are proteins or glycoproteins that range in molecular weight from 5000 to 100,000 Da, although polysaccharides and low molecular weight substances also may be allergenic. Common allergens include pollens, fungal spores, house-dust mites, and animal epithelial materials but can also include drugs, biological products, and insect venoms. The allergic response is dependent on the route of exposure. If exposure is to an inhaled aeroallergen, the allergic response will be a respiratory reaction in nature. Ingested or injected exposure gives rise to gastrointestinal, cutaneous, or anaphylactic reactions. Size of pollen determines clinical manifestation of allergy. For example, particles between 20 and 60 μm in diameter can be carried in the wind and cause nasal and ocular symptoms (allergic rhinoconjunctivitis). Particles <7 μm can deposit in the airways and cause symptoms of asthma. Animals produce allergens in forms unique to each species. Cat allergen, most importantly Fel d 1, is found mainly in cat saliva, sebaceous glands in the skin, and in urine of male cats. It is buoyant and "sticky," which means it easily remains airborne and may last in a home for up to 6-9 months after the source is removed. Cat allergen adheres to clothes and can be found in public places such as schools. Dog allergen, particularly Can f 1, is present in dander, saliva, urine, and serum. There are allergens specific to dog breeds, but all breeds produce allergenic proteins (even poodles and "hairless" dogs).

  6. 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.

  7. Extracts of Morus nigra L. Leaves Standardized in Chlorogenic Acid, Rutin and Isoquercitrin: Tyrosinase Inhibition and Cytotoxicity

    Science.gov (United States)

    Fontes, Pedro Ribeiro; Souza, Paula Monteiro; William Fagg, Christopher; Neves Silva Guerra, Eliete; de Medeiros Nóbrega, Yanna Karla; Silveira, Damaris; Fonseca-Bazzo, Yris; Simeoni, Luiz Alberto; Homem-de-Mello, Maurício; Oliveira Magalhães, Pérola

    2016-01-01

    Melanogenesis is a process responsible for melanin production, which is stored in melanocytes containing tyrosinase. Inhibition of this enzyme is a target in the cosmetics industry, since it controls undesirable skin conditions such as hyperpigmentation due to the overproduction of melanin. Species of the Morus genus are known for the beneficial uses offered in different parts of its plants, including tyrosinase inhibition. Thus, this project aimed to study the inhibitory activity of tyrosinase by extracts from Morus nigra leaves as well as the characterization of its chromatographic profile and cytotoxicity in order to become a new therapeutic option from a natural source. M. nigra leaves were collected, pulverized, equally divided into five batches and the standardized extract was obtained by passive maceration. There was no significant difference between batches for total solids content, yield and moisture content, which shows good reproducibility of the extraction process. Tyrosinase enzymatic activity was determined for each batch, providing the percentage of enzyme inhibition and IC50 values obtained by constructing dose-response curves and compared to kojic acid, a well-known tyrosinase inhibitor. High inhibition of tyrosinase activity was observed (above 90% at 15.625 μg/mL). The obtained IC50 values ranged from 5.00 μg/mL ± 0.23 to 8.49 μg/mL ± 0.59 and were compared to kojic acid (3.37 μg/mL ± 0.65). High Performance Liquid Chromatography analysis revealed the presence of chlorogenic acid, rutin and, its major compound, isoquercitrin. The chromatographic method employed was validated according to ICH guidelines and the extract was standardized using these polyphenols as markers. Cytotoxicity, assessed by MTT assay, was not observed on murine melanomas, human keratinocytes and mouse fibroblasts in tyrosinase IC50 values. This study demonstrated the potential of M. nigra leaf extract as a promising whitening agent of natural source against skin

  8. Extracts of Morus nigra L. Leaves Standardized in Chlorogenic Acid, Rutin and Isoquercitrin: Tyrosinase Inhibition and Cytotoxicity.

    Directory of Open Access Journals (Sweden)

    Marcela Medeiros de Freitas

    Full Text Available Melanogenesis is a process responsible for melanin production, which is stored in melanocytes containing tyrosinase. Inhibition of this enzyme is a target in the cosmetics industry, since it controls undesirable skin conditions such as hyperpigmentation due to the overproduction of melanin. Species of the Morus genus are known for the beneficial uses offered in different parts of its plants, including tyrosinase inhibition. Thus, this project aimed to study the inhibitory activity of tyrosinase by extracts from Morus nigra leaves as well as the characterization of its chromatographic profile and cytotoxicity in order to become a new therapeutic option from a natural source. M. nigra leaves were collected, pulverized, equally divided into five batches and the standardized extract was obtained by passive maceration. There was no significant difference between batches for total solids content, yield and moisture content, which shows good reproducibility of the extraction process. Tyrosinase enzymatic activity was determined for each batch, providing the percentage of enzyme inhibition and IC50 values obtained by constructing dose-response curves and compared to kojic acid, a well-known tyrosinase inhibitor. High inhibition of tyrosinase activity was observed (above 90% at 15.625 μg/mL. The obtained IC50 values ranged from 5.00 μg/mL ± 0.23 to 8.49 μg/mL ± 0.59 and were compared to kojic acid (3.37 μg/mL ± 0.65. High Performance Liquid Chromatography analysis revealed the presence of chlorogenic acid, rutin and, its major compound, isoquercitrin. The chromatographic method employed was validated according to ICH guidelines and the extract was standardized using these polyphenols as markers. Cytotoxicity, assessed by MTT assay, was not observed on murine melanomas, human keratinocytes and mouse fibroblasts in tyrosinase IC50 values. This study demonstrated the potential of M. nigra leaf extract as a promising whitening agent of natural source

  9. EXTRACT

    DEFF Research Database (Denmark)

    Pafilis, Evangelos; Buttigieg, Pier Luigi; Ferrell, Barbra

    2016-01-01

    The microbial and molecular ecology research communities have made substantial progress on developing standards for annotating samples with environment metadata. However, sample manual annotation is a highly labor intensive process and requires familiarity with the terminologies used. We have...

  10. Computational analysis of the relationship between allergenicity and digestibility of allergenic proteins in simulated gastric fluid

    Directory of Open Access Journals (Sweden)

    Ni Ting

    2007-10-01

    Full Text Available Abstract Background Safety assessment of genetically modified (GM food, with regard to allergenic potential of transgene-encoded xenoproteins, typically involves several different methods, evaluation by digestibility being one thereof. However, there are still debates about whether the allergenicity of food allergens is related to their resistance to digestion by the gastric fluid. The disagreements may in part stem from classification of allergens only by their sources, which we believe is inadequate, and the difficulties in achieving identical experimental conditions for studying digestion by simulated gastric fluid (SGF so that results can be compared. Here, we reclassify allergenic food allergens into alimentary canal-sensitized (ACS and non-alimentary canal-sensitized (NACS allergens and use a computational model that simulates gastric fluid digestion to analyze the digestibilities of these two types. Results The model presented in this paper is as effective as SGF digestion experiments, but more stable and reproducible. On the basis of this model, food allergens are satisfactorily classified as ACS and NACS types by their pathways for sensitization; the former are relatively resistant to gastric fluid digestion while the later are relatively labile. Conclusion The results suggest that it is better to classify allergens into ACS and NACS types when understanding the relationship between their digestibility and allergenicity and the digestibility of a target foreign protein is a parameter for evaluating its allergenicity during safety assessments of GM food.

  11. Identification of a 62-kDa major allergen from Artemisia pollen as a putative galactose oxidase.

    Science.gov (United States)

    Fu, W; Gao, Z; Gao, L; Jin, J; Liu, M; Sun, Y; Wu, S; Wu, L; Ma, H; Dong, Y; Wang, X; Gao, B; Wang, H; Akkerdaas, J H; Versteeg, S A; van Ree, R

    2017-12-08

    Around 20 years ago, a 60- to 70-kDa protein was reported as a major allergen of mugwort (Artemisia vulgaris) pollen. This study was to identify and characterize its molecular properties. Sera from 113 Chinese and 20 Dutch Artemisia-allergic/sensitized subjects (and pools thereof) were used to identify the 60- to 70-kDa allergen. Pollen extracts of seven Artemisia species were compared by immunoblotting. Transcriptomics and proteomics (mass spectrometry) of A. annua pollen were used to identify the putative 60- to 70-kDa Artemisia allergen. Both the natural purified and recombinant allergens were evaluated for IgE reactivity by ImmunoCAP. Fourteen Chinese Artemisia-allergic patients were tested intradermally with purified natural allergen. Immunoblots revealed two major bands at 12 and 25 kDa, and a weak band at 70 kDa for all seven Artemisia species. Using a combined transcriptomic and proteomic approach, the high molecular mass allergen in A. annua pollen was shown to be a 62-kDa putative galactose oxidase, with a putative N-glycosylation site. More than 94% of Artemisia pollen-allergic patients had IgE response to this allergen. Although recognition of a nonglycosylated recombinant version was only confirmed in a minority (16%) and at much lower IgE levels, this discrepancy cannot be explained simply by reactivity to the carbohydrate moiety on the natural allergen. Intradermal testing with the natural allergen was positive in five of nine sensitized patients. The previously reported 60- to 70-kDa allergen of Artemisia pollen is most likely a 62-kDa putative galactose oxidase here designated Art an 7. © 2017 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.

  12. 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

    . 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...... 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...

  13. Monoclonal antibodies to the major Lolium perenne (rye grass) pollen allergen Lol p I (Rye I).

    Science.gov (United States)

    Kahn, C R; Marsh, D G

    1986-12-01

    Thirteen monoclonal antibodies (MAbs) were produced against Lol p I (Rye I), the major Lolium perenne (rye grass) pollen allergen. Spleen cells from A/J and SJL mice immunized with highly purified Lol p I (Lol I) were allowed to fuse with cells from the non-secreting Sp2/0-Ag14 myeloma cell line. Each MAb was analyzed for antigenic specificity by radioimmunoassay (RIA) using 125I-Lol I. The epitope specificities of seven of the MAbs were examined by competitive binding against a labelled standard MAb for the Lol I antigen (Ag). The dissociation constant, Kd, of one MAb (No. 3.2) that was studied most extensively was determined by double Ab RIA to be 3.5 X 10(-6) L/M. This MAb recognized the related 27,000-30,000 Group I glycoproteins found in the pollens of nine other species of grass pollens tested, including weak binding to Bermuda grass Group I (Cyn d I), which by conventional analysis using polyclonal anti-Lol I serum shows no detectable binding. Monoclonal antibody No. 3.2 was coupled covalently to Sepharose 4B and used to prepare highly purified Lol I from a partially purified rye pollen extract. Finally, an RIA was developed which permitted the analysis of the Group I components in rye grass and nine other grass pollen species. The latter assay is likely to prove useful in the standardization of grass pollen extracts according to their Group I contents.

  14. Application of Enzymatic Extracts from a CALB Standard Strain as Biocatalyst within the Context of Conventional Biodiesel Production Optimization

    Directory of Open Access Journals (Sweden)

    Carlos Luna

    2017-11-01

    Full Text Available The application of biocatalysts in the transesterification process of triglycerides (TG allows integrating the glycerol in the form of monoglyceride (MG, sharply increasing the yield and the environmental sustainability of the conventional biodiesel production process. This is known as Ecodiesel. To overcome the inconvenient of the high cost of the currently employed highly purified commercial enzymes, the use of scarcely purified extracts obtained from standard strains of the same species of commercial lipases currently applied in this process is being investigated. Thus, Candida antarctica type B (CALB was chosen to determine the optimal conditions of culture of this yeast. The standard strain was obtained from the Spanish Type Microbial Cultures Collection (CECT and has been used to carry out several studies to elucidate its optimum growth conditions. Through a process of lyophilization with prior dialysis of the liquid cultures, the enzymatic extracts were obtained. The different obtained cultures have been applied as biocatalysts in the 1,3-selective transesterification reaction of sunflower oil with ethanol to obtain Ecodiesel (FAEE + MG. Selectivity and reaction yields were obtained by gas chromatography. Acceptable yields are obtained during the reaction time as well as in successive reactions, demonstrating the feasibility of using these CALB enzymatic extracts as biocatalysts.

  15. 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

  16. A Dietary supplement containing standardized Phaseolus vulgaris extract influences body composition of overweight men and women.

    Science.gov (United States)

    Celleno, Leonardo; Tolaini, Maria Vittoria; D'Amore, Alessandra; Perricone, Nicholas V; Preuss, Harry G

    2007-01-24

    More than one billion human adults worldwide are overweight and, therefore, are at higher risk of developing cardiovascular diseases, diabetes, and a variety of other chronic perturbations. Many believe that use of natural dietary supplements could aid in the struggle against obesity. So-called "starch blockers" are listed among natural weight loss supplements. Theoretically, they may promote weight loss by interfering with the breakdown of complex carbohydrates thereby reducing, or at least slowing, the digestive availability of carbohydrate-derived calories and/or by providing resistant starches to the lower gastrointestinal tract. The present research study examines a dietary supplement containing 445 mg of Phaseolus vulgaris extract derived from the white kidney bean, previously shown to inhibit the activity of the digestive enzyme alpha amylase, on body composition of overweight human subjects. A randomized, double-blinded, placebo-controlled study was conducted on 60 pre-selected, slightly overweight volunteers, whose weight had been essentially stable for at least six months. The volunteers were divided into two groups, homogeneous for age, gender, and body weight. The test product containing Phaseolus vulgaris extract and the placebo were taken one tablet per day for 30 consecutive days before a main meal rich in carbohydrates. Each subject's body weight, fat and non-fat mass, skin fold thickness, and waist/hip/thigh circumferences were measured. After 30 days, subjects receiving Phaseolus vulgaris extract with a carbohydrate-rich, 2000- to 2200-calorie diet had significantly (preduction of body weight, BMI, fat mass, adipose tissue thickness, and waist,/hip/ thigh circumferences while maintaining lean body mass compared to subjects receiving placebo. The results indicate that Phaseolus vulgaris extract produces significant decrements in body weight and suggest decrements in fat mass in the face of maintained lean body mass.

  17. Presenting a New Standard Drug Model for Turmeric and Its Prized Extract, Curcumin

    OpenAIRE

    Franco Cavaleri

    2018-01-01

    Various parts of the turmeric plant have been used as medicinal treatment for various conditions from ulcers and arthritis to cardiovascular disease and neuroinflammation. The rhizome’s curcumin extract is the most studied active constituent, which exhibits an expansive polypharmacology with influence on many key inflammatory markers. Despite the expansive reports of curcucmin’s therapeutic value, clinical reliability and research repeatability with curcumin treatment are still poor. The phar...

  18. Novel cytosolic allergens of Aspergillus fumigatus identified from germinating conidia.

    Science.gov (United States)

    Singh, Bharat; Sharma, Gainda L; Oellerich, Michael; Kumar, Ram; Singh, Seema; Bhadoria, Dharam P; Katyal, Anju; Reichard, Utz; Asif, Abdul R

    2010-11-05

    Aspergillus fumigatus is the common cause of allergic broncho-pulmonary aspergillosis (ABPA) and most of the allergens have been described from its secreted fraction. In the present investigation, germinating conidial cytosolic proteins of A. fumigatus were extracted from a 16 h culture. The proteome from this fraction was developed, and immuno-blots were generated using pooled ABPA patients' sera. Well separated Immunoglobulin-E (IgE) and Immunoglobulin-G (IgG) reactive spots were picked from corresponding 2DE gels and subjected to mass spectrometric analysis. As a result, 66 immuno-reactive proteins were identified from two geographically different strains (190/96 and DAYA) of A. fumigatus. Only 3 out of 66 proteins reacted with IgG, and the remaining 63 proteins were found to be IgE reactive. These 63 IgE-reactive cytosolic proteins from germinating conidia included 2 already known (Asp f12 and Asp f22) and 4 predicted allergens (Hsp88, Hsp70, malate dehydrogenase, and alcohol dehydrogenase) based on their homology with other known fungal allergens. In view of this, the panel of presently identified IgE-reactive novel proteins holds the potential of providing a basis for the wider diagnostic application in assay for allergic aspergillosis. We could demonstrate that recombinantly expressed proteins from this panel showed consistent reactivity with IgE of individual sera of ABPA patients. The recombinantly expressed proteins may also be useful in desensitization therapy of allergic disorders including ABPA.

  19. Chinese Guideline on allergen immunotherapy for allergic rhinitis

    Science.gov (United States)

    Bao, Yixiao; Chen, Jianjun; Cheng, Lei; Guo, Yinshi; Hong, Suling; Kong, Weijia; Lai, He; Li, Houyong; Li, Huabin; Li, Jing; Li, Tianying; Lin, Xiaoping; Liu, Shixi; Liu, Zheng; Lou, Hongfei; Meng, Juan; Qiu, Qianhui; Shen, Kunling; Tang, Wei; Tao, Zezhang; Wang, Chengshuo; Wang, Xiangdong; Wei, Qingyu; Xiang, Li; Xie, Hua; Xu, Yu; Zhang, Gehua; Zhang, Yuan; Zheng, Yiwu; Zhi, Yuxiang; Chen, Dehua; Hong, Haiyu; Li, Quansheng; Liu, Lin; Meng, Yifan; Wang, Nan; Wang, Yihui; Zhou, Yue

    2017-01-01

    The present document is based on a consensus reached by a panel of experts from Chinese Society of Allergy (CSA) and Chinese Allergic Rhinitis Collaborative Research Group (C2AR2G). Allergen immunotherapy (AIT), has increasingly been used as a treatment for allergic rhinitis (AR) globally, as it has been shown to provide a long-term effect in improving nasal and ocular symptoms, reducing medication need, and improving quality of life. AIT is currently the only curative intervention that can potentially modify the immune system in individuals suffering from AR and prevent the development of new sensitization and the progression of disease from AR to asthma. Although the use of AIT is becoming more acceptable in China, to date no AR immunotherapy guideline from China is available for use by the international community. This document has thus been produced and covers the main aspects of AIT undertaken in China; including selection of patients for AIT, the allergen extracts available on the Chinese market, schedules and doses of allergen employed in different routes of AIT, assessment of effect and safety, patients’ administration and follow-up, and management of adverse reactions. The Chinese guideline for AR immunotherapy will thus serve as a reference point by doctors, healthcare professionals and organizations involved in the AIT of AR in China. Moreover, this guideline will serve as a source of information for the international community on AIT treatment strategies employed in China. PMID:29268533

  20. Chloroatranol, an extremely potent allergen hidden in perfumes

    DEFF Research Database (Denmark)

    Johansen, Jeanne Duus; Andersen, Klaus Ejner; Svedman, Cecilia

    2003-01-01

    Oak moss absolute is a long-known, popular natural extract widely used in perfumes. It is reported as the cause of allergic reactions in a significant number of those with perfume allergy. Oak moss absolute has been the target of recent research to identify its allergenic components. Recently...... an open test simulating the use of perfumes on the volar aspect of the forearms in a randomized and double-blinded design. A solution with 5 p.p.m. chloroatranol was used for 14 days, and, in case of no reaction, the applications were continued for another 14 days with a solution containing 25 p.p.m. All....... The dose eliciting a reaction in 50% of the test subjects at patch testing was 0.2 p.p.m. In conclusion, the hidden exposure to a potent allergen widely used in perfumes has caused a highly sensitized cohort of individuals. Judged from the elicitation profile, chloroatranol is the most potent allergen...

  1. Extraction of Indigo from Some Isatis species and Dyeing Standardization Using Low-technology Methods

    Directory of Open Access Journals (Sweden)

    Nazan Comlekcioglu

    2015-02-01

    Full Text Available Fresh leaves of four Isatis species culture form of I. tinctoria L and wild forms of I. buschiana Schischkin, I. candolleana Boiss. (endemic and I. tinctoria L. subsp. corymbosa. (Boiss. were used for indigo production. Dyes were extracted by fermentation and hot water application. The extracted dyes were optimized with different pH and reducing agents. Results showed that the dye from hot water application produced the desired dying quality at pH 11. Reducing agent concentrations had no significant effect on color quality. Dark blue and blue colors were obtained from I. tinctoria and I. candolleana extracts although I. tinctoria subsp. corymbosa and I. buschiana produced mostly yellow-gray colors. Light, dry and wet rubbing fastness values varied between 3 and 3/4 while washing fastness was between 2 and 4/5. The highest indigo amounts were determined spectrophotometrically as 4.19 mg/g and 2.53 mg/g in I. tinctoria and I. candolleana, respectively. Results also showed that harvesting season was important for indigo production and the highest indigo amount was observed in mid-June.

  2. Identification of IgE-mediated food allergy and allergens in older children and adults with asthma and allergic rhinitis.

    Science.gov (United States)

    Kumar, Raj; Kumari, Dolly; Srivastava, Prakriti; Khare, Vishal; Fakhr, Hena; Arora, Naveen; Gaur, S N; Singh, B P

    2010-01-01

    Prevalence of immunoglobulin (Ig) E-mediated food allergy is primarily reported for certain pediatric populations and adults. The present study was aimed to investigate the relative prevalence of food allergy and allergens in older children and adults with asthma and allergic rhinitis. Patients (12-62 years) were screened using standard questionnaire and skin prick-test (SPT) with common foods and aeroallergens. Specific IgE level was determined by enzyme linked immunosorbent assay (ELISA) and allergy was established by blinded food challenges. Of 1860 patients screened, 1097 (58.9%) gave history of food allergy. Of the history positive patients skin tested (n=470), 138 (29.3%) showed a marked positive reaction to food extracts. Rice elicited positive SPT reaction in maximum number of cases 29 (6.2%) followed by blackgram 28 (5.9%), lentil 26 (5.5%), citrus fruits 25 (5.3%), pea 18 (3.8%), maize 18 (3.8%) and banana 17 (3.6%). The SPT positive patients showed elevated specific IgE levels (range: 0.8-79 IU/mL) against respective food allergens than normal controls (0.73 IU/mL, mean +/- 2 SD). Food allergy was confirmed in 21/45 (46.6%) of the patients by blinded controlled food challenges. The prevalence of food allergy was estimated to be 4.5% (2.6%-6.34%) at 95% confidence interval (95% CI) in test population (n=470). Sensitisation to food was significantly associated with asthma (p = 0.0065) while aeroallergens were strongly related to rhinitis (p Food allergy is estimated to be 4.5% in adolescents and adults with asthma, rhinitis or both. Rice, citrus fruits, blackgram and banana are identified as major allergens for inducing allergic symptoms.

  3. Simultaneous determination of four food allergens using compact disc immunoassaying technology.

    Science.gov (United States)

    Badran, Ahmed Ali; Morais, Sergi; Maquieira, Ángel

    2017-03-01

    A multiplex competitive microimmunoassay for the simultaneous determination of gliadin, casein, β-lactoglobulin, and ovalbumin is presented. The assay in microarray format is performed on a DVD where the allergens are physisorbed on the polycarbonate surface of the disc. The immunointeraction is detected using a mixture of specific gold-labeled antibodies and the signal amplified with the silver enhancement method. The optical density of the precipitate, read by a DVD drive, is related to the concentration of the four allergens in sample. An optimized protocol for the simultaneous extraction of the allergen proteins from food samples is also addressed. The suitability of the method is demonstrated for the simultaneous quantitative extraction and determination of the targeted allergens in spiked baby foods, juices, and beers. The sensitivity (EC50) of the multiplexed assay was 0.04, 0.40, 0.08, and 0.16 mg L -1 for gliadin, casein, β-lactoglobulin, and ovalbumin, respectively, and the recovery results from the analysis of food samples ranged from 72 to 117%. A portable, easy-to-use, array-based bioanalytical method is developed for quantification of food allergens with a limit of detection below the accepted levels of the international legislations, which allows promotion of food safety and quality. Graphical abstract GLI Gliadin, CAS Casein, β-LAC β-lactoglobulin, OVA Ovalbumin.

  4. Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen.

    Science.gov (United States)

    Ohgiya, Yoko; Arakawa, Fumihiro; Akiyama, Hiroshi; Yoshioka, Yasuo; Hayashi, Yoshiki; Sakai, Shinobu; Ito, Sumio; Yamakawa, Yuko; Ohgiya, Satoru; Ikezawa, Zenro; Teshima, Reiko

    2009-05-01

    Carmine is a natural red pigment obtained from dried gravid female cochineal insects (Dactylopius coccus or Coccus cacti). There have been several reports of allergies to carmine, but the major allergens responsible have not been identified. To identify the major allergenic proteins in cochineal. Immunoblots of purified cochineal extract were probed with sera from 3 patients with allergy. Partial amino acid sequences were determined for the proteins bound by IgE, and the corresponding cDNA, containing a complete coding region, was cloned by 5' and 3' rapid cDNA extension and PCR. The recombinant protein was expressed in yeast and subjected to immunoblotting. We identified a full-length cDNA encoding a protein, which we named CC38K, with 335 amino acids and a molecular mass calculated as 38 kd. This amino acid sequence included all the partial amino acid sequences obtained from the purified proteins identified by IgE from patients with allergy. Recombinant CC38K protein was recognized by patients' sera, indicating that this is a major allergen present in carmine. The CC38K sequence showed homology to phospholipases. We have, for the first time, identified the major allergen in cochineal extract. This protein may be a phospholipase or related enzyme, both of which are known to be allergens in other insects.

  5. 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

  6. New Horizons in Allergen Immunotherapy

    DEFF Research Database (Denmark)

    Backer, Vibeke

    2016-01-01

    to and continued throughout the pollen season, as opposed to the year-round injections requiredwith SCIT. The main disadvantage of SLIT is that it requires adherence with daily dosing, and studies have consistently shown lowrates of long-term adherence. For example, a review of published studies found that 55%to......, these adherence rates are similar to those seen with SCIT and even with long-term pharmacotherapy for allergic disease and asthma.17 SLIT is currently approved for numerous allergens by the European regulatory authorities and is inwide use throughout Europe. In 2014, the US Food and Drug Administration announced...

  7. 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.

  8. Skin Test Reactivity to Indoor Allergens Correlates with Asthma Severity in Jeddah, Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Koshak Emad A

    2006-03-01

    Full Text Available Abstract Background There is increased emphasis on the role of indoor allergens in asthma. Objective To examine the spectrum of skin test reactivity (sensitization to indoor allergens and its correlation with asthma severity in Jeddah, Saudi Arabia. Methods Asthmatic patients referred to the allergy clinic at King Abdulaziz University Hospital (KAUH in Jeddah were studied. Measures of clinical severity were adopted from national and international asthma guidelines. The degree of sensitization was assessed by the wheal size (positive ≥ 3 mm from standard skin-prick tests for the following common indoor inhalant allergens: house dust mites (Dermatophagoides pteronyssinus [Dp] and Dermatophagoides farinae [Df], cat, and cockroach. Results Skin test results from 113 of 151 (74.8% asthmatic patients were positive for one or more allergens. The patients' ages ranged between 9 and 63 years (mean, 30 ± 13 years, and females constituted 65.5%. The predominant asthma severity level was moderate persistent (55.8%, followed by mild persistent (33.6%. The prevalences of sensitization to indoor allergens were as follows: Dp, 87% (3-25 mm [mean, 7 mm]; Df, 84% (3-20 mm [mean, 7 mm]; cat, 44% (3-15 mm [mean, 6 mm]; and cockroach, 33% (3-12 mm [mean, 4 mm]. Higher asthma severity levels were significantly correlated with the number of allergens with positive sensitization (R = 0.3, p Dp [degrees of freedom {df} = 16, p Df [df = 17, p df = 10, p df = 8, p Conclusions Immunoglobulin E-mediated skin test reactivity to indoor allergens, particularly to house dust mites, was common in asthmatic patients from Jeddah at KAUH. Increased sensitization was associated with higher levels of asthma severity, which is compatible with the literature. This emphasizes the importance of identifying sensitization to relevant indoor allergens in the clinical evaluation of asthmatic persons.

  9. Biomarkers of acute respiratory allergen exposure: Screening for sensitization potential

    International Nuclear Information System (INIS)

    Pucheu-Haston, Cherie M.; Copeland, Lisa B.; Vallanat, Beena; Boykin, Elizabeth; Ward, Marsha D.W.

    2010-01-01

    Effective hazard screening will require the development of high-throughput or in vitro assays for the identification of potential sensitizers. The goal of this preliminary study was to identify potential biomarkers that differentiate the response to allergens vs non-allergens following an acute exposure in naive individuals. Female BALB/c mice received a single intratracheal aspiration exposure to Metarhizium anisopliae crude antigen (MACA) or bovine serum albumin (BSA) in Hank's Balanced Salt Solution (HBSS) or HBSS alone. Mice were terminated after 1, 3, 6, 12, 18 and 24 h. Bronchoalveolar lavage fluid (BALF) was evaluated to determine total and differential cellularity, total protein concentration and LDH activity. RNA was isolated from lung tissue for microarray analysis and qRT-PCR. MACA administration induced a rapid increase in BALF neutrophils, lymphocytes, eosinophils and total protein compared to BSA or HBSS. Microarray analysis demonstrated differential expression of genes involved in cytokine production, signaling, inflammatory cell recruitment, adhesion and activation in 3 and 12 h MACA-treated samples compared to BSA or HBSS. Further analyses allowed identification of ∼ 100 candidate biomarker genes. Eleven genes were selected for further assessment by qRT-PCR. Of these, 6 demonstrated persistently increased expression (Ccl17, Ccl22, Ccl7, Cxcl10, Cxcl2, Saa1), while C3ar1 increased from 6-24 h. In conclusion, a single respiratory exposure of mice to an allergenic mold extract induces an inflammatory response which is distinct in phenotype and gene transcription from the response to a control protein. Further validation of these biomarkers with additional allergens and irritants is needed. These biomarkers may facilitate improvements in screening methods.

  10. Standardized Sophora pachycarpa Root Extract Enhances Osteogenic Differentiation in Adipose-derived Human Mesenchymal Stem Cells.

    Science.gov (United States)

    Mollazadeh, Samaneh; Neshati, Vajiheh; Fazly Bazzaz, Bibi Sedigheh; Iranshahi, Mehrdad; Mojarrad, Majid; Naderi-Meshkin, Hojjat; Kerachian, Mohammad Amin

    2017-05-01

    Bone defect is an important topic in public health. Novel therapies are based on osteogenic induction by natural antiosteoporotic compounds including plant-derived estrogens. In the current study, the osteogenic potential of Sophora pachycarpa root extract (SPRE) was explored on human adipose-derived mesenchymal stem cells. Herein, adipose-derived mesenchymal stem cells were osteoinducted in the presence of increased concentrations of the extract for 21 days. Then, cell viability was evaluated by MTT assay, and the differentiated cells were stained by Alizarin Red S for calcium deposition and subjected to alkaline phosphatase (ALP) assay for enzymatic activity. To assess the expression of bone-related genes, treated cells were evaluated by real-time polymerase chain reaction. The MTT test demonstrated that SPRE had no toxic effects on the cell viability. Treating the cells with SPRE noticeably promoted ALP activity, mineralization, and mRNA expression of runt-related transcription factor 2 (RUNX2), bone gamma-carboxyglutamate protein (BGLAP), secreted phosphoprotein 1 (SPP1), and collagen type I alpha 1 (COL1A1). Additionally, cells subjected to 0.1 μg/mL SPRE showed the highest osteogenic effects. According to high-performance liquid chromatography fingerprinting of SPRE, the osteoprotective effects of SPRE is probably due to presence of phytochemicals with estrogen-like activity in the extract. Thus, SPRE might be a suitable therapeutic agent for bone defects therapy in the future research. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  11. First-principles calibration of 40Ar/39Ar mineral standards and complete extraction of 40Ar* from sanidine

    Science.gov (United States)

    Morgan, L. E.; Kuiper, K.; Mark, D.; Postma, O.; Villa, I. M.; Wijbrans, J. R.

    2010-12-01

    40Ar/39Ar geochronology relies on comparing argon isotopic data for unknowns to those for knowns. Mineral standards used as neutron fluence monitors must be dated by the K-Ar method (or at least referenced to a mineral of known K-Ar age). The commonly used age of 28.02 ± 0.28 Ma for the Fish Canyon sanidine (FCs) (Renne et al., 1998) is based upon measurements of radiogenic 40Ar in GA1550 biotite (McDougall and Roksandic, 1974), but underlying full data were not published (these measurements were never intended for use as an international standard), so uncertainties are difficult to assess. Recent developments by Kuiper et al. (2008) and Renne et al. (2010) are limited by their reliance on the accuracy of other systems. Modern technology should allow for more precise and accurate calibration of primary K-Ar and 40Ar/39Ar standards. From the ideal gas law, the number of moles of 40Ar in a system can be calculated from measurements of pressure, volume, and temperature. Thus we have designed and are proceeding to build a pipette system to introduce well-determined amounts of 40Ar into noble gas extraction lines and mass spectrometers. This system relies on components with calibrations traceable to SI unit prototypes, including a diaphragm pressure gauge (MKS Instruments), thermocouples, and a “slug” of an accurately determined volume to be inserted into the reservoir for volume determinations of the reservoir and pipette. The system will be renewable, with a lifetime of ca. 1 month for gas in the reservoir, and portable, to permit interlaboratory calibrations. The quantitative extraction of 40Ar* from the mineral standard is of highest importance; for sanidine standards this is complicated by high melt viscosity during heating. Experiments adding basaltic “zero age glass” (ZAG) to decrease melt viscosity are underway. This has previously been explored by McDowell (1983) with a resistance furnace, but has not been quantitatively addressed with laser heating

  12. Contact sensitization to cosmetic series of allergens in a general population in Beijing.

    Science.gov (United States)

    Zhao, Jian; Li, Lin-Feng

    2014-03-01

    Cosmetic allergic contact dermatitis (CACD) due to common cosmetic allergens in standard series has been extensively studied; however, the prevalence of contact allergy to other cosmetic allergens other than those in standard series is largely unknown. In this study, the frequency of contact sensitization to a European cosmetic series of allergens (Chemotechnique Diagnostics, Vellinge, Sweden) in healthy university student volunteers were detected in Beijing. Of 201 students studied, fifty-eight exhibited positive results, and 9 of them reported had cosmetics related dermatitis previously. The total positivity rate was not correlated to gender. The leading allergens were thimerosal (19.4%), shellac (3.0%), cocamidopropyl betaine (2.0%), hexamethylenetetramine (1.5%), dodecyl gallate (1.5%), hexahydro-1,3,5-tris-(2-hydroxyethyl)triazine (1.0%) and methyldibromo glutaronitrile (1.0%). The positivity rate of thimerosal patch test in men (9.8%) was lower than that of women (23.6%, P cosmetic allergens in men and women (P > 0.05, Chi square test). These results suggested that some cosmetic-related contact allergies may be missed by just testing patients with the European standard series or T.R.U.E. test system only, we recommend shellac, cocamidopropyl betaine, hexamethylenetetramine and dodecyl gallate as the additionally candidates for patch testing in patients with suspected CACD. © 2014 Wiley Periodicals, Inc.

  13. Peanut allergen Ara h 1 interacts with proanthocyanidins into higher molecular weight complexes

    NARCIS (Netherlands)

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

    2007-01-01

    Mildly extracted peanut allergen Ara h 1 was previously reported to occur as an oligomeric complex. In this paper we describe how the protein in this oligomeric complex interacts noncovalently with phenolic compounds of the proanthocyanidin type. These interactions are being disrupted during anion

  14. Peanut Allergen Ara h 1 Interacts with Proanthocyanidins into Higher Molecular Weight Complexes

    NARCIS (Netherlands)

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

    2007-01-01

    Mildly extracted peanut allergen Ara h 1 was previously reported to occur as an oligomeric complex. In this paper we describe how the protein in this oligomeric complex interacts noncovalently with phenolic compounds of the proanthocyanidin type. These interactions are being disrupted during anion

  15. The standardized Ginkgo biloba extract Egb-761 protects vascular endothelium exposed to oxidized low density lipoproteins.

    Science.gov (United States)

    Pierre, S V; Lesnik, P; Moreau, M; Bonello, L; Droy-Lefaix, M-T; Sennoune, S; Duran, M-J; Pressley, T A; Sampol, J; Chapman, J; Maixent, J-M

    2008-10-26

    Dietary antioxidants are frequently proposed as protective agents for the vascular endothelium during the onset of atherosclerosis. This protection may occur at two distinct levels. First, they prevent oxidative modification of atherogenic lipoproteins (LDL). Second, they can provide a cellular protection against oxidized LDL-mediated endothelium dysfunction, although this mechanism remains poorly considered in many instances. To gain insight into the mechanism underlying such cellular protection against oxidized LDL, we examined the impact of a popular traditional medicine, an extract from Ginkgo biloba with well-known antioxidant properties, on two endothelial cells properties: cell adhesion and ionic homeostasis. Cellular lipoperoxides levels were also measured as a marker of cellular oxidative stress. Human umbilical-vein endothelial cells were exposed to native (nat-) or oxidized (ox-) LDL, the latter prepared to be compatible with clinically observed levels of oxidation. Although nat-LDL had little effect, ox-LDL increased endothelial adhesive properties (35%, pGinkgo biloba EGb-761 totally protected adhesive properties and endothelial lipoperoxide levels. Moreover, it limited the decrease in Na,K-ATPase activity induced by ox-LDL to levels similar to nat-LDL. This suggests that EGb-761 protects endothelial adhesive properties and helps prevent the disruption of ionic homeostasis. The EGb-761-mediated inhibition of ox-LDL-induced lipoperoxide levels in endothelial cells appears to be an important mechanism by which Ginkgo biloba extract protects endothelial properties.

  16. In vivo toxicity evaluation of a standardized extract of Syzygium aqueum leaf

    Directory of Open Access Journals (Sweden)

    Thamilvaani Manaharan

    2014-01-01

    Full Text Available In this study, the acute and subchronic toxicity effect of the Syzygium aqueum leaf extract (SA was evaluated. For the acute toxicity study, a single dose of 2000 mg/kg of the SA was given by oral-gavage to male Sprague-Dawley (SD rats. The rats were observed for mortality and toxicity signs for 14 days. In the subchronic toxicity study the SA was administered orally at doses of 50, 100, and 200 mg/kg per day for 28 days to male SD rats. The animals were sacrificed at the end of the experiment. The parameters measured including food and water intake, body weight, absolute and relative organ weight, blood biochemical tests and histopathology observation. In both the acute and subchronic toxicity studies, SA did not show any visible signs of toxicity. There were also no significant differences between the control and SA treated rats in terms of their food and water intake, body weight, absolute and relative organ weight, biochemical parameters or gross and microscopic appearance of the organs. There were no acute or subchronic toxicity observed and our results indicate that this extract could be devoid of any toxic risk. This is the first in vivo study reported the safety and toxicity of SA.

  17. In silico assessment of the potential allergenicity of transgenes used for the development of GM food crops.

    Science.gov (United States)

    Mishra, Ankita; Gaur, S N; Singh, B P; Arora, Naveen

    2012-05-01

    Genetically modified (GM) crops require allergenicity and toxicity assessment of the novel protein(s) to ensure complete safety to the consumers. These assessments are performed in accordance with the guidelines proposed by Codex (2003) and ICMR (2008). The guidelines recommend sequence homology analysis as a preliminary step towards allergenicity prediction, later in vitro experiments may be performed to confirm allergenicity. In the present study, an in silico approach is employed to evaluate the allergenic potential of six transgenes routinely used for the development of GM food crops. Among the genes studied, manganese superoxide dismutase (MnSOD) and osmotin shares greater than 90% identity with Hev b 10 and Cap a 1w, respectively. Chitinase shares greater than 70% identity with allergens namely Pers a 1 and Hev b 11, and fungal chitinase showed significant IgE binding with 7 of 75 patients' sera positive to different food extracts. Glucanases (alfalfa, wheat) and glycine betaine aldehyde dehydrogenase gene share 50% homology with allergens like - Ole e 9, Cla h 10 and Alt a 10. The results demonstrate the allergenic potential of six genes and can serve as a guide for selection of transgenes to develop GM crops. Copyright © 2012 Elsevier Ltd. All rights reserved.

  18. 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...

  19. Chloroatranol, an extremely potent allergen hidden in perfumes

    DEFF Research Database (Denmark)

    Johansen, J.D.; Andersen, Klaus Ejner; Svedman, C.

    2003-01-01

    Oak moss absolute is a long-known, popular natural extract widely used in perfumes. It is reported as the cause of allergic reactions in a significant number of those with perfume allergy. Oak moss absolute has been the target of recent research to identify its allergenic components. Recently...... and chloroatranol were included, together with a control group of 10 patients without sensitization to either of the 2 materials. A serial dilution patch test was performed on the upper back with concentrations ranging from 200 to 0.0063 p.p.m. of chloroatranol in ethanol. Simultaneously, the participant performed...

  20. Characterization of the soluble allergenic proteins of cashew nut (Anacardium occidentale L.).

    Science.gov (United States)

    Teuber, Suzanne S; Sathe, Shridhar K; Peterson, W Rich; Roux, Kenneth H

    2002-10-23

    The allergens associated with cashew food allergy have not been well-characterized. We sought to identify the major allergens in cashew nut by performing IgE immunoblots to dissociated and reduced or nonreduced cashew protein extracts, followed by sequencing of the peptides of interest. Sera from 15 subjects with life-threatening reactions to cashews and 8 subjects who tolerate cashews but have life-threatening reactions to other tree nuts were compared. An aqueous cashew protein extract containing albumin/globulin was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and subjected to IgE immunoblotting using patient sera. Selected IgE reactive bands were subjected to N-terminal amino acid sequencing. Each of the 15 sera from cashew-allergic subjects showed IgE binding to the cashew protein extract. The dominant IgE-binding antigens in the reduced preparations included peptides in the 31-35 kD range, consistent with the large subunits of the major storage 13S globulin (legumin-like protein). Low-molecular-weight polypeptides of the 2S albumin family, with similarity to the major walnut allergen Jug r 1, also bound IgE. The sera from eight patients who tolerate cashew but displayed allergies to other tree nuts showed only minimal or no IgE binding to cashew. Cashew food allergy is associated with the presence of IgE directed against the major seed storage proteins in cashew, including the 13S globulin (legumin group) and 2S albumins, both of which represent major allergen classes in several plant seeds. Thus, the legumin-group proteins and 2S albumins are again identified as major food allergens, which will help further research into seed protein allergenicity.

  1. The hammock: a reservoir of allergens

    Directory of Open Access Journals (Sweden)

    Francisca X. M. Rego

    2011-01-01

    Full Text Available INTRODUCTION: Asthma affects approximately 10% of the world's population. Sensitization to allergens is an important risk factor, and exposure to allergens is associated with disease severity. METHODS: We performed skin tests to evaluate allergen sensitization to mites, cockroaches, cats, dogs, and molds in 73 asthmatic patients. Enzyme Linked Immunosorbent Assay was used to assay the mite and cockroach allergens found in dust from the bedding, hammocks, bedroom floors, living rooms, and kitchens of 29 patients and 14 controls. RESULTS: Fifty patients (68.5% had positive skin test responses. There were positive responses to D. pteronyssinus (52.0%, B. tropicalis (53.4%, T. putrescentiae (15.0%, E. maynei (12.3%, L. destructor (8.2%, B. germanica (20.5%, P. americana (21.9%, Felis catus (10.9%, C. herbarium (2.7%, A. alternata (4.1%, and P. notatun (1.3%. The exposure to mite and cockroach allergens was similar in the patients and the controls. The Dermatophagoides pteronyssinus Group 1 levels were highest in the beds and hammocks. The Blattella germanica Group 1 levels were highest in the kitchens, living rooms and hammocks. DISCUSSION: The positive skin tests to mites, cockroaches and cats were consistent with previous studies. D pteronyssinus was the most prevalent home dust mite, and hammocks were a source of allergens. To improve asthma prophylaxis, it is important to determine its association with mite allergen exposure in hammocks.

  2. Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    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

  3. Characterisation of potential novel allergens in the fish parasite Anisakis simplex

    Directory of Open Access Journals (Sweden)

    Christiane Kruse Fæste

    2014-09-01

    Full Text Available The parasitic nematode Anisakis simplex occurs in fish stocks in temperate seas. A. simplex contamination of fish products is unsavoury and a health concern considering human infection with live larvae (anisakiasis and allergic reactions to anisakid proteins in seafood. Protein extracts of A. simplex produce complex band patterns in gel electrophoresis and IgE-immunostaining. In the present study potential allergens have been characterised using sera from A. simplex-sensitised patients and proteome data obtained by mass spectrometry. A. simplex proteins were homologous to allergens in other nematodes, insects, and shellfish indicating cross-reactivity. Characteristic marker peptides for relevant A. simplex proteins were described.

  4. Critical appraisal of Timothy grass pollen extract GRAZAX® in the management of allergic rhinitis

    Directory of Open Access Journals (Sweden)

    Scaparrotta A

    2015-11-01

    Full Text Available Alessandra Scaparrotta, Marina Attanasi, Marianna I Petrosino, Paola Di Filippo, Sabrina Di Pillo, Francesco Chiarelli Department of Pediatrics, University of Chieti, Chieti, Italy Abstract: Allergic rhinitis is one of the most common diseases of adult and pediatric age, associated with grass pollen (GP allergy in >50% cases, with a consistent impact on quality of life of affected patients. A grass allergen tablet, containing standardized extract derived from Timothy grass (Phleum pratense pollen and ~15 µg major allergen P. pratense (rPhl p 5, may be the future of allergen-specific immunotherapy (IT for GP allergy. The aim of this review was to critically evaluate the role of Timothy GP extract IT for the management of allergic rhinitis. For this purpose, we have tried to analyze potential mechanisms of action at the basis of Timothy GP extract, we have reviewed efficacy studies to establish potential benefits and clinical response, and we have also evaluated safety and tolerability profiles and patient focus perspective, such as quality of life, satisfaction and acceptability, and compliance to this IT. Keywords: Timothy grass pollen extract, allergic rhinitis, Grazax, efficacy, safety, compliance

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

    Directory of Open Access Journals (Sweden)

    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.

  6. Identification of two metallothioneins as novel inhalative coffee allergens cof a 2 and cof a 3.

    Science.gov (United States)

    Peters, Ulrike; Frenzel, Karsten; Brettschneider, Reinhold; Oldenburg, Marcus; Bittner, Cordula

    2015-01-01

    Dust of green coffee beans is known to be a relevant cause for occupational allergic disorders in coffee industry workers. Recently, we described the first coffee allergen (Cof a 1) establishing an allergenic potential of green coffee dust. Our aim was to identify allergenic components of green coffee in order to enhance inhalative coffee allergy diagnosis. A Coffea arabica pJuFo cDNA phage display library was created and screened for IgE binding with sera from allergic coffee workers. Two further coffee allergens were identified by sequence analysis, expressed in E. coli, and evaluated by Western blots. The prevalence of sensitization to recombinant Cof a 1, Cof a 2, and Cof a 3 and to commercially available extract was investigated by ELISA (enzyme-linked immunosorbent assay) respectively CAP (capacity test) screening in 18 sera of symptomatic coffee workers. In addition to the previously described chitinase Cof a 1, two Coffea arabica cysteine-rich metallothioneins of 9 and 7 kDa were identified and included in the IUIS Allergen Nomenclature as Cof a 2 and Cof a 3. Serum IgE antibodies to at least one of the recombinant allergens were found in 8 out of 18 symptomatic coffee workers (44%). Only 2 of the analysed sera (11%) had reacted previously to the commercial allergy test. In addition to the previously described Cof a 1 we have identified two further coffee proteins to be type I coffee allergens (Cof a 2 and Cof a 3) which may have a relevant potential for the specific diagnosis and/or therapy of coffee allergy.

  7. Identification of two metallothioneins as novel inhalative coffee allergens cof a 2 and cof a 3.

    Directory of Open Access Journals (Sweden)

    Ulrike Peters

    Full Text Available Dust of green coffee beans is known to be a relevant cause for occupational allergic disorders in coffee industry workers. Recently, we described the first coffee allergen (Cof a 1 establishing an allergenic potential of green coffee dust.Our aim was to identify allergenic components of green coffee in order to enhance inhalative coffee allergy diagnosis.A Coffea arabica pJuFo cDNA phage display library was created and screened for IgE binding with sera from allergic coffee workers. Two further coffee allergens were identified by sequence analysis, expressed in E. coli, and evaluated by Western blots. The prevalence of sensitization to recombinant Cof a 1, Cof a 2, and Cof a 3 and to commercially available extract was investigated by ELISA (enzyme-linked immunosorbent assay respectively CAP (capacity test screening in 18 sera of symptomatic coffee workers.In addition to the previously described chitinase Cof a 1, two Coffea arabica cysteine-rich metallothioneins of 9 and 7 kDa were identified and included in the IUIS Allergen Nomenclature as Cof a 2 and Cof a 3. Serum IgE antibodies to at least one of the recombinant allergens were found in 8 out of 18 symptomatic coffee workers (44%. Only 2 of the analysed sera (11% had reacted previously to the commercial allergy test.In addition to the previously described Cof a 1 we have identified two further coffee proteins to be type I coffee allergens (Cof a 2 and Cof a 3 which may have a relevant potential for the specific diagnosis and/or therapy of coffee allergy.

  8. Chemical characterization and pharmacological assessment of polysaccharide free, standardized cashew gum extract (Anacardium occidentale L.).

    Science.gov (United States)

    da Silva, Daiany Priscilla Bueno; Florentino, Iziara Ferreira; da Silva Moreira, Lorrane Kelle; Brito, Adriane Ferreira; Carvalho, Verônica Vale; Rodrigues, Marcella Ferreira; Vasconcelos, Géssica Adriana; Vaz, Boniek Gontijo; Pereira-Junior, Marcus Antônio; Fernandes, Kátia Flávia; Costa, Elson Alves

    2018-03-01

    The cashew gum (Anacardium occidentale L.) is used in traditional Brazilian medicine in the treatment of inflammatory conditions, asthma, diabetes, and gastrointestinal disturbances. In the present study, we aimed at forming a chemical characterization and investigation of the antinociceptive and anti-inflammatory activities of the aqueous extract of cashew gum without the presence of polysaccharides in its composition (CGE). The CGE was obtained after the precipitation and removal of polysaccharides through the use of acetone. After, the acetone was removed by rotaevaporation, and the concentrated extract was lyophilized. The chemical characterization of CGE was performed by liquid chromatography mass spectrometry (LC-MS) and tandem mass spectrometry (MS/MS) analyses. Mice were used for the evaluation of the antinociceptive and anti-inflammatory activities. CGE was analyzed via the Irwin test, acetic acid-induced writhing test, formalin-induced pain test, and carrageenan-induced paw edema test. The motor activity or probable sedation was verified through the chimney, open-field, and sodium pentobarbital-induced sleep tests. We investigated if the analgesic and anti-inflammatory effects of CGE depend of reduction in PGE 2 levels, were performed the carrageenan or PGE 2 -induced hyperalgesia tests. The chemical characterization of CGE showed the presence of anacardic acids as the predominant phytoconstituents. The treatment with CGE (75, 150, and 300mg/kg, p.o.) inhibited the number of writhing in a dose-dependent manner. With an intermediate dose, CGE did not cause motor impairment with the chimney test or alterations in either the open-field or sodium pentobarbital-induced sleep. In the formalin-induced pain test, CGE (150mg/kg, p.o.) produced an antinociceptive effect only in the first phase of the test, suggesting anti-inflammatory activity. With the same dosage, CGE also reduced the carrageenan-induced paw edema at all hours of the test, confirming its anti

  9. Standardized Bacopa monnieri extract ameliorates acute paraquat-induced oxidative stress, and neurotoxicity in prepubertal mice brain.

    Science.gov (United States)

    Hosamani, Ravikumar; Krishna, Gokul; Muralidhara

    2016-12-01

    Bacopa monnieri (BM), an ayurvedic medicinal plant, has attracted considerable interest owing to its diverse neuropharmacological properties. Epidemiological studies have shown significant correlation between paraquat (PQ) exposure and increased risk for Parkinson's disease in humans. In this study, we examined the propensity of standardized extract of BM to attenuate acute PQ-induced oxidative stress, mitochondrial dysfunctions, and neurotoxicity in the different brain regions of prepubertal mice. To test this hypothesis, prepubertal mice provided orally with standardized BM extract (200 mg/kg body weight/day for 4 weeks) were challenged with an acute dose (15 mg/kg body weight, intraperitoneally) of PQ after 3 hours of last dose of extract. Mice were sacrificed after 48 hours of PQ injection, and different brain regions were isolated and subjected to biochemical determinations/quantification of central monoamine (dopamine, DA) levels (by high-performance liquid chromatography). Oral supplementation of BM for 4 weeks resulted in significant reduction in the basal levels of oxidative markers such as reactive oxygen species (ROS), malondialdehyde (MDA), and hydroperoxides (HP) in various brain regions. PQ at the administered dose elicited marked oxidative stress within 48 hours in various brain regions of mice. However, BM prophylaxis significantly improved oxidative homeostasis by restoring PQ-induced ROS, MDA, and HP levels and also by attenuating mitochondrial dysfunction. Interestingly, BM supplementation restored the activities of cholinergic enzymes along with the restoration of striatal DA levels among the PQ-treated mice. Based on these findings, we infer that BM prophylaxis renders the brain resistant to PQ-mediated oxidative perturbations and thus may be better exploited as a preventive approach to protect against oxidative-mediated neuronal dysfunctions.

  10. 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.

  11. Subchronic Toxicity Study of Standardized Methanolic Extract of Mitragyna Speciosa Korth in Sprague-Dawley Rats

    Directory of Open Access Journals (Sweden)

    Mohd Ulul Ilmie Ahmad Nazri

    2015-06-01

    Full Text Available Mitragyna speciosa Korth, or better known as ketum, has long been used by traditional folk around Southeast Asia to prevent fatigue from working under hot tropical weather and as a replacement of opium, which can then cause addiction. To date, no findings have been reported of the toxic effect of ketum subchronically (28 days. Hence, the aim of this study was to investigate the toxicity of subchronic effect of standardised methanolic extract of ketum (SMEMS in Sprague-Dawley rats. Rats were orally administered with 100, 200, and 500 mg/kg of SMEMS for 28 days. Body weights were recorded daily. They were terminated at day 28 to obtain data for haematology, biochemistry, and histopathology of the brain, liver, kidney, lung, heart, sciatic nerve, and spinal cord. The SMEMS affected body weight compared to control group. Biochemistry findings showed that liver and kidney were affected with the abnormal values in AST, creatinine, globulin, glucose, total protein, and urea. However, SMEMS produced toxic effect more to liver, kidney, and lung than other organs as observed histopathologically. The results suggested subchronic exposure of ketum is toxic to the physiology of the animals.

  12. Enteric-coated, highly standardized cranberry extract reduces risk of UTIs and urinary symptoms during radiotherapy for prostate carcinoma

    Science.gov (United States)

    Bonetta, Alberto; Di Pierro, Francesco

    2012-01-01

    Background Cranberry (Vaccinium macrocarpon) proanthocyanidins can interfere with adhesion of bacteria to uroepithelial cells, potentially preventing lower urinary tract infections (LUTIs). Because LUTIs are a common side effect of external beam radiotherapy (EBRT) for prostate cancer, we evaluated the clinical efficacy of enteric-coated tablets containing highly standardized V. msacrocarpon (ecVM) in this condition. Methods A total of 370 consecutive patients were entered into this study. All patients received intensity-modulated radiotherapy for prostate cancer; 184 patients were also treated with ecVM while 186 served as controls. Cranberry extract therapy started on the simulation day, at which time a bladder catheterization was performed. During EBRT (over 6–7 weeks), all patients underwent weekly examination for urinary tract symptoms, including regular urine cultures during the treatment period. Results Compliance was excellent, with no adverse effects or allergic reactions being observed, apart from gastric pain in two patients. In the cranberry cohort (n = 184), 16 LUTIs (8.7%) were observed, while in the control group (n = 186) 45 LUTIs (24.2%) were recorded. This difference was statistically significant. Furthermore, lower rates of nocturia, urgency, micturition frequency, and dysuria were observed in the group that received cranberry extract. Conclusion Cranberry extracts have been reported to reduce the incidence of LUTIs significantly in women and children. Our data extend these results to patients with prostate cancer undergoing irradiation to the pelvis, who had a significant reduction in LUTIs compared with controls. These results were accompanied by a statistically significant reduction in urinary tract symptoms (dysuria, nocturia, urinary frequency, urgency), suggesting a generally protective effect of cranberry extract on the bladder mucosa. PMID:22977312

  13. Enteric-coated, highly standardized cranberry extract reduces risk of UTIs and urinary symptoms during radiotherapy for prostate carcinoma

    International Nuclear Information System (INIS)

    Bonetta, Alberto; Di Pierro, Francesco

    2012-01-01

    Cranberry (Vaccinium macrocarpon) proanthocyanidins can interfere with adhesion of bacteria to uroepithelial cells, potentially preventing lower urinary tract infections (LUTIs). Because LUTIs are a common side effect of external beam radiotherapy (EBRT) for prostate cancer, we evaluated the clinical efficacy of enteric-coated tablets containing highly standardized V. msacrocarpon (ecVM) in this condition. A total of 370 consecutive patients were entered into this study. All patients received intensity-modulated radiotherapy for prostate cancer; 184 patients were also treated with ecVM while 186 served as controls. Cranberry extract therapy started on the simulation day, at which time a bladder catheterization was performed. During EBRT (over 6–7 weeks), all patients underwent weekly examination for urinary tract symptoms, including regular urine cultures during the treatment period. Compliance was excellent, with no adverse effects or allergic reactions being observed, apart from gastric pain in two patients. In the cranberry cohort (n = 184), 16 LUTIs (8.7%) were observed, while in the control group (n = 186) 45 LUTIs (24.2%) were recorded. This difference was statistically significant. Furthermore, lower rates of nocturia, urgency, micturition frequency, and dysuria were observed in the group that received cranberry extract. Cranberry extracts have been reported to reduce the incidence of LUTIs significantly in women and children. Our data extend these results to patients with prostate cancer undergoing irradiation to the pelvis, who had a significant reduction in LUTIs compared with controls. These results were accompanied by a statistically significant reduction in urinary tract symptoms (dysuria, nocturia, urinary frequency, urgency), suggesting a generally protective effect of cranberry extract on the bladder mucosa

  14. Enteric-coated, highly standardized cranberry extract reduces risk of UTIs and urinary symptoms during radiotherapy for prostate carcinoma

    Directory of Open Access Journals (Sweden)

    Bonetta A

    2012-08-01

    Full Text Available Alberto Bonetta,1 Francesco Di Pierro21Unità Operativa Radioterapia Oncologica, Istituti Ospedalieri di Cremona, Cremona; 2Velleja Research, Milan, ItalyBackground: Cranberry (Vaccinium macrocarpon proanthocyanidins can interfere with adhesion of bacteria to uroepithelial cells, potentially preventing lower urinary tract infections (LUTIs. Because LUTIs are a common side effect of external beam radiotherapy (EBRT for prostate cancer, we evaluated the clinical efficacy of enteric-coated tablets containing highly standardized V. msacrocarpon (ecVM in this condition.Methods: A total of 370 consecutive patients were entered into this study. All patients received intensity-modulated radiotherapy for prostate cancer; 184 patients were also treated with ecVM while 186 served as controls. Cranberry extract therapy started on the simulation day, at which time a bladder catheterization was performed. During EBRT (over 6–7 weeks, all patients underwent weekly examination for urinary tract symptoms, including regular urine cultures during the treatment period.Results: Compliance was excellent, with no adverse effects or allergic reactions being observed, apart from gastric pain in two patients. In the cranberry cohort (n = 184, 16 LUTIs (8.7% were observed, while in the control group (n = 186 45 LUTIs (24.2% were recorded. This difference was statistically significant. Furthermore, lower rates of nocturia, urgency, micturition frequency, and dysuria were observed in the group that received cranberry extract.Conclusion: Cranberry extracts have been reported to reduce the incidence of LUTIs significantly in women and children. Our data extend these results to patients with prostate cancer undergoing irradiation to the pelvis, who had a significant reduction in LUTIs compared with controls. These results were accompanied by a statistically significant reduction in urinary tract symptoms (dysuria, nocturia, urinary frequency, urgency, suggesting a generally

  15. Prevalence of sensitization to Artemisia allergens Art v 1, Art v 3 and Art v 60 kDa. Cross-reactivity among Art v 3 and other relevant lipid-transfer protein allergens.

    Science.gov (United States)

    Lombardero, M; García-Sellés, F J; Polo, F; Jimeno, L; Chamorro, M J; García-Casado, G; Sánchez-Monge, R; Díaz-Perales, A; Salcedo, G; Barber, D

    2004-09-01

    Artemisia vulgaris is a widespread weed in the Mediterranean area and several allergens have been detected in its pollen. One of them, Art v 3, belongs to the lipid-transfer protein (LTP) family and its prevalence in Artemisia-sensitized patients or its relationship with other LTP allergens is not clear. To assess the pattern of sensitization to an array of mugwort allergens in a Mediterranean population, and to study the cross-reactivity of Art v 3 with Pru p 3 and Par j 1, relevant LTP allergens in the area. Skin prick test was performed with whole extracts (A. vulgaris, Parietaria judaica and peach) and pure natural allergens Art v 1, Art v 3, Art v 60 kDa and Par j 1 in 24 mugwort-allergic patients from a Mediterranean area. In vitro assays included measurement of specific IgE and ELISA inhibition among LTP allergens. The three Artemisia allergens elicited a positive skin response in 70-80% of the patients. Seven patients were clearly sensitized to Par j 1 and 11 to Pru p 3. There was no correlation between Par j 1 and Pru p 3 sensitization, but a highly significant correlation was found between peach extract and Art v 3 as regards the skin response. No IgE cross-reactivity was observed between Art v 3/Par j 1 or Pru p 3/Par j 1. In contrast, Art v 3 significantly inhibited the binding to Pru p 3 of IgE from three patients' sera out of six studied, but Pru p 3 was not able to inhibit the IgE binding to Art v 3. Art v 3 is a major mugwort allergen and in some patients with IgE to both Art v 3 and Pru p 3, Art v 3 behaves as the primary sensitizing agent.

  16. Examining database persistence of ISO/EN 13606 standardized electronic health record extracts: relational vs. NoSQL approaches.

    Science.gov (United States)

    Sánchez-de-Madariaga, Ricardo; Muñoz, Adolfo; Lozano-Rubí, Raimundo; Serrano-Balazote, Pablo; Castro, Antonio L; Moreno, Oscar; Pascual, Mario

    2017-08-18

    The objective of this research is to compare the relational and non-relational (NoSQL) database systems approaches in order to store, recover, query and persist standardized medical information in the form of ISO/EN 13606 normalized Electronic Health Record XML extracts, both in isolation and concurrently. NoSQL database systems have recently attracted much attention, but few studies in the literature address their direct comparison with relational databases when applied to build the persistence layer of a standardized medical information system. One relational and two NoSQL databases (one document-based and one native XML database) of three different sizes have been created in order to evaluate and compare the response times (algorithmic complexity) of six different complexity growing queries, which have been performed on them. Similar appropriate results available in the literature have also been considered. Relational and non-relational NoSQL database systems show almost linear algorithmic complexity query execution. However, they show very different linear slopes, the former being much steeper than the two latter. Document-based NoSQL databases perform better in concurrency than in isolation, and also better than relational databases in concurrency. Non-relational NoSQL databases seem to be more appropriate than standard relational SQL databases when database size is extremely high (secondary use, research applications). Document-based NoSQL databases perform in general better than native XML NoSQL databases. EHR extracts visualization and edition are also document-based tasks more appropriate to NoSQL database systems. However, the appropriate database solution much depends on each particular situation and specific problem.

  17. Skin Barrier Function and Allergens

    DEFF Research Database (Denmark)

    Engebretsen, Kristiane Aasen; Thyssen, Jacob Pontoppidan

    2016-01-01

    and skin barrier status. Psoriasis has traditionally been regarded a Th1-dominated disease, but the discovery of Th17 cells and IL-17 provides new and interesting information regarding the pathogenesis of the disease. Research suggests an inverse relationship between psoriasis and CA, possibly due......The skin is an important barrier protecting us from mechanical insults, microorganisms, chemicals and allergens, but, importantly, also reducing water loss. A common hallmark for many dermatoses is a compromised skin barrier function, and one could suspect an elevated risk of contact sensitization...... to increased levels of Th17 cells and its associated cytokines. As for AD, a positive association to CS has been established in epidemiological studies, but is still unresolved. Experimental studies show, however, an inverse relationship between AD and CS. The opposing and antagonistic influences of Th1 (CS...

  18. Bet v 1 homologues in strawberry identified as IgE-binding proteins and presumptive allergens.

    Science.gov (United States)

    Karlsson, A L; Alm, R; Ekstrand, B; Fjelkner-Modig, S; Schiött, A; Bengtsson, U; Björk, L; Hjernø, K; Roepstorff, P; Emanuelsson, C S

    2004-12-01

    No strawberry allergen has so far been identified and characterized. Serum samples were collected from patients with a suggestive case history of adverse reactions to strawberry and other fruits. Extracts from fresh and frozen strawberries were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and mass spectrometry. Patient blood samples were analysed for inhibition of IgE binding and basophil degranulation. Several IgE-binding proteins could be detected. In more than half of the patient sera, a 20/18-kDa doublet band was observed in Western blotting. These two bands were excised and analysed by mass spectrometry showing the presence of proteins belonging to the Bet v 1 family of allergens. Inhibition of the IgE binding to the 20/18-kDa doublet was obtained by addition of two recombinantly expressed allergens belonging to the Bet v 1 family (Bet v 1 and Mal d 1) and strawberry protein extract. In a cell-based assay of patient blood samples, basophil degranulation could be induced by strawberry protein extract and by Bet v 1 and Mal d 1. We conclude that strawberry homologues to Bet v 1 may be allergens of importance for adverse reactions to strawberry.

  19. Production and characterization of a monoclonal antibody against an Ascaris suum allergenic component

    Directory of Open Access Journals (Sweden)

    R.R. Pires

    2001-08-01

    Full Text Available Ascaris suum allergenic components (PIII separated by gel filtration chromatography of an adult worm extract were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells using polyethylene glycol (MW 1450 as fusogen. The hybridomas were cultured in HAT-containing medium and cloned at limiting dilutions. Supernatants from the growing hybrids were screened by ELISA using plates coated with PIII or the A. suum crude extract. The monoclonal antibody obtained, named MAC-3 (mouse anti-A. suum allergenic component, is an IgG1 kappa mouse immunoglobulin that specifically recognizes a 29,000 molecular weight protein (called allergenic protein with an affinity constant of 1.7 x 10(9 M-1. The A. suum components recognized by MAC-3 induce specific IgE antibody production in immunized BALB/c mice. Ascitic fluid induced in Swiss mice by injecting ip the hybridoma cells and incomplete Freund's adjuvant was purified by affinity chromatography using a protein A-Sepharose column. The purified monoclonal antibody was then coupled to activated Sepharose beads in order to isolate the A. suum allergenic component from the whole extract by affinity chromatography.

  20. Inactivation of allergens and toxins.

    Science.gov (United States)

    Morandini, Piero

    2010-11-30

    Plants are replete with thousands of proteins and small molecules, many of which are species-specific, poisonous or dangerous. Over time humans have learned to avoid dangerous plants or inactivate many toxic components in food plants, but there is still room for ameliorating food crops (and plants in general) in terms of their allergens and toxins content, especially in their edible parts. Inactivation at the genetic rather than physical or chemical level has many advantages and classical genetic approaches have resulted in significant reduction of toxin content. The capacity, offered by genetic engineering, of turning off (inactivating) specific genes has opened up the possibility of altering the plant content in a far more precise manner than previously available. Different levels of intervention (genes coding for toxins/allergens or for enzymes, transporters or regulators involved in their metabolism) are possible and there are several tools for inactivating genes, both direct (using chemical and physical mutagens, insertion of transposons and other genetic elements) and indirect (antisense RNA, RNA interference, microRNA, eventually leading to gene silencing). Each level/strategy has specific advantages and disadvantages (speed, costs, selectivity, stability, reversibility, frequency of desired genotype and regulatory regime). Paradigmatic examples from classical and transgenic approaches are discussed to emphasize the need to revise the present regulatory process. Reducing the content of natural toxins is a trade-off process: the lesser the content of natural toxins, the higher the susceptibility of a plant to pests and therefore the stronger the need to protect plants. As a consequence, more specific pesticides like Bt are needed to substitute for general pesticides. Copyright © 2010 Elsevier B.V. All rights reserved.

  1. Standardized Passiflora incarnata L. Extract Reverts the Analgesia Induced by Alcohol Withdrawal in Rats.

    Science.gov (United States)

    Schunck, Rebeca Vargas Antunes; Macedo, Isabel Cristina; Laste, Gabriela; de Souza, Andressa; Valle, Marina Tuerlinckx Costa; Salomón, Janaína L O; Nunes, Ellen Almeida; Campos, Andreia Cristina Wildner; Gnoatto, Simone Cristina Baggio; Bergold, Ana Maria; Konrath, Eduardo L; Dallegrave, Eliane; Arbo, Marcelo Dutra; Torres, Iraci L S; Leal, Mirna Bainy

    2017-08-01

    Passiflora incarnata L. (Passifloraceae) has been traditionally used for treatment of anxiety, insomnia, drug addiction, mild infections, and pain. The aim of this study was to investigate the effect of a commercial extract of P. incarnata in the analgesia induced by alcohol withdrawal syndrome in rats. In addition, brain-derived neurotrophic factor and interleukin-10 levels were evaluated in prefrontal cortex, brainstem, and hippocampus. Male adult rats received by oral gavage: (1: water group) water for 19 days, 1 day interval and water (8 days); (2: P. incarnata group) water for 19 days, 1 day interval and P. incarnata 200 mg/kg (8 days); (3: alcohol withdrawal group) alcohol for 19 days, 1 day interval and water (8 days); and (4: P. incarnata in alcohol withdrawal) alcohol for 19 days, 1 day interval and P. incarnata 200 mg/kg (8 days). The tail-flick and hot plate tests were used as nociceptive response measures. Confirming previous study of our group, it was showed that alcohol-treated groups presented an increase in the nociceptive thresholds after alcohol withdrawal, which was reverted by P. incarnata, measured by the hot plate test. Besides, alcohol treatment increased brain-derived neurotrophic factor and interleukin-10 levels in prefrontal cortex, which was not reverted by P. incarnata. Considering these results, the P. incarnata treatment might be a potential therapy in the alcohol withdrawal syndrome. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  2. Inactivation of dust mites, dust mite allergen, and mold from carpet.

    Science.gov (United States)

    Ong, Kee-Hean; Lewis, Roger D; Dixit, Anupma; MacDonald, Maureen; Yang, Mingan; Qian, Zhengmin

    2014-01-01

    Carpet is known to be a reservoir for biological contaminants, such as dust mites, dust mite allergen, and mold, if it is not kept clean. The accumulation of these contaminants in carpet might trigger allergies or asthma symptoms in both children and adults. The purpose of this study is to compare methods for removal of dust mites, dust mite allergens, and mold from carpet. Carpets were artificially worn to simulate 1 to 2 years of wear in a four-person household. The worn carpets were inoculated together with a common indoor mold (Cladosporium species) and house dust mites and incubated for 6 weeks to allow time for dust mite growth on the carpet. The carpets were randomly assigned to one of the four treatment groups. Available treatment regimens for controlling carpet contaminants were evaluated through a literature review and experimentation. Four moderately low-hazard, nondestructive methods were selected as treatments: vacuuming, steam-vapor, Neem oil (a natural tree extract), and benzalkonium chloride (a quaternary ammonium compound). Steam vapor treatment demonstrated the greatest dust mite population reduction (p mite allergens (p = 0.084), but have higher efficacy when compared to the chemical method on dust mite allergens (p = 0.002). There is no statistically significant difference in the efficacy for reducing mold in carpet (p > 0.05) for both physical and chemical methods. The steam-vapor treatment effectively killed dust mites and denatured dust mite allergen in the laboratory environment.

  3. Allergens and β-glucans in dutch homes and schools: characterizing airborne levels.

    Directory of Open Access Journals (Sweden)

    Esmeralda J M Krop

    Full Text Available BACKGROUND: Indoor air quality has an effect on respiratory health. Children are more vulnerable to a decreased indoor air quality as their lungs are still developing. We measured levels of allergens and β-(1,3-glucans in 19 school buildings and determined whether measured levels could be reproduced. School levels were compared to those in 169 homes and the effect of building characteristics on both home and school exposure was explored. METHODS: Electrostatic Dust fall Collectors were placed in school buildings for 8 weeks and in homes for 2 weeks to collect settled airborne dust. Cat, dog, and mouse allergen levels, domestic mite antigen levels and β-(1,3-glucans were measured in the extracts from the collectors. Results were corrected for sampling duration. Using questionnaire data, relations between measured levels and building and classroom characteristics were explored. RESULTS: In schools, exposure levels were highest in classrooms and were influenced by the socioeconomic status of the children, the season measurements were performed, moisture status of the building and pet ownership. Repeated measurements in different seasons and over the years showed significantly different levels. Home exposure was influenced by socioeconomic status, occupancy and pet ownership. Domestic mite antigen was found in higher levels in extracts from homes compared to schools while pet allergen levels were 13 times higher in schools compared to homes without pets. For mouse allergen overall levels of exposure were low but still two times higher in schools compared to homes. Levels of β-(1,3-glucans were also approximately two times higher in schools than in homes. CONCLUSION: Exposure levels of several allergens and β-(1,3-glucans in schools differ over time and are higher than in homes. For children, exposure levels measured at school could contribute to their total exposure as especially animal allergen levels can be much higher in schools compared to

  4. Allergen immunotherapy for allergic respiratory diseases

    Science.gov (United States)

    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

  5. Allergen management in the food industry

    National Research Council Canada - National Science Library

    Boye, Joyce I; Godefroy, Samuel Benrejeb

    2010-01-01

    "This book comprehensively addresses the sources of allergenic contaminants in foods, their fate during processing, and the specific measures that need to be taken to minimize their occurrence in foods...

  6. Associations between baseline allergens and polysensitization

    DEFF Research Database (Denmark)

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

    2008-01-01

    sulfate, epoxy resin, primin, and nickel sulfate showed statistically significant negative associations to polysensitization. For the allergens with the strongest associations, only every second individual with these particular allergies had two or more additional allergies. CONCLUSIONS: No common...

  7. Gastroprotective effect of standardized leaf extract from Argyreia speciosa on experimental gastric ulcers in rats.

    Science.gov (United States)

    Jaiswal, Sunil K; Rao, Chandana V; Sharma, Brijesh; Mishra, Pritee; Das, Sanjib; Dubey, Mukesh K

    2011-09-01

    Argyreia speciosa (L.f), Sweet (Family Convolvulaceae) is used traditionally in Indian System of Medicine as aphrodisiac, rejuvenating agent, intellect promoting agent, brain tonic and in the therapy of hepatomegaly, diabetes and chronic ulcer. To study the gastroprotective effect of standardized butanol fraction of Argyreia speciosa leaf (ASE). The butanol fraction of Argyreia speciosa leaf (ASE; 50, 100 and 200mg/kg body weight) was administered orally, twice daily for 5 days for prevention from Aspirin (ASP)-, ethanol (EtOH)-, cold-restraint stress (CRS) - and pylorus ligation (PL)-induced ulcers. Estimation of antioxidant enzymes activity was carried out in CRS-induced ulcer model, and various gastric secretion parameters like volume of gastric juice, acid output, and pH value were estimated in PL-induced ulcer model. ASE showed dose-dependent ulcer protective effect in ASP 23.64-58.76% (plevel (plevel (pcompared with CRS-induced group. A gradual and significant increase in CAT values were observed at 100 and 200mg/kg dose levels (pstudy revealed that Argyreia speciosa possess significant dose dependent gastroprotective activity, probably due to its free radical scavenging activity. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  8. Quantification of Pla or 3, aPlatanus orientalisAllergen, Grown under Different Environmental Conditions, by Sandwich ELISA.

    Science.gov (United States)

    Sedghy, Farnaz; Sankian, Mojtaba; Moghadam, Maliheh; Varasteh, Abdol-Reza

    2016-10-01

    Platanus species are widely cultured around the world and considered an important cause of allergic reactions. In the present study, we developed a sandwich ELISA to quantify Pla or 3 allergen in P. orientalis pollen extracts grown near high-traffic roads and compared it to pollen extracts collected from rural areas as control. Pollen samples were collected from three polluted and two unpolluted sites in Mashhad, northeast Iran. Recombinant Pla or 3 was expressed and used for polyclonal antibody production in rabbit. A sandwich ELISA was developed and validated to quantify Pla or 3 levels in pollen extracts from the different sites. The coefficients of variation (CVs) for the intra- and inter- day assays were less than 5 and 18%, respectively. The working range of the standard curve was between 0.1 and 25 ng/ml, with the detection limit being 0.037 ng/ml. The recovery percentage was 88-106.4% at working concentrations from 0.31 to 26.5 ng/ml. Pla or 3 levels were significantly greater in pollens grown near high-traffic roads than in those grown in rural regions (p Pla or 3 in pollen extracts. Using this validated ELISA, we showed a substantial difference between the amounts of Pla or 3 in pollens grown in different environments. This finding should be considered in developing public policies to reduce traffic pollution, which leads to reduced allergic reactions in atopic subjects.

  9. Soybean flour asthma: detection of allergens by immunoblotting

    International Nuclear Information System (INIS)

    Bush, R.K.; Schroeckenstein, D.; Meier-Davis, S.; Balmes, J.; Rempel, D.

    1988-01-01

    A 43-year-old woman developed asthma 6 years after beginning work in a food-processing plant in which soybean flour was used as a protein extender. Symptoms of sneezing, coughing, and wheezing would begin within minutes of exposure to soybean flour and resolve 2 hours after exposure ceased. Skin tests were positive to a soy extract prepared from the flour. Airway hyperreactivity was confirmed by a positive bronchial challenge to methacholine. Bronchial challenge with soybean flour produced an immediate increase in specific airway resistance from 5.0 to 22.7 L. cm of H2O/L/sec. There was no response to challenge with lactose. The patient's allergic response to soy-flour extract was further characterized by several immunologic methods. IgE binding to soy-flour protein by direct RAST was 5.98 times that of a normal control serum. The soy-flour extract was separated by dodecyl sulfate-polyacrylamide gel electrophoresis. Twenty-four protein bands were detected in the crude soy-flour extract. After immunoblotting and subsequent autoradiography, nine proteins with molecular weights ranging from 54,500 to 14,875 were found. Cross-reactivity studies with other legumes demonstrated apparent immunologic identity between a component in green pea extract and a soybean protein with a molecular weight of 17,000. The clinical significance of this cross-reactivity is not known. We conclude that in this case of occupational asthma to soybean flour, multiple allergens were involved. Immunoblotting may be useful in identifying the allergens involved in occupational asthma

  10. Allergen homologs in the Euroglyphus maynei draft genome.

    Directory of Open Access Journals (Sweden)

    S Dean Rider

    Full Text Available Euroglyphus maynei is a house dust mite commonly found in homes worldwide and is the source of allergens that sensitize and induce allergic reactions in humans. It is the source of species-specific allergens as well as allergens that are cross-reactive with the allergens from house dust mites Dermatophagoides farinae and D. pteronyssinus, and the ectoparasitic scabies mite Sarcoptes scabiei. The genomics, proteomics and molecular biology of E. maynei and its allergens have not been as extensively investigated as those of D. farinae, D. pteronyssinus, and S. scabiei where natural and recombinant allergens from these species have been characterized. Until now, little was known about the genome of E. maynei and it allergens but this information will be important for producing recombinant allergens for diagnostic and therapeutic purposes and for understanding the allergic response mechanism by immune effector cells that mediate the allergic reaction. We sequenced and assembled the 59 Mb E. maynei genome to aid the identification of homologs for known allergenic proteins. The predicted proteome shared orthologs with D. farinae and S. scabiei, and included proteins with homology to more than 30 different groups of allergens. However, the majority of allergen candidates could not be assigned as clear orthologs to known mite allergens. The genomic sequence data, predicted proteome, and allergen homologs identified from E. maynei provide insight into the relationships among astigmatid mites and their allergens, which should allow for the development of improved diagnostics and immunotherapy.

  11. Psychomotor performance in relation to acute oral administration of Delta9-tetrahydrocannabinol and standardized cannabis extract in healthy human subjects.

    Science.gov (United States)

    Roser, Patrik; Gallinat, Jürgen; Weinberg, Gordon; Juckel, Georg; Gorynia, Inge; Stadelmann, Andreas M

    2009-08-01

    Abnormalities in psychomotor performance are a consistent finding in schizophrenic patients as well as in chronic cannabis users. The high levels of central cannabinoid (CB(1)) receptors in the basal ganglia, the cerebral cortex and the cerebellum indicate their implication in the regulation of motor activity. Based on the close relationship between cannabis use, the endogenous cannabinoid system and motor disturbances found in schizophrenia, we expected that administration of cannabinoids may change pattern of psychomotor activity like in schizophrenic patients. This prospective, double-blind, placebo-controlled cross-over study investigated the acute effects of cannabinoids on psychomotor performance in 24 healthy right-handed volunteers (age 27.9 +/- 2.9 years, 12 male) by comparing Delta(9)-tetrahydrocannabinol (Delta(9)-THC) and standardized cannabis extract containing Delta(9)-THC and cannabidiol. Psychomotor performance was assessed by using a finger tapping test series. Cannabis extract, but not Delta(9)-THC, revealed a significant reduction of right-hand tapping frequencies that was also found in schizophrenia. As to the pure Delta(9)-THC condition, left-hand tapping frequencies were correlated with the plasma concentrations of the Delta(9)-THC metabolite 11-OH-THC. These effects are thought to be related to cannabinoid actions on CB(1) receptors in the basal ganglia, the cerebral cortex and the cerebellum. Our data further demonstrate that acute CB(1) receptor activation under the cannabis extract condition may also affect intermanual coordination (IMC) as an index of interhemispheric transfer. AIR-Scale scores as a measure of subjective perception of intoxication were dose-dependently related to IMC which was shown by an inverted U-curve. This result may be due to functional changes involving GABAergic and glutamatergic neurotransmission within the corpus callosum.

  12. Protective effect of Convolvulus pluricaulis standardized extract and its fractions against 3-nitropropionic acid-induced neurotoxicity in rats.

    Science.gov (United States)

    Malik, Jai; Choudhary, Sunayna; Kumar, Puneet

    2015-01-01

    Convolvulus pluricaulis Chois. (Convolvulaceae), a well-known Ayurvedic "Medhya Rasayana" (nervine tonic), is extensively used for different central nervous system (CNS) disorders. The objective of this study was to evaluate the protective effect of standardized hydro-methanol extract of C. pluricaulis (CPE) and its fractions, namely ethyl acetate (EAE), butanol (BE), and aqueous (AE), against 3-nitropropionic acid (3-NP) induced neurotoxicity in rats. The extract of the whole plant was standardized on the basis of scopoletin content (0.014%) using thin layer chromatography densitometric analysis. CPE (100 and 200 mg/kg) and its fractions, namely EAE (15 and 30 mg/kg), BE (25 and 50 mg/kg), and AE (50 and 100 mg/kg) were administered orally for 20 d. Their protective effect against 3-NP (10 mg/kg, i.p. for 14 d) was assessed by the effect on various behavioral parameters, namely body weight, locomotor activity, grip strength, gait pattern, and the effect on cognitive dysfunction. Biochemical parameters for oxidative damage were also assessed in the striatum and cortex regions of the brain. Administration of 3-NP induced HD-like symptoms that were indicated by reduced body weight, locomotor activity, memory, grip strength, and oxidative defense. CPE (200 mg/kg), EAE (30 mg/kg), and BE (50 mg/kg) significantly (p < 0.001) attenuated 3-NP induced reduction in locomotor activity, grip strength, memory, body weight, and oxidative defense in comparison with 3-NP-treated animals on 10 and 15 d. The present study suggested that CPE has a protective action against 3-NP-induced neurotoxicity and can be further explored for its efficacy against Huntington's disease.

  13. Immunological comparison of allergen immunotherapy tablet treatment and subcutaneous immunotherapy against grass allergy

    DEFF Research Database (Denmark)

    Aasbjerg, K; Backer, V; Lund, G

    2014-01-01

    BACKGROUND: IgE-mediated allergic rhinitis to grass pollen can successfully be treated with either allergen immunotherapy tablets (SLIT tablet) or SQ-standardized subcutaneous immunotherapy (SCIT). The efficacy of these two treatment modalities for grass allergy is comparable, but the immunological...

  14. Highly Sensitive Matrix-Independent Quantification of Major Food Allergens Peanut and Soy by Competitive Real-Time PCR Targeting Mitochondrial DNA.

    Science.gov (United States)

    Ladenburger, Eva-Maria; Dehmer, Markus; Grünberg, Ruben; Waiblinger, Hans-Ulrich; Stoll, Dieter; Bergemann, Jörg

    2018-01-01

    The development of two competitive real-time PCR assays for the quantitative detection of trace amounts of two major food allergens, peanut and soybean, is reported. In order to achieve very low detection levels for both allergens, we established PCR primers and probes targeting mitochondrial DNA sequences. We were able to demonstrate that this approach led to an increase in detection sensitivity in the range of at least 1 order of magnitude compared with published assays targeting nuclear DNA. Furthermore, we generated corresponding competitor molecules, which were used as internal standards to compete with matrix effects that are evident during DNA extraction and PCR amplification in heterogeneous analytical matrixes like food. According to the recently described competitive quantitative PCR method published by Holzhauser et al. (2014), we performed threshold calibration against milk powder spiked with 10 ppm peanut and soy. Matrix-independent quantitative determination of peanut and soy could be demonstrated for three different calibrated food matrix standards in a range between 1 and 100 ppm. The data presented indicate that both assay concepts are powerful analytical tools for the quantitative detection of trace amounts of peanut and soy in commercial food products.

  15. Comparison of allergen responses based on the TRUE Test and IQ Chamber system in Korean patients.

    Science.gov (United States)

    Kwon, Won Joo; Ko, Joo Yeon; Ro, Young Suck; Kim, Myung Hwa; Kim, Byung Soo; Kim, Hye One; Nam, Jae Hui; Lee, Ga-Young; Park, Mi Youn; Son, Sang Wook; Yang, Jun Mo; Lee, Dong Hun; Lee, Ai Young; Lee, Jun Young; Chang, Sung Eun; Choi, Hae Young; Park, Eun Joo

    2017-12-01

    A comparison between responses to allergens based on the TRUE Test ® (TT) and IQ ® Chamber (IQC) in Europeans has been previously reported, however, no such study has been performed in Asians. To compare allergen responses using the TT and IQC (using the Korean standard series) in order to gather more information regarding the positive response rates for each allergen and the clinical value of IQC. Suspected contact dermatitis patients were enrolled and tested with 18 allergens using the TT and IQC. The test was performed in 214 patients. Simultaneous positive results for both tests were recorded in 242 cases (positive concordance rate: 66.7%). IQC yielded more positive results. Allergens with a high positive concordance rate were nickel sulphate (82.1%), thimerosal (78.6%), and p-phenylenediamine (73.3%). IQC mostly showed similar or higher positive rates than TT, with high concordance. We recommend the IQC method as a screening test in patients with suspected contact dermatitis. To compensate for possible false positive results, careful history taking and a different patch test should be performed when appropriate.

  16. 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.

  17. Accurate quantification of creatinine in serum by coupling a measurement standard to extractive electrospray ionization mass spectrometry

    Science.gov (United States)

    Huang, Keke; Li, Ming; Li, Hongmei; Li, Mengwan; Jiang, You; Fang, Xiang

    2016-01-01

    Ambient ionization (AI) techniques have been widely used in chemistry, medicine, material science, environmental science, forensic science. AI takes advantage of direct desorption/ionization of chemicals in raw samples under ambient environmental conditions with minimal or no sample preparation. However, its quantitative accuracy is restricted by matrix effects during the ionization process. To improve the quantitative accuracy of AI, a matrix reference material, which is a particular form of measurement standard, was coupled to an AI technique in this study. Consequently the analyte concentration in a complex matrix can be easily quantified with high accuracy. As a demonstration, this novel method was applied for the accurate quantification of creatinine in serum by using extractive electrospray ionization (EESI) mass spectrometry. Over the concentration range investigated (0.166 ~ 1.617 μg/mL), a calibration curve was obtained with a satisfactory linearity (R2 = 0.994), and acceptable relative standard deviations (RSD) of 4.6 ~ 8.0% (n = 6). Finally, the creatinine concentration value of a serum sample was determined to be 36.18 ± 1.08 μg/mL, which is in excellent agreement with the certified value of 35.16 ± 0.39 μg/mL.

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

    DEFF Research Database (Denmark)

    Rastogi, Suresh Chandra; Johansen, J D; Menné, T

    1999-01-01

    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...... 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...... 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....

  19. Sensitization to green coffee bean (GCB) and castor bean (CB) allergens among dock workers.

    Science.gov (United States)

    De Zotti, R; Patussi, V; Fiorito, A; Larese, F

    1988-01-01

    Dock workers (n = 218) occupationally exposed to green coffee beans (GCB) were studied, using a specific questionnaire for allergic symptoms and skin tests for common and occupational allergens. Thirty-one workers (14.3%) complained of allergic symptoms of the eye, nose and bronchial system at the workplace. The prick tests, using both commercial allergens and specific extracts prepared from the most common types of coffee and their corresponding sacks, confirmed a sensitization in 21 workers (9.6%). A positive skin reaction to castor beans (CB) was found in nearly all these cases; in ten workers there was also a positive reaction to GCB allergens and in 14 cases prick tests were positive to extracts of sacks. There was a good concordance between prick tests and specific IgE for CB (95.0%) and also, but to a lesser extent, for GCB. The authors concluded that there is a significant risk of sensitization to CB and GCB allergens in dock workers occupied in handling green coffee bean, despite the fact that the exposure is not continuous. CB emerged as a common contaminant of GCB from various countries. For effective prevention, a modification of the methods of transport is required to avoid CB contamination to other products. Eliminating environmental dust during shipping operations is the most important preventive measure and it can be achieved by the use of containers, as some exporting countries are already doing.

  20. Pru p 3, a marker allergen for lipid transfer protein sensitization also in Central Europe.

    Science.gov (United States)

    Mothes-Luksch, N; Raith, M; Stingl, G; Focke-Tejkl, M; Razzazi-Fazeli, E; Zieglmayer, R; Wöhrl, S; Swoboda, I

    2017-09-01

    In the Mediterranean area, lipid transfer proteins (LTPs) are important causes of plant-food allergies often associated with severe allergic reactions. There, peach LTP (Pru p 3) seems to be the primary sensitizer, whereas in Central Europe, little is known about the importance of LTP sensitization. In this region, allergen extract-based diagnosis is often complicated by co-sensitization to Bet v 1, the major birch pollen allergen, its cross-reactive food allergens, and profilins. We investigated the role of LTP sensitization in Central European patients displaying strong allergic reactions to plant-derived food. Analysis of IgE reactivity revealed that ten of thirteen patients were sensitized to Pru p 3, nine to Bet v 1, and two to profilin. Our results showed that LTP sensitization represents a risk factor for severe allergic symptoms in Central Europe. Furthermore, the strong IgE reactivity detected in immunoblots of plant-food extracts indicated that Pru p 3 can be used as a marker allergen for LTP sensitization also in Central European patients. © 2017 The Authors Allergy Published by John Wiley & Sons Ltd.

  1. Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds

    International Nuclear Information System (INIS)

    Fernandes, K.V.; Deus-de-Oliveira, N.; Godoy, M.G.; Guimarães, Z.A.S.; Nascimento, V.V.; Melo, E.J.T. de; Freire, D.M.G.; Dansa-Petretski, M.; Machado, O.L.T.

    2012-01-01

    Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 × 10 5 cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH) 2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained

  2. Allergic respiratory disease: Different allergens, different symptoms.

    Science.gov (United States)

    Valero, A; Quirce, S; Dávila, I; Delgado, J; Domínguez-Ortega, J

    2017-09-01

    Spanish population is rather homogeneous in its genetic and sociocultural characteristics, but allergen sensitization shows wide geographical variations. We aimed at assessing whether sensitization to different allergens in the diverse geographical areas induced different clinical and quality-of-life characteristics in adult patients with a first-time diagnosis of rhinitis and/or asthma. Two sequential, identically designed studies were carried out to evaluate such associations (PERFILAR I and II). PERFILAR II was an extension of PERFILAR I with additional allergens being included. Both phases were epidemiological, descriptive, cross-sectional, nonintervention multicenter studies. Participants were required to have lived for at least the last 2 years in the geographical zone. Asthma control assessment was based on ACQ-5. Health-related quality of life was evaluated with validated scales for rhinitis (ESPRINT-15) and asthma (Mini-AQLQ). Skin prick tests were used to identify sensitization to involved allergens. A total of 301 physicians recruited 2711 patients for PERFILAR I+II. Sensitization to allergens was significantly different in patients with rhinitis with/without asthma. Seasonal allergens were associated with rhinitis, a longer time to asthma development, and more severe and commonly intermittent rhinitis. HDM were associated with more common rhinitis, and Alternaria was associated with asthma. The study confirms an association of geographical areas with relevant allergens and allergic clinical picture. Different types of aeroallergens and specific sensitization profiles are associated with different allergic clinical pictures (rhinitis with/without asthma), different clinical symptoms, and different levels of severity. This could have implications to predict later clinical course and to select appropriate management approaches. © 2017 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.

  3. Allergenic Proteins in Foods and Beverages

    Directory of Open Access Journals (Sweden)

    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.

  4. Simultaneous Determination of the Main Peanut Allergens in Foods Using Disposable Amperometric Magnetic Beads-Based Immunosensing Platforms

    Directory of Open Access Journals (Sweden)

    Víctor Ruiz-Valdepeñas Montiel

    2016-06-01

    Full Text Available In this work, a novel magnetic beads (MBs-based immunosensing approach for the rapid and simultaneous determination of the main peanut allergenic proteins (Ara h 1 and Ara h 2 is reported. It involves the use of sandwich-type immunoassays using selective capture and detector antibodies and carboxylic acid-modified magnetic beads (HOOC-MBs. Amperometric detection at −0.20 V was performed using dual screen-printed carbon electrodes (SPdCEs and the H2O2/hydroquinone (HQ system. This methodology exhibits high sensitivity and selectivity for the target proteins providing detection limits of 18.0 and 0.07 ng/mL for Ara h 1 and Ara h 2, respectively, with an assay time of only 2 h. The usefulness of the approach was evaluated by detecting the endogenous content of both allergenic proteins in different food extracts as well as trace amounts of peanut allergen (0.0001% or 1.0 mg/kg in wheat flour spiked samples. The developed platform provides better Low detection limits (LODs in shorter assay times than those claimed for the allergen specific commercial ELISA kits using the same immunoreagents and quantitative information on individual food allergen levels. Moreover, the flexibility of the methodology makes it readily translate to the detection of other food-allergens.

  5. Potential Allergens in Disposable Diaper Wipes, Topical Diaper Preparations, and Disposable Diapers: Under-recognized Etiology of Pediatric Perineal Dermatitis.

    Science.gov (United States)

    Yu, JiaDe; Treat, James; Chaney, Keri; Brod, Bruce

    2016-01-01

    Allergic contact dermatitis in young children may be an under-recognized cause of perineal dermatitis. The diapered infant skin is uniquely susceptible to allergic contact dermatitis because of more permeable neonatal skin, a moist environment, frequent contact with irritants and resultant skin barrier breakdown, and exposure to topical products such as diaper wipes, diaper preparations, and disposable diapers. To our knowledge, potential allergens in these products have not been thoroughly catalogued or studied. We explore and review potential allergenic ingredients in diaper wipes, topical diaper preparations, and disposable diapers. We analyzed 63 diaper wipes, 41 topical diaper preparations, and the 3 top selling diaper brands available from two of the largest retailers in the United States. Each potential allergen is discussed, and epidemiologic studies of rates of sensitization to potential allergens in children are also reported. Botanical extracts, including members of the Compositae family, were the most commonly represented potential allergen in both diaper wipes and topical preparations. Other potential allergens identified with high frequency include α-tocopherol, fragrances, propylene glycol, parabens, iodopropynyl butylcarbamate, and lanolin. Frequent culprits such as formaldehyde releasers and methylchloroisothiazolinone/methylisothiazolinone were not prevalent in our analyzed products.

  6. 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.

  7. 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.

  8. House-dust mite and mite-and pet-allergens in indoor environment; Dani oyobi dani-allergen, pet-allergen no jittai

    Energy Technology Data Exchange (ETDEWEB)

    Irie, T. [Shinshu Univ., Matsumoto, Nagano (Japan)

    1998-05-31

    In this paper, the actual state of mite and mite-allergen, and the actual state of pet-allergen that the researches thereof proceed gradually in recent years are described. The following findings are obtained as the results of the actual state investigations. In respect to the mite-allergen, airborne mite, particularly I with high allergen activity shows high concentration from fall to winter. The reasons thereof are considered from two aspects, I.e., the fragmentation and suspension of mite-allergen, and the decrease of ventilation frequency, while it can also be explained from a fact that the fit of infantile asthma is most frequent during a period from October to November. Mite-allergen contained in futon dust is about 10 times more than that contained in floor dust. In respect to pet-allergen, although the indoor accumulative amount and airborne concentration of main pet-allergen such as cat-, and dog-allergen varies greatly depending on whether pet is kept indoor or outdoor, the quantities of dog- and cat-allergen are almost of the same order, but 2 to 3-order higher than the quantity of mite-allergen. 9 refs., 9 figs.

  9. Carrier-bound Alt a 1 peptides without allergenic activity for vaccination against Alternaria alternata allergy

    Science.gov (United States)

    Twaroch, T. E.; Focke, M.; Fleischmann, K.; Balic, N.; Lupinek, C.; Blatt, K.; Ferrara, R.; Mari, A.; Ebner, C.; Valent, P.; Spitzauer, S.; Swoboda, I.; Valenta, R.

    2017-01-01

    Background The mould Alternaria alternata is a major elicitor of allergic asthma. Diagnosis and specific immunotherapy (SIT) of Alternaria allergy are often limited by the insufficient quality of natural mould extracts. Objective To investigate whether recombinant Alt a 1 can be used for reliable diagnosis of Alternaria alternata allergy and to develop a safe, non-allergenic vaccine for SIT of Alternaria allergy. Methods The qualitative sensitization profile of 80 Alternaria-allergic patients from Austria and Italy was investigated using an allergen micro-array and the amount of Alternaria-specific IgE directed to rAlt a 1 was quantified by ImmunoCAP measurements. Peptides spanning regions of predicted high surface accessibility of Alt a 1 were synthesized and tested for IgE reactivity and allergenic activity, using sera and basophils from allergic patients. Carrier-bound peptides were studied for their ability to induce IgG antibodies in rabbits which recognize Alt a 1 and inhibit allergic patients’ IgE reactivity to Alt a 1. Results rAlt a 1 allowed diagnosis of Alternaria allergy in all tested patients, bound the vast majority (i.e. >95%) of Alternaria-specific IgE and elicited basophil activation already at a concentration of 0.1 ng/mL. Four non-allergenic peptides were synthesized which, after coupling to the carrier protein keyhole limpet hemocyanin, induced Alt a 1-specific IgG and inhibited allergic patients’ IgE binding to Alt a 1. Conclusions and clinical relevance rAlt a 1 is a highly allergenic molecule allowing sensitive diagnosis of Alternaria allergy. Carrier-bound non-allergenic Alt a 1 peptides are candidates for safe SIT of Alternaria allergy. PMID:22909168

  10. Development of Nordic Standard for analysis of oil and fat in water based on supercritical fluid extraction. Preliminary study, part 2

    International Nuclear Information System (INIS)

    Jenssen, L.

    1994-06-01

    This report describes a preliminary study of a method of determining oil in water. The method is based on solid phase extraction and supercritical fluid extraction (SPE-SFE). The oil is extracted from the water by absorption to extraction disks from which it is then desorbed by supercritical carbon dioxide and detected by means of infrared spectrophotometry or gas chromatography. The results of the study will indicate if the method is suitable as a future substitute for the present Norwegian Standard, NS 9803 (Swedish Standard, SS 02 8145). The method has been validated using water samples with addition of real oil to 1-100 ppm. The accuracy is almost 70%, and the method has good repeatability and is linear in the 1-100 ppm range. 5 refs., 6 figs., 10 tabs

  11. ROLE OF ENVIRONMENTAL ALLERGENS ON ATOPIC DERMATITIS

    Directory of Open Access Journals (Sweden)

    M. Wardhana

    2014-01-01

    Full Text Available Background: Atopic dermatitis (AD is a chronic eczematous skin disease that develops in a patient with atopic diathesis, which is characterized by an increased liability to produce IgE antibodies for allergens mostly derived from environmental or inhalant allergens and food allergens. They are produced by cell-mediated allergic contact reactions, and recently contact sensitivity to various environmental allergens has been demonstrated in patients with AD. Atopic patients are recognized by their ability to produce large amounts of specific IgE antibodies to common substances as environmental allergens, i.e. house dust mites, grass pollens, animal danders, molds, food, etc. These antibodies can be detected by skin prick test. The aim of this study was to identify the sensitization against environmental or inhalants allergens through skin prick tests in the patients with atopic dermatitis. Material and Methods: This is a retrospective, descriptive study. We revised all medical records of patients with AD since January 2002 to December 2004 in the Out Patients Unit of Sanglah General Hospital, Bali-Indonesia. The variables studied were: gender, age, work related, diagnosis associates to AD, and prick test of environmental allergens. Results: In 3 years periods we had revised 46 of patients with AD that was done skin prick tests. The median age was 38 years (range 29-54 years, 34/46 (73.9 % of these were male and 12 (26.1 % female. Twenty nine patients presented pure AD, and 17 patients had AD with asthma and allergic rhinitis. Only 16 (34.7% of patients had no history of allergic disease. Thirsty six of 46 (78.20% of all tested AD patients had a positive skin prick tests against inhalant (aeroallergens 16 patients and food allergens 21 patients. Sixteen patients with positive of skin test include; dust mite in 12 patients, animal dander in 10 patients, grass pollen in 9 patients and cockroach in 6 patients. Conclusion: We concluded that

  12. Involvement of monoaminergic systems in anxiolytic and antidepressive activities of the standardized extract of Cocos nucifera L.

    Science.gov (United States)

    Lima, Eliane Brito Cortez; de Sousa, Caren Nádia Soares; Meneses, Lucas Nascimento; E Silva Pereira, Yuri Freitas; Matos, Natália Castelo Branco; de Freitas, Rayanne Brito; Lima, Nycole Brito Cortez; Patrocínio, Manoel Cláudio Azevedo; Leal, Luzia Kalyne Almeida Moreira; Viana, Glauce Socorro Barros; Vasconcelos, Silvânia Maria Mendes

    2017-01-01

    Extracts from the husk fiber of Cocos nucifera are used in folk medicine, but their actions on the central nervous system have not been studied. Here, the anxiolytic and antidepressant effects of the standardized hydroalcoholic extract of C. nucifera husk fiber (HECN) were evaluated. Male Swiss mice were treated with HECN (50, 100, or 200 mg/kg) 60 min before experiments involving the plus maze test, hole-board test, tail suspension test, and forced swimming test (FST). HECN was administered orally (p.o.) in acute and repeated-dose treatments. The forced swimming test was performed with dopaminergic and noradrenergic antagonists, as well as a serotonin release inhibitor. Administration of HECN in the FST after intraperitoneal (i.p.) pretreatment of mice with sulpiride (50 mg/kg), prazosin (1 mg/kg), or p-chlorophenylalanine (PCPA, 100 mg/kg) caused the actions of these three agents to be reversed. However, this effect was not observed after pretreating the animals with SCH23390 (15 µg/kg, i.p.) or yohimbine (1 mg/kg, i.p.) The dose chosen for HECN was 100 mg/kg, p.o., which increased the number of entries as well as the permanence in the open arms of the maze after acute and repeated doses. In both the forced swimming and the tail suspension tests, the same dose decreased the time spent immobile but did not disturb locomotor activity in an open-field test. The anxiolytic effect of HECN appears to be related to the GABAergic system, while its antidepressant effect depends upon its interaction with the serotoninergic, noradrenergic (α1 receptors), and dopaminergic (D2 dopamine receptors) systems.

  13. Pollensomes as Natural Vehicles for Pollen Allergens.

    Science.gov (United States)

    Prado, Noela; De Linares, Concepción; Sanz, María L; Gamboa, Pedro; Villalba, Mayte; Rodríguez, Rosalía; Batanero, Eva

    2015-07-15

    Olive (Olea europaea) pollen constitutes one of the most important allergen sources in the Mediterranean countries and some areas of the United States, South Africa, and Australia. Recently, we provided evidence that olive pollen releases nanovesicles of respirable size, named generically pollensomes, during in vitro germination. Olive pollensomes contain allergens, such as Ole e 1, Ole e 11, and Ole e 12, suggesting a possible role in allergy. The aim of this study was to assess the contribution of pollensomes to the allergic reaction. We show that pollensomes exhibit allergenic activity in terms of patients' IgE-binding capacity, human basophil activation, and positive skin reaction in sensitized patients. Furthermore, allergen-containing pollensomes have been isolated from three clinically relevant nonphylogenetically related species: birch (Betula verrucosa), pine (Pinus sylvestris), and ryegrass (Lolium perenne). Most interesting, pollensomes were isolated from aerobiological samples collected with an eight-stage cascade impactor collector, indicating that pollensomes secretion is a naturally occurring phenomenon. Our findings indicate that pollensomes may represent widespread vehicles for pollen allergens, with potential implications in the allergic reaction. Copyright © 2015 by The American Association of Immunologists, Inc.

  14. 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).

  15. Domestic cat allergen and allergic sensitisation in young children

    NARCIS (Netherlands)

    Chen, Chih-Mei; Gehring, Ulrike; Wickman, Magnus; Hoek, Gerard; Giovannangelo, Mariella; Nordling, Emma; Wijga, Alet; de Jongste, Johan; Pershagen, Goeran; Almqvist, Catarina; Kerkhof, Marjan; Bellander, Tom; Wichmann, H. -Erich; Brunekreef, Bert; Heinrich, Joachim

    Studies have presented conflicting associations between cat allergen exposure and sensitisation and atopic disease. We therefore investigated the association between the observed domestic cat allergen level and cat sensitisation in young children in four study populations from three European

  16. Threshold dose distributions for 5 major allergenic foods in children

    NARCIS (Netherlands)

    Blom, W.M.; Vlieg-Boerstra, B.J.; Kruizinga, A.G.; Heide, S. van der; Houben, G.F.; Dubois, A.E.J.

    2013-01-01

    Background: For most allergenic foods, insufficient threshold dose information within the population restricts the advice on levels of unintended allergenic foods which should trigger precautionary labeling on prepackaged foods. Objective: We wanted to derive threshold dose distributions for major

  17. Threshold dose distributions for 5 major allergenic foods in children

    NARCIS (Netherlands)

    Blom, W. Marty; Vlieg-Boerstra, Berber J.; Kruizinga, Astrid G.; van der Heide, Sicco; Houben, Geert F.; Dubois, Anthony E. J.

    Background: For most allergenic foods, insufficient threshold dose information within the population restricts the advice on levels of unintended allergenic foods which should trigger precautionary labeling on prepackaged foods. Objective: We wanted to derive threshold dose distributions for major

  18. A study of standardized extracts of Picrorhiza kurroa Royle ex Benth in experimental nonalcoholic fatty liver disease

    Directory of Open Access Journals (Sweden)

    Sapna N Shetty

    2010-01-01

    Full Text Available As a major organ of intermediary metabolism, the liver is exposed to a variety of metabolic insults due to diseases and xenobiotics viz., insulin resistance (IR drugs, toxins, microbial products, etc. One of the consequences of these metabolic insults including obesity and type 2 diabetes mellitus is the development of non-alcoholic fatty liver disease (NAFLD. The recent alarming increase in the prevalence of NAFLD compels the need to develop an appropriate animal model of the disease so as to evolve effective interventions. In this study, we have developed, in the rat, a new model of NAFLD showing several key features akin to the disease in humans. Male Wistar rats were challenged with 30% high fat diet (HFD - butter, for 2 weeks to induce NAFLD. A hydroalcoholic extract of Picrorhiza kurroa was administered to study the possible reversal of fatty changes in the liver. The extract was given in two doses viz., 200mg/kg and 400 mg/kg b.i.d., p.o. for a period of 4 weeks. There were three control groups (n = 6/group - vehicle with a regular diet, vehicle with HFD, and HFD with silymarin - a known hepatoprotective. Histopathology showed that the P. kurroa extract brought about a reversal of the fatty infiltration of the liver (mg/g and a lowering of the quantity of hepatic lipids (mg/g compared to that in the HFD control group (38.33 ± 5.35 for 200mg/kg; 29.44 ± 8.49 for 400mg/kg of P. kurroa vs.130.07 ± 6.36mg/g of liver tissue in the HFD control group; P<0.001. Compared to the standard dose of the known hepatoprotective silymarin, P. kurroa reduced the lipid content (mg/g of the liver more significantly at the dose of 400mg/kg (57.71 ± 12.45mg/kg vs. 29.44 ± 8.49 for the silymarin group vs. 400mg/kg of P. kurroa, P<0.001. In view of the increasing prevalence of metabolic syndrome and NAFLD, P. kurroa should be investigated by the reverse pharmacology path as a potential drug for the treatment of NAFLD, and essential safety studies and

  19. A study of standardized extracts of Picrorhiza kurroa Royle ex Benth in experimental nonalcoholic fatty liver disease.

    Science.gov (United States)

    Shetty, Sapna N; Mengi, Sushma; Vaidya, Rama; Vaidya, Ashok D B

    2010-07-01

    As a major organ of intermediary metabolism, the liver is exposed to a variety of metabolic insults due to diseases and xenobiotics viz., insulin resistance (IR) drugs, toxins, microbial products, etc. One of the consequences of these metabolic insults including obesity and type 2 diabetes mellitus is the development of non-alcoholic fatty liver disease (NAFLD). The recent alarming increase in the prevalence of NAFLD compels the need to develop an appropriate animal model of the disease so as to evolve effective interventions. In this study, we have developed, in the rat, a new model of NAFLD showing several key features akin to the disease in humans. Male Wistar rats were challenged with 30% high fat diet (HFD) - butter, for 2 weeks to induce NAFLD. A hydroalcoholic extract of Picrorhiza kurroa was administered to study the possible reversal of fatty changes in the liver. The extract was given in two doses viz., 200mg/kg and 400 mg/kg b.i.d., p.o. for a period of 4 weeks. There were three control groups (n = 6/group) - vehicle with a regular diet, vehicle with HFD, and HFD with silymarin - a known hepatoprotective.Histopathology showed that the P. kurroa extract brought about a reversal of the fatty infiltration of the liver (mg/g) and a lowering of the quantity of hepatic lipids (mg/g) compared to that in the HFD control group (38.33 ± 5.35 for 200mg/kg; 29.44 ± 8.49 for 400mg/kg of P. kurroa vs.130.07 ± 6.36mg/g of liver tissue in the HFD control group; P<0.001). Compared to the standard dose of the known hepatoprotective silymarin, P. kurroa reduced the lipid content (mg/g) of the liver more significantly at the dose of 400mg/kg (57.71 ± 12.45mg/kg vs. 29.44 ± 8.49 for the silymarin group vs. 400mg/kg of P. kurroa, P<0.001). In view of the increasing prevalence of metabolic syndrome and NAFLD, P. kurroa should be investigated by the reverse pharmacology path as a potential drug for the treatment of NAFLD, and essential safety studies and preformulation

  20. Efficacy of standardized extract of Hibiscus sabdariffa L. (Malvaceae) in improving iron status of adults in malaria endemic area: A randomized controlled trial.

    Science.gov (United States)

    Peter, Emanuel L; Rumisha, Susan F; Mashoto, Kijakazi O; Minzi, Omary Ms; Mfinanga, Sayoki

    2017-09-14

    Indigenous community of Mkuranga district have been using aqueous extract of H. sabdariffa L. for treating anemia. However, there have been neither safety nor efficacy studies to validate this medicinal product in anemia. The purpose of this study was to establish efficacy and safety of standardized aqueous extract of H. sabdariffa L. in anemic adults. This was a randomized controlled clinical trial in which 130 adults' men and women aged 18-50 years were involved after meeting the inclusion criteria. Initially, standardized aqueous extract of H. sabdariffa L. was prepared using optimized extraction parameters. Stratified randomization was used to randomize participants into four fixed dose groups. The first group received oral dose of 1000ml while the 2nd group was randomized to receive 1500ml orally. The last two groups were given a dose of 2000ml of extract and 200mg ferrous sulphate tablet respectively. Primary endpoint was the actual change of iron status indicators at the end of 30 days follow up period as compared to those recorded at baseline. Adverse effects were assessed at every 10th day scheduled visit. In all arms, HB and hematopoietic parameters were measured using HemoCue hemoglobinometer® (HemoCue, Ängelholm, Sweden) and hematology analyzer® respectively at the trial site. Follow up was done for 30 days. A total of 82 participants were included for analysis. A standardized aqueous extract of H. sabdariffa L. did not improve iron status in anemic adults in malaria endemic region (P>0.005). However, there was evidence to support the safety of the extract for human consumptions as herbal supplement. Iron and organic acids contents of H. sabdariffa L. extract showed the potential of improving hematopoietic parameters. Studies with bigger sample size are therefore needed to establish the efficacy of the extract when concurrently used with malaria chemoprophylaxis in malaria endemic areas. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

  1. Animal Allergens and Their Presence in the Environment

    OpenAIRE

    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 ...

  2. Origin and Functional Prediction of Pollen Allergens in Plants.

    Science.gov (United States)

    Chen, Miaolin; Xu, Jie; Devis, Deborah; Shi, Jianxin; Ren, Kang; Searle, Iain; Zhang, Dabing

    2016-09-01

    Pollen allergies have long been a major pandemic health problem for human. However, the evolutionary events and biological function of pollen allergens in plants remain largely unknown. Here, we report the genome-wide prediction of pollen allergens and their biological function in the dicotyledonous model plant Arabidopsis (Arabidopsis thaliana) and the monocotyledonous model plant rice (Oryza sativa). In total, 145 and 107 pollen allergens were predicted from rice and Arabidopsis, respectively. These pollen allergens are putatively involved in stress responses and metabolic processes such as cell wall metabolism during pollen development. Interestingly, these putative pollen allergen genes were derived from large gene families and became diversified during evolution. Sequence analysis across 25 plant species from green alga to angiosperms suggest that about 40% of putative pollen allergenic proteins existed in both lower and higher plants, while other allergens emerged during evolution. Although a high proportion of gene duplication has been observed among allergen-coding genes, our data show that these genes might have undergone purifying selection during evolution. We also observed that epitopes of an allergen might have a biological function, as revealed by comprehensive analysis of two known allergens, expansin and profilin. This implies a crucial role of conserved amino acid residues in both in planta biological function and allergenicity. Finally, a model explaining how pollen allergens were generated and maintained in plants is proposed. Prediction and systematic analysis of pollen allergens in model plants suggest that pollen allergens were evolved by gene duplication and then functional specification. This study provides insight into the phylogenetic and evolutionary scenario of pollen allergens that will be helpful to future characterization and epitope screening of pollen allergens. © 2016 American Society of Plant Biologists. All rights reserved.

  3. Indoor Environmental Interventions for Furry Pet Allergens, Pest Allergens, and Mold: Looking to the Future.

    Science.gov (United States)

    Ahluwalia, Sharon K; Matsui, Elizabeth C

    Over the last 2 to 3 decades, significant advances have been made in understanding the role that indoor allergen exposures play with regard to respiratory health. Multiple studies have confirmed that sensitization and exposure to indoor allergens can be a risk factor for asthma morbidity. Environmental interventions targeting key indoor allergens have been evaluated with the aims of examining their causal effects on asthma-related outcomes and identifying clinically efficacious interventions to incorporate into treatment recommendations. Historically, it appeared that the most successful intervention, as performed in the Inner-City Asthma Study, was individually tailored, targeting multiple allergens in a predominantly low-income, minority, and urban pediatric population. Recent studies suggest that single-allergen interventions may be efficacious when targeting the most clinically relevant allergen for a population. In this article, we review recent literature on home environmental interventions and their effects on specific indoor allergen levels and asthma-related outcomes. Copyright © 2017 American Academy of Allergy, Asthma & Immunology. All rights reserved.

  4. Optimal conditions for the removal of house dust mite, dog dander, and pollen allergens using mechanical laundry.

    Science.gov (United States)

    Choi, Soo-Young; Lee, In-Yong; Sohn, Jung-Ho; Lee, Yong-Won; Shin, Yoo-Seob; Yong, Tae-Soon; Hong, Chein-Soo; Park, Jung-Won

    2008-06-01

    Mechanical laundry is an effective tool for the environmental control of allergens, but the optimal conditions for removing allergens are not yet clear. To evaluate the optimal conditions of mechanical laundry for the removal of house dust mite (HDM), dog dander, and pollen allergens. The 4 washing modes of 30 degrees C (86 degrees F), 40 degrees C (104 degrees F), 60 degrees C (140 degrees F), and steam water (SW) with detergent were evaluated. Allergen removal performance was assayed using a 2-site enzyme-linked immunosorbent assay (ELISA) or an ELISA inhibition test. Using the 30 degrees C and 40 degrees C washing modes, only 6.5% and 9.6% of Dermatophagoides farinae, respectively, were killed. However, using the 60 degrees C and SW washing modes, all HDMs were killed. The amounts of Der f 1 remaining after the 30 degrees C, 40 degrees C, 60 degrees C, and SW washing modes were 26.8%, 2.4%, 1.3%, and 0.6%, respectively, with unmanipulated contaminated sheets. The effects of rinse on Der f 1 levels after the 30 degrees C washing were greater compared with those after the 40 degrees C, 60 degrees C, and SW modes. The amounts of Can f 1 in the extractions after washing were 0.3% to 1.3% for all modes, and all extracts, even without a rinse, did not inhibit specific IgE binding to dog allergens according to ELISA. The remaining pollen allergen levels after washing were lower in the 60 degrees C and SW modes than in the lower temperature modes. However, the levels did not differ among the various washing modes after rinsing once. Water temperature and number of rinses are critical factors for the removal of HDM, dog dander, and pollen allergens.

  5. Prevention of birch pollen-related food allergy by mucosal treatment with multi-allergen-chimers in mice.

    Directory of Open Access Journals (Sweden)

    Elisabeth Hoflehner

    Full Text Available Among birch pollen allergic patients up to 70% develop allergic reactions to Bet v 1-homologue food allergens such as Api g 1 (celery or Dau c 1 (carrot, termed as birch pollen-related food allergy. In most cases, specific immunotherapy with birch pollen extracts does not reduce allergic symptoms to the homologue food allergens. We therefore genetically engineered a multi-allergen chimer and tested if mucosal treatment with this construct could represent a novel approach for prevention of birch pollen-related food allergy.BALB/c mice were poly-sensitized with a mixture of Bet v 1, Api g 1 and Dau c 1 followed by a sublingual challenge with carrot, celery and birch pollen extracts. For prevention of allergy sensitization an allergen chimer composed of immunodominant T cell epitopes of Api g 1 and Dau c 1 linked to the whole Bet v 1 allergen, was intranasally applied prior to sensitization.Intranasal pretreatment with the allergen chimer led to significantly decreased antigen-specific IgE-dependent β-hexosaminidase release, but enhanced allergen-specific IgG2a and IgA antibodies. Accordingly, IL-4 levels in spleen cell cultures and IL-5 levels in restimulated spleen and cervical lymph node cell cultures were markedly reduced, while IFN-γ levels were increased. Immunomodulation was associated with increased IL-10, TGF-β and Foxp3 mRNA levels in NALT and Foxp3 in oral mucosal tissues. Treatment with anti-TGF-β, anti-IL10R or anti-CD25 antibodies abrogated the suppression of allergic responses induced by the chimer.Our results indicate that mucosal application of the allergen chimer led to decreased Th2 immune responses against Bet v 1 and its homologue food allergens Api g 1 and Dau c 1 by regulatory and Th1-biased immune responses. These data suggest that mucosal treatment with a multi-allergen vaccine could be a promising treatment strategy to prevent birch pollen-related food allergy.

  6. Prevention of birch pollen-related food allergy by mucosal treatment with multi-allergen-chimers in mice.

    Science.gov (United States)

    Hoflehner, Elisabeth; Hufnagl, Karin; Schabussova, Irma; Jasinska, Joanna; Hoffmann-Sommergruber, Karin; Bohle, Barbara; Maizels, Rick M; Wiedermann, Ursula

    2012-01-01

    Among birch pollen allergic patients up to 70% develop allergic reactions to Bet v 1-homologue food allergens such as Api g 1 (celery) or Dau c 1 (carrot), termed as birch pollen-related food allergy. In most cases, specific immunotherapy with birch pollen extracts does not reduce allergic symptoms to the homologue food allergens. We therefore genetically engineered a multi-allergen chimer and tested if mucosal treatment with this construct could represent a novel approach for prevention of birch pollen-related food allergy. BALB/c mice were poly-sensitized with a mixture of Bet v 1, Api g 1 and Dau c 1 followed by a sublingual challenge with carrot, celery and birch pollen extracts. For prevention of allergy sensitization an allergen chimer composed of immunodominant T cell epitopes of Api g 1 and Dau c 1 linked to the whole Bet v 1 allergen, was intranasally applied prior to sensitization. Intranasal pretreatment with the allergen chimer led to significantly decreased antigen-specific IgE-dependent β-hexosaminidase release, but enhanced allergen-specific IgG2a and IgA antibodies. Accordingly, IL-4 levels in spleen cell cultures and IL-5 levels in restimulated spleen and cervical lymph node cell cultures were markedly reduced, while IFN-γ levels were increased. Immunomodulation was associated with increased IL-10, TGF-β and Foxp3 mRNA levels in NALT and Foxp3 in oral mucosal tissues. Treatment with anti-TGF-β, anti-IL10R or anti-CD25 antibodies abrogated the suppression of allergic responses induced by the chimer. Our results indicate that mucosal application of the allergen chimer led to decreased Th2 immune responses against Bet v 1 and its homologue food allergens Api g 1 and Dau c 1 by regulatory and Th1-biased immune responses. These data suggest that mucosal treatment with a multi-allergen vaccine could be a promising treatment strategy to prevent birch pollen-related food allergy.

  7. Diet influences growth rates and allergen and endotoxin contents of cultured Dermatophagoides farinae and Dermatophagoides pteronyssinus house dust mites.

    Science.gov (United States)

    Avula-Poola, Swetha; Morgan, Marjorie S; Arlian, Larry G

    2012-01-01

    The house dust mites Dermatophagoides farinae and Dermatophagoides pteronyssinus are cultured to obtain material for the production of allergen extracts for research, diagnostic and immunotherapeutic purposes. We cultured mites on two different diets that supported thriving populations and determined the population growth rates, dynamics of allergen accumulation, and endotoxin concentrations in extracts made from mites harvested from the cultures. D. farinae populations grew faster on a diet of rodent chow/yeast than on an egg/yeast diet but a larger peak population size was achieved on the egg/yeast diet. Diet influenced the dynamics of the production of groups 1 and 2 allergens and the group 1/2 ratios for both species. To population peak, Der f 1 was produced at a faster rate on the chow/yeast diet but greater amounts of Der f 1 were produced by mites grown on the egg/yeast diet. D. pteronyssinus populations grew faster and achieved greater density on the egg/yeast diet compared to the chow/yeast diet. D. pteronyssinus produced more Der p 1 than Der p 2 when grown on chow/yeast while more Der p 2 than Der p 1 was produced on egg/yeast. Endotoxin concentrations in extracts made from whole cultures for both species at maximum population density were very different in the two diets. Washing the mites resulted in the loss of up to 88% of the allergen. Mite-culturing diet directly effects population growth, the dynamics of allergen accumulation, the group 1/2 allergen ratio and the endotoxin contents in extracts of cultured house dust mites. Copyright © 2012 S. Karger AG, Basel.

  8. A mutant of the major apple allergen, Mal d 1, demonstrating hypo-allergenicity in the target organ by double-blind placebo-controlled food challenge

    NARCIS (Netherlands)

    Bolhaar, S. T. H. P.; Zuidmeer, L.; Ma, Y.; Ferreira, F.; Bruijnzeel-Koomen, C. A. F. M.; Hoffmann-Sommergruber, K.; van Ree, R.; Knulst, A. C.

    2005-01-01

    BACKGROUND: Allergen-specific immunotherapy for food allergy has been hindered by severe side-effects in the past. Well-characterized hypo-allergenic recombinant food allergens potentially offer a safe solution. OBJECTIVE: To demonstrate hypo-allergenicity of a mutated major food allergen from

  9. In vitro assessment of the synergism between extracts of Cocos nucifera husk and some standard antibiotics

    Directory of Open Access Journals (Sweden)

    Taiwo Adesola Akinyele

    2017-04-01

    Conclusions: This investigation suggests the crude extracts of C. nucifera to be a potential broad spectrum antimicrobial compound. Therefore, further study is needed to isolate the pure compounds from these crude extracts.

  10. Regulation of proliferation and gene expression in cultured human aortic smooth muscle cells by resveratrol and standardized grape extracts

    International Nuclear Information System (INIS)

    Wang Zhirong; Chen Yan; Labinskyy, Nazar; Hsieh Tzechen; Ungvari, Zoltan; Wu, Joseph M.

    2006-01-01

    Epidemiologic studies suggest that low to moderate consumption of red wine is inversely associated with the risk of coronary heart disease; the protection is in part attributed to grape-derived polyphenols, notably trans-resveratrol, present in red wine. It is not clear whether the cardioprotective effects of resveratrol can be reproduced by standardized grape extracts (SGE). In the present studies, we determined, using cultured human aortic smooth muscle cells (HASMC), growth and specific gene responses to resveratrol and SGE provided by the California Table Grape Commission. Suppression of HASMC proliferation by resveratrol was accompanied by a dose-dependent increase in the expression of tumor suppressor gene p53 and heat shock protein HSP27. Using resveratrol affinity chromatography and biochemical fractionation procedures, we showed by immunoblot analysis that treatment of HASMC with resveratrol increased the expression of quinone reductase I and II, and also altered their subcellular distribution. Growth of HASMC was significantly inhibited by 70% ethanolic SGE; however, gene expression patterns in various cellular compartments elicited in response to SGE were substantially different from those observed in resveratrol-treated cells. Further, SGE also differed from resveratrol in not being able to induce relaxation of rat carotid arterial rings. These results indicate that distinct mechanisms are involved in the regulation of HASMC growth and gene expression by SGE and resveratrol

  11. Parvalbumin--the major tropical fish allergen.

    Science.gov (United States)

    Lim, Dawn Li-Chern; Neo, Keng Hwee; Yi, Fong Cheng; Chua, Kaw Yan; Goh, Denise Li-Meng; Shek, Lynette Pei-Chi; Giam, Yoke Chin; Van Bever, Hugo P S; Lee, Bee Wah

    2008-08-01

    Fish allergy is common in countries where consumption is high. Asian nations are amongst the world's largest consumers of fish but the allergen profiles of tropical fish are unknown. This study sought to evaluate the allergenicity of four commonly consumed tropical fish, the threadfin (Polynemus indicus), Indian anchovy (Stolephorus indicus), pomfret (Pampus chinensis) and tengirri (Scomberomorus guttatus). Immunoglobulin E (IgE) cross-reactivity with parvalbumin of cod fish (Gad c 1), the major fish allergen, was also studied. Detection of tropical fish and cod specific-IgE was performed by UniCap assay, and skin prick tests were also carried out. The IgE-binding components of tropical fish were identified using IgE immunoblot techniques, and cross-reactivity with Gad c 1 was assessed by ELISA inhibition and IgE immunoblot inhibition. Clinically, nine of 10 patients studied were allergic to multiple fish. All patients exhibited detectable specific-IgE to cod fish (10 of 10 skin prick test positive, eight of 10 UniCap assay positive) despite lack of previous exposure. The major allergen of the four tropical fish was the 12-kDa parvalbumin. IgE cross-reactivity of these allergens to Gad c 1 was observed to be moderate to high in the tropical fish studied. Parvalbumins are the major allergens in commonly consumed tropical fish. They are cross-reactive with each other as well as with Gad c 1. Commercial tests for cod fish appear to be sufficient for the detection of tropical fish specific-IgE.

  12. Allergic contact dermatitis and changes in the frequency of the causative allergens demonstrated with patch testing in 2008-2012.

    Science.gov (United States)

    Tichy, Martin; Karlova, Iva

    2015-09-01

    The frequency of contact allergens is often regionally different and hence it regional identification is important for the prevention of allergic contact dermatitis (ACD). The aim of the study was to determine the most common contact allergens in the etiology of ACD from 2008-2012 and evaluate their dynamics in comparison with similar studies in the Czech Republic and internationally carried out in previous years. A total of 1941 patients with suspected ACD were examined using the patch tests from the European Standard Series. The data obtained were subjected to statistical analysis. The most common contact allergy was to nickel (15.4%), the second most frequent allergen was Balsam of Peru (11.6%). The top five allergens in the whole sample also included, in descending order, fragrance-mix I (7.1%), cobalt chloride (6.4%), and Kathon CG (4.0%). While the order of contact allergens causing sensitisation in women is consistent with the results of the whole sample, in men instead of cobalt and Kathon CG, epoxy resin and chromium appeared among the five most common allergens. In the Czech Republic, despite existing EU legislation, action to reduce the frequency of sensitization to nickel failed in contrast to other EU countries. In addition to nickel, consistent preventive measures should be focused mainly on sensitization to Balsam of Peru, cobalt, epoxy resins, chromium and aromatic substances and preservatives used in the cosmetic industry.

  13. Ana o 2, a major cashew (Anacardium occidentale L.) nut allergen of the legumin family.

    Science.gov (United States)

    Wang, Fang; Robotham, Jason M; Teuber, Suzanne S; Sathe, Shridhar K; Roux, Kenneth H

    2003-09-01

    We recently cloned and described a vicilin and showed it to be a major cashew allergen. Additional IgE-reactive cashew peptides of the legumin group and 2S albumin families have also been reported. Here, we attempt to clone, express and characterize a second major cashew allergen. A cashew cDNA library was screened with human IgE and rabbit IgG anti-cashew extract antisera, and a reactive nonvicilin clone was sequenced and expressed as a fusion protein in Escherichia coli. Immunoblotting was used to screen for reactivity with patients' sera, and inhibition of immunoblotting was used to identify the corresponding native peptides in cashew nut extract. The identified allergen was subjected to linear epitope mapping using SPOTs solid-phase synthetic peptide technology. Sequence analysis showed the selected clone, designated Ana o 2, to encode for a member of the legumin family (an 11S globulin) of seed storage proteins. By IgE immunoblotting, 13 of 21 sera (62%) from cashew-allergic patients were reactive. Immunoblot inhibition data showed that the native Ana o 2 constitutes a major band at approximately 33 kD and a minor band at approximately 53 kD. Probing of overlapping synthetic peptides with pooled human cashew-allergic sera identified 22 reactive peptides, 7 of which gave strong signals. Several Ana o 2 epitopes were shown to overlap those of the peanut legumin group allergen, Ara h 3, in position but with little sequence similarity. Greater positional overlap and identity was observed between Ana o 2 and soybean glycinin epitopes. We conclude that this legumin-like protein is a major allergen in cashew nut. Copyright 2003 S. Karger AG, Basel

  14. A Standardized Composition Comprised of Extracts from Rosmarinus officinalis, Annona squamosa and Zanthoxylum clava-herculis for Cellulite

    Science.gov (United States)

    Yimam, Mesfin; Lee, Young-Chul; Jiao, Ping; Hong, Mei; Brownell, Lidia; Jia, Qi

    2017-01-01

    cellulite onset, continuation, or exacerbations. SUMMARY Cellulite represents one of the main esthetic concerns of women with a likely cause of psychological insecurities. Its pathophysiology involves multiple pathways that include vascular, adipose tissues, inflammation, structural and physiological.Treatment strategies for cellulite comprises increasing microcirculation flow, reducing lipogenesis, promoting lipolysis, free radicals scavenging or formation reduction, anti-inflammation and other invasive procedures.We screened our plant library for extracts that reduces cellular lipid accumulation, improves microcirculation, possesses high total antioxidant capacity, inhibits platelet aggregation, and moderates inflammation.Botanical extracts from Rosmarinus officinalis, Annona squamosa and Zanthoxylum clava-herculis were identified as leads and formulated to yield a standardized composition designated as UP1307 and suggested its usage for cellulite. Abbreviations Used: GMP: Good Manufacturing Practice; CA: Carnosic acid; NF-kB: nuclear factor-kB; HPLC: high-performance liquid chromatography; EtOH: Ethanol; DMEM: Dulbecco's modified Eagle's medium; FBS: fetal bovine serum; SD: Sprague Dawley; RAC: recto-anus coefficient; LPS: Lipopolysaccharide; DPPH: 1,1-diphenyl-2-picryl-hydrazyl; TNF-α: tumor necrosis factor; NO: Nitric oxide PMID:29263624

  15. A Standardized Composition Comprised of Extracts fromRosmarinus officinalis,Annona squamosaandZanthoxylum clava-herculisfor Cellulite.

    Science.gov (United States)

    Yimam, Mesfin; Lee, Young-Chul; Jiao, Ping; Hong, Mei; Brownell, Lidia; Jia, Qi

    2017-01-01

    represents one of the main esthetic concerns of women with a likely cause of psychological insecurities. Its pathophysiology involves multiple pathways that include vascular, adipose tissues, inflammation, structural and physiological.Treatment strategies for cellulite comprises increasing microcirculation flow, reducing lipogenesis, promoting lipolysis, free radicals scavenging or formation reduction, anti-inflammation and other invasive procedures.We screened our plant library for extracts that reduces cellular lipid accumulation, improves microcirculation, possesses high total antioxidant capacity, inhibits platelet aggregation, and moderates inflammation.Botanical extracts from Rosmarinus officinalis , Annona squamosa and Zanthoxylum clava-herculis were identified as leads and formulated to yield a standardized composition designated as UP1307 and suggested its usage for cellulite. Abbreviations Used: GMP: Good Manufacturing Practice; CA: Carnosic acid; NF-kB: nuclear factor-kB; HPLC: high-performance liquid chromatography; EtOH: Ethanol; DMEM: Dulbecco's modified Eagle's medium; FBS: fetal bovine serum; SD: Sprague Dawley; RAC: recto-anus coefficient; LPS: Lipopolysaccharide; DPPH: 1,1-diphenyl-2-picryl-hydrazyl; TNF-α: tumor necrosis factor; NO: Nitric oxide.

  16. Allergen immunotherapy as a drug: the new deal of grass allergen tablets from clinical trials to current practice.

    Science.gov (United States)

    Manzotti, G; Lombardi, C

    2013-04-01

    Currently, sublingual immunotherapy (SLIT) may be performed by a number of allergen extract in different preparations but in a near future only products fulfilling the requirements from the regulatory agencies, that make mandatory a pharmaceutical quality, will be authorized. Indeed, two products with such characteristics are already available for SLIT in grass pollen allergic patients, Grazax from Alk-Abellò and Oralair from Stallergenes. The data from registrative trials as well as from postmarketing studies provide evidence of efficacy and safety of such products. This articles reviews the similarities and the differences of Grazax and Oralair, both designed as drugs for the treatment of grass pollen allergy with the aim, which is exclusive of allergen immunotherapy, to work on the natural history of allergy and not only on symptoms as rescue medications do. The aim of this paper is to evaluate the available trials with Grazax and Oralair in terms of pre-seasonal schedule approach to support their use in clinical practice. Such kind of treatment makes possible a continuous dialogue between clinical investigators and clinical practitioners, and is the only way for scientific progress that puts the patient's health at the first place.

  17. Citral a fragrance allergen and irritant

    DEFF Research Database (Denmark)

    Heydorn, S; Menné, T; Andersen, Klaus Ejner

    2003-01-01

    Citral is a well known contact allergen and a contact irritant. Routine patch testing in the past may have been restricted because of possible irritant (IR) patch test responses. 586 consecutive patients, with hand eczema, were patch tested with a selection of fragrances including citral 2...... and positive patch test reactions to other fragrances compared with IR reactions (n = 82) was established. The difference regarding fragrance history found between those with IR and positive reactions to citral was not significant. Citral could be an allergen and/or irritant, worthy of further more extensive...

  18. Comparison of elicitation potential of chloroatranol and atranol - 2 allergens in oak moss absolute

    DEFF Research Database (Denmark)

    Johansen, J.D.; Bernard, G.; Gimenez-Arnau, E.

    2006-01-01

    Chloroatranol and atranol are degradation products of chloroatranorin and atranorin, respectively, and have recently been identified as important contact allergens in the natural fragrance extract, oak moss absolute. Oak moss absolute is widely used in perfumery and is the cause of many cases...... of fragrance allergic contact dermatitis. Chloroatranol elicits reactions at very low levels of exposure. In oak moss absolute, chloroatranol and atranol are present together and both may contribute to the allergenicity and eliciting capacity of the natural extract. In this study, 10 eczema patients with known...... sensitization to chloroatranol and oak moss absolute were tested simultaneously to a serial dilution of chloroatranol and atranol in ethanol, in equimolar concentrations (0.0034-1072 microM). Dose-response curves were estimated and analysed by logistic regression. The estimated difference in elicitation potency...

  19. Effects of the recombinant allergen rDer f 2 on neuro-endocrino-immune network in asthmatic mice.

    Science.gov (United States)

    Jiang, Yong-Qian; Zhou, Zhi-Xiang; Ji, You-Lin

    2014-01-01

    Severe and life-threatening side effects can occur in patients receiving allergen-specific immunotherapy (SIT), and recombinant allergens made from cDNA have been used in clinical trials for ten years and appear promising for SIT. The aim of this study is to explore the effects of the recombinant allergen Der f 2 (the group 2 allergen from Dermatophagoides farinae) on the neuro-endocrino-immune network in asthmatic mice. Twenty-eight mice were divided into four groups - A, B, C and D. To induce asthma, a crude extract of D. farinae was injected intraperitoneally into the mice in groups B, C and D. Later, the crude extract or recombinant allergen rDer f 2 was given to groups C and D, respectively. Normal saline was given to groups A and B. Serum corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), corticosterone (CORT), interleukin 4 (IL-4), and interferon γ (IFN-γ) were detected by immunoassay and the pathological change of lung tissue was observed by hematoxylin and eosin (HE) staining. Serum CRH, ACTH, CORT, and IFN-γ were highest in healthy group A but lowest in asthma group B. Treatment with the crude extract or recombinant allergen rDer f 2 significantly attenuated this response in asthmatic mice, but there was no difference between the two treatments (p > 0.05). Serum IL-4 was elevated in asthma group B but lowest in healthy group A. Treatment with the crude extract or recombinant allergen rDer f 2 significantly attenuated this response in asthmatic mice, but there was no significant difference between the two treatments (p > 0.05). However, lung pathology as measured histologically (Underwood Score) showed that rDer f 2-treatment was significantly better than crude extract treatment (p < 0.05). In brief, recombinant allergen Der f 2 can strengthen the function of hypothalamus-pituitary-adrenal (HPA) axis, affect the balance of Th1 and Th2 cytokines, and reduce pulmonary inflammation in asthmatic mice.

  20. 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.

  1. Determination of Ergot Alkaloids: Purity and Stability Assessment of Standards and Optimization of Extraction Conditions for Cereal Samples

    DEFF Research Database (Denmark)

    Krska, R.; Berthiller, F.; Schuhmacher, R.

    2008-01-01

    considerably above 98% apart from ergocristinine (94%), ergosine (96%), and ergosinine (95%). Also discussed is the optimization of extraction conditions presented in a recently published method for the quantitation of ergot alkaloids in food samples using solid-phase extraction with primary secondary amine...... (PSA) before LC/MS/MS. Based on the results obtained from these optimization studies, a mixture of acetonitrile with ammonium carbonate buffer was used as extraction solvent, as recoveries for all analyzed ergot alkaloids were significantly higher than those with the other solvents. Different sample......-solvent ratios and extraction times showed just minor influences in extraction efficacy. Finally, the stability of the ergot alkaloids in both raw cereals and cereal-based processed food extracts was studied. According to these studies, extracts should be prepared and analyzed the same day or stored below...

  2. The TLR5 ligand flagellin promotes asthma by priming allergic responses to indoor allergens

    Science.gov (United States)

    Wilson, Rhonda H.; Maruoka, Shuichiro; Whitehead, Gregory S.; Foley, Julie F.; Flake, Gordon P.; Sever, Michelle L.; Zeldin, Darryl C.; Kraft, Monica; Garantziotis, Stavros; Nakano, Hideki; Cook, Donald N.

    2012-01-01

    Allergic asthma is a complex disease characterized by eosinophilic pulmonary inflammation, mucus production and reversible airway obstruction1. Exposure to indoor allergens is a clear risk factor for asthma, but this disease is also associated with high household levels of total and Gram-negative bacteria2. The ability of bacterial products to act as adjuvants3 suggests they might promote asthma by priming allergic sensitization to inhaled allergens. In support of this idea, house dust extracts (HDEs) can activate antigen presenting dendritic cells (DC) in vitro and promote allergic sensitization to inhaled innocuous proteinsin vivo4. It is unknown which microbial products provide most of the adjuvant activity in HDEs. A screen of microbial products for their adjuvant activity in the airway revealed that the bacterial protein, flagellin (FLA) stimulated strong allergic responses to an innocuous inhaled protein. Moreover, toll-like receptor (TLR)5, the mammalian receptor for FLA5,6, was required for priming strong allergic responses to natural indoor allergens present in HDEs. In addition, the incidence of human asthma was associated with high serum levels of FLA-specific antibodies. Together, these findings suggest that household FLA promotes the development of allergic asthma by TLR5-dependent priming of allergic responses to indoor allergens. PMID:23064463

  3. Allergens involved in the cross-reactivity of Aedes aegypti with other arthropods.

    Science.gov (United States)

    Cantillo, Jose Fernando; Puerta, Leonardo; Lafosse-Marin, Sylvie; Subiza, Jose Luis; Caraballo, Luis; Fernandez-Caldas, Enrique

    2017-06-01

    Cross-reactivity between Aedes aegypti and mites, cockroaches, and shrimp has been previously suggested, but the involved molecular components have not been fully described. To evaluate the cross-reactivity between A aegypti and other arthropods. Thirty-four serum samples from patients with asthma and/or allergic rhinitis were selected, and specific IgE to A aegypti, Dermatophagoides pteronyssinus, Dermatophagoides farinae, Blomia tropicalis, Periplaneta americana. and Litopenaeus vannamei was measured by enzyme-linked immunosorbent assay. Cross-reactivity was investigated using pooled serum samples from allergic patients, allergenic extracts, and the recombinant tropomyosins (Aed a 10.0201, Der p 10, Blo t 10, Lit v 1, and Per a 7). Four IgE reactive bands were further characterized by matrix-assisted laser desorption/ionization tandem time of flight. Frequency of positive IgE reactivity was 82.35% to at least one mite species, 64.7% to A aegypti, 29.4% to P americana, and 23.5% to L vannamei. The highest IgE cross-reactivity was seen between A aegypti and D pteronyssinus (96.6%) followed by L vannamei (95.4%), B tropicalis (84.4%), and P americana (75.4%). Recombinant tropomyosins from mites, cockroach, or shrimp inhibited the IgE reactivity to the mosquito at a lower extent than the extracts from these arthropods. Several bands of A aegypti cross-reacted with arthropod extracts, and 4 of them were identified as odorant binding protein, mitochondrial cytochrome C, peptidyl-prolyl cis-trans isomerase, and protein with hypothetical magnesium ion binding function. We identified 4 novel cross-reactive allergens in A aegypti allergenic extract. These molecules could influence the manifestation of allergy to environmental allergens in the tropics. Copyright © 2017 American College of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

  4. 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...... or as recombinant forms, within the EU-funded EuroPrevall project. These allergens have been characterized by a battery of diagnostic tests demonstrating that they constitute an authentic, well-defined library of comparable quality. The review summarizes the applications, potentials and limitations of key...

  5. 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)

  6. Real-Life Study for the Diagnosis of House Dust Mite Allergy - The Value of Recombinant Allergen-Based IgE Serology.

    Science.gov (United States)

    Becker, Sven; Schlederer, Thomas; Kramer, Matthias F; Haack, Mareike; Vrtala, Susanne; Resch, Yvonne; Lupinek, Christian; Valenta, Rudolf; Gröger, Moritz

    2016-01-01

    Dermatophagoides pteronyssinus is one of the most important perennial allergen sources worldwide. Molecular diagnostics using the commercially available major allergens (Der p 1 and Der p 2) in combination with Der p 10 do not detect house dust mite (HDM) sensitization in a number of cases when used alone. The objective was to evaluate the IgE reactivity profiles of these patients using an experimental immunoassay biochip. Sera of HDM-allergic patients (positive skin prick test, CAP class ≥1 for allergen extract, and positive intranasal provocation) were tested for IgE antibodies against Der p 1, Der p 2, and Der p 10 by ImmunoCAP fluorescence enzyme immunoassay. Negatively tested sera were examined by an experimental chip containing 13 microarrayed HDM allergens. Of 97 patients tested, 16 showed negative results to Der p 1, Der p 2, and Der p 10. MeDALL chip evaluation revealed 5 patients monosensitized to Der p 23, and 11 patients were negative for all HDM MeDALL chip components. Seven sera were available for further testing, and 3 of them showed IgE reactivity to dot-blotted nDer p 1, and 2 reacted with high-molecular weight components (>100 kDa) in nitrocellulose-blotted HDM extract when tested with 125I-labeled anti-IgE in a RAST-based assay. The HDM extract-specific IgE levels of the 11 patients were <3.9 kU/l. Recombinant allergen-based IgE serology is of great value when conventional IgE diagnostics fails. Der p 23 is an important HDM allergen, especially when major allergens are negative. Therefore, it would be desirable to have Der p 23 commercially available. Further research concerning the prevalence and clinical significance of different HDM allergens is needed. © 2016 S. Karger AG, Basel.

  7. The anti-inflammatory activity of standard aqueous stem bark extract of Mangifera indica L. as evident in inhibition of Group IA sPLA2.

    Science.gov (United States)

    Dhananjaya, Bhadrapura Lakkappa; Shivalingaiah, Sudharshan

    2016-03-01

    The standard aqueous stem bark extract is consumed as herbal drink and used in the pharmaceutical formulations to treat patients suffering from various disease conditions in Cuba. This study was carried out to evaluate the modulatory effect of standard aqueous bark extract of M. indica on Group IA sPLA2. M. indica extract, dose dependently inhibited the GIA sPLA2 (NN-XIa-PLA2) activity with an IC50 value 8.1 µg/ml. M. indica extract effectively inhibited the indirect hemolytic activity up to 98% at ~40 µg/ml concentration and at various concentrations (0-50 µg/ml), it dose dependently inhibited the edema formation. When examined as a function of increased substrate and calcium concentration, there was no relieve of inhibitory effect on the GIA sPLA2. Furthermore, the inhibition was irreversible as evidenced from binding studies. It is observed that the aqueous extract ofM. indica effectively inhibits sPLA2 and it is associated inflammatory activities, which substantiate their anti-inflammatory properties. The mode of inhibition could be due to direct interaction of components present in the extract, with sPLA2 enzyme. Further studies on understanding the principal constituents, responsible for the anti-inflammatory activity would be interesting to develop this into potent anti-inflammatory agent.

  8. Allergen-loaded strontium-doped hydroxyapatite spheres improve allergen-specific immunotherapy in mice

    OpenAIRE

    Garbani, M; Xia, W; Rhyner, C; Prati, M; Scheynius, A; Malissen, B; Engqvist, H; Maurer, M; Crameri, R; Terhorst, D

    2017-01-01

    Background Immunomodulatory interventions play a key role in the treatment of infections and cancer as well as allergic diseases. Adjuvants such as micro- and nanoparticles are often added to immunomodulatory therapies to enhance the triggered immune response. Here, we report the immunological assessment of novel and economically manufactured microparticle adjuvants, namely strontium-doped hydroxyapatite porous spheres (SHAS), which we suggest for the use as adjuvant and carrier in allergen-s...

  9. Allergen-specific immunotherapy of Hymenoptera venom allergy

    DEFF Research Database (Denmark)

    Schiener, Maximilian; Graessel, Anke; Ollert, Markus

    2017-01-01

    Stings of hymenoptera can induce IgE-mediated hypersensitivity reactions in venom-allergic patients, ranging from local up to severe systemic reactions and even fatal anaphylaxis. Allergic patients' quality of life can be mainly improved by altering their immune response to tolerate the venoms...... by injecting increasing venom doses over years. This venom-specific immunotherapy is highly effective and well tolerated. However, component-resolved information about the venoms has increased in the last years. This knowledge is not only able to improve diagnostics as basis for an accurate therapy......, but was additionally used to create tools which enable the analysis of therapeutic venom extracts on a molecular level. Therefore, during the last decade the detailed knowledge of the allergen composition of hymenoptera venoms has substantially improved diagnosis and therapy of venom allergy. This review focuses...

  10. Allergen immunotherapy: routes, safety,efficacy, and mode of action

    Directory of Open Access Journals (Sweden)

    Hochfelder JL

    2013-07-01

    Full Text Available Jillian Leigh Hochfelder, Punita PondaDivision of Allergy and Immunology, North Shore–Long Island Jewish Health System, New Hyde Park, NY, USAAbstract: Allergic rhinitis, allergic conjunctivitis, and allergic asthma have been steadily increasing in prevalence in recent years. These allergic diseases have a major impact on quality of life and are a major economic burden in the US. Although allergen avoidance and pharmacotherapy are currently the mainstays of therapy, they are not always successful in treating patients’ symptoms effectively. If a patient fails allergen avoidance and medical therapy, immunotherapy may be indicated. Furthermore, immunotherapy is the only therapy that may change the course of the disease and induce long-term remission. Though subcutaneous administration has been the standard route for immunotherapy for many decades, there are several other routes of administration that have been and are currently being studied. The goal of utilizing alternative routes of immunotherapy is to improve safety without decreasing the efficacy of treatment. This paper will review the novel routes of immunotherapy, including sublingual, oral, local nasal, epicutaneous, and intralymphatic.Keywords: immunotherapy, allergic rhinitis, allergic asthma, sublingual, intralymphatic

  11. Effects of Maillard reaction on allergenicity of buckwheat allergen Fag t 3 during thermal processing.

    Science.gov (United States)

    Yang, Zhen-Huang; Li, Chen; Li, Yu-Ying; Wang, Zhuan-Hua

    2013-04-01

    Fag t 3 is a major allergenic protein in tartary buckwheat. The Maillard reaction commonly occurs in food processing, but few studies have been conducted on the influence of thermal processing on the allergenic potential of buckwheat allergen. The aim of the present study was to investigate the effects of autologous plant polysaccharides on the immunoreactivity of buckwheat Fag t 3 (11S globulin) following the Maillard reaction. Fag t 3 and crude polysaccharides were prepared from tartary buckwheat (Fagopyrum tataricum) flour. After heating, the polysaccharides were covalently linked to Fag t 3 via a Maillard reaction, and the IgE/IgG-binding properties of Fag t 3 decreased dramatically, with significant changes also being observed in the electrophoretic mobility, secondary structure and solubility of the glycated Fag t 3. The great influence of glycation on IgE/IgG binding to Fag t 3 was correlated with a significant change in the structure and epitopes of the allergenic protein. These data indicated that conjugation of polysaccharides to Fag t 3 markedly reduced the allergen's immunoreactivity. Glycation that occurs via the Maillard reaction during the processing of buckwheat food may be an efficient method to reduce Fag t 3 allergenicity. © 2012 Society of Chemical Industry.

  12. Inhibition of serum naphthylamidases by atopic allergens

    NARCIS (Netherlands)

    Berrens, L.

    1968-01-01

    Human serum hydrolyses the 2-naphthylamides of -lysine and -arginine. The enzymatic activity of the serum naphthylamidases is inhibited by atopic allergens and by protein-sugar conjugates carrying 1-deoxy-2-ketose residues attached to the -amino groups of lysine residues in their peptide moieties.

  13. Chemicals and proteins as allergens and adjuvants

    NARCIS (Netherlands)

    Kapsenberg, M. L.

    1996-01-01

    Small molecular weight chemicals may irritate tissues via the induction of the production of various proinflammatory and chemotactic molecules. The structure of these irritants is heterogenous, as is the pattern of their effects. Chemicals are potentially allergenic (i.e. haptens) when they are able

  14. Consumer-friendly food allergen detection

    NARCIS (Netherlands)

    Ross, Georgina M.S.; Bremer, Monique G.E.G.; Nielen, Michel W.F.

    2018-01-01

    In this critical review, we provide a comprehensive overview of immunochemical food allergen assays and detectors in the context of their user-friendliness, through their connection to smartphones. Smartphone-based analysis is centered around citizen science, putting analysis into the hands of the

  15. The carbohydrate moiety of house dust allergen

    NARCIS (Netherlands)

    Morris, J.H.; Berrens, L.; Young, E.

    1965-01-01

    Purified house dust allergen has been found to contain monosaccharides of plant and animal origin, and has been partially fractionated by DEAE-Sephadex column chromatography. The carbohydrate is highly branched, suffering little attack during periodate oxidation. Furanose configuration for a

  16. Zinc diethyldithiocarbamate allergenicity: potential haptenation mechanisms.

    Science.gov (United States)

    Chipinda, Itai; Hettick, Justin M; Simoyi, Reuben H; Siegel, Paul D

    2008-08-01

    Zinc diethyldithiocarbamate (ZDEC) and its disulfide, tetraethylthiuram disulfide (TETD), are rubber accelerators and contact allergens that cross-react in some individuals. This study explored potential protein haptenation mechanisms of ZDEC and its oxidation products. ZDEC oxidation/reduction products and sites of protein binding were assessed using high-performance liquid chromatography and mass spectrometry. The murine local lymph node assay (LLNA) was employed to probe haptenation mechanisms of ZDEC by examining its allergenicity along with its oxidation products and through elimination of oxidation and chelation mechanisms by substituting cobalt for zinc [cobalt (II) dithiocarbamate, CoDEC]. Oxidation of ZDEC by hypochlorous acid (bleach, HOCl), iodine, or hydrogen peroxide resulted in production of TETD, tetraethylthiocarbamoyl disulfide, and tetraethyldicarbamoyl disulfide (TEDCD). Albumin thiols reduced TETD with subsequent mixed disulfide formation/haptenation. ZDEC directly chelated the copper ion on the active site of the superoxide dismutase, whereas CoDEC did not bind to Cu proteins or form mixed disulfides with free thiols. ZDEC, sodium diethyldithiocarbamate, TEDCD, and TETD were all positive in the LLNA except CoDEC, which was non-allergenic. The thiol is the critical functional group in ZDEC's allergenicity, and haptenation is predominantly through chelation of metalloproteins and formation of mixed disulfides.

  17. Isoeugenol is an important contact allergen

    DEFF Research Database (Denmark)

    White, I R; Johansen, J D; Gimenéz-Arnau, Elena

    1999-01-01

    The prevalence of contact allergy to the fragrance mix in individuals with eczema is up to 10%. Within the mix, isoeugenol (CAS 97-54-1) is an important individual allergen. Until May 1998, the IFRA (International Fragrance Association) guidelines suggested that isoeugenol could safely be used at...

  18. Update on allergen detection in childhood asthma.

    African Journals Online (AJOL)

    False-negative results occur in patients who have true IgE mediated disease as confirmed by skin testing or allergen challenge. The sensitivity of blood allergy testing is approximately 25% to 30% lower than that of skin testing, based on comparative studies.13,14. Limitations of blood testing of specific IgE. Levels of specific ...

  19. Innate immune responses to environmental allergens

    NARCIS (Netherlands)

    Kauffman, HF

    Aero-allergens, including plant pollens, house dust mite particles, fungal spores, and mycelium fragments, are continuously inhaled and deposited on the airway mucosa. These particles and their soluble components actively interact with innate recognition systems present in the mucosal layer (e.g.,

  20. Inhibition of secretary PLA₂--VRV-PL-VIIIa of Russell's viper venom by standard aqueous stem bark extract of Mangifera indica L.

    Science.gov (United States)

    Dhananjaya, B L; Sudarshan, S

    2015-03-01

    The aqueous extract of Mangifera indica is known to possess anti-snake venom activities. However, its inhibitory potency and mechanism of action on multi-toxic phospholipases A2s, which are the most toxic and lethal component of snake venom is still unknown. Therefore, this study was carried out to evaluate the modulatory effect of standard aqueous bark extract of M. indica on VRV-PL-VIIIa of Indian Russells viper venom. Mangifera indica extract dose dependently inhibited the GIIB sPLA2 (VRV-PL-VIIIa) activity with an IC50 value of 6.8±0.3 μg/ml. M. indica extract effectively inhibited the indirect hemolytic activity up to 96% at ~40 μg/ml concentration. Further, M. indica extract at different concentrations (0-50 μg/ml) inhibited the edema formed in a dose dependent manner. It was found that there was no relieve of inhibitory effect of the extract when examined as a function of increased substrate and calcium concentration. The inhibition was irreversible as evident from binding studies. The in vitro inhibition is well correlated with in situ and in vivo edema inducing activities. As the inhibition is independent of substrate, calcium concentration and was irreversible, it can be concluded that M. indica extracts mode of inhibition could be due to direct interaction of components present in the extract with PLA2 enzyme. In conclusion, the aqueous extract of M. indica effectively inhibits svPLA2 (Snake venom phospholipase A2) enzymatic and its associated toxic activities, which substantiate its anti-snake venom properties. Further in-depth studies are interesting to known on the role and mechanism of the principal inhibitory constituents present in the extract, so as to develop them into potent anti-snake venom and as an anti-inflammatory agent.

  1. Improvement in botanical standardization of commercial freeze-dried herbal extracts by using the combination of antioxidant capacity and constituent marker concentrations.

    Science.gov (United States)

    Ninfali, Paolino; Gennari, Lorenzo; Biagiotti, Enrica; Cangi, Francesca; Mattoli, Luisa; Maidecchi, Anna

    2009-01-01

    Botanical extracts are standardized to > or = 1 marker compounds (MCs). This standardization provides a certain level of quality control, but not complete quality assurance. Thus, industries are looking for other satisfactory systems to improve standardization. This study focuses on the standardization of herbal medicines by combining 2 parameters: the concentration of the MC and antioxidant capacity. Antioxidant capacity was determined with the oxygen radical absorbance capacity (ORAC) method and the concentrations of the MCs, by high-performance liquid chromatography. Total phenols were also determined by the Folin-Ciocolteau method. The ORAC values, expressed as micromol Trolox equivalents/100 g (ORAC %), of 12 commercial herbal extracts were related to the ORAC values of the respective pure MCs at the concentrations at which the MCs occur in products (ORAC-MC %). The ORAC % values of 11 extracts were higher than those of the respective MCs and the ratios ORAC-MC %/ORAC % ranged from 0.007 to 0.7, whereas in the case of Olea europaea leaves, the same ratio was 1.36. The ORAC parameters and their ratios, as well as the linear relationship between ORAC-MC % and ORAC %, are described and discussed as tools for improving the standardization of herbal products and detecting modifications due to herb processing and storage.

  2. 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)

  3. Immunochemical estimations of allergenic activities from outdoor aero-allergens, collected by a high-volume air sampler

    DEFF Research Database (Denmark)

    Jensen, J; Poulsen, L K; Mygind, K

    1989-01-01

    To quantify airborne allergens in amorphus and morphological particles, a survey with collection of aero-allergens on glass fibre filters by means of a high-volume air-sampler (HIVOL) was conducted. In preliminary laboratory experiments we compared various filter elution techniques, and the pulve......To quantify airborne allergens in amorphus and morphological particles, a survey with collection of aero-allergens on glass fibre filters by means of a high-volume air-sampler (HIVOL) was conducted. In preliminary laboratory experiments we compared various filter elution techniques...... method for quantification of airborne allergens was validated, as a high recovery of timothy grass pollen allergens was eluted from air filters, and eluates were shown specific by RAST inhibition. The amount of immunochemically measured airborne timothy and birch allergens collected by means of the HIVOL...

  4. Human serum substitution by artificial sera of scalable allergen reactivity based on polyclonal antibodies and chimeras of human FcγRI and IgE domains.

    Science.gov (United States)

    Offermann, N; Plum, M; Hübner, U; Rathloff, K; Braren, I; Fooke, M; Spillner, E

    2016-12-01

    Human sera are the first choice as controls for diagnostic applications such as immunoassays, but are limited regarding availability, varying quality, and high costs. In this study, we aimed to circumvent these limitations by the use of a chimeric adaptor molecule comprising the extracellular domains of the human FcγRI (CD64) fused with human IgE Fc domains (CD64-IgE Fc). Allergen-specific antibodies were produced in rabbits using eight different allergens, extracts, and allergen mixtures including mites, pollen, drugs, and food. Preincubation of polyclonal IgG with CD64-IgE Fc established allergen-specific artificial sera that showed comparable results for more than 20 allergens and allergen extracts in three diagnostic systems for the determination of specific IgE. The agreement for these artificial sera is within ±1 radioallergosorbent test (RAST) class. Hence, rabbit IgG complexed with the IgG-specific CD64-IgE Fc adaptor molecule could provide a substitute for human reference sera with specificity for virtually any protein of interest. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. The potential of papain and alcalase enzymes and process optimizations to reduce allergenic gliadins in wheat flour.

    Science.gov (United States)

    Li, Ying; Yu, Jianmei; Goktepe, Ipek; Ahmedna, Mohamed

    2016-04-01

    The objectives of this study were to select effective enzymes that catalyze the hydrolysis of allergenic proteins, gliadins, in wheat flour and to optimize the enzymatic treatment conditions. Six proteases were tested. Hydrolyzed samples were tested for residual gliadin concentrations and in vitro allergenicity. The hydrolysis conditions of wheat protein by the effective enzymes were optimized by central composite design. Results showed that alcalase from Bacillus licheniformis, and papain from latex of papaya fruit had greater ability to reduce gliadin content of wheat flour than flavourzyme, pepsin, trypsin or α-chymotrypsin. The sequential-treatment of wheat flour by alcalase-papain was more effective in reducing gliadin content than single enzyme treatment. Under the optimal conditions of sequential enzymatic treatment, gliadin was almost completely removed, resulting in the flour extract showing lowest IgE-binding. Therefore, this could be a promising biotechnology for preparing low allergenic wheat products. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Identification and characterization of DC-SIGN-binding glycoproteins in allergenic foods.

    Science.gov (United States)

    Kamalakannan, M; Chang, L M; Grishina, G; Sampson, H A; Masilamani, M

    2016-08-01

    DC-SIGN (dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin) is a C-type lectin receptor expressed on macrophages and dendritic cells. DC-SIGN has high affinity for fucosylated glycans in several plant glycoproteins and pathogens. DC-SIGN is thought to be crucial for the development of allergic sensitization. However, the precise role of DC-SIGN in food allergy pathogenesis is not yet understood. We sought to characterize DC-SIGN-binding glycoproteins in a panel of allergenic and non-allergenic foods. Fluorescent-labeled peanut and soy extracts were used to test protein binding to human monocyte-derived dendritic cells (DCs) by flow cytometry. DC-SIGN-blocking assays were performed by incubating DCs with food extracts followed by staining with anti-DC-SIGN antibody. Using a DC-SIGN-Fc chimera, food extracts were tested for binding by ELISA and autoradiography. IgE immunoblotting was performed with pooled sera from food-allergic subjects. DC activation and maturation were assessed by flow cytometry. We demonstrate that peanut agglutinin, a minor peanut allergen, is a novel ligand for DC-SIGN. Peanut agglutinin activates DCs to induce the expression of costimulatory molecules in vitro. We present a comprehensive report on the characterization of DC-SIGN-binding proteins in common allergenic foods such as peanut, soy, tree nuts, egg, and milk. Foods that rarely induce allergy, such as pine nuts, chickpea, and corn, showed no binding to DC-SIGN. Several DC-SIGN-binding proteins show reactivity in serum IgE immunoblots. We have also identified novel non-IgE-binding proteins that interact with DC-SIGN; these proteins may be important for regulating immune responses to these foods. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  7. A standardized Humulus lupulus (L.) ethanol extract partially prevents ovariectomy-induced bone loss in the rat without induction of adverse effects in the uterus.

    Science.gov (United States)

    Keiler, Annekathrin M; Helle, Janina; Bader, Manuela I; Ehrhardt, Tino; Nestler, Kristin; Kretzschmar, Georg; Bernhardt, Ricardo; Vollmer, Günter; Nikolić, Dejan; Bolton, Judy L; Pauli, Guido F; Chen, Shao-Nong; Dietz, Birgit M; van Breemen, Richard B; Zierau, Oliver

    2017-10-15

    Hops (Humulus lupulus (L.)) dietary supplements are of interest as herbal remedies to alleviate menopausal symptoms, such as hot flushes, depression and anxiety. So far, the evidence regarding estrogenic and related properties of hops preparations has been considered insufficient for a market authorization for menopausal indications. The study aims to investigate a chemically standardized hops extract regarding its safety in the uterus, as wells as its efficacy to prevent bone loss in the ovariectomized rat model. Female Wistar rats were ovariectomized and divided into a control group receiving phytoestrogen-free diet, a group treated with E 2 benzoate (0.93 mg/kg body weight/d) and a group treated with the standardized hops extract (60 mg/kg body weight/d) for 8 weeks. Micro-computed tomography of the tibiae and vertebrae, as wells as histological changes in the uterus and tibia were analyzed. Neither uterotrophic nor proliferative effects were observed in the endometrium in response to the oral 8-week administration of the hops extract. However, site-dependent skeletal effects were observed. The hops extract significantly decreased the number of osteoclasts in the tibial metaphysis and prevented reduction of the trabecular thickness that resulted from estradiol depletion. In contrast, the hops extract did not prevent the ovariectomy-induced micro-architectural changes in the lumbar vertebra. Certain parameters (e.g. thickness and number of trabeculae) were even found to be below the values determined in the ovariectomized control group. Taken together, the results provide evidence for the safety of the standardized hops extract and point to a weak bone type-specific, protective effect on bone loss following estradiol depletion. Copyright © 2017. Published by Elsevier GmbH.

  8. Fragrance allergy could be missed without patch testing with 26 individual fragrance allergens.

    Science.gov (United States)

    Vejanurug, Patnapa; Tresukosol, Poohglin; Sajjachareonpong, Praneet; Puangpet, Pailin

    2016-04-01

    In 2003, the EU Cosmetics Directive stated that 26 fragrance substances must be listed on the cosmetic product ingredient labels. Not all of these 26 fragrance substances are detected by the usual screening markers comprising fragrance mix I, fragrance mix II, and Myroxylon pereirae. To evaluate the usefulness of testing with the 26 individual fragrance substances in addition to the standard fragrance screening markers. Three hundred and twelve consecutive patients were patch tested with our baseline series and the 26 specific fragrance substances required to be declared on cosmetic product ingredient labels in accordance with the EU Cosmetics Directive. Positive reactions to at least either one of the 26 individual fragrance substances or the usual fragrance screening markers were seen in 84 of 312 patients (26.9%). Fifteen of these 84 patients (17.8%) reacted negatively to the fragrance screening markers. The most common individual fragrance allergens were cinnamyl alcohol (11.2%), cinnamal (9%), and hydroxycitronellal (3.8%). Sixty-two of 312 patients (19.8%) had at least one positive reaction to the fragrance screening markers. Additional patch testing with the 26 individual fragrance allergens, or with the commonest fragrance allergens identified within these 26, should be performed to optimize the detection of fragrance allergy. Cinnamyl alcohol and cinnamal are important fragrance allergens in Thailand. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  9. Impacts of air pollution exposure on the allergenic properties of Arizona cypress pollens

    Science.gov (United States)

    Shahali, Y.; Pourpak, Z.; Moin, M.; Zare, A.; Majd, A.

    2009-02-01

    Epidemiological studies have demonstrated that urbanization and high levels of vehicle emissions correlated with the increasing trend of pollen-induced respiratory allergies. Numerous works have investigated the role of pollutants in the pathogenesis of respiratory diseases but impacts of anthropogenic pollution on pollen allergenic properties are still poorly understood. The objective of this survey was to evaluate impacts of the traffic-related pollution on the structure and allergenic protein content of Arizona cypress (Cupressus arizonica, CA) pollens, recognized as a rising cause of seasonal allergy in various regions worldwide. According to our results, traffic-related air pollution by its direct effects on the elemental composition of pollens considerably increased the fragility of the pollen exine, causing numerous cracks in its surface and facilitating pollen content liberation. Pollen grains were also covered by numerous submicronic orbicules which may act as effective vectors for pollen-released components into the lower regions of respiratory organs. On the other hand, this study provides us reliable explications about the low efficiency of standard commercial allergens in the diagnosis of the Arizona cypress pollen allergy in Tehran. Although traffic related pollution affects the allergenic components of CA pollens, the repercussions on the respiratory health of urban populations have yet to be clarified and need further investigations.

  10. The human skin/chick chorioallantoic membrane model accurately predicts the potency of cosmetic allergens.

    Science.gov (United States)

    Slodownik, Dan; Grinberg, Igor; Spira, Ram M; Skornik, Yehuda; Goldstein, Ronald S

    2009-04-01

    The current standard method for predicting contact allergenicity is the murine local lymph node assay (LLNA). Public objection to the use of animals in testing of cosmetics makes the development of a system that does not use sentient animals highly desirable. The chorioallantoic membrane (CAM) of the chick egg has been extensively used for the growth of normal and transformed mammalian tissues. The CAM is not innervated, and embryos are sacrificed before the development of pain perception. The aim of this study was to determine whether the sensitization phase of contact dermatitis to known cosmetic allergens can be quantified using CAM-engrafted human skin and how these results compare with published EC3 data obtained with the LLNA. We studied six common molecules used in allergen testing and quantified migration of epidermal Langerhans cells (LC) as a measure of their allergic potency. All agents with known allergic potential induced statistically significant migration of LC. The data obtained correlated well with published data for these allergens generated using the LLNA test. The human-skin CAM model therefore has great potential as an inexpensive, non-radioactive, in vivo alternative to the LLNA, which does not require the use of sentient animals. In addition, this system has the advantage of testing the allergic response of human, rather than animal skin.

  11. Impacts of air pollution exposure on the allergenic properties of Arizona cypress pollens

    International Nuclear Information System (INIS)

    Shahali, Y; Pourpak, Z; Moin, M; Zare, A; Majd, A

    2009-01-01

    Epidemiological studies have demonstrated that urbanization and high levels of vehicle emissions correlated with the increasing trend of pollen-induced respiratory allergies. Numerous works have investigated the role of pollutants in the pathogenesis of respiratory diseases but impacts of anthropogenic pollution on pollen allergenic properties are still poorly understood. The objective of this survey was to evaluate impacts of the traffic-related pollution on the structure and allergenic protein content of Arizona cypress (Cupressus arizonica, CA) pollens, recognized as a rising cause of seasonal allergy in various regions worldwide. According to our results, traffic-related air pollution by its direct effects on the elemental composition of pollens considerably increased the fragility of the pollen exine, causing numerous cracks in its surface and facilitating pollen content liberation. Pollen grains were also covered by numerous submicronic orbicules which may act as effective vectors for pollen-released components into the lower regions of respiratory organs. On the other hand, this study provides us reliable explications about the low efficiency of standard commercial allergens in the diagnosis of the Arizona cypress pollen allergy in Tehran. Although traffic related pollution affects the allergenic components of CA pollens, the repercussions on the respiratory health of urban populations have yet to be clarified and need further investigations.

  12. Quantitative analysis of the 26 allergens for cosmetic labeling in fragrance raw materials and perfume oils.

    Science.gov (United States)

    Leijs, Hans; Broekhans, Joost; van Pelt, Leon; Mussinan, Cynthia

    2005-07-13

    The adoption of the 7th amendment of the European Cosmetic Directive 76/768/EEC requires any cosmetic product containing any of 26 raw materials identified by the Scientific Committee on Cosmetic Products and Non-Food Products intended for Consumers as likely to cause a contact allergy when present above certain trigger levels to be declared on the package label. Of these 26, 24 are volatile and can be analyzed by GC. This paper describes a method for the quantitative analysis of these volatile raw materials in perfume ingredients as well as complex perfume compositions. The method uses sequential dual-column GC-MS analysis. The full-scan data acquired minimize the false-positive and false-negative identifications that can be observed with alternate methods based on data acquired in the SIM mode. For each sample, allergen levels are determined on both columns sequentially, leading to two numerical results for each allergen. Quantification limits for each allergen in a perfume mixture based on the analysis of a standard are 0.999) and stable for multiple days. Studies on perfumes spiked with multiple allergens at 30, 50, and 70 mg/kg show recoveries close to nominal values.

  13. Magnetophoretic immunoassay of allergen-specific IgE in an enhanced magnetic field gradient.

    Science.gov (United States)

    Hahn, Young Ki; Jin, Zongwen; Kang, Joo H; Oh, Eunkeu; Han, Min-Kyu; Kim, Hak-Sung; Jang, Jung-Tak; Lee, Jae-Hyun; Cheon, Jinwoo; Kim, Seung Hyun; Park, Hae-Sim; Park, Je-Kyun

    2007-03-15

    We demonstrate a novel magnetophoretic immunoassay of allergen-specific immunoglobulin E (IgE) based on the magnetophoretic deflection velocity of a microbead that is proportional to the associated magnetic nanoparticles under enhanced magnetic field gradient in a microchannel. In this detection scheme, two types of house dust mites, Dermatophagoides farinae (D. farinae) and Dermatophagoides pteronyssinus (D. pteronyssinus), were used as the model allergens. Polystyrene microbeads were conjugated with each of the mite extracts followed by incubation with serum samples. The resulting mixture was then reacted with magnetic nanoparticle-conjugated anti-human IgE for detection of allergen-specific IgE by using sandwich immuno-reactions. A ferromagnetic microstructure combined with a permanent magnet was employed to increase the magnetic field gradient ( approximately 10(4) T/m) in a microfluidic device. The magnetophoretic velocities of microbeads were measured in a microchannel under applied magnetic field, and the averaged velocity was well correlated with the concentration of allergen-specific IgE in serum. From the analysis of pooled sera obtained from 44 patients, the detection limits of the allergen-specific human IgEs for D. farinae and D. pteronyssinus were determined to be 565 (0.045 IU/mL) and 268 fM (0.021 IU/mL), respectively. These values are 1 order of magnitude lower than those by a conventional CAP system. For evaluation of reproducibility and accuracy, unknown sera were subjected to a blind test by using the developed assay system, and they were compared with the CAP system. As a result, coefficient of variance was less than 10%, and the developed method enabled a fast assay with a tiny amount of serum ( approximately 10 microL).

  14. 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.

  15. Sensitization to cat and dog allergen molecules in childhood and prediction of symptoms of cat and dog allergy in adolescence: A BAMSE/MeDALL study.

    Science.gov (United States)

    Asarnoj, Anna; Hamsten, Carl; Wadén, Konrad; Lupinek, Christian; Andersson, Niklas; Kull, Inger; Curin, Mirela; Anto, Josep; Bousquet, Jean; Valenta, Rudolf; Wickman, Magnus; van Hage, Marianne

    2016-03-01

    Sensitization to individual cat and dog allergen molecules can contribute differently to development of allergy to these animals. We sought to investigate the association between sensitization patterns to cat and dog allergen molecules during childhood and symptoms to these furry animals up to age 16 years. Data from 779 randomly collected children from the Barn/Children Allergy/Asthma Milieu Stockholm Epidemiologic birth cohort at 4, 8, and 16 years were used. IgE levels to cat and dog were determined by using ImmunoCAP, and levels to allergen molecules were determined by using an allergen chip based on ISAC technology (Mechanisms for the Development of Allergy chip). Allergy was defined as reported rhinitis, conjunctivitis, or asthma at exposure to cat or dog. Cross-sectionally, IgE to Fel d 1 and cat extract had similar positive predictive values for cat allergy. IgE to Can f 1 showed a higher positive predictive value for dog allergy than dog extract IgE. Sensitizations to Fel d 1 and Can f 1 in childhood were significantly associated with symptoms to cat or dog at age 16 years. Polysensitization to 3 or more allergen molecules from cat or dog was a better longitudinal predictor of cat or dog symptoms than results of IgE tests with cat or dog allergen extract, respectively. Cross-sectionally, cat/dog-polysensitized children had higher IgE levels and more frequent symptoms to cat and dog than monosensitized children. Sensitization to Fel d 1 and Can f 1 in childhood and polysensitization to either cat or dog allergen molecules predict cat and dog allergy cross-sectionally and longitudinally significantly better than IgE to cat or dog extract. Copyright © 2015 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

  16. Inhibitory activity of a standardized elderberry liquid extract against clinically-relevant human respiratory bacterial pathogens and influenza A and B viruses

    Directory of Open Access Journals (Sweden)

    Domann Eugen

    2011-02-01

    Full Text Available Abstract Background Black elderberries (Sambucus nigra L. are well known as supportive agents against common cold and influenza. It is further known that bacterial super-infection during an influenza virus (IV infection can lead to severe pneumonia. We have analyzed a standardized elderberry extract (Rubini, BerryPharma AG for its antimicrobial and antiviral activity using the microtitre broth micro-dilution assay against three Gram-positive bacteria and one Gram-negative bacteria responsible for infections of the upper respiratory tract, as well as cell culture experiments for two different strains of influenza virus. Methods The antimicrobial activity of the elderberry extract was determined by bacterial growth experiments in liquid cultures using the extract at concentrations of 5%, 10%, 15% and 20%. The inhibitory effects were determined by plating the bacteria on agar plates. In addition, the inhibitory potential of the extract on the propagation of human pathogenic H5N1-type influenza A virus isolated from a patient and an influenza B virus strain was investigated using MTT and focus assays. Results For the first time, it was shown that a standardized elderberry liquid extract possesses antimicrobial activity against both Gram-positive bacteria of Streptococcus pyogenes and group C and G Streptococci, and the Gram-negative bacterium Branhamella catarrhalis in liquid cultures. The liquid extract also displays an inhibitory effect on the propagation of human pathogenic influenza viruses. Conclusion Rubini elderberry liquid extract is active against human pathogenic bacteria as well as influenza viruses. The activities shown suggest that additional and alternative approaches to combat infections might be provided by this natural product.

  17. Immunoproteomic characterization of Ambrosia artemisiifolia pollen allergens in canine atopic dermatitis.

    Science.gov (United States)

    Ognjenovic, Jana; Milcic-Matic, Natalija; Smiljanic, Katarina; Vuckovic, Olga; Burazer, Lidija; Popovic, Nikola; Stanic-Vucinic, Dragana; Velickovic, Tanja Cirkovic

    2013-09-01

    Canine atopic dermatitis (CAD) is an immune system disorder that affects 10-15% of the canine population. Short ragweed (Ambrosia artemisiifolia) pollen represents one of the major seasonal sources of allergenic pollen proteins in Europe, particularly in the Pannonian valley of the Balkan region. In Serbia, about 66% of atopic dogs showed a positive intradermal skin test with its pollen extract, which is second to house dust mites. Therefore, characterization of Ambrosia artemisiifolia pollen components, in terms of defining major and minor allergens that induce clinically manifested allergic reaction in dogs, is important for valid diagnosis and efficient therapy. This study has, for the first time, characterized and identified major Ambrosia artemisiifolia allergens in CAD, using an immunoproteomic approach. To assess the prevalence of specific IgE in electrophoretically separated ragweed pollen proteins, individual reactivity of sera from dogs with CAD was analyzed and compared to the reactivity of sera from healthy dogs in the non-reducing conditions, which were found optimal for specific canine IgE detection. A specific IgE band (38 kDa) was recognized as the most dominant allergen in CAD, occurring in 81% of positive dog's sera. 2-D immunoblotting followed by a mass spectrometry peptide fingerprint analyses with pooled canine and human atopic sera, revealed that 38 kDa major Ambrosia atremisiifolia allergens in CAD were all five isoallergens of the Amb a 1 group (antigen E), including the previously named Amb a 2 (antigen K). In contrast to canine sera, human atopic sera also recognized lower mass allergens such as the β fragment of Amb a 1 and profilins (Amb a 8 variants). The most prominent ragweed proteins in CAD, represent, as in humans, variants of all five isoallergens of the Amb a 1 group (pectate lyase): Amb a 1.0101 and its natural variant E1XUL2, Amb a 1.0202, 1.0304, 1.0402 and the natural variant of Amb a 1.0501, E1XUM0, as well as the

  18. Platelets Play a Central Role in Sensitization to Allergen.

    Science.gov (United States)

    Amison, Richard T; Cleary, Simon; Riffo-Vasquez, Yanira; Bajwa, Maidda; Page, Clive P; Pitchford, Simon C

    2018-01-24

    Platelet activation occurs in patients with allergic inflammation, and platelets can be activated directly by allergen via an IgE-dependent process. Platelets activate antigen-presenting cells (APCs) such as CD11c+ dendritic cells (DCs) in vitro. Whilst CD11c+ DCs are requisite for allergen sensitization, the role of platelets in this process is unknown. We investigated whether platelets were necessary for allergen sensitization. Balb/c mice sensitised to ovalbumin (OVA), were exposed to subsequent aerosolized allergen (OVA challenge). We analysed lung CD11c+ cell activation, co-localization with platelets, and some other indices of inflammation. The role of platelets at the time of allergen sensitization was assessed through platelet depletion experiments restricted to the period of sensitization. Platelets co-localized with airway CD11c+ cells, and this association increased after allergen sensitization, and after subsequent allergen exposure. Temporary platelet depletion (>95%) at the time of allergen sensitization led to a suppression of IgE and IL-4 synthesis, and a decrease in the pulmonary recruitment of eosinophils, macrophages, and lymphocytes after subsequent allergen exposure. Furthermore, in mice previously depleted of platelets at the time of sensitization, the recovered platelet population were shown to have reduced expression of FcɛRI. Pulmonary CD11c+ cell recruitment was suppressed in these mice after allergen challenge, suggesting the migration of CD11c+ cells in vivo may be dependent on direct platelet recognition of allergen. Platelets are necessary for efficient host sensitization to allergen. This propagates the subsequent inflammatory response during secondary allergen exposure, and increases platelet association with airway CD11c+ cells.

  19. Allergy to guinea pigs: II Identification of specific allergens in guinea pig dust by crossed radio-immunoelectrophoresis and investigation of the possible origin.

    Science.gov (United States)

    Walls, A F; Newman Taylor, A J; Longbottom, J L

    1985-11-01

    An extract of dust from the air-vent filters of a room housing guinea pigs was analysed by quantitative immunoelectrophoretic procedures and compared with extracts of various materials derived from guinea pigs. Crossed radio-immunoelectrophoresis (CRIE) of the dust, performed with sera from twenty asthmatic patients who were positive by skin testing and RAST to guinea pig extracts, identified fourteen IgE-binding constituents. Although responses varied, most sera reacted with four particular allergens, antigens 2, 3, 10 and Sl. The numbers of allergens recognized by individual patients correlated with the RAST score, but not with total serum IgE. All seventeen dust constituents detected by crossed immunoelectrophoresis (and all four major allergens), were also present in extracts of guinea pig dander, fur, saliva and urine; several of these components were absent in an epithelial extract, and there were even less in preparations of shaved pelt, serum or faeces. None of the dust extract antigens were detected in materials used in animal husbandry, dust samples from rooms without guinea pigs, or a D. pteronyssinus extract. These findings suggest that inhalant allergens may be derived predominantly from material shed from the guinea pig coat after contamination with saliva, and possibly to a lesser extent, urine.

  20. Pin p 1 is a major allergen in pine nut and the first food allergen described in the plant group of gymnosperms.

    Science.gov (United States)

    Cabanillas, Beatriz; Crespo, Jesus F; Maleki, Soheila J; Rodriguez, Julia; Novak, Natalija

    2016-11-01

    This study aimed to report the complete sequence of a 2S albumin purified from pine nut and to analyze its allergenic properties. Individual recognition of this protein by serum IgE from pine nut-allergic patients was assessed. IgE cross-linking capacity was analyzed in a basophil activation test. Inhibition of IgE-binding and stability to heating was also assessed. The complete nucleotide sequence was obtained and a phylogenetic study was carried out. 2S albumin from pine nut (registered as Pin p 1.0101) was recognized by IgE of 75% of sera. The allergen was heat-stable and had a robust capacity to inhibit IgE-binding to whole pine nut extract. The IgE cross-linking capacity of Pin p 1 on basophils was also demonstrated. Despite the low homology of Pin p 1 sequence with other allergenic 2S albumins from angiosperms, Pin p 1 contains the typical skeleton of 8 cysteine residues, important for its α-helixes enriched structure. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. From classical taxonomy to genome and metabolome: towards comprehensive quality standards for medicinal herb raw materials and extracts.

    Science.gov (United States)

    Govindaraghavan, Suresh; Hennell, James R; Sucher, Nikolaus J

    2012-09-01

    Fundamental to herbal medicine quality is the use of 'authentic' medicinal herb species. Species, however, 'represent more or less arbitrary and subjective man-made units'. Against this background, we discuss, with illustrative examples, the importance of defining species boundaries by accommodating both the fixed (shared) diagnostic and varying (within-species) traits in medicinal herb populations. We emphasize the role of taxonomy, floristic information and genomic profiling in authenticating medicinal herb species, in addition to the need to include within species phytochemical profile variations while developing herbal extract identification protocols. We outline the application of species-specific genomic and phytochemical markers, chemoprofiling and chemometrics as additional tools to develop qualifying herbal extract references. We list the diagnostic traits available subsequent to each step during the medicinal herb extract manufacturing process and delineate limits to qualification of extract references. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Variation in allergen content in sublingual allergen immunotherapy with house dust mites.

    Science.gov (United States)

    Moreno Benítez, F; Espinazo Romeu, M; Letrán Camacho, A; Mas, S; García-Cózar, F J; Tabar, A I

    2015-11-01

    Allergen immunotherapy is a treatment modality which can be applied using different vaccines. The aim of this study was to quantify and compare the allergen content of different house dust mites (HDM)' sublingual treatments and to review the evidence on their efficacy. Five sublingual allergen immunotherapy (SLIT) products were ordered and purchased at an ordinary pharmacy and masked for blinding before the study was started. Detection of Dermatophagoides pteronyssinus and Dermatophagoides farinae allergens Der p 1, Der f 1, Der p 2 and Der f 2 was carried out by immunoblotting and fluorescent multiplex. A literature search for meta-analyses and systematic reviews that included SLIT-HDM products was performed. Der p 1 concentrations ranged from 0.6 to 14.5 μg/ml; similar figures were found for Der f 1 that ranged from 0.2 to 12.4 μg/ml. Der p 2+ Der f 2 ranged from 0.2 to 1.5 μg/ml. Data on efficacy are scarce for most of the five products. Substantial variations regarding allergen content were found among these five SLIT-HDM products. Therefore, it can be necessary to guarantee the quality of the SLIT-HDM products and to demonstrate their effectiveness before they are marketed. It seems necessary, for the moment, to take into account these characteristics of the products before prescribing. © 2015 The Authors. Allergy Published by John Wiley & Sons Ltd.

  3. Pharmacognostical Analysis and Protective Effect of Standardized Extract and Rizonic Acid fromErythrina velutinaagainst 6-Hydroxydopamine-Induced Neurotoxicity in SH-SY5Y Cells.

    Science.gov (United States)

    Silva, Aline H; Fonseca, Francisco Noé; Pimenta, Antônia T A; Lima, MaryAnne S; Silveira, Edilberto Rocha; Viana, Glauce S B; Vasconcelos, Silvânia M M; Leal, Luzia Kalyne A M

    2016-01-01

    Erythrina velutina is a tree common in the northeast of Brazil extensively used by traditional medicine for the treatment of central nervous system disorders. To develop a standardized ethanol extract of E. velutina (EEEV) and to investigate the neuroprotective potential of the extract and rizonic acid (RA) from E. velutina on neuronal cells. The plant drug of E. velutina previously characterized was used for the production of EEEV. Three methods were evaluated in order to obtain an extract with higher content of phenols. The neuroprotective effect of standardized EEEV (HPLC-PDA) and RA was investigated on SH-SY5Y cell exposure to the neurotoxin 6-hydroxydopamine (6-OHDA). The powder of the plant drug was classified as moderately coarse and several quality control parameters were determined. EEEV produced by percolation gave the highest phenol content when related to others extractive methods, and its HPLC-PDA analysis allowed to identify four flavonoids and RA, some reported for the first time for the species. EEEV and RA reduced significantly the neurotoxicity induced by 6-OHDA in SH-SY5Y cells determined by the MTT assay and the nitrite concentration. EEEV also showed a free radical scavenging activity. This is the first pharmacological study about E. velutina which used a controlled standardized extract since the preparation of the herbal drug. This extract and RA, acting as an antioxidant, presents a neuroprotective effect suggesting that they have potential for future development as a therapeutic agent in neurodegenerative disease as Parkinson. The powder of Erythrina velutina was classified as moderately coarse and several quality-control parameters were determined.Ethanolic extract from E. velutina (EEEV) produced by percolation gave the highest phenol content when related to others extractive methods and its HPLC-PDA analysis of EEEV allowed to identify four flavonoids and rizonic acid (RA), some reported for the first time for the species.The EEEV and RA

  4. An alternative allergen risk management approach.

    Science.gov (United States)

    Manning, Louise; Soon, Jan Mei

    2017-12-12

    Protein components in food can trigger immune-mediated response in susceptible individuals. International law requires risk assessment to be undertaken by competent individuals to minimize food safety risk to consumers. Historically, allergen control legislation has been food-focused and on the requirement for on pack labeling, and the need for formal food recalls in the event of misleading or inappropriate labeling. In order to develop a mechanism for decision makers when assessing allergenic risk from plant-derived materials, the aim of this research was to consider a more holistic risk assessment method whereby rather than just using the food-based approach, an additive element in terms of considering the families of proteins is included. This approach reflects the need for food professionals to fully understand the role of proteins in triggering an allergic response to plant material and the health risk to individuals who show cross-reactivity to such proteins.

  5. Allergen Immunotherapy: History and Future Developments.

    Science.gov (United States)

    Passalacqua, Giovanni; Canonica, Giorgio Walter

    2016-02-01

    Allergen immunotherapy (AIT) was introduced in clinical practice more than 100 years ago. The clinical effectiveness in allergic rhinitis (and asthma) and in hymenoptera allergy was apparent early on but it was not until the mid-1900s that randomized placebo-controlled trials proved its efficacy. In the 1980s, sublingual immunotherapy (SLIT) was accepted in official guidelines. The availability of safer routes, such as SLIT, prompted increasing investigation of AIT for food allergy. The introduction of molecular-based diagnosis introduced the possibility of better targeted prescription of AIT. Other approaches are being explored, such as immunogenic peptides, recombinant allergens, and adjuvants. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. 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.

  7. Respiratory Allergens from Furred Mammals: Environmental and Occupational Exposure.

    Science.gov (United States)

    Zahradnik, Eva; Raulf, Monika

    2017-08-04

    Furry mammals kept as pets, farm and laboratory animals are important allergen sources. The prevalence of sensitization to furred mammals appears to be increasing worldwide. Several mammalian allergens from diverse species are well characterized with regard to their molecular structure and immunogenicity, and some are already available for component-resolved allergy diagnostics. The distribution of various mammalian allergens has been extensively studied during the past few decades. Animal allergens were found to be ubiquitous in the human environment, even in places where no animals reside, with concentrations differing considerably between locations and geographical regions. This review presents an overview of identified mammalian respiratory allergens classified according to protein families, and compiles the results of allergen exposure assessment studies conducted in different public and occupational environments.

  8. Purification, crystallization and initial crystallographic characterization of brazil-nut allergen Ber e 2

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Feng; Jin, Tengchuan; Howard, Andrew; Zhang, Yu-Zhu, E-mail: zhangy@iit.edu [Department of Biology, Illinois Institute of Technology, Chicago, IL 60616 (United States)

    2007-11-01

    The crystallization of the brazil nut allergen Ber e 2 is reported. Peanut and tree-nut allergies have attracted considerable attention because of their frequency and their lifelong persistence. Brazil-nut (Bertholletia excelsa) allergies have been well documented and the 11S legumin-like seed storage protein Ber e 2 (excelsin) is one of the two known brazil-nut allergens. In this study, Ber e 2 was extracted from brazil-nut kernels and purified to high purity by crystalline precipitation and gel-filtration chromatography. Well diffracting single crystals were obtained using the hanging-drop vapour-diffusion method. A molecular-replacement structural solution has been obtained. Refinement of the structure is currently under way.

  9. 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

  10. A role for Waldeyer's ring in immunological response to allergens.

    Science.gov (United States)

    Masieri, Simonetta; Trabattoni, Daria; Incorvaia, Cristoforo; De Luca, Maria Cristina; Dell'Albani, Ilaria; Leo, Gualtiero; Frati, Franco

    2014-02-01

    Adenoids, tubal tonsil, palatine tonsil, and lingual tonsil are immunological organs included in the Waldeyer's ring, the basic function of which is the antibody production to common environmental antigens. Adenoidal hypertrophy (AH) is a major medical issue in children, and adenoidectomy is still the most used treatment worldwide. The response of adenoids to allergens is a good model to evaluate their immunological function. This report assessed the immunological changes in adenoid tissues from children with allergic rhinitis (AR) undergoing sublingual immunotherapy (SLIT). Adenoid samples from 16 children (seven males, nine females, mean age 7.12 years) with AH and clinical indication to adenoidectomy were collected. Of them, five children were not allergic and 11 had house dust mite and grass pollen-induced AR. Among allergic children, in four AR was treated by antihistamines while in seven AR was treated by high-dose SLIT during 4-6 months. The evaluation addressed the T helper 1 (Th1), Th2, and Th3 cells by performing a PCR array on mRNA extracted from adenoid samples. In non-allergic children, a typical Th1 pattern was found. SLIT induced a strong down-regulation of genes involved in Th2 and Th1 activation and function. In particular, in SLIT-treated allergic children IL-4, CCR2, CCR3, and PTGDR2 (Th2 related genes) and CD28, IL-2, and INHA (Th1 related genes) expression was reduced, compared with children treated with antihistamines. These preliminary findings warrant investigation in trials including larger numbers of patients, but indicate that hypertrophic adenoids of allergic children have the typical response to the specific allergen administered by SLIT. This should suggest that one should reconsider the immunological role of adenoids.

  11. The activity of pomegranate extract standardized 40% ellagic acid during the healing process of incision wounds in albino rats (Rattus norvegicus

    Directory of Open Access Journals (Sweden)

    Wiwik Misaco Yuniarti

    2018-03-01

    Full Text Available Aim: This research aimed to evaluate the effects of pomegranate extract standardized to 40% ellagic acid on the incised wound in albino rats. Materials and Methods: Fifty albino rats were divided into 10 treatment groups. The five groups were sacrificed on the 8th day, while the others were sacrificed on the 15th day. Two groups of albino rats with incised wound were not treated at all (P0, the other two groups of albino rats with incised wound were treated with Betadine® (P1 ointment, and the rest of the groups were treated with pomegranate extract standardized to 40% ellagic acid with a concentration of 2.5% (P2, 5% (P3, and 7.5% (P4. The treatments were carried out twice a day with an interval of 12 h for 7 and 14 days. At the end of the research, the skin tissue of those albino rats had been taken for histopathologic preparations before H and E staining was performed. Results: Collagen deposition, polymorphonuclear neutrophils (PMN infiltration, angiogenesis, and fibrosis degree in Group P4 treated with 7.5% pomegranate extract standardized to 40% ellagic acid for 14 days were significantly different from those in Groups P0, P1, P2, and P3, especially in the case of PMN inflammation (p<0.05. Conclusion: The administration of 7.5% pomegranate extract standardized to 40% ellagic acid for 14 days on incised wounds of those albino rats can accelerate the wound healing process characterized by collagen deposition improvement, PMN infiltration in the wound area, angiogenesis, and fibrosis degree.

  12. Immunoglobulin G4 antibodies to rat urinary allergens, sensitization and symptomatic allergy in laboratory animal workers.

    Science.gov (United States)

    Portengen, L; de Meer, G; Doekes, G; Heederik, D

    2004-08-01

    We have previously reported that high rat urinary allergen (RUA) exposure was not associated with increased risk of rat allergy in long-term-exposed laboratory animal (LA) workers. We aimed to assess whether strong allergen-specific IgG4 responses could explain the absence of a dose response in these subjects. We investigated whether IgG4 was associated with allergen exposure and prevalence of sensitization or respiratory symptoms to rats. The longitudinal relation between IgG4 and rat allergy was studied using data obtained during 2 years of follow-up. Five hundred and twenty-nine LA workers answered a questionnaire on respiratory symptoms and occupational history and participated in skin prick testing. Blood samples were analysed for specific IgG4 and IgE to RUA. Exposure to RUA was estimated based on personal air samples. The relation between IgG4 and newly occurring sensitization or rat allergy was studied in workers who were not sensitized or did not report respiratory symptoms to rats. IgG4 titres were higher in atopic than in non-atopic subjects, and increased with higher allergen exposure. Titres were highest in subjects who were sensitized and reported respiratory symptoms to rats when compared with those who were not (geometric mean [geometric standard deviation] = 202 [5.7] vs. 8.4 [18.3] AU). The association between IgG4 and sensitization or symptomatic rat allergy was independent of estimated allergen exposure. IgG4 was a strong predictor of newly occurring sensitization and symptomatic rat allergy during follow-up in atopic and rat-sensitized subjects. High exposure to RUA is associated with a strong allergen-specific IgG4 antibody response. High anti-RUA IgG4 is a strong predictor of prevalent and incident sensitization and symptomatic rat allergy in atopic and rat-sensitized subjects. IgG4 can therefore not explain the absence of a dose response between allergen exposure and allergy in long-term-exposed workers. We consider anti-RUA IgG4 to be a

  13. Recombinant Fusion Allergens, Cry j 1 and Cry j 2 from Japanese Cedar Pollen, Conjugated with Polyethylene Glycol Potentiate the Attenuation of Cry j 1-Specific IgE Production in Cry j 1-Sensitized Mice and Japanese Cedar Pollen Allergen-Sensitized Monkeys.

    Science.gov (United States)

    Fujimura, Takashi; Fujinami, Koji; Ishikawa, Ryosuke; Tateno, Minoru; Tahara, Yoshio; Okumura, Yasushi; Ohta, Hisashi; Miyazaki, Hiroyuki; Taniguchi, Masaru

    2015-01-01

    Japanese cedar (Cryptomeria japonica) pollinosis is the most prevalent seasonal rhinitis in Japan. A standardized Japanese cedar pollen extract (CPE) containing 1.5-4.2 μg of Cry j 1 is currently the highest-concentration extract available for allergen-specific immunotherapy (SIT) against this pollinosis. Therefore, we developed a PEGylated fusion protein as a more effective SIT vaccine against Japanese cedar pollinosis. The fusion protein of major allergens for Japanese cedar pollen, Cry j 1 and Cry j 2, was expressed in Escherichia coli and conjugated with polyethylene glycol (PEG). The purified PEGylated Cry j 1/2 fusion protein (PEG-fusion) was subcutaneously injected four times into Cry j 1- sensitized mice and CPE-sensitized monkeys. The mice were then subcutaneously challenged with Cry j 1 and serum levels of Cry j 1-specific immunoglobulin, and the proliferation and cytokine production of splenocytes were analyzed. The monkeys were intranasally challenged with CPE and analyzed for Cry j 1-specific immunoglobulin levels in plasma. Cry j 1-specific IgE was significantly attenuated in the PEG-fusion-treated group after Cry j 1-challenge and Cry j 1-specific IgG was significantly increased following PEG-fusion treatment in mice and monkeys. Proliferation and Th2-type cytokine production in splenocytes stimulated with Cry j 1 were also reduced in PEG-fusion-treated mice. IL10 and IL2 production were reduced, but not significantly, while IFN-x03B3; was significantly increased in the PEG-fusion-treated group. A high-dose injection of PEG-fusion appears to be a valid candidate for a safer and more effective vaccine than the conventional SIT extract for Japanese cedar pollinosis. © 2015 S. Karger AG, Basel.

  14. A two-centre evaluation of the human organotypic skin explant culture model for screening contact allergens

    NARCIS (Netherlands)

    Lehé, Cynthia L.; Jacobs, John J. L.; Elliott, Graham R.; Das, Pranab K.

    2003-01-01

    Animal models are considered to be the "gold standard" for determining the potential contact allergenicity of low molecular weight chemicals. However, governmental regulations and ethical considerations limit the use of animals for such purposes. There is therefore a need for in vitro alternative

  15. Biotransformation Strategy To Reduce Allergens in Propolis

    Science.gov (United States)

    Gardana, Claudio; Barbieri, Andrea; Simonetti, Paolo

    2012-01-01

    Propolis (bee glue) is a resinous, sticky, dark-colored material produced by honeybees. Propolis today, due to its medicinal properties, is increasingly popular and is extensively used in food, beverages, and cosmetic products. Besides its numerous positive properties, propolis may also have adverse effects, such as, principally, allergic eczematous contact dermatitis in apiarists and in consumers with an allergic predisposition. In this study, we found appropriate conditions for removing caffeate esters, which are the main allergenic components, from raw propolis. The proposed method consists of the resuspension of propolis in a food grade solvent, followed by a biotransformation based on the cinnamoyl esterase activity of Lactobacillus helveticus. We showed that the reduction of caffeate esters by L. helveticus did not affect the content of flavonoids, which are the main bioactive molecules of propolis. Furthermore, we verified that the biotransformation of propolis did not cause a loss of antimicrobial activity. Finally, we demonstrated that the ability of L. helveticus to hydrolyze caffeate esters in propolis is strain specific. In conclusion, the proposed strategy is simple, employs food grade materials, and is effective in selectively removing allergenic molecules without affecting the bioactive fraction of propolis. This is the first study demonstrating that the allergenic caffeate esters of propolis can be eliminated by means of a bacterial biotransformation procedure. PMID:22522681

  16. The Level of Sensitivity of Food Allergens

    Directory of Open Access Journals (Sweden)

    Iris Rengganis

    2011-06-01

    Full Text Available In recent years, the occurrence of allergy continues to increase rapidly both domestically and globally. World Allergy Organization (WAO revealed that 22% of the world population suffers from allergies, and this number increases every year. Food allergy is a condition caused by the reaction of IgE against substances (chemicals in food. Food allergy can interfere with brain function and body organ systems as well as affect the quality of life. The purpose of this study is to know the level of sensitivity of food allergens in the Immunology Allergy Poly RSCM in 2007. Data were collected from 208 patients who have medical records and went through skin prick tests in the Immunology Allergy Clinic RSCM in 2007. Univariate analysis was performed to describe the types of food allergens within groups of children and adults. Around 49% of the respondents were sensitive to food allergens. The types of foods that caused the most allergies for children and adults are respectively shrimp, egg white and cornstarch. Cow's milk and wheat flour are the types of food that caused most allergies for children only, whereas for adults, the food that caused the most allergies is crab.

  17. Lack of detectable allergenicity of transgenic maize and soya samples

    OpenAIRE

    Batista, Rita; Nunes, Baltazar; Carmo, Manuela; Cardoso, Carlos; Helena, São José; Almeida, António Bugalho de; Manique, Alda; Bento, Leonor; Ricardo, Cândido Pinto; Oliveira, Maria Margarida

    2005-01-01

    Background: The safety issues regarding foods derived from genetically modified (GM) plants are central to their acceptance into the food supply. The potential allergenicity of proteins newly introduced in GM foods is a major safety concern. Objective: We sought to monitor, in potentially sensitive human populations, the allergenicity effects of 5 GM materials obtained from sources with no allergenic potential and already under commercialization in the European Union. ...

  18. 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...

  19. Liquid chromatography-particle beam electron ionization mass spectrometry method for analysis of botanical extracts: evaluation of ephedrine alkaloids in standard reference materials.

    Science.gov (United States)

    Castro, Joaudimir; Krishna, M V Balarama; Marcus, R Kenneth

    2010-01-01

    The preliminary validation of a high-performance liquid chromatography particle beam mass spectrometry method (HPLC-PB/MS) with electron impact ionization source for analysis of botanical extracts is presented. The LC-PB/MS system was evaluated for the analysis of ephedrine alkaloids using ephedra-containing National Institute of Standards and Technology dietary supplement standard reference materials (SRMs) 3241 Ephedra Sinica Stapf Native Extract and 3242 Ephedra Sinica Stapf Commercial Extract. The ephedrine alkaloids were separated by reversed-phase chromatography using a phenyl column at room temperature. A linear gradient method with a mobile phase composition varying from 5:95 [MeOH:0.1% trifluoroacetic acid (TFA) in water] to 20:80 (MeOH:0.1% TFA in water) at a flow rate of 1.0 ml/min, with an analysis time of less than 20 min, was used. The source block temperature was evaluated to determine the optimal operating conditions by monitoring the intensities and fragmentation patterns of the ephedrine alkaloids. Ephedrine and N-methylephedrine were taken as a representative of the test alkaloids. The LODs on the sub-nanogram level were achieved, with ephedrine, pseudoephedrine, and methylephedrine in the SRMs quantified by a standard addition method with recoveries of > or = 86% and RSDs of < or = 14% (n = 3).

  20. Allergen and Epitope Targets of Mouse-Specific T Cell Responses in Allergy and Asthma

    Directory of Open Access Journals (Sweden)

    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.

  1. Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds.

    Science.gov (United States)

    Fernandes, K V; Deus-de-Oliveira, N; Godoy, M G; Guimarães, Z A S; Nascimento, V V; Melo, E J T de; Freire, D M G; Dansa-Petretski, M; Machado, O L T

    2012-11-01

    Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5) cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained.

  2. Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds

    Directory of Open Access Journals (Sweden)

    K.V. Fernandes

    2012-11-01

    Full Text Available Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin and allergenic (2S albumin proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5 cells/well. Solid-state fermentation (SSF and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained.

  3. Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds

    Energy Technology Data Exchange (ETDEWEB)

    Fernandes, K.V.; Deus-de-Oliveira, N. [Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Rio de Janeiro, RJ (Brazil); Godoy, M.G. [Laboratório de Biotecnologia Microbiana, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ (Brazil); Guimarães, Z.A.S. [Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Rio de Janeiro, RJ (Brazil); Nascimento, V.V. [Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Rio de Janeiro, RJ (Brazil); Melo, E.J.T. de [Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Rio de Janeiro, RJ (Brazil); Freire, D.M.G. [Laboratório de Biotecnologia Microbiana, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ (Brazil); Dansa-Petretski, M.; Machado, O.L.T. [Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Rio de Janeiro, RJ (Brazil)

    2012-08-24

    Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 × 10{sup 5} cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH){sub 2} or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained.

  4. Immunological cross-reactivity between four distant parvalbumins-Impact on allergen detection and diagnostics.

    Science.gov (United States)

    Sharp, Michael F; Stephen, Juan N; Kraft, Lukas; Weiss, Thomas; Kamath, Sandip D; Lopata, Andreas L

    2015-02-01

    Fish are the largest and most diverse group of vertebrates. Fish are also a part of the eight food groups that cause the majority of IgE mediated food reactions. Detection tools for fish allergens are however limited due to the great diversity of fish species, despite fish allergy and its major allergen parvalbumin being well documented. The most commonly studied fish are frequently consumed in North America and Europe. However, much less is known about fish allergens in the Australasian region although fish is widely consumed in this region. A comprehensive phylogenetic analysis was performed of known parvalbumin amino acid sequences to determine possible candidate antigens for new cross-reactive antibodies to be used to detect most fish parvalbumins. Polyclonal rabbit antibodies were raised against parvalbumins from frequently consumed barramundi (Lates calcarifer), basa (Pangasius bocourti), pilchard (Sardinops sagax) and Atlantic salmon (Salmo salar). These were evaluated for cross-reactivity against a panel of 45 fish extracts (raw, heated and canned fish). Anti-barramundi parvalbumin proved to be the most cross-reactive antibody, detecting 87.5% of the 40 species analyzed, followed by anti-pilchard and anti-basa antibody. In contrast the anti-salmon antibody was very specific and only reacted to salmonidae and a few other fish. All analyzed fish species, except mahi mahi, swordfish, yellowfin tuna and all 5 canned fish had parvalbumin detected in raw extracts. However antibody reactivity to many fish was heat liable or susceptible to denaturation, demonstrating that some parvalbumins have most likely conformational epitopes, which lose antibody reactivity after heat treatment. We have demonstrated the generation of highly cross-reactive anti-parvalbumin antibodies that could be used for the detection of allergenic fish parvalbumin in contaminated food products. This cross-reactivity study thus shows processing of fish, especially canning, can have on impact

  5. Lafoensia pacari A. St. -Hil: wound healing activity and mechanism of action of standardized hydroethanolic leaves extract.

    Science.gov (United States)

    Pereira, Lucas Olivo Martins; Vilegas, Wagner; Tangerina, Marcelo Marucci Pereira; Arunachalam, Karuppusamy; Balogun, Sikiru Olaitan; Matos, Paulo Eduardo Orlandi; Martins, Domingos Tabajara de Oliveira

    2018-02-28

    Lafoensia pacari A. St.-Hil., (Lythraceae) is a native tree of Brazilian Cerrado and commonly known in Brazil as "mangava-brava". Its leaves are used in Brazilian folk medicine in wound healing, cutaneous mycoses, and in the treatment of gastritis and ulcers. The present study was designed to evaluate the wound healing activity and mechanism of action of the hydroethanolic extract of Lafoensia pacari A. St. -Hil leaves (HELp), and to advance in its chemical profiling. HELp was prepared by maceration in 70% hydroethanolic solution (1:10, w/v). The phytochemical analyses were investigated using colorimetry and electrospray ionization/mass spectrometric detection (ESI-MS n ). Its in vitro cytotoxicity was evaluated in CHO-K1 and L929 cells, while the in vivo acute toxicity was performed in mice. The potential in vivo wound healing activity was assessed using excision and incision rat models and histopathology of the wounded skin (excision model) was carried out. The in vitro wound healing activity of HELp was demonstrated by scratch assay in L-929 cells, by measuring proliferation/migration rate and p-ERK 1/2 protein expression using western blot analysis. HELp's in vivo anti-inflammatory activity was evaluated by lipopolysaccharide (LPS) induced peritonitis in mice, along with the determination of nitric oxide (NO) and cytokines (TNF-α and IL-10) in the peritoneal lavages. Its potential in vitro antibacterial activity was performed using microbroth dilution assay, while in vitro antioxidant activities was by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, and ferric reducing antioxidant power (FRAP)) were assessed by using in vitro assays. The phytochemical analysis of HELp revealed the presence of polyphenols with ellagic acid, punicalagin, punicalin, kaempferol, quercetin-3-O-xylopyranoside and quercetin-3-O-rhamnopyranoside being the most prominent. HELp showed no toxicity on CHO-k1 and L929 cell lines. Topical treatment with HELp (10 and 30mg/g of gel) presented

  6. Evaluation of the allergenicity of tropical pollen and airborne spores in Singapore.

    Science.gov (United States)

    Chew, F T; Lim, S H; Shang, H S; Dahlia, M D; Goh, D Y; Lee, B W; Tan, H T; Tan, T K

    2000-04-01

    Sensitization to pollen and spores of the Southeast Asian tropical region is not well documented. This study evaluated the allergenicity of the tropical airspora in Singapore. On the basis of the results of an aerobiologic survey of the airspora profile of Singapore, crude extracts of 23 main spore (fungal and fern) and pollen types were prepared. A total of 231 patients with asthma and/or allergic rhinitis and 76 healthy controls were evaluated by skin prick test (SPT). Total and specific IgE levels were also quantified by the fluorescence allergosorbent test (FAST). All 23 allergenic extracts tested elicited positive SPT responses. Among the patients with atopic diseases, extracts of oil-palm pollen (Elaeis guineensis) were observed to have the highest frequency of positive reactions (40%), followed by extracts of resam-fern spores (Dicranopteris linearis) (34%) and sea-teak pollen (Podocarpus polystachyus) (33.8%). Fungal spores with the highest SPT responses were Curvularia spp. (26-32%) and Drechslera-like spores (31%). Positive responses to these extracts correlated with total serum IgE levels of the subjects and were significantly associated with the presence of atopic disease. We have documented sensitization to tropical pollen and spores in our population. Its association with atopy suggests that it has a role in allergic diseases in the tropics.

  7. Inhibitory Effects of Standardized Extracts of Phyllanthus amarus and Phyllanthus urinaria and Their Marker Compounds on Phagocytic Activity of Human Neutrophils

    Directory of Open Access Journals (Sweden)

    Yuandani

    2013-01-01

    Full Text Available The standardized methanol extracts of Phyllanthus amarus and P. urinaria, collected from Malaysia and Indonesia, and their isolated chemical markers, phyllanthin and hypophyllanthin, were evaluated for their effects on the chemotaxis, phagocytosis and chemiluminescence of human phagocytes. All the plant extracts strongly inhibited the migration of polymorphonuclear leukocytes (PMNs with the Malaysian P. amarus showing the strongest inhibitory activity (IC50 value, 1.1 µg/mL. There was moderate inhibition by the extracts of the bacteria engulfment by the phagocytes with the Malaysian P. amarus exhibiting the highest inhibition (50.8% of phagocytizing cells. The Malaysian P. amarus and P. urinaria showed strong reactive oxygen species (ROS inhibitory activity, with both extracts exhibiting IC50 value of 0.7 µg/mL. Phyllanthin and hypophyllanthin exhibited relatively strong activity against PMNs chemotaxis, with IC50 values slightly lower than that of ibuprofen (1.4 µg/mL. Phyllanthin exhibited strong inhibitory activity on the oxidative burst with an IC50 value comparable to that of aspirin (1.9 µg/mL. Phyllanthin exhibited strong engulfment inhibitory activity with percentage of phagocytizing cells of 14.2 and 27.1% for neutrophils and monocytes, respectively. The strong inhibitory activity of the extracts was due to the presence of high amounts of phyllanthin and hypophyllanthin although other constituents may also contribute.

  8. 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....

  9. Studies of the allergens of Olea europea pollen.

    Science.gov (United States)

    Blanca, M; Boulton, P; Brostoff, J; Gonzalez-Reguera, I

    1983-09-01

    Allergens of Olea europea have been prepared by isoelectrofocusing and tested by RAST, RAST inhibition and skin tests. The major allergen was found to be an acidic protein of pI 6, consisting of two polypeptide chains of molecular weight 15000 and 17000 daltons, possibly non-covalently associated into a dimer. Most other allergens were acidic proteins, but basic proteins were also observed with allergenic activity. The RAST inhibition studies show a considerable degree of allergic cross-reactivity between the isoelectrofocusing fractions.

  10. Allergen manufacturing and quality aspects for allergen immunotherapy in Europe and the United States

    DEFF Research Database (Denmark)

    Bonertz, A; Roberts, G; Slater, J E

    2018-01-01

    Adequate quality is essential for any medicinal product to be eligible for marketing. Quality includes verification of the identity, content and purity of a medicinal product in combination with a specified production process and its control. Allergen products derived from natural sources require...... particular considerations to ensure adequate quality. Here, we describe key aspects of the documentation on manufacturing and quality aspects for allergen immunotherapy products in the European Union and the United States. In some key parts, requirements in these areas are harmonized while other fields...

  11. The standard aqueous stem bark extract of Mangifera indica L. inhibits toxic PLA2 - NN-XIb-PLA2 of Indian cobra venom.

    Science.gov (United States)

    Dhananjaya, Bhadrapura Lakkappa; Sudarshan, Shivalingaiah; Dongol, Yashad; More, Sunil S

    2016-05-01

    The aqueous extract of Mangifera indica is known to possess diverse medicinal properties, which also includes anti-snake venom activities. However, its inhibitory potency and mechanism of action on multi-toxic snake venom phospholipases A2s are still unknown. Therefore, the objective of this study was to evaluate the modulatory effect of standard aqueous bark extract of M. indica on NN-XIb-PLA2 of Indian cobra venom. The in vitro sPLA2, in situ hemolytic and in vivo edema inhibition effect were carried out as described. Also the effect of substrate and calcium concentration was carried out. M. indica extract dose dependently inhibited the GIA sPLA2 (NN-XIb-PLA2) activity with an IC50 value of 7.6 μg/ml. M. indica extract effectively inhibited the indirect hemolytic activity up to 98% at ∼40 μg/ml concentration. Further, M. indica extract (0-50 μg/ml) inhibited the edema formed in a dose dependent manner. When examined as a function of increased substrate and calcium concentration, there was no relieve of inhibitory effect of M. indica extract on the NN-XIb-PLA2. Further, the inhibition was irreversible as evident from binding studies. The in vitro inhibition is well correlated with in situ and in vivo edema inhibiting activities of M. indica. As the inhibition is independent of substrate and calcium and was irreversible, it can be concluded that M. indica extract mode of inhibition could be due to direct interaction of components present in the extract with the PLA2 enzyme. The aqueous extract of M. indica effectively inhibits svPLA2 enzymatic and its associated toxic activities, which substantiate their anti-snake venom properties. Further in-depth studies on the role and mechanism of the principal constituents present in the extract, responsible for the anti-PLA2 activity will be interesting to develop them into potent antisnake component and also as an anti-inflammatory agent.

  12. Prevalence of IgE antibodies to an extract from rubber tree (Hevea brasiliensis latex and recombinant pollen allergens (Phl P 1, Phl P 2, Phl P 5, Bet v 1 and Bet v 2 in the sera of Italian atopic patients

    Directory of Open Access Journals (Sweden)

    Renato E. Rossi

    2001-01-01

    Conclusions: The findings of the present study may support the concept that a high proportion of sera containing IgE to rBet v 2, rPhl p 1 and rPhl p 5 simultaneously contain antilatex IgE. Therefore, patients with specific IgE to these recombinant allergens with no history of current latex exposure may need additional evaluation.

  13. 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.

  14. 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.

  15. In house validation of a high resolution mass spectrometry Orbitrap-based method for multiple allergen detection in a processed model food.

    Science.gov (United States)

    Pilolli, Rosa; De Angelis, Elisabetta; Monaci, Linda

    2018-02-13

    In recent years, mass spectrometry (MS) has been establishing its role in the development of analytical methods for multiple allergen detection, but most analyses are being carried out on low-resolution mass spectrometers such as triple quadrupole or ion traps. In this investigation, performance provided by a high resolution (HR) hybrid quadrupole-Orbitrap™ MS platform for the multiple allergens detection in processed food matrix is presented. In particular, three different acquisition modes were compared: full-MS, targeted-selected ion monitoring with data-dependent fragmentation (t-SIM/dd2), and parallel reaction monitoring. In order to challenge the HR-MS platform, the sample preparation was kept as simple as possible, limited to a 30-min ultrasound-aided protein extraction followed by clean-up with disposable size exclusion cartridges. Selected peptide markers tracing for five allergenic ingredients namely skim milk, whole egg, soy flour, ground hazelnut, and ground peanut were monitored in home-made cookies chosen as model processed matrix. Timed t-SIM/dd2 was found the best choice as a good compromise between sensitivity and accuracy, accomplishing the detection of 17 peptides originating from the five allergens in the same run. The optimized method was validated in-house through the evaluation of matrix and processing effects, recoveries, and precision. The selected quantitative markers for each allergenic ingredient provided quantification of 60-100 μg ingred /g allergenic ingredient/matrix in incurred cookies.

  16. Effect of standardized aqueous extract of Withania somnifera on tests of cognitive and psychomotor performance in healthy human participants

    Science.gov (United States)

    Pingali, Usharani; Pilli, Raveendranadh; Fatima, Nishat

    2014-01-01

    Background: Withania somnifera is an herbal medicine that has been known to possess memory-enhancing properties. The current study involved an assessment of cognitive and psychomotor effects of Withania somnifera extract in healthy human participants. Materials and Methods: In this prospective, double-blind, multi-dose, placebo-controlled, crossover study, 20 healthy male participants were randomized to receive 250 mg two capsules twice daily of an encapsulated dried aqueous extract of roots and leaves of Withania somnifera or a matching placebo for a period of 14 days. Cognitive and psychomotor performance was assessed pre-dose (day 1) and at 3 hrs post-dose on day 15 using a battery of computerized psychometric tests. After a washout period of 14 days, the subjects crossed-over to receive the other treatment for a further period of 14 days as per prior randomization schedule. Same battery of test procedures were performed to assess cognitive and psychomotor performance. Results: Significant improvements were observed in reaction times with simple reaction, choice discrimination, digit symbol substitution, digit vigilance, and card sorting tests with Withania somnifera extract compared to placebo. However, no effect can be seen with the finger tapping test. Conclusion: These results suggest that Withania somnifera extract can improve cognitive and psychomotor performance and may, therefore, be a valuable adjunct in the treatment of diseases associated with cognitive impairment. PMID:24497737

  17. Effect of the standardized Cimicifuga foetida extract on Hsp 27 expression in the MCF-7 cell line

    Directory of Open Access Journals (Sweden)

    Maritza C Soler

    2011-01-01

    Full Text Available Cimicifuga foetida, an Asian Cimicifuga species, has been employed as a cooling and detoxification agent in traditional Chinese medicine since ancient times. For this herb, two cycloartane triterpene glycosides isolated from the rhizomes have demonstrated cytotoxicity on rat tumor and human cancer cell lines. Since human Hsp27 is increased in various human cancers and exhibits cytoprotective activity that affects tumorigenesis and the susceptibility of tumours to cancer treatment, the purpose of this research was to study the expression of this protein in MCF-7 breast cancer cells. To accomplish this aim, MCF-7 cells were exposed to different concentrations of Cimicifuga foetida extract showing a reduction in cell number measured by the sulforhodamine assay. In addition, the expression of Hsp-27 mRNA detected by RT-PCR and Hsp-27 protein detected by immnofluorescence was present in all conditions, except when using the highest concentration of Cimicifuga foetida extract (2,000 jig /L. We conclude that Hsp 27 expression at 2,000 jig /L Cimicifuga foetida extract is diminished. This is the first report showing the Hsp-27 expression after exposure to Cimicifuga foetida extract in MCF-7 cells.

  18. Effect of the standardized Cimicifuga foetida extract on Hsp 27 expression in the MCF-7 cell line.

    Science.gov (United States)

    Soler, Maritza C; Molina, Jessica L; Díaz, Hugo A; Pinto, Vivian C; Barrios, Yasenka L; He, Kan; Roller, Marc; Weinstein-Oppenheimer, Caroline R

    2011-01-01

    Cimicifuga foetida, an Asian Cimicifuga species, has been employed as a cooling and detoxification agent in traditional Chinese medicine since ancient times. For this herb, two cycloartane triterpene glycosides isolated from the rhizomes have demonstrated cytotoxicity on rat tumor and human cancer cell lines. Since human Hsp27 is increased in various human cancers and exhibits cytoprotective activity that affects tumorigenesis and the susceptibility of tumours to cancer treatment, the purpose of this research was to study the expression of this protein in MCF-7 breast cancer cells. To accomplish this aim, MCF-7 cells were exposed to different concentrations of Cimicifuga foetida extract showing a reduction in cell number measured by the sulforhodamine assay. In addition, the expression of Hsp-27 mRNA detected by RT-PCR and Hsp-27 protein detected by immnofluorescence was present in all conditions, except when using the highest concentration of Cimicifuga foetida extract (2,000 jig /L). We conclude that Hsp 27 expression at 2,000 jig /L Cimicifuga foetida extract is diminished. This is the first report showing the Hsp-27 expression after exposure to Cimicifuga foetida extract in MCF-7 cells.

  19. 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.

  20. B cells play key roles in th2-type airway immune responses in mice exposed to natural airborne allergens.

    Directory of Open Access Journals (Sweden)

    Li Yin Drake

    Full Text Available Humans are frequently exposed to various airborne allergens. In addition to producing antibodies, B cells participate in immune responses via various mechanisms. The roles of B cells in allergic airway inflammation and asthma have been controversial. We examined the functional importance of B cells in a mouse model of asthma, in which mice were exposed repeatedly to common airborne allergens. Naïve wild-type BALB/c mice or B cell-deficient JH-/- mice were exposed intranasally to a cocktail of allergen extracts, including Alternaria, Aspergillus, and house dust mite, every other day for two weeks. Ovalbumin was included in the cocktail to monitor the T cell immune response. Airway inflammation, lung pathology, and airway reactivity were analyzed. The airway exposure of naïve wild type mice to airborne allergens induced robust eosinophilic airway inflammation, increased the levels of Th2 cytokines and chemokines in the lung, and increased the reactivity to inhaled methacholine. These pathological changes and immune responses were attenuated in B cell-deficient JH-/- mice. The allergen-induced expansion of CD4+ T cells was impaired in the lungs and draining lymph nodes of JH-/- mice. Furthermore, lymphocytes from JH-/- mice failed to produce Th2 cytokines in response to ovalbumin re-stimulation in vitro. Our results suggest that B cells are required for the optimal development of Th2-type immune responses and airway inflammation when exposed to common airborne allergens. The therapeutic targeting of B cells may be beneficial to treat asthma in certain patients.

  1. Development of a strategy for the total chemical synthesis of an allergenic protein: the peach LTP Pru p 3.

    Science.gov (United States)

    Buhler, Sofie; Akkerdaas, Jaap H; A Pertinhez, Thelma; Van Ree, Ronald; Dossena, Arnaldo; Sforza, Stefano; Tedeschi, Tullia

    2017-04-01

    The possibility to obtain allergenic proteins by means of total chemical synthesis would be a big step forward in the development of cures to food allergy and in the study of the mechanism of allergic reactions, because this would allow to achieve control at the molecular level over the structure of the product and to study its relationship with the allergenic activity in fine details. This is instead not possible by using allergens produced by extraction from natural sources or by recombinant DNA techniques. In this work, we aimed to test for the first time the feasibility of the total chemical synthesis of an allergenic protein. Pru p 3, the most studied member of the family of lipid transfer proteins, relevant plant food pan-allergens, was used as model target. Strategies for the convergent assembly of the target protein, starting from five peptide fragments to be bound by means of either native chemical ligation or peptide hydrazide ligation, followed by desulfurization, to achieve ligations at alanine, were developed and tested. All the reaction conditions were set up and optimized. Two large peptides covering the two halves of the protein sequence were synthesized and structurally characterized by means of circular dichroism, and their immunogenicity was proved by means of immunoblot, using antibodies against Pru p 3, and immunoCAP inhibition tests. Finally, the five peptides were bound together to produce the whole protein stretch. The obtained results demonstrate the feasibility of total chemical synthesis as a new way to obtain pure allergens. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

  2. [Epidemiology of contact dermatitis: prevalence of sensitization to different allergens and associated factors].

    Science.gov (United States)

    Bordel-Gómez, Ma T; Miranda-Romero, A; Castrodeza-Sanz, J

    2010-01-01

    In clinical practice, contact dermatitis is a relatively common skin complaint, whose prevalence has increased in recent years. Study by patch testing is essential for diagnosis of contact sensitization. To study the prevalence of sensitization to different allergens in a standard battery and observe the influence of different epidemiological and clinical variables on contact sensitization. A large number of allergens were included in our battery in order to detect new sensitizations whose prevalence might justify further study. This was a retrospective, observational, epidemiological study of 1092 patients, conducted in our skin allergy unit between January 1, 2000, and December 31, 2005. All patients were studied with a battery of 51 allergens. We assessed the following variables: sex, age, type of referral, occupation, site and course of skin lesions, personal and family history of atopy, positive patch tests, clinical significance, diagnosis, source of sensitization, and occupational relationship. At least 1 positive result was found in 55% of the patients, and 55.7% presented atopic dermatitis in one of its clinical variants: allergic contact dermatitis (28.2%), irritant contact dermatitis (20.1%), photoallergic contact dermatitis (2.2%), and phototoxic contact dermatitis (1.2%). The most prevalent allergens were nickel sulfate (29.3%), palladium chloride (11.7%), cobalt chloride (10.8%), potassium dichromate (7.5%), fragrance blends (6.3%), and p-phenylenediamine (6.1%). A positive occupational relationship was found in 41.1%, and 21.3% of the patients studied were diagnosed with occupational contact dermatitis. Metal workers, construction workers, and professional hairdressers were the most strongly represented groups. The most common source of sensitization was contact with metallic objects, followed by drugs, cosmetics, and rubber items. Female sex was the only independent variable that had a significant influence on the risk of contact sensitization in

  3. Enteric-coated and highly standardized cranberry extract reduces antibiotic and nonsteroidal anti-inflammatory drug use for urinary tract infections during radiotherapy for prostate carcinoma

    Directory of Open Access Journals (Sweden)

    Bonetta A

    2017-04-01

    Full Text Available Alberto Bonetta,1 Giandomenico Roviello,2,3 Daniele Generali,3,4 Laura Zanotti,3 Maria Rosa Cappelletti,3 Chiara Pacifico,5 Francesco Di Pierro6 1Oncological Radiotherapy Operative Unit, ASST, Cremona, 2Department of Molecular and Translational Medicine, University of Brescia, Brescia, 3Molecular Therapy and Pharmacogenomics Unit, ASST, Cremona, 4Department of Medical, Surgery and Health Sciences, University of Trieste, Trieste, 5Department of Medical, Surgical and Neurological Sciences, University Hospital of Siena, Siena, 6Velleja Research Scientific Department, Milan, Italy Introduction: Worldwide, bacterial resistance to antibiotic therapy is a major concern for the medical community. Antibiotic resistance mainly affects Gram-negative bacteria that are an important cause of lower urinary tract infections (LUTIs. Pelvic irradiation for prostate cancer is a risk factor for LUTIs. Cranberry extract is reported to reduce the incidence of LUTIs. The prophylactic role of an enteric-coated, highly standardized cranberry extract (VO370® in reducing LUTI episodes, urinary discomfort, and nonsteroidal anti-inflammatory drug (NSAID and antibiotic use during radiotherapy for prostate carcinoma was evaluated. Methods: A total of 924 patients with prostate carcinoma treated by radiotherapy to the prostatic and pelvic areas were randomized to receive (n=489 or not (n=435 the enteric-coated, highly standardized cranberry extract for 6–7 weeks concurrently with irradiation. Outcomes were analyzed by using Mann–Whitney U test and Pearson’s X2 test. Primary endpoint was the number of patients with LUTI; secondary endpoints were incidence of recurrence, days of treatment with antibiotics and number of subjects treated with NSAIDs, and incidence of dysuria. Results: The treatment was very well tolerated, and there were no serious side effects. All enrolled patients completed the study. Urinary infections were detected in 53 of the 489 patients (10

  4. A novel liquid-liquid extraction for the determination of naphthalene by GC-MS with deuterated anthracene as internal standard.

    Science.gov (United States)

    Erarpat, Sezin; Özzeybek, Gözde; Chormey, Dotse Selali; Erulaş, Fatih; Turak, Fatma; Bakırdere, Sezgin

    2017-09-30

    Polycyclic aromatic hydrocarbons are known for their carcinogenic and mutagenic effects on human health. This therefore calls for the regulation of their concentrations in air, water, and soil. Naphthalene as the simplest in structure of the polycyclic aromatic hydrocarbons is mainly used as a starter material for other chemicals but also has impacts on human health. A method is therefore proposed for the determination of naphthalene in water samples by gas chromatography mass spectrometry after liquid-liquid extraction. The extraction method was optimized to improve the extraction output, thereby lowering the limit of detection. The limits of detection and quantification obtained for naphthalene were 4.4 and 14.6 ng mL -1 , respectively. Deuterated anthracene was used as internal standard to enhance the precision of the method, for which a relative standard deviation of 4.3% was obtained. The percent recovery of naphthalene obtained from tap water was ranged between 93.8 and 102.2.

  5. Proteomic and immunochemical characterization of glutathione transferase as a new allergen of the nematode Ascaris lumbricoides.

    Science.gov (United States)

    Acevedo, Nathalie; Mohr, Jens; Zakzuk, Josefina; Samonig, Martin; Briza, Peter; Erler, Anja; Pomés, Anna; Huber, Christian G; Ferreira, Fatima; Caraballo, Luis

    2013-01-01

    Helminth infections and allergy have evolutionary and clinical links. Infection with the nematode Ascaris lumbricoides induces IgE against several molecules including invertebrate pan-allergens. These antibodies influence the pathogenesis and diagnosis of allergy; therefore, studying parasitic and non-parasitic allergens is essential to understand both helminth immunity and allergy. Glutathione transferases (GSTs) from cockroach and house dust mites are clinically relevant allergens and comparative studies between them and the GST from A. lumbricoides (GSTA) are necessary to evaluate their allergenicity. We sought to analyze the allergenic potential of GSTA in connection with the IgE response to non-parasitic GSTs. IgE to purified GSTs from Ascaris (nGSTA and rGSTA), house dust mites (rDer p 8, nBlo t 8 and rBlo t 8), and cockroach (rBla g 5) was measured by ELISA in subjects from Cartagena, Colombia. Also, multidimensional proteomic approaches were used to study the extract of A. lumbricoides and investigate the existence of GST isoforms. We found that among asthmatics, the strength of IgE levels to GSTA was significantly higher than to mite and cockroach GSTs, and there was a strong positive correlation between IgE levels to these molecules. Specific IgE to GSTA was found in 13.2% of controls and 19.5% of asthmatics. In addition nGSTA induced wheal and flare in skin of sensitized asthmatics indicating that it might be of clinical relevance for some patients. Frequency and IgE levels to GSTA were higher in childhood and declined with age. At least six GST isoforms in A. lumbricoides bind human IgE. Four isoforms were the most abundant and several amino acid substitutions were found, mainly on the N-terminal domain. In conclusion, a new allergenic component of Ascaris has been discovered; it could have clinical impact in allergic patients and influence the diagnosis of mite and cockroach allergy in tropical environments.

  6. Proteomic and Immunochemical Characterization of Glutathione Transferase as a New Allergen of the Nematode Ascaris lumbricoides

    Science.gov (United States)

    Acevedo, Nathalie; Mohr, Jens; Zakzuk, Josefina; Samonig, Martin; Briza, Peter; Erler, Anja; Pomés, Anna; Huber, Christian G.; Ferreira, Fatima; Caraballo, Luis

    2013-01-01

    Helminth infections and allergy have evolutionary and clinical links. Infection with the nematode Ascaris lumbricoides induces IgE against several molecules including invertebrate pan-allergens. These antibodies influence the pathogenesis and diagnosis of allergy; therefore, studying parasitic and non-parasitic allergens is essential to understand both helminth immunity and allergy. Glutathione transferases (GSTs) from cockroach and house dust mites are clinically relevant allergens and comparative studies between them and the GST from A. lumbricoides (GSTA) are necessary to evaluate their allergenicity. We sought to analyze the allergenic potential of GSTA in connection with the IgE response to non-parasitic GSTs. IgE to purified GSTs from Ascaris (nGSTA and rGSTA), house dust mites (rDer p 8, nBlo t 8 and rBlo t 8), and cockroach (rBla g 5) was measured by ELISA in subjects from Cartagena, Colombia. Also, multidimensional proteomic approaches were used to study the extract of A. lumbricoides and investigate the existence of GST isoforms. We found that among asthmatics, the strength of IgE levels to GSTA was significantly higher than to mite and cockroach GSTs, and there was a strong positive correlation between IgE levels to these molecules. Specific IgE to GSTA was found in 13.2% of controls and 19.5% of asthmatics. In addition nGSTA induced wheal and flare in skin of sensitized asthmatics indicating that it might be of clinical relevance for some patients. Frequency and IgE levels to GSTA were higher in childhood and declined with age. At least six GST isoforms in A. lumbricoides bind human IgE. Four isoforms were the most abundant and several amino acid substitutions were found, mainly on the N-terminal domain. In conclusion, a new allergenic component of Ascaris has been discovered; it could have clinical impact in allergic patients and influence the diagnosis of mite and cockroach allergy in tropical environments. PMID:24223794

  7. Proteomic and immunochemical characterization of glutathione transferase as a new allergen of the nematode Ascaris lumbricoides.

    Directory of Open Access Journals (Sweden)

    Nathalie Acevedo

    Full Text Available Helminth infections and allergy have evolutionary and clinical links. Infection with the nematode Ascaris lumbricoides induces IgE against several molecules including invertebrate pan-allergens. These antibodies influence the pathogenesis and diagnosis of allergy; therefore, studying parasitic and non-parasitic allergens is essential to understand both helminth immunity and allergy. Glutathione transferases (GSTs from cockroach and house dust mites are clinically relevant allergens and comparative studies between them and the GST from A. lumbricoides (GSTA are necessary to evaluate their allergenicity. We sought to analyze the allergenic potential of GSTA in connection with the IgE response to non-parasitic GSTs. IgE to purified GSTs from Ascaris (nGSTA and rGSTA, house dust mites (rDer p 8, nBlo t 8 and rBlo t 8, and cockroach (rBla g 5 was measured by ELISA in subjects from Cartagena, Colombia. Also, multidimensional proteomic approaches were used to study the extract of A. lumbricoides and investigate the existence of GST isoforms. We found that among asthmatics, the strength of IgE levels to GSTA was significantly higher than to mite and cockroach GSTs, and there was a strong positive correlation between IgE levels to these molecules. Specific IgE to GSTA was found in 13.2% of controls and 19.5% of asthmatics. In addition nGSTA induced wheal and flare in skin of sensitized asthmatics indicating that it might be of clinical relevance for some patients. Frequency and IgE levels to GSTA were higher in childhood and declined with age. At least six GST isoforms in A. lumbricoides bind human IgE. Four isoforms were the most abundant and several amino acid substitutions were found, mainly on the N-terminal domain. In conclusion, a new allergenic component of Ascaris has been discovered; it could have clinical impact in allergic patients and influence the diagnosis of mite and cockroach allergy in t