Lack of detectable allergenicity in genetically modified maize containing "Cry" proteins as compared to native maize based on in silico & in vitro analysis.

BACKGROUND: Genetically modified, (GM) crops with potential allergens must be evaluated for safety and endogenous IgE binding pattern compared to native variety, prior to market release. OBJECTIVE: To compare endogenous IgE binding proteins of three GM maize seeds containing Cry 1Ab,1Ac,1C transgenic proteins with non GM maize. METHODS: An integrated approach of in silico & in vitro methods was employed. Cry proteins were tested for presence of allergen sequence by FASTA in allergen databases. Biochemical assays for maize extracts were performed. Specific IgE (sIgE) and Immunoblot using food sensitized patients sera (n = 39) to non GM and GM maize antigens was performed. RESULTS: In silico approaches, confirmed for non sequence similarity of stated transgenic proteins in allergen databases. An insignificant (p> 0.05) variation in protein content between GM and non GM maize was observed. Simulated Gastric Fluid (SGF) revealed reduced number of stable protein fractions in GM then non GM maize which might be due to shift of constituent protein expression. Specific IgE values from patients showed insignificant difference in non GM and GM maize extracts. Five maize sensitized cases, recognized same 7 protein fractions of 88-28 kD as IgE bindng in both GM and non-GM maize, signifying absence of variation. Four of the reported IgE binding proteins were also found to be stable by SGF. CONCLUSION: Cry proteins did not indicate any significant similarity of >35% in allergen databases. Immunoassays also did not identify appreciable differences in endogenous IgE binding in GM and non GM maize.

Effect of thermal processing and γ-irradiation on allergenicity of legume proteins.

Legumes are implicated in IgE mediated food allergy in different countries. The present study aimed to investigate the effect of different processing methods on allergenicity of legume proteins. The extracts were processed by boiling, γ-irradiation or by combination of both. The changes in soluble protein content, specific IgE binding and allergenic potential of legume proteins were assessed. Thermal processing resulted in a 3- to 4-fold reduction in soluble protein. Specific IgE binding was reduced 74±6.5%, 83±11.6% and 62±7.2% in the soluble protein of kidney bean, black gram and peanut, respectively, after boiling (p<0.01) whereas there was 34±5.2%, 74±15.6% and 44±11.1% IgE binding reduction in the insoluble protein fraction of respective legumes. Boiling followed by γ-irradiation reduced IgE binding significantly (p<0.05). Biopotency of soluble protein of kidney bean, black gram and peanut was reduced 7-, 3- and 26-folds (p<0.001), respectively, and that of insoluble protein decreased 6-, 4- and 8-folds (p<0.001), respectively, after boiling. Combination treatment was effective in reducing the potency of both soluble and insoluble protein significantly as compared to boiling alone (p<0.001). However, γ-irradiation alone did not bring any change in allergenicity. In conclusion, boiling followed by γ-irradiation is effective in attenuating allergenicity of legume proteins.

World allergy organization study on aerobiology for creating first pollen and mold calendar with clinical significance in islamabad, pakistan;: a project of world allergy organization and pakistan allergy, asthma & clinical immunology centre of islamabad.

Pollen and mold allergies are highly problematic in Islamabad. This study was conducted to investigate the type and concentration of airborne pollens/molds causing allergic diseases in susceptible individuals. A volumetric spore trap (Burkard) was placed at the height of 11 m and ran continuously for 3 years. Once a week, the collecting drum was prepared by affixing Melinex tape with a double-sided adhesive that was coated with a thin layer of silicone grease. Every Sunday at 9:00 AM the drum was replaced by another drum and the pollen/mold spores were removed and permanently mounted on slides. Using a microscope, the trapped particles were identified and recorded as counts per cubic meter of air per hour. From these data, the pollen and mold calendars were constructed and expressed as counts per cubic meter of air per day. Skin prick tests were performed on more than 1000 patients attending the Pakistan Allergy, Asthma & Clinical Immunology Centre of Islamabad. The results indicated that there were 2 main pollen plants that contributed to seasonal allergies. These were Broussonetia papyrifera and Cannabis sativa during the March/April season and the July/September season, respectively. Although mold spores were continuously detected throughout the year, the most prominent mold was undetected mold and unconfirmed mold species similar to Stachybotrys species, which was high from July to September/October. Two additional molds contributing to allergic reactions were Pithomyces species and Cladosporium species, which were active during January and April, with the latter also being detected between October and November. These results may prove beneficial to both patients and physicians in planning a therapeutic protocol for avoidance and amelioration.

Association between indoor fungi in Delhi homes and sensitization in children with respiratory allergy.

Indoor fungi are potential sensitizing agents in children and their detection and quantification in indoor air are important in the diagnosis and environmental management of fungal allergies. The objective of this investigation was to assess the prevalence of fungal allergies in children in Delhi and to study the association between mold counts in the homes of children and their sensitization to respective fungal extracts. Fungal concentrations and seasonality were studied at two-week intervals for one year using Andersen Volumetric and Burkard Slide samplers. Sensitization to fungi frequently encountered in patients' homes was assessed by Skin Prick Tests (SPTs). Total fungal specific IgE was measured by ELISA in the sera of patients positive to fungal extracts. Skin Prick Tests revealed that 39.3% (33/84) of patients were markedly positive (2 + and above) to one or the other fungal allergens. Raised serum IgE to predominant indoor fungal species was observed in patients with marked SPT results. Highest marked skin reactivity (2 + and above) was obtained with Alternaria alternata allergens in 17.9% of the children, which was followed by the response to fungal antigens of Aspergillus fumigatus and Penicillium citrinum (15.5%). Exposure to high fungal counts of some dominant fungi (Penicillium, A. nidulans and A. fumigatus) was found associated with increased fungal sensitization in the patients. Total serum IgE level was revealed to be significantly linked with the intensity of skin reactions, as well as with skin index (r(2) = 0.052; P < 0.05). We concluded that children in Delhi are exposed to high concentrations of fungi in the indoor environment and that respiratory allergies were connected with higher prevalence of skin sensitization.

Kidney bean: a major sensitizer among legumes in asthma and rhinitis patients from India.

BACKGROUND: The prevalence of IgE mediated food allergies has increased over the last two decades. Food allergy has been reported to be fatal in highly sensitive individuals. Legumes are important food allergens but their prevalence may vary among different populations. The present study identifies sensitization to common legumes among Indian population, characterizes allergens of kidney bean and establishes its cross reactivity with other legumes. METHODOLOGY: Patients (n = 355) with history of legume allergy were skin prick tested (SPT) with 10 legumes. Specific IgE (sIgE) and total IgE were estimated in sera by enzyme-linked immunosorbent assay. Characterization of kidney bean allergens and their cross reactivity was investigated by immunobiochemical methods. Identification of major allergens of kidney bean was carried out by mass spectrometry. PRINCIPAL FINDINGS: Kidney bean exhibited sensitization in 78 (22.0%) patients followed by chickpea 65 (18.0%) and peanut 53 (15%). SPT positive patients depicted significantly elevated sIgE levels against different legumes (r = 0.85, p<0.0001). Sera from 30 kidney bean sensitive individuals exhibited basophil histamine release (16-54%) which significantly correlated with their SPT (r = 0.83, p<0.0001) and sIgE (r = 0.99, p<0.0001). Kidney bean showed eight major allergens of 58, 50, 45, 42, 40, 37, 34 and 18 kDa on immunoblot and required 67.3±2.51 ng of homologous protein for 50% IgE inhibition. Inhibition assays revealed extensive cross reactivity among kidney bean, peanut, black gram and pigeon pea. nLC-MS/MS analysis identified four allergens of kidney bean showing significant matches with known proteins namely lectin (phytohemagglutinin), phaseolin, alpha-amylase inhibitor precursor and group 3 late embryogenesis abundant protein. CONCLUSION/SIGNIFICANCE: Among legumes, kidney bean followed by chick pea and peanut are the major allergic triggers in asthma and rhinitis patients in India. Kidney bean showed eight major allergens and cross reacted with other legumes. A combination of SPT, sIgE and histamine release assay is helpful in allergy diagnosis.

Indoor fungal concentration in the homes of allergic/asthmatic children in Delhi, India.

Allergy to fungi has been linked to a wide range of illnesses, including rhinitis and asthma. Therefore, exposure to fungi in home environment is an important factor for fungal allergy. The present study was aimed to investigate types of airborne fungi inside and outside the homes of asthmatic children and control subjects (nonasthmatic children). The dominant fungi were evaluated for their quantitative distribution and seasonal variation. The air samples were collected from indoors and immediate outdoors of 77 selected homes of children suffering from bronchial asthma/allergic rhinitis using Andersen volumetric air sampler. The isolated fungal genera/species were identified using reference literature, and statistical analysis of the dominant fungi was performed to study the difference in fungal concentration between indoor and immediate outdoor sites as well as in between different seasons. A total of 4423 air samples were collected from two indoor and immediate outdoor sites in a 1-year survey of 77 homes. This resulted in the isolation of an average of 110,091 and 107,070 fungal colonies per metric cube of air from indoor and outdoor sites, respectively. A total of 68 different molds were identified. Different species of Aspergillus, Alternaria, Cladosporium, and Penicillium were found to be the most prevalent fungi in Delhi homes, which constituted 88.6% of the total colonies indoors. Highest concentration was registered in autumn and winter months. Total as well as dominant fungi displayed statistically significant differences among the four seasons (p < 0.001). The largest number of isolations were the species of Aspergillus (>40% to total colony-forming units in indoors as well as outdoors) followed by Cladosporium spp. Annual concentration of Aspergillus spp. was significantly higher (p < 0.05) inside the homes when compared with outdoors. Most of the fungi also occurred at a significantly higher (p < 0.001) rate inside the homes when compared with immediate outdoors. Asthmatic children in Delhi are exposed to a substantial concentration of mold inside their homes as well as immediate outdoor air. The considerable seasonal distributions of fungi provide valuable data for investigation of the role of fungal exposure as a risk for respiratory disorders among patients suffering from allergy or asthma in Delhi.

O que é um alergista? / What is an allergist?

Introdução: O alergista é o médico que concluiu com êxito um período de treinamento especializado em alergia e imunologia e um período de treinamento em medicina interna e/ou pediatria. Os alergistas também são imunologistas clínicos especializados, devido à base imunológica das doenças que diagnosticam e tratam. Na maioria dos países, o período aprovado de formação na especialidade em alergia e imunologia é de dois a três anos de treinamento intenso e específico. Dependendo dos sistemas de credenciamento nacionais, a conclusão desse treinamento será reconhecida por um certificado de treinamento especializado em alergia, em alergia e imunologia ou em alergia e imunologia clínica, outorgado por uma comissão diretiva. Em alguns países, isso acompanha a conclusão bem-sucedida de um exame de qualificação e, em outros, as competências apresentadas por um supervisor de treinamento. Os alergistas totalmente treinados fazem uma importante contribuição para o delineamento dos sistemas de atendimento local e proporcionam o atendimento necessário aos pacientes com doenças alérgicas. Os alergistas agem como defensores do paciente, e apóiam e questionam o caso para melhorar a educação dos médicos de atendimento primário e secundário, assim como de outros profissionais de saúde que também atendem pacientes alérgicos. Os alergistas devem estar disponíveis para fazer o atendimento dos casos mais complicados, que estão além do campo de ação de médicos de atendimento primário e secundário e de outros profissionais de saúde com bom treinamento. As principais características que definem um alergista são a apreciação da importância dos desencadeantes externos que causam a doença e o conhecimento de como identificar e tratar essas doenças, juntamente com a experiência nas terapias imunológicas e fármacos apropriados. Essa conduta no diagnóstico e na terapia é um valor essencial do especialista em alergia, e destaca o alergista entre muitos especialistas cujas bases de pacientes podem sobrepor-se com a especialidade...

Requisitos para treinamento médico em alergia: competências clínicas essenciais para o tratamento de pacientes com doenças alérgicas ou imunológicas: declaração da posição provisória da word allergy organization (WAO) / Requirements for physician training in allergy: key clinical competencies appropriate for the care of patients with allergic or immunologic diseases: a provisional position statement of the world allergy organization

Introdução: As doenças alérgicas têm prevalência extraordinária em todo o mundo, e a incidência de alergia é crescente em to¬dos os lugares!-7. Como os processos alérgicos e imunol¬gicos sobrepõem todos os sistemas orgânicos, nem sempre a alergia é ensinada nas escolas de medicina como uma disciplina separada. Realmente, a falta de reconhecimento da especialidade e da necessidade de ensinar as doenças alérgicas e imunológicas resulta no fato de a alergia não ser incluída em certos currículos de medicinas. Com a estimativa de que 22 pt por cento da população global tem doenças alérgicas e imunológicas, está na hora de reconhecer e fortalecer a educação em alergia e imunologias. A World Aflergy Organization, uma aliança de 74 sociedades nacionais e regionais de alergia, criou este documento consensual para estabelecer diretrizes educacionais que aplicadas mundialmente, para identificar e corrigir as deficiências do treinamento em alergia e para definir metas de treinamento apropriadas. Ao criar este consenso, é reconhecido que cada país tem seus próprios princípios e metas de educação médica nos níveis de graduação e pós-graduação. Este documento define o que um médico deve saber para tratar pacientes alérgicos.

Aeroallergens in clinical practice of allergy in India- ARIA Asia Pacific Workshop report.

Allergic diseases such as bronchial asthma, allergic rhinitis and atopic dermatitis are dramatically increasing all over the world including developing countries like India. Today, more than 30% of the population is known to suffer from one or other allergic ailment. Major causative agents implicated are pollen grains, fungal spores, dust mites, insect debris, animal epithelia, etc. Several aerobiological studies have been conducted in different parts of the country to ascertain aerial concentration and seasonality of pollen grains and fungi. Recently, an "All India Coordinated Project on Aeroallergens and Human Health" was undertaken by us to discover the quantitative and qualitative prevalence of aerosols at 18 different centers in the country. Allergenically important airborne pollen identified by clinico-immunologic evaluation are Alnus, Amaranthus, Argemone, Brassica, Cannabis, Cassia, Cedrus, Chenopodium, Cocos, Holoptelia, Mallotus, Morus, Parthenium, Prosopis juliflora, Quercus, Ricinus communis, and grasses such as Cenchrus, Cynodon, Imperata, Pennisetum etc. Cross-reactivity of the IgE antibodies is a common phenomenon among various pollen allergens. Ricinus communis pollen a commonly growing weed/shrub in India, cross-reacts with latex (Hevea brasiliensis), Mercurialis annua and also with seeds of Ricinus communis--all belonging to family Euphorbiaceae but geographically distantly located. Areca catechu cross-reacts with other members of Arecaceae such as Phoenix sylvestris, Cocos nucifera and Borassus flabelifer while pollen of Holoptelia integrifolia from India cross reacts with pollen of Parietaria judaica from Mediterranean Europe, both of which are members of family Urticaceae. Several reports on pollen and fruit syndrome have been analyzed. Experiments conducted by us revealed that pollutants (NO2 and SO2) not only affect pollen morphology but also changes its allergenic potency.

Soluble and nonsoluble protein assay for antigenic extracts from pollen and seeds of mustard (Brassica spp.).

A study on heterogeneity in water-soluble and non-water-soluble protein profiles of different species of Brassica pollen, seeds, and industrial flour for efficient allergy detection and immunotherapy has been lacking in India. The purpose of this study was to examine heterogeneity in the protein profile of antigenic extracts of different species of pollen, seeds, and seed flour of Brassica. A comparison of water-soluble and non-water-soluble protein profiles of seed extracts was studied. Water-soluble and non-water-soluble proteins were extracted, concentration was estimated by Lowry's method, and biochemical characterization by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was analyzed. Pollen from four species, four species of Brassica seeds comprising 25 varieties, and industrial seed flour were included for water-soluble studies, whereas four species of seeds were included for non-water-soluble protein extraction. Significant variation in protein content was observed among four different species of pollen, seeds, and industrial seed flour, respectively. No significant variation was observed in non-water-soluble extracts of four species or among water-soluble and non-water-soluble content of seeds. Heterogeneity in the protein profile of different species of pollen was not observed. However, variation in banding pattern of water-soluble as well as non-water-soluble protein extracts among four different species of seeds was observed. Interestingly, the industrial seed flour also showed a rich protein banding pattern. Variation in protein content as well as protein profile among different species of seeds of Brassica is recorded.

Allergy practice worldwide: a report by the World Allergy Organization Specialty and Training Council.

In 2004 the World Allergy Organization's Specialty and Training Council conducted a survey of World Allergy Organization (WAO) member societies to obtain information about the status of the specialty of allergy worldwide. Responses were received from 33 countries, representing a population of 1.39 billion people, of whom it was estimated that 22% may suffer from some form of allergic disease. Allergy was reported by 23 respondents to be a certified or accredited specialty in their country, and the number of certified allergists per head of population ranged from 1:25 million to 1:16,000. Allergists were ranked as the fifth most likely clinicians to see cases of allergic asthma, third most likely to see allergic rhinitis, and fourth most likely to see eczema or sinusitis. Nine countries only reported that children with allergic diseases would be seen by a pediatrician with appropriate training. The survey results highlight a pressing need for the development of allergy services worldwide.

Aeroallergens in clinical practice of allergy in India. An overview.

Allergic diseases such as bronchial asthma, allergic rhinitis and atopic dermatitis are dramatically increasing all over the world including developing countries like India. Today, more than 30 % of the population is known to suffer from one or other allergic ailment. Major causative agents implicated are pollen grains, fungal spores, dust mites, insect debris, animal epithelia, etc. Several aerobiological studies have been conducted in different parts of the country to ascertain aerial concentration and seasonality of pollen grains and fungi. Recently, an "All India Coordinated Project on Aeroallergens and Human Health" was undertaken to discover the quantitative and qualitative prevalence of aerosols at 18 different centres in the country. Predominant airborne pollen are Holoptelea, Poaceae, Asteraceae, Eucalyptus, Casuarina, Putanjiva, Cassia, Quercus, Cocos, Pinus, Cedrus, Ailanthus, Cheno/Amaranth, Cyperus, Argemone, Xanthium, Parthenium and others. Clinical and immunological evaluations have revealed allergenically important texa - some of them for the first time. Allergenically important pollen are Prosopis juliflora, Ricinus communis, Morus, Mallotus, Alnus, Querecus, Cedrus, Argemone, Amaranthus, Chenopodium, Holoptelea, Brassica, Cocos, Cannabis, Parthenium, Cassia and grasses. Further cross-reactivity of the IgE antibodies is a common phenomenon among various pollen allergens. Ricinus communis pollen from commonly growing weeds in India, cross-reacts with latex (Hevea brasiliensis), Mercurialis annua and also with seeds of Ricinus communis - all belonging to family Euphorbiaceae. Areca catechu cross-reacts with other members of Arecaceae such as Phoenix sylvestris, Cocos nucifera and Borassus flabelifer. Several reports on pollen and fruit syndrome have been analyzed. Experiments conducted by us revealed that pollutants (NO(2) and SO(2)) not only affect pollen morphology but also changes their allergenic potency. Immunotherapy with recombinant proteins having similar epitopes from different allergens have been advocated, besides allergen avoidance.

Antigenic and allergenic properties of Amaranthus Spinosus pollen--a commonly growing weed in India.

Amaranthus spinosus (Fam. Amaranthaceae) is an important aeroallergen in India and grows commonly in different parts of the country. In spite of its clinical significance in Type I hypersensitivity disorders, the antigenic and the allergenic properties of the pollen have not been systematically resolved. We investigated antigenic and allergenic properties of 5 pollen samples of Amaranthus spinosus collected from the Delhi area at fortnightly intervals. The protein content did not exhibit statistically significant variability. However, samples collected during the peak flowering season showed higher protein content. Biochemical characterization of samples showed multiple protein fractions by IEF and SDS-PAGE analysis. Samples collected during peak season showed a slightly higher number of bands (22) in the mw range of 14-70 kD. Seven protein fractions of 70, 66, 60, 50, 40, 30 and 14 kD were observed to have IgE binding capabilities and 9 were treated as allergenic. The observations will be helpful in standardizing pollen antigens for diagnosis and immunotherapy in India.

Association of IFNG gene polymorphism with asthma in the Indian population.

Epidemiologic studies in India show that the prevalence of asthma is increasing, but no genetic studies have been reported on the Indian population thus far. We selected the IFNG locus on 12q21 as a candidate gene for asthma on the basis of its role in pathophysiology and positive linkage demonstrated in other populations. The aim of this study was to investigate association of a CA-repeat marker in this gene with asthma and total serum IgE levels in the North Indian population. The repeat region was PCR-amplified from patients and control subjects and analyzed through use of GeneScan. The distributions of allele sizes were found to be significantly different between patients and control subjects (Kolmogorov-Smirnov test, P < 10(-6)). Alleles 10 and 11 were found to be overrepresented in individuals with asthma, whereas alleles 13 and 15 were less likely in asthmatic individuals. We found that the CA-repeat polymorphism in the IFNG gene was significantly associated with total serum IgE levels (ANOVA, P < 10(-4) for control subjects and P =.0036 for patients). Furthermore, a previously reported promoter polymorphism at the -333 base pair position was not detected in our population. This is the first report on the association of a candidate gene with asthma from the Indian subcontinent.

Allergen cross-reactivity between proteins of the latex from Hevea brasiliensis, seeds and pollen of Ricinus communis, and pollen of Mercurialis annua, members of the Euphorbiaceae family.

Allergen cross-reactions among three strongly sensitizing Euphorbiaceae species, i.e., the rubber tree (Hevea brasiliensis), castor bean (Ricinus communis), and the Mediterranean weed Mercurialis annua were studied in Finnish patients (n = 25) allergic to natural rubber latex (NRL), but with no known exposure to castor bean or M. annua, and French patients allergic to castor bean (n = 26) or to M. annua (n = 9), but not to NRL. In immunoglobulin E (IgE)-immunoblotting, 28% of NRL-allergic patient sera recognized castor bean seed and 48% reacted to castor bean pollen proteins. Likewise, 35% of the NRL-allergic patient sera bound to M. annua pollen allergens. Nineteen percent of castor bean-allergic patients showed IgE to NRL and 8% to M. annua proteins. Sera from patients allergic to M. annua reacted in 44% to NRL, in 56% to castor bean seed, and in 78% to castor bean pollen proteins. In immunoblotting, castor bean seed extract inhibited the binding of NRL-reactive IgE to 20 kDa, 30 kDa of NRL, and 55 kDa of proteins; NRL extract, in turn, inhibited the binding of castor bean-reactive IgE to 14, 21-22, 29, and 32-34 kDa of castor bean proteins. In ELISA inhibition, NRL extract inhibited 33% of the binding of M. annua--reactive IgE of pooled sera to M. annua pollen. In conclusion, allergen cross-reactivity in vitro was observed among three botanically related Euphorbiaceae members, H. brasiliensis, R. communis, and M. annua, but the molecular specificity of the observed cross-reactions as well as their clinical significance remains to be elucidated. Allergen cross-reactivity should be taken into account in diagnostic work.