Kung, Wing Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 92-115). / Abstracts in English and Chinese. / Abstract --- p.ii / Acknowledgements --- p.vii / Table of contents --- p.ix / List of Tables --- p.xii / List of Figures --- p.xiii / List of Abbreviations --- p.xv / Chapter Chapter 1. --- General introduction --- p.1 / Chapter Chapter 2. --- Literature review --- p.4 / Chapter 2.1 --- Food allergy and its prevalence --- p.4 / Chapter 2.2 --- Mechanism and clinical symptoms of food allergy --- p.6 / Chapter 2.3 --- Tropomyosin as the major allergen in shellfish --- p.15 / Chapter 2.4 --- Cross reactivity and epitope mapping of tropomyosin --- p.21 / Chapter 2.5 --- Novel approaches for the treatment of food allergy --- p.29 / Chapter Chapter 3. --- Expression of shrimp recombinant tropomyosin and sensitization of mice --- p.36 / Chapter 3.1 --- Introduction --- p.36 / Chapter 3.2 --- Materials and Methods --- p.40 / Chapter 3.2.1 --- "Recovery of E, coli with tropomyosin-carrying plasmid" --- p.40 / Chapter 3.2.2 --- Preparation of tropomyosin-carrying plasmid --- p.41 / Chapter 3.2.3 --- Confirmation of DNA sequence of the tropomyosin --- p.41 / Chapter 3.2.4 --- Identification of the recombinant protein --- p.43 / Chapter 3.2.5 --- Purification of the recombinant protein --- p.43 / Chapter 3.2.6 --- Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.44 / Chapter 3.2.7 --- Concentration measurement of the recombinant tropomyosin --- p.45 / Chapter 3.2.8 --- Mice --- p.46 / Chapter 3.2.9 --- Mice sensitization and challenging --- p.46 / Chapter 3.2.10 --- Tropomyosin-specific IgE level in blood --- p.47 / Chapter 3.2.11 --- Statistical analysis --- p.49 / Chapter 3.3 --- Results --- p.52 / Chapter 3.3.1 --- DNA sequence of the cloned tropomyosin --- p.52 / Chapter 3.3.2 --- Expression and purification of tropomyosin --- p.52 / Chapter 3.3.3 --- Hypersensitivity symptoms after challenge --- p.53 / Chapter 3.3.4 --- Blood tropomyosin-specific IgE level --- p.53 / Chapter 3.4 --- Discussion --- p.62 / Chapter Chapter 4. --- Identification of T cell epitopes --- p.67 / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.2 --- Materials and methods --- p.67 / Chapter 4.2.1 --- Soluble epitope peptide synthesis --- p.68 / Chapter 4.2.2 --- Isolation of spleen cells from mice --- p.69 / Chapter 4.2.3 --- T cell proliferation assay --- p.70 / Chapter 4.3 --- Results --- p.71 / Chapter 4.3.1 --- Splenocyte proliferation to synthetic peptide --- p.72 / Chapter 4.3.2 --- Splenocyte proliferation to synthetic peptides pool --- p.72 / Chapter 4.4 --- Discussion --- p.77 / Chapter Chapter5 --- General conclusion --- p.89 / References --- p.92

Bakgrund: Eksem förekommer hos 10-20% av barn i hela världen. En tredjedel av barnen med eksem har födoämnesallergi. Hos de flesta växer födoämnesallergin bort innan skolåldern. Förbättrat kliniskt omhändertagande och bättre förståelse av hur klinisk tolerans uppkommer är viktiga mål för forskning inom barnmedicin. Studieupplägg: Denna doktorsavhandling baseras på studier av två grupper av barn. Den första är en stor grupp med misstänkt allergi som undersökts med pricktest vid ett tillfälle. Den andra gruppen består av små barn med eksem och misstänkt födoämnesallergi. Barnen påbörjade studien innan två års ålder och har sedan följts över tid till fyra och ett halvt års ålder. Säkerhet vid pricktest: 5908 barn med en medelålder på 6 år och 5 månader, undersöktes med pricktest (SPT). Sju barn (0,12 %) reagerade med generaliserad allergisk reaktion (GAR), och behövde antiallergisk medicinering. Sju barn reagerade vasovagalt (VVR) med svimning eller ”nära-svimning”. Riskfaktorer för GAR var ålder <1 år (RR 6,28) och aktivt eksem (RR 16,98). Risken för VVR var högst hos tonårsflickor och barn/ungdomar undersökta med många allergen (många prickar) samtidigt, oavsett om de var positiva eller inte. Effekt av lokalbehandling och födoämneselimination hos spädbarn med eksem: 123 barn, 52 flickor och 71 pojkar deltog i studien. Åldern varierade mellan 1-24 månader, med en medelålder på 8,4 månader vid studiens början. Kraven för att få delta var eksem och/eller misstänkt födoämnesallergi. Diagnos av eksem gjordes med stöd av Hanifin och Rajkas kriterier. Eksemgrad bedömdes med instrumentet SCORAD. Barnen bedömdes vid två tillfällen med ca sex veckors mellanrum. 62 % av barnen hade positiv pricktest för födoämnen. SCORAD-värdena i gruppen med positiv pricktest var högre än i gruppen med negativ pricktest, barnen som var födoämnessensibiliserade hade alltså svårare eksem. Efter sex veckors behandling; födoämneselimination+ lokalbehandling hos SPTpositiva barn; endast lokalbehandling hos SPT-negativa barn; var det ingen skillnad i eksemens svårighetsgrad mellan de två grupperna. Både födoämnessensibiliserade och icke födoämnessensibiliserade förbättrades signifikant av behandling. En grupp med negativ pricktest, men med påvisade antikroppar mot födoämnen i blodet (analyserade först i efterhand), som behandlades enbart med lokalbehandling förbättrade sina eksem lika mycket som de barn som också ställts på eliminationskost. Antikroppar i blod och saliv i relation till toleransutveckling: Serumnivåer av total- samt ägg- och mjölkspecifika antikroppar av IgE, IgG1 och IgG4 analyserades. I saliv analyserades totalnivåer av sekretoriskt IgA samt specifikt IgA mot mjölk och ägg. Prover togs vid studiens början, efter sex veckor samt vid 4,5 års ålder. Barn som var sensibiliserade mot mjölk och/eller ägg, men som tålde dessa födoämnen vid 4,5 års ålder hade högre IgG4 nivåer och högre IgG4/IgE-kvot vid studiens början, än de barn som ej uppnått tolerans. De högsta IgG4/IgE-kvoterna sågs hos barnen med negativt pricktest men positivt specifikt IgE i blod. Under den första korta observationsperioden på sex veckor sågs ingen påverkan på barnens antikroppsnivåer. Recept/metodutvecklande och resultat av öppna och dubbel-blinda placebokontrollerade födoämnesprovokationer: Efter recept och metodutveckling för födoämnesprovokationer med mjölk och ägg, utfördes 52 provokationer på 39 barn. Fyra barn, alla provocerade blint, reagerade på provokationen. Generella slutsatser: Risken för generaliserade allergiska reaktioner vid pricktest är liten hos barn och tonåringar, men den finns. Riskfaktorer är låg ålder och aktivt eksem. Vasovagala reaktioner är lika vanliga som generaliserade allergiska reaktioner. Lokalbehandling/smörjning ger signifikant förbättring av eksem. Elimination av födoämnen kanske inte är nödvändigt hos eksembarn med sensibilisering för mjölk och ägg under förutsättning att hudvården sköts noga. Höga IgG4/IgE-kvoter av specifika antikroppar mot födoämnen kan vara associerat med snabbare toleransutveckling, och kan stödja idén med fortsatt allergenexponering hos födoämnessensibiliserade barn. Recept på beredningar som väl maskerar komjölk och ägg, vid öppna och blindade födoämnesprovokationer, är en god hjälp vid provokationer av små barn som ofta är misstänksamma mot nya smaker och konsistenser av mat. / Background: Eczema affects at least 20 % of children worldwide, and 1/3 of them also have food allergy. In most children, the food allergy is temporary. Improved clinical management and better understanding of etiological mechanisms underlying the tolerance development are target issues in paediatric research. Study design: The thesis is based on two study groups. The first is a large group of children with suspected allergy investigated with skin prick test in a cross-sectional study. The second group is a cohort of infants with eczema and/or suspected food allergy before 2 years of age, investigated prospectively with follow-up to 4.5 years of age. Safety of skin prick test (SPT): 5908 children with a mean age of 6.4 years (range: 1 month – 18 years) were investigated with SPT. Seven children, i.e. 0.12%, displayed a generalized allergic reaction (GAR), necessitating pharmacological treatment. Seven children showed a vasovagal reaction (VVR). Risk factors for GAR were age < 1 year (RR 6.28) and eczema (RR 16.98). The risk for VVR was highest among female adolescents, and children investigated with multiple skin pricks. The effect of skin care and food elimination on eczema in infants: 123 children, 52 girls and 71 boys, with a mean age of 8.4 months (range: 1-24 months) were recruited due to eczema and/or suspected food allergy. For diagnosis of eczema, the Hanifin and Rajka criteria were used, and for scoring of eczema severity SCORAD. The infants were investigated twice with an interval of 6 weeks. 62% showed positive SPTs. The SCORAD was higher among the sensitized children before treatment compared to not sensitized children. After treatment, i.e. skin care for all and elimination diet for sensitized children, there was no difference regarding eczema severity. Both SPT-positive and SPT-negative children decreased their SCORAD values significantly after treatment. A SPT-negative subgroup, with circulating specific IgE to milk/egg, was only treated with skin care, but these children improved their eczema to the same extent as those also treated with an elimination diet. Serum and salivary antibodies and achievement of tolerance Analyses were performed regarding: serum levels of total and egg- and milk-specific IgE antibodies, IgG1 and IgG4 antibodies to β-lactoglobulin (BLG) and ovalbumin (OVA); and salivary levels of total IgA, total SIgA and salivary IgA antibodies to OVA and BLG. Samples were drawn at inclusion, after 6 weeks of intervention (skin care, elimination diet), and at 4.5 years of age. Children sensitized to egg and/or milk who had developed tolerance at 4 ½ years of age had higher levels of IgG4 antibodies to OVA and BLG and also higher IgG4/IgE ratios on inclusion in the study, than those who remained non-tolerant. The highest IgG4/IgE ratios were found in children with circulating IgE antibodies to egg and/or milk but negative SPT on inclusion. The six-week treatment period did not significantly affect the levels of serum and salivary antibodies. Recipes and outcomes of open and double-blinded food challenges in children: After development of recipes for open and blinded challenge with cow’s milk and egg, 52 challenges were performed in 39 children. 4 children, challenged blindly, had a positive outcome of the challenge. General conclusions: The risk for generalized allergic reactions at SPT is low among children and teenagers, but allergic reactions do occur, and low age and eczema are risk factors. Vasovagal reactions occur as often as allergic reactions. Skin care gives significant improvement of eczema severity. Elimination diet may not be needed in infants with sensitization to milk and/or egg, provided that the skin care is adequate. High ratios of serum IgG4/IgE antibodies to food allergens may be associated with faster achievement of clinical tolerance, and may support the concept of benefit from continuing allergen exposure in sensitized children. Recipes for masking of cow’s milk and egg in open or blinded food challenges may help to accomplish challenges in young children, often suspicious to unfamiliar tastes or textures.

Food allergies are an important health problem and affect up to 2% of the adult population and 8% of children worldwide. Under the Food Allergen Labeling and Consumer Protection Act (FALCPA) of 2004, foods that contain or derive from the "Big 8" allergens (milk, egg, finfish, crustacean shellfish, tree nuts, peanuts, wheat, and soybeans) must be declared and the "common or usual name" of the allergen source must be printed on the label of the food product. Currently, the most common used detection methods for food allergens are enzyme-linked immunosorbent assay (ELISA) based. ELISA, a protein-based method, targets specific allergen(s) and detects by colorimetric reaction following binding with a specific-enzyme labeled antibody. However, studies have demonstrated that matrix interference and heat treatment can interfere with the detectability of commercial ELISA kits. An alternative approach to targeting the allergen in soy is to use deoxyribonucleic acid (DNA) as a unique marker that can be used to indicate the presence of soy in food. According to FALCPA the source of an allergen should be declared on the label, therefore identifying an allergen, such as soy, by DNA detection could be a valid means of meeting FALCPA requirements. Real-time polymerase chain reaction (real-time PCR), a DNA-based method, can identify the presence of soy through amplification of specific sequences of DNA through the use of primers. However, the sensitivity of real-time PCR can be influenced by the amplification protocol, primer design and DNA extraction methods. Thus, the main objectives of this study were to 1) verify the specificity of primers designed to detect soy DNA from different soy products, 2) optimize the previously developed real-time PCR protocol to detect soy DNA, 3) investigate the application of two commercially available DNA recovery systems (column and magnetic beads) to recover soy DNA from different forms of soy products using real-time PCR and 4) determine the effect of food matrices and thermal processing on soy detection using DNA and ELISA methods. In this study, Wizard Magnetic DNA Purification system kit (Promega, Madison, WI) was selected as the column DNA recovery system while DNeasy mericon Food Kit (Qiagen, Valencia, CA) was selected as the magnetic beads system. Neogen Veratox for soy allergen was selected as the ELISA system. The evaluations of both DNA recovery systems were conducted on soy protein isolates (SPI), powdered soybean and soymilk. The effect of thermal processing in soy detection was conducted on four different food matrices (protein, fat, carbohydrate and water). Each food matrix was spiked with 10% soy protein isolates and heated at 95ºC for an hour. Both DNA (column and magnetic beads DNA recovery system) and ELISA detection methods were used to detect soy in heated and non-heated food matrices. The limit of detection for column DNA recovery method in soybean, SPI and soymilk can be as low as 20 ppm, while magnetic beads DNA method was matrix dependent. The magnetic beads methods demonstrated a lower detection for soybean sample (1.33 ppm) but higher for soymilk (133.3 ppm). The soy percent recovery for non-heated food matrices was higher in ELISA methods and lower in magnetic beads DNA method. For heated food matrices, percent recovery for both DNA methods was higher than ELISA method. Overall, heat treatment can significantly reduce the ability of the ELISA method to detect soy in all food matrices. However, for DNA methods (column and magnetic beads), water and ranch matrices were the only two that were significantly affected by thermal processing. In terms of food matrices, water matrix (heated and non-heated) has the highest percent recovery of soy for all detection methods. However, percent recovery of soy in flour matrix (non-heated) was the lowest using both DNA methods. / Graduation date: 2013

published_or_final_version / Paediatrics and Adolescent Medicine / Master / Master of Medical Sciences

Orientador: Flavia Maria Netto / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-20T14:16:39Z (GMT). No. of bitstreams: 1 VillasBoas_MarianaBattaglin_D.pdf: 3357506 bytes, checksum: 10d878e81440a9db2f379bd1dd93146f (MD5) Previous issue date: 2012 / Resumo: Tratamento térmico, alta pressão ou hidrólise enzimática, utilizados em conjunto ou separadamente, podem alterar determinantes antigênicos (epítopos) presentes nas proteínas, e têm sido estudados como estratégia para obtenção de produtos com menor potencial alergênico, porém ainda com sucesso limitado. A enzima transglutaminase (TG), que catalisa a reação de ligação cruzada inter ou intramolecular em diversas proteínas, também tem sido utilizada com este propósito. A associação da hidrólise enzimática com a polimerização por TG é uma estratégia ainda pouco explorada, mas que oferece boas perspectivas em relação à redução da antigenicidade de proteínas. Diante disso, o presente estudo teve como objetivos obter e caracterizar a ß-Lactoglobulina (ß-Lg) polimerizada por TG e a ß-Lg polimerizada pré e pós hidrólise enzimática com as proteases Alcalase, Neutrase ou bromelina, e avaliar o efeito destes processos na antigenicidade da proteína. A melhor condição de hidrólise da ß-Lg, com cada uma das proteases estudadas foi definida a partir de um delineamento composto central rotacional (DCCR), tendo como variáveis independentes a concentração de proteína (2,2 -7,8% p/v) e relação enzima : substrato (E:S), 6,9 - 28 U g-1 proteína e, como variável dependente o grau de hidrólise (GH), determinado pelo método pH-stat. A condição selecionada, 3% de proteína e E:S 25 U g-1, foi a utilizada para a hidrólise da ß-Lg pré ou pós polimerização com a enzima TG. As condições experimentais foram: (1) hidrólise da ß-Lg com as três enzimas, seguida de polimerização com TG (10 ou 25 U g-1) ou (2) polimerização da ß-Lg (7%p/v) com TG (10U TG g-1) realizada pós tratamento térmico ou na presença de agente redutor Cys (0,1 mol L-1), seguido por hidrólise com as diferentes proteases. As amostras hidrolisadas e as polimerizadas pré ou pós hidrólise foram caracterizadas por eletroforese SDS-PAGE/tricina, cromatografia líquida de alta eficiência de fase reversa (CLAE-FR), cromatografia líquida acoplada à espectrometria de massas (MS) e ensaios imunoquímicos (ELISA e Imunoblote). Os hidrolisados obtidos com Alcalase apresentaram 12,7% de GH e com Neutrase e bromelina aproximadamente 4%. Os perfis eletroforéticos da ß-Lg polimerizada pós tratamento térmico (ß-Lg TT TG) ou na presença de Cys (ß-Lg Cys TG), apresentaram bandas de massa molecular (MM) alta, porém a polimerização pós hidrólise, avaliada por eletroforese e cromatografia, não resultou em aumento da MM aparente dos hidrolisados. A ligação isopeptídica e-(?-Gln)Lys, catalisada pela TG, foi detectada em concentração mais elevada no material polimerizado pós hidrólise com alcalase, seguido do tratamento de polimerização da presença de Cys. No estudo da polimerização pré hidrólise, a alcalase foi a enzima que hidrolisou mais eficientemente tanto a ß-Lg como seus polímeros. Os polimerizados pre ou pós hidrólise com neutrase ou bromelina apresentaram ß-Lg residual, observada no perfil eletroforético e cromatográfico, sendo que para os hidrolisados com neutrase, a fração correspondente à ß-Lg residual apresentou maior intensidade. O tratamento isolado de hidrólise, principalmente com alcalase ou bromelina, ou a associação deste processo com a TG foram capazes de reduzir significativamente a resposta antigênica da ß-Lg. Os hidrolisados da ß-Lg com neutrase apresentaram reação antígeno-anticorpo apenas na região correspondente ao monômero da ß-Lg, sendo que a reação foi menos intensa após a polimerização do hidrolisado com a TG. As amostras foram submetidas à digestão in vitro e avaliadas por espectrometria de massas (MS/MS) quanto à identificação de epítopos lineares. Verificou-se que tanto a ß-Lg polimerizada após tratamento térmico ou na presença de Cys como a amostras polimerizadas pré ou pós hidrólise com bromelina apresentaram alguns fragmentos correspondentes aos epítopos, embora em menor número quando comparados aos epítopos identificados na ß-Lg nativa. Para o tratamento com a alcalase, isolado ou associado à TG, não foram identificados fragmentos correspondentes aos epítopos após digestão gastrointestinal das amostras. Os resultados obtidos indicaram que a hidrólise, principalmente com alcalase, associada ou não à polimerização com TG, foi capaz de reduzir a antigenicidade da ß-Lg, mostrando-se uma alternativa para a produção de alimentos hipoalergênicos / Abstract: Heat treatment, high pressure and enzymatic hydrolysis have been studied aiming at obtaining products with low allergenic potential since they can alter antigenic determinants (epitopes) in the protein. The transglutaminase (TG), an enzyme that catalyzes inter or intramolecular cross-link in different proteins has also been used to reduce the protein antigenicity. The association of enzymatic hydrolysis and polymerization by TG is another strategy for reducing the antigenicity of food proteins, promising that needs further research. Therefore, the present study aimed to obtain and characterize the ß-Lg polymerized by TG before or after enzymatic hydrolysis with the proteases alcalase, neutrase or bromelain and evaluate the effect of these processes on the protein antigenic response. The optimum conditions for hydrolysis of ß-Lg, obtained with the three different enzymes, were defined using a central composite rotatable design (CCRD), where the independent variables were protein concentration (2.2% -7.8 w / v) and enzyme: substrate ratio (E:S), 6.9 to 28 U g-1 protein, and the dependent variable (response) was the degree of hydrolysis (DH) determined by the pH-stat method. The hydrolysis condition defined by CCRD was 3% of protein (w/v) and E:S ratio 25 U g-1. The experimental conditions were: (1) hydrolysis of ß-Lg with the three enzymes used separately, followed by polymerization by TG (10 or 25 U g-1) or (2) polymerization of ß-Lg (7% w/v) by TG (10U g TG-1) after heat-treatment (80 °C/60 min) or in presence of reducing agent Cys (0.1 mol L-1), followed by hydrolysis. The samples were characterized by SDS-PAGE/tricine, reverse phase high-performance liquid chromatography (RP-HPLC), liquid chromatography coupled to mass spectrometry (HPLC-MS) and by immunochemical assays (ELISA and Imunoblote). The highest DH of the ß-Lg was obtained with alcalase (12.6%), while with neutrase or bromelain a DH of 4% approximately was achieved. The electrophoresis profiles of ß-Lg polymerized after heat treatment (ß-Lg TT TG) or in the presence of Cys (ß-Lg Cys TG) showed bands with high molecular mass (MM). On the other hand, the post-hydrolysis polymerization, analyzed by SDS-PAGE and RP-HPLC, resulted in no increase in the MM of the hydrolysates. The highest concentration of the cross-link, by formation of the isopeptide bound e-(?-Gln) Lys, was found in the sample polymerized after hydrolysis with alcalase, followed by the sample ß-Lg Cys TG. The polymerized pre or pos hydrolysis with neutrase or with bromelain showed residual ß-Lg, observed in electrophoretic and chromatographic profiles, and for hydrolysates with neutrase, the band corresponding to ß-Lg showed higher residual intensity. Alcalase was the enzyme which hydrolysed more efficiently both untreated and polymerized ß-Lg. Using neutrase or bromelain to hydrolyse polymerized ß-Lg, a residual ß-Lg (18.3 kDa) was observed in the electrophoretic and chromatographic profiles. The avaluation of antigenicity showed that hydrolysis treatment used alone, especially with alcalase or bromelain, or the combination of this process with TG was capable to reduce significantly the antigenic response of ß-Lg. The ß-Lg hydrolysed with neutrase showed antigen-antibody reaction only in the region corresponding to the ß-Lg monomer, and this reaction was less intense after the association of hydrolysis with neutrase and polymerization by TG. The samples were submitted to in vitro digestion and then analyzed by mass spectrometry (MS / MS) to identification of linear epitopes in the protein. It was verified that the polymerized samples (ß-Lg TT TG or ß-Lg Cys TG) as well as those polymerized before or after hydrolysis with bromelain, retained some fragments corresponding to the epitopes, although to a lesser extent when compared to the epitopes identified in the untreated ß-Lg. Interestingly that for the treatment with alcalase, associated or not with the TG, no epitopes were identified after gastrointestinal digestion. These results indicated that the hydrolysis, especially with alcalase, and the association of hydrolysis with polymerization, were capable to reduce the ß-Lg antigenicity and could be an alternative for elaboration of hypoallergenic products / Doutorado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Doutor em Alimentos e Nutrição

Orientador: Nilton Carlos Machado / Resumo: Introdução. As alergias alimentares são um problema crescente no mundo, e a única maneira de tratar continua sendo a exclusão de alimentos com a proteína implicada. A ANVISA estabeleceu requisitos para a rotulagem dos principais alimentos que causam alergias alimentares. Portanto a leitura dos rótulos dos alimentos embalados deve ser praticada por todo cuidador de crianças alérgicas. Objetivo. Qualificar os rótulos apresentados pelas indústrias de alimentos com base na resolução atual e propor medidas que beneficiem a leitura de rótulos pelo público alérgico. Métodos. Estudo observacional transversal para avaliar a rotulagem de alérgenos de alimentos embalados. No primeiro momento foi realizada uma busca em supermercados de diferentes marcas de gêneros alimentícios. No segundo momento mediante sorteio eletrônico foi obtida amostra para analise, composta do mínimo de 50% de marcas de alimentos embalados de cada gênero alimentício. Posteriormente, foi realizada fotografia digital de cada produto, em todas as suas dimensões (painel principal, laterais e fundos). Os alimentos embalados foram divididos com base nos Grupos da Pirâmide alimentar brasileira. Foram analisados: os ingredientes presentes e as características da rotulagem. Dados apresentados de forma descritiva. A classificação dos rótulos foi apresentada em escore baseado na resolução vigente, gerando pontuação de 0 a 10 aos diferentes alimentos dos Grupos da Pirâmide alimentar brasileira. Resultados. Os alimentos embal... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction. Food allergies are a growing problem worldwide, and the only way to treat them is to exclude foods with the protein involved. ANVISA has established requirements for the labelling of major foods that cause food allergies. Therefore, the reading of packaged food labels should be practiced by all caregivers of allergic children. Aims. Qualify the labels presented by the food industries based on the current resolution and propose measures that benefit the reading of labels by the allergic public. Methods. Cross-sectional observational study to evaluate allergen labelling of packaged foods. At first, a search was done in supermarkets of different brands of foodstuffs. In the second moment, utilizing an electronic draw, a sample was obtained for analysis, composed of at least 50% of packaged food brands of each foodstuff. Subsequently, digital photography of each product was performed, in all its dimensions (main panel, sides and bottoms). The packaged foods were divided based on the Brazilian Food Pyramid Groups. Were analyzed: the present ingredients and the characteristics of the labelling. Data are presented descriptively. The classification of the labels was presented in a score based on the current resolution, generating a score from 0 to 10 to the different foods of the Brazilian Food Pyramid Groups. Results. Packaged foods have quality information, with labeling scores close to the maximum in all groups. Regarding the indication of the presence of allergens: ... (Complete abstract click electronic access below) / Mestre

Food allergies are an increasing health concern in the United States, affecting nearly 6 million children under the age of 18 years. Research has suggested that 18% of school-age children will have their first allergic reactions at school. Life-threatening allergic reactions experienced by children in the school setting are on the rise; however, little is known about how schools implement policies and practices in response to this issue. The purpose of this quantitative cross-sectional study was to narrow the knowledge gap by examining teachers' knowledge, ability, and confidence level caring for students with food allergies. Bandura's social cognitive theory, which holds that education and experience influence confidence implementing tasks, served as the framework that guided this research. The electronic survey was distributed to a convenience sample of 300 elementary school teachers; 93 respondents completed it. Eighty completed surveys were used in the analysis. Multiple linear regression models were constructed to analyze the relationships among confidence, education, and training related to food allergies. Results showed that teachers who lacked knowledge of food allergies also lacked confidence implementing food allergy plans. School personnel responsible for planning or revising food allergy response protocols can use these findings. The potential for positive social change includes identifying training opportunities, developing policies to sustain food allergy knowledge, and building the capacity of all school staff to implement life-saving measures when children are experiencing allergic reactions.

La régulation de l'homéostasie intestinale est de la plus haute importance en raison de la constante exposition de l'intestin aux antigènes alimentaires et à la flore commensale. La perturbation de la flore intestinale est souvent associée à diverses maladies telles que l'allergie, l'obésité et certaines maladies inflammatoires. La plupart des individus sont tolérant aux antigènes alimentaires et ne développe pas de réponse immunitaire sauf en cas de prédisposition génétique ou d'exposition à un environnement défavorable. La réponse allergique se caractérise par la production d'IgE stimulé par les lymphocytes Th2. Les symptômes allergiques sont très variés et affectent plusieurs parties de l'organisme. La plupart des travaux de recherche se sont focalisé jusqu'à présent sur le rôle des cellules de l'immunité adaptative dans le développement de l'allergie en sous-estimant le rôle majeur des cellules épithéliales et des cellules de l'immunité innée. L'objectif de ce projet est de comprendre comment les cellules épithéliales intestinales modulent la réponse immunitaire à distance vers la muqueuse pulmonaire ou la peau après stimulation allergique. L'ingestion de l'antigène associé à l'adjuvant de la toxine cholérique permet d'étudier la réponse allergique chez l'animal. Nous avons démontré sur ce modèle animal que l'absence de la kinase inhibitrice IKKb dans la voie de signalisation du facteur de transcription NF-kB altère la composition de la flore intestinal d'une part et transforme la réponse immunitaire inflammatoire au niveau pulmonaire et de la peau grâce à la présence d'IgA et de lymphocyte Th17 d'autre part. En adéquation avec les observations cliniques rapportées chez les patients allergiques (allergies alimentaires, asthme, dermatite atopique), nos résultats identifient IKKb dans la cellule épithéliale intestinale comme cible potentielle pour traiter les allergies alimentaires. De futurs efforts devront être faits pour développer de nouvelles stratégies thérapeutiques qui considèrent la muqueuse intestinale, la production d'IgA et l'importance des bactéries commensales dans le traitement des allergies. / Immune homeostasis is of paramount importance in the gastrointestinal tract, which is constantly exposed to ingested antigens and commensal microbiota. The gut microbiota can be perturbed by endogenous or exogenous factors and it is now established that microbial dysbiosis is associated with allergy, obesity, and inflammatory diseases. Ingestion of food antigens generally fails to promote brisk immune responses but rather results in a state of immune tolerance. However, aberrant immune responses can develop in individuals with a genetic predisposition. Food allergies are generally regarded as pathologic responses to food antigens mediated by excessive Th2 responses and antigen-specific IgE antibody responses. Clinical manifestations of food allergies are very broad and symptoms can affect different organs. While past research on allergy focused on the role of cells and molecules involved in adaptive immunity, epithelial cells lining the sites of antigen entry and innate immune responses have recently emerged as important players in allergy. This project was undertaken to understand the mechanisms employed by intestinal epithelial cells (IECs) to shape immune responses to allergens and influence allergic manifestations in distant mucosal sites such as the airways or the skin. Oral administration of food antigen with cholera toxin as adjuvant in experimental animals is a well-accepted model to study allergic sensitization to food antigens. Using this model, we show that a localized impairment of the canonical NF-κB pathway through deletion of IkB kinase (IKKβ) in IECs alters the gut microbiota during oral allergic sensitization and regulates the magnitude of allergic inflammatory responses at distant sites of the airway and the skin through enhancement of IgA Abs and Th17 responses. Consistent with the clinical observations linking atopic diseases (food allergy, allergic asthma, atopic dermatitis), our results identify IKKβ in IECs as a potential therapeutic target for treatment of food allergies and subsequent disease. They also suggest that future efforts for controlling allergic responses in the airways and the skin could include strategies that use the gut microbiota and promote IgA Ab responses and prevent IL-17 responses.

A hipersensibilidade a alimentos é comum em crianças e adultos. O camarão, apesar de muito apreciado, é um alimento potencialmente capaz de desencadear reações alérgicas em indivíduos geneticamente predispostos. O principal alérgeno em camarão é a tropomiosina. O objetivo desse experimento foi avaliar os efeitos da radiação gama na reatividade IgE a tropomiosina de camarão da espécie Litopenaeus vannamei e avaliar as modificações físico-químicas e sensoriais no produto, induzidas por esse processo. As amostras foram constituídas de camarões sem o cefalotórax, descongelados e liofilizados, submetidos a diferentes doses de radiação gama: 0 (test.), 5, 10 e 15 kGy. Utilizou-se para a irradiação um irradiador multipropósito, semi-industrial de 60 Co. A taxa de dose empregada foi de 7,5 kGy/hora. Determinou-se a reatividade IgE utilizando as técnicas: ELISA e \"Imunobloting\". Foram realizados testes físico-químicos e sensoriais, para verificar a viabilidade do produto após a irradiação. Os dados obtidos foram submetidos à análise de variância (ANOVA) e teste de Tukey (p<=0,05). As doses de radiação gama aplicadas nesse experimento não alteraram de forma significativa a reatividade IgE, a cor, o colesterol, teor de ácidos graxos e a aceitação dos camarões. Concluiu-se que a radiação gama nessas doses, apesar de não causar modificações que comprometam a qualidade do produto, não pode ser indicada para o controle de alérgenos em camarões, por ter apresentado baixo impacto na modificação da estrutura protéica alergênica / The hypersensitivity to foods is common in children and adults. The shrimp in spite of very appreciated is a food potentially capable to unchain allergic reactions genetically in individuals per disposed. The major allergen in shrimp is the tropomiosin. The aim of this study was to evaluate the effects of gamma irradiation on IgE reactivity to shrimp tropomyosin (Litopenaeus vannamei) and evaluate the physical and chemical changes and sensory product, induced by this process. The samples will be constituted of thawed and freeze-dried shrimp submitted to different radiation doses: 0 (test.), 5, 10 and 15 kGy. Was used for irradiating an irradiation multipurpose semi-industrial sector. 60 Co. The dose rate used was 7.5 kGy / hr. The samples, later to the irradiation, they will go by analyses for allergens determination using the techniques: ELISA and Imunobloting. Acceptance test will be done to verify physic-chemical and sensorial attributes after irradiation. Datas will be evaluated by ANOVA and Tukey test (p<=0.05). The gamma radiation doses applied in this experiment did not change significantly IgE reactivity, color, cholesterol, fatty acid content and the acceptance of shrimp studied. It was concluded that the gamma radiation in these doses, although not causing changes that compromise the quality of the product cannot be indicated for control of allergens in shrimp because it presented a low impact on allergenic protein structure modification

As alergias alimentares afetam 6% das crianças e 3% a 4% dos adultos e é uma reação adversa que o organismo produz com a ingestão de alimentos, a qual envolvem mecanismos imunológicos, mediados ou não pela imunoglobulina E (IgE) ou reações mistas. Ante a falta de norma específica de regulamentação da rotulagem da presença de alimentos alérgenos em medicamentos, cosméticos, perfumes e saneantes domissanitários, este estudo apresenta uma análise da necessidade de se publicar norma que regulamente a rotulagem desses produtos, à luz dos direitos à informação, à saúde, à vida e do princípio da dignidade da pessoa humana, uma vez que os rótulos de produtos de beleza, higiene e limpeza, bem como as bulas de medicamentos não trazem expressos todos os ingredientes envolvidos que os compõem. Como objetivos específicos, foram feitas considerações acerca do direito à saúde, com ênfase no princípio da dignidade da pessoa humana; redigir acerca do direito à vida e à saúde do consumidor, bem como o direito à informação; apontar a alergia alimentar, com demonstração dos tipos de reações e os principais alimentos alérgenos, bem como expor as normas existentes acerca da rotulagem de alimentos. Em relação às escolhas metodológicas foram utilizadas as pesquisas explicativa, qualitativa e bibliográfica. Em decorrência da inexistência de norma específica, como produto final desta dissertação, foi elaborada uma minuta de resolução a qual regulamenta a presença de alimentos alérgenos nos produtos mencionados, a ser encaminhada à ANVISA como proposta de publicação. / Food allergies affect 6% of children and 3% to 4% of adults and are an adverse reaction that the organism produces with the food intake, which involves immunological mechanisms, whether or not mediated by immunoglobulin E (IgE) or mixed reactions. In view of the lack of specific rules on the labeling of the presence of food allergens in medicinal products, cosmetics, perfumes and sanitizing products, this study presents an analysis of the need to publish a standard that regulates the labeling of these products, based on the right to information, health, life and the principle of human dignity, since the labels of beauty, hygiene and cleaning products as well as the package inserts do not express all the ingredients that compose them. As specific objectives, conclusions were made on the right to health, with emphasis on the principle of human dignity; to draw up the right to life and health of the consumer, as well as the right to information; to point up the food allergy, with demonstration of the types of reactions and the main allergenic foods, as well as to expose the rules on the labeling of foods. Regarding the methodological choices, there were used explanatory, qualitative and bibliographic researches. Due to the lack of a specific standard, as a final product of this dissertation, a draft resolution was prepared which regulates the presence of allergens in the mentioned products, to be sent to ANVISA as a proposal for publication.