O Teste de Provocação Oral Duplo Cego Placebo Controlado (TPODCPC) é considerado um método diagnóstico de extrema importância na alergia alimentar, entretanto não existe, em nosso meio, uma padronização em relação aos materiais e métodos para a sua execução, especialmente na faixa etária pediátrica. O objetivo deste estudo foi adaptar TPODCPC para o diagnóstico de alergia às proteínas do leite de vaca (APLV) mediada por IgE, em crianças e adolescentes. O objetivo secundário foi descrever a relação entre os antecedentes de atopia associados a dados laboratoriais e os resultados dos TPODCPC. Foram incluídos 58 pacientes que se dividiram em dois grupos. O grupo 1 foi composto por 39 pacientes (mediana de idade: 5,3 anos; 1,6M:1F) com história sugestiva de APLV IgE mediada, sem relato de história de anafilaxia recente e com pesquisa positiva de IgE específica para leite de vaca (LV) e/ou frações. No grupo 2 foram incluídos 19 pacientes (mediana de idade: 8,3 anos; 1,4 M:1F) sem história sugestiva de APLV. Os itens avaliados na adaptação deste método diagnóstico incluíram: escolha do local, materiais a serem utilizados e operacionalização do teste. O hospital dia mostrou-se, por suas características, adequado à realização do exame. Em relação aos materiais escolhidos para a oferta do veículo, os recipientes opacos e vedados revelaram-se mais adaptados às necessidades do teste, os veículos de maior aceitação foram sopa de legumes e bebida à base de soja e o LV a ser oferecido de maneira mais apropriada foi na forma líquida e com baixo teor de lactose. As dificuldades observadas na realização do TPODCPC não comprometeram a sua execução, entretanto houve dificuldade na interpretação dos resultados especialmente nos pacientes com sintomas como pápulas periorais ou achados clínicos inespecíficos. Observou-se concordância entre os critérios clínico-laboratoriais adotados para a definição dos grupos 1 e 2 com os resultados do TPODCPC. Em conclusão, o teste mostrou-se exeqüível e seguro e a adaptação deste para o diagnóstico de APLV IgE mediada nesta faixa etária pediátrica foi possível, sendo necessária a avaliação da sua reprodutibilidade em outras populações. / The Double Blind Placebo Controlled Food Challenge (DBPCFC) is considered an important method for food allergy diagnosis, nevertheless it is not standardized among us. The aim of this study was to adapt DBPCFC for cows milk allergy (CMA) diagnosis in children and adolescents. A secondary aim was to describe the relation between history of atopy and laboratorial findings with DBPCFC results. It was included 58 patients that were distributed in two groups. Group 1 was composed of 39 patients (median age: 5,3 years; 1,6M:1F) with suggestive history of CMA without recent anaphylaxis, and specific IgE to cow´s milk (CM) and/or its fractions. In group 2 was included 19 patients (median age 8,3 years; 1,4 M:1F) where the CMA diagnosis was excluded based on clinical findings. The items evaluated in the adaptation of this method were: setting choice, kind of milk processing and vehicles, besides material and test performance. Day hospital was considered adequate for this test. Regarding to elected materials to offer the vehicle, opaque and sealed recipients showed more suitable to test demands. Vehicles more accepted were legumes soup and soy beverage and the most adequate CM to be offered was liquid and with low lactose concentration. The difficult observed in performance of DBPCFC do not compromise its execution, however the interpretation of the results was very difficult specially in patients with perioral wheals and unspecific clinical findings. There is an agreement Comparing adopted clinical and laboratorial criterion for definition of group 1 and 2 with DBPCFC results. In conclusion, the test proved to be feasible and safe and its adaptation is possible for IgE mediated CMA diagnosis, being necessary to evaluate its reproducibility in other populations.

O objetivo deste estudo foi avaliar, in vitro, quantitativamente e qualitativamente, a estabilidade de um híbrido experimental em duas concentrações diferentes (concentrado e diluído) aplicado sobre a simulação de uma dentina sensível. Dentes molares humanos foram selecionados e tiveram suas coroas seccionadas abaixo do sulco oclusal de forma a obter espécimes de discos de dentina que foram planificados e polidos até atingirem a espessura de 1,0 milímetro. Os espécimes foram divididos em 4 grupos (n=9) de acordo com os tratamentos de superfície propostos: saliva artificial (SAL), adesivo dentinário autocondicionante (AD), híbrido experimental concentrado (TC) e híbrido experimental diluído na proporção de 1:3 (TD). Dois métodos foram empregados para avaliar a estabilidade: condutância hidráulica (permeabilidade dentinária) e microscopia eletrônica de varredura. Foi realizada a leitura da permeabilidade dentinária em 6 tempos experimentais: Mínima (sem tratamento), Máxima (com túbulos abertos), Tratamento (após aplicação dos respectivos tratamentos), Erosão (após 5 minutos de imersão em ácido cítrico 0,05M pH 3,8), Escovação (após escovação de 3900 ciclos) e Erosão Pós (repetição da erosão após o processo de escovação). A microscopia eletrônica de varredura foi realizada em espécimes de dentina com área central de aplicação dos tratamentos e tecido natural nas laterais para evidenciar as características da película aplicada. As leituras foram feitas após a aplicação dos tratamentos, após a erosão, após a escovação e após a erosão pós escovação, para todos os 4 tratamentos propostos. O teste de análise de variância (ANOVA) de Medidas Repetidas com 2 fatores de variação foi aplicado juntamente com o teste de comparações múltiplas pareadas (Tukey). Para permeabilidade dentinária todos os tratamentos reduziram a condutância hidráulica (Lp) em relação à Máxima. TC e TD apresentaram os menores valores (24% e 15%) respectivamente. O TD continuou apresentando valores semelhantes estatisticamente após a Erosão (36%), sendo estatisticamente semelhante ao TC (55%).. No tempo Escovação o TD apresentou Lp estatisticamente semelhante aos tempos Tratamento e Erosão. Todos os grupos apresentaram-se estatisticamente semelhantes entre os tratamentos nos tempos Escovação e Erosão Pós. A análise das MEVs evidencia túbulos dentinários com conteúdo no seu interior nos grupos TC e TD, mantendo-se durante todos os tempos experimentais. O AD apresentou uma película evidente, que começou a se destacar e exibir falhas a partir do tempo Erosão. Conclui-se que o TD apresentou o melhor comportamento sendo capaz de diminuir a permeabilidade dentinária, formando uma película fina, transparente, imperceptível, capaz de vedar (totalmente ou parcialmente) e penetrar dentro dos túbulos dentinários, resistindo aos desafios erosivos e abrasivos. / The aim of this study was to evaluate, in vitro, quantitatively and qualitatively, the stability of an experimental hybrid with two different concentrations (concentrated and diluted) applied at a simulation of a sensitive dentin. Human molar teeth were selected and their crowns were sectioned below the occlusal groove in order to obtain specimens of dentine disks that were ground flat and polished to achieve a thickness of 1.0 millimeter. The specimens were divided into 4 groups (n = 9) in accordance with proposed surface treatments: Artificial saliva (SAL) dentinal self-etching adhesive (AD), concentrated experimental hybrid (TC) and experimental hybrid diluted in the ratio 1: 3 (TD). Two methods were used to assess the stability: hydraulic conductance (dentin permeability) and scanning electron microscopy. The dentin permeability in 6 experimental times was carried out: Minimum (no treatment), Maximum (with open tubules), treatment (after application of their treatments), erosion (after 5 minutes of immersion in citric acid 0.05M pH 3, 8), brushing (brushing after 3900 cycles) and Post erosion (erosion was repeated after brushing). The scanning electron microscopy was performed on dentin specimens with a central area of application of treatments and natural tissues on the sides to show the film characteristics applied. Readings were made after application of treatments, after erosion, after brushing and after erosion after brushing, for all 4 treatments proposed. The analysis of variance (ANOVA) for repeated measures with two variation factors was applied with the multiple comparisons paired test (Tukey). For dentin all treatments reduced hydraulic conductance (Lp) in relation to Maximum. TC and TD showed the lowest values (24% and 15%) respectively. The TD continued to show statistically similar values after erosion (36%), being statistically similar to TC (55%). In brushing time the TD Lp was statistically similar to Treatment and erosion times. All groups were statistically similar between treatments in brushing and Post Erosion times. The analysis of SEM shows dentinal tubules with content inside on the TC and TD groups, remaining during all experimental period. AD presented a clear film, which began to stand out and show failure from erosion time. It follows that the TD had better behavior being able to decrease permeability of dentin by forming a thin film transparent, imperceptible, capable of sealing (fully or partially) and penetrate within the dentine tubules, resisting the erosive and abrasive challenges.

Asma é uma doença inflamatória pulmonar crônica usualmente associada com imunidade do tipo 2, eosinofilia pulmonar, hiperreatividade brônquica (AHR), hiper-produção de muco e altos níveis de IgE. Indíviduos asmáticos podem responder aos alérgenos por duas distintas fases: uma fase imediata (EPR) e uma fase tardia (LPR), ambas não reproduzidas na maioria dos modelos murinos de asma. No presente trabalho utilizamos camundongos AIRmax e AIRmin. Verificamos que camundongos AIRmin respondem com uma AHR intrínseca a metacolina. Esta resposta intensa correlacionou-se com uma menor expressão de receptores muscarínicos do tipo 2. Camundongos AIRmax sensibilizados e desafiados com OVA, ao contrário dos AIRmin, desenvolveram LPR e AHR a MCh. Os AIRmax apresentam um denso infiltrado inflamatório com predominância de eosinófilos, uma elevada produção de muco, citocinas (IL-5 e IL-13) e anticorpos anafiláticos IgE e IgG1. De forma surpreendente animais AIRmax desenvolvem quadro alérgico pulmonar crônico que cursa com AHR a MCh e uma inflamação pulmonar com infiltrado de eosinófilos com deposição de colágeno no tecido pulmonar além de uma produção elevada de anticorpos anafiláticos. Em conclusão desenvolvemos um novo modelo murino de asma. / Asthma is a chronic inflammatory lung disease usually associated to Type 2 T helper cells, lung eosinophilia, airway hyper-reactivity (AHR), mucus hyper-secretion and increased titers of IgE. Asthmatic individuals may react to allergens by two distinct phases: an immediate phase (EPR) and a late phase (LPR) both phases were not reproduced in classical murine models of asthma. In the present study we use AIRmax AIRmin mice. We found that AIRmin mice exposed to methacholine presented intense intrinsic AHR. This intense reaction was related to a lower expression of muscarinic type 2 receptors. AIRmax mice sensitized and challenged with OVA, unlike AIRmin developed LPR and AHR to MCh. The AIRmax also developed a dense inflammatory infiltrate containing predominantly eosinophils, hyper-secretion of mucus, cytokines (IL-5 and IL-13) and anaphylactic antibodies (IgE and IgG1). Surprisingly AIRmax mice develop chronic pulmonary allergic framework that leads to AHR to MCh and lung inflammation with eosinophilic infiltration with collagen deposition in lung tissue and a high production of anaphylactic antibodies. In conclusion, we developed a new murine model of asthma.

As alergias podem ser desencadeadas por substancias químicas, medicamentos, proteínas de origem vegetal e animal como, por exemplo, ácaros, alimentos, fungos e venenos. Reações alérgicas desenvolvidas em resposta a venenos de animais marinhos vêm sendo pouco estudadas. Este é o primeiro estudo que descreve a indução de reação anafilática em camundongos pelo veneno de um peixe brasileiro, acompanhado da caracterização detalhada dos mediadores solúveis e celulares envolvidos no processo. Nossos resultados mostram que o veneno do peixe T. nattereri possui proteínas alergênicas capazes de desencadear um processo alérgico, caracterizado por anafilaxia mediada por IgE e IgG1 e inflamação eosinofílica de fase tardia dependente de citocinas Th2. Ainda demonstramos a regulação da resposta pela positiva pela citocina IL-4 e participação da IL-12 e IFN-<font face=\"Symbol\">g na resposta. / Allergies are a significant and widespread public health problem. Anaphylaxis reactions are inducing by foods, medications and venoms and are IgE mediated. In mice, allergy can be caused also by IgG1. The results presented here describe for the first time anaphylaxis induced by Brazilian fish venom, accompanied by detailed characterization of soluble and cellular mediators involved in the process. Together our results demonstrated that the venom of T. natereri has allergenic proteins that can trigger allergic process, a phenomenon IgE-IgG1 dependent, IL-4 mediated and regulated by IFN-<font face=\"Symbol\">g. Furthermore, we observed a positive participation of the cytokines IL-12 and IFN-<font face=\"Symbol\">g in the exacerbation of the late phase reaction.

Tang Chi-Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 89-112). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.vi / Table of contents --- p.viii / List of Tables --- p.xi / List of Figures --- p.xii / List of Abbreviations --- p.xiv / Chapter Chapter 1. --- General introduction --- p.1 / Chapter Chapter 2. --- Literature review --- p.4 / Chapter 2.1 --- History and prevalence of food allergy --- p.4 / Chapter 2.2 --- Mechanism and clinical symptoms of food allergy --- p.6 / Chapter 2.3 --- Tropomyosin as a major shellfish allergen --- p.13 / Chapter 2.4 --- Use of animal model in the studies of food allergy --- p.22 / Chapter 2.5 --- Future approaches for treatment of food allergy --- p.27 / Chapter Chapter 3. --- Cloning and expression of recombinant tropomyosin --- p.30 / Chapter 3.1 --- Introduction --- p.30 / Chapter 3.2 --- Materials and Methods --- p.31 / Chapter 3.2.1 --- Design of PCR primers for amplification of tropomyosin gene --- p.31 / Chapter 3.2.2 --- Cloning of PCR-amplified cDNA into vector --- p.32 / Chapter 3.2.3 --- Transformation of competent E. coli Ml5 cells --- p.34 / Chapter 3.2.4 --- Confirmation of DNA sequence of the cloned vector --- p.34 / Chapter 3.2.5 --- Induction of the recombinant protein --- p.35 / Chapter 3.2.6 --- Purification and storage of the recombinant protein under native condition --- p.36 / Chapter 3.2.7 --- Concentration measurement and storage of the recombinant protein --- p.37 / Chapter 3.2.8 --- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.38 / Chapter 3.2.9 --- Regeneration of the Ni-NTA column --- p.40 / Chapter 3.3 --- Results and discussion --- p.42 / Chapter 3.3.1 --- DNA sequence of the cloned vector --- p.42 / Chapter 3.3.2 --- Expression of the recombinant protein --- p.42 / Chapter 3.3.3 --- Sodium dodecyl sulfate polyacrylamide gel eletrophoresis (SDS-PAGE) --- p.43 / Chapter Chapter 4. --- Induction of hypersensitive response to shrimp tropomyosin in mice --- p.47 / Chapter 4.1 --- Introduction --- p.47 / Chapter 4.2 --- Materials and methods --- p.52 / Chapter 4.2.1 --- Mice and reagents --- p.52 / Chapter 4.2.2 --- Animal sensitization and challenge --- p.53 / Chapter 4.2.3 --- Morphological and behavioral changes --- p.54 / Chapter 4.2.4 --- Tropomyosin-specific IgE level --- p.55 / Chapter 4.2.5 --- Passive cutaneous anaphylaxis (PCA) reaction --- p.56 / Chapter 4.2.6 --- Tropomyosin-specific cellular proliferation level of splenocytes --- p.56 / Chapter 4.2.7 --- Cytokine profiles of splenoctyes --- p.58 / Chapter 4.2.8 --- Histological examination of small intestine --- p.59 / Chapter 4.2.9 --- Statistical analysis --- p.59 / Chapter 4.3 --- Results --- p.63 / Chapter 4.3.1 --- Morphological and behavioral changes after challenge --- p.63 / Chapter 4.3.2 --- Tropomyosin-specific IgE level --- p.63 / Chapter 4.3.3 --- Passive cutaneous anaphylaxis (PCA) --- p.64 / Chapter 4.3.4 --- Tropomyosin-specific cellular proliferation level of splenocytes --- p.68 / Chapter 4.3.5 --- Cytokine profiles of splenocytes --- p.70 / Chapter 4.3.6 --- Histology of small intestines --- p.76 / Chapter 4.4 --- Discussion --- p.79 / Chapter Chapter 5. --- General conclusion --- p.88 / References --- p.89

O estudo in vitro avaliou os agentes dessensibilizantes sobre a evolução do processo erosivo em dentina radicular e o estudo in vivo avaliou o tratamento dessensibilizante na progressão das lesões erosivas e na hipersensibilidade dentinária (HD). 90 espécimes foram imersos em ácido cítrico 6% (erosão inicial) e observados através da Microscopia Confocal a Laser (MCL) (avaliação inicial). Os espécimes tiveram metade de sua superfície isolada (área controle) e então divididos aleatoriamente em 6 grupos (n=15): G1(sem tratamento); G2(Oxagel); G3(Desensibilize Nano P); G4(MI Paste); G5(pasta experimental); G6(laser de diodo, 970nm-0,7W/10Hz, 70mJ). Logo após foram submetidos à ciclagem erosiva (ácido cítrico 0,3%, 9 dias) e aplicação dos tratamentos (a cada 3 dias). As avaliações do perfil, rugosidade, degrau, perda de volume e morfologia da área tratada foram realizadas pelo MCL em diferentes tempos da ciclagem. Após exposição da área controle, a análise final foi realizada no 9º dia. O teste de permeabilidade foi realizado ao final do experimento (microscopia óptica). Para análise superficial, os tratamentos foram comparados por ANOVA e o fator tempo foi analisado internamente pelo teste de Friedman (&alpha;=5%). Para a análise morfológica e da permeabilidade dentinária realizou-se ANOVA e o teste de Tukey (&alpha;=5%). Para o estudo clínico, 20 pacientes foram selecionados com lesões cervicais erodidas em dois diferentes quadrantes sendo estas divididas aleatoriamente em dois grupos experimentais: grupo controle (agente placebo) e grupo tratado (Desensibilize Nano P). Foram realizadas 4 sessões de tratamento (intervalo de 7 dias) e 3 sessões de acompanhamento pós-tratamento (1, 3 e 6 meses). Para avaliar a progressão das lesões, em cada sessão, foi realizada a moldagem e réplicas, feitas para obtenção de imagens e análise no MCL. A avaliação da HD foi realizada por meio da aplicação de jato de ar na lesão e pela escala de estimativa numérica. Realizou-se ANOVA, os testes de Friedman e Kruskall-Wallis e Tukey (&alpha;=5%). Para análise da HD, os dados foram submetidos ao teste não-paramétrico Wilcoxon (&alpha;=5%). Avaliação in vitro do desgaste: observou-se diferença significante para todas as variáveis de resposta; G3 apresentou os menores valores para degrau de desgaste e volume perdido. Análise morfológica: G5 apresentou o menor número e área de 14 15 túbulos expostos durante toda ciclagem. Permeabilidade dentinária: G3 apresentou os maiores valores e G5 os menores; os demais grupos foram semelhantes entre si e ao G3 e G5. Avaliação clínica do desgaste: não houve diferença significante entre os grupos. Com relação ao tamanho das lesões também não houve diferença, porém, comparando os valores iniciais e finais, houve um aumento similar em ambos os grupos. HD: não houve diferença significante entre os grupos somente entre inicial e final. In vitro, o Desensililize Nano P foi capaz de controlar o processo erosivo na dentina radicular e a pasta experimental demonstrou resultados positivos na oclusão dos túbulos dentinários e na permeabilidade dentinária. Clinicamente o Desensililize Nano P não foi capaz de controlar a progressão das lesões erosivas e demonstrou resultados positivos para o tratamento da HD. / The in vitro study evaluated the desensitizing agents in the evolution of the erosive process on root dentin and the in vivo study evaluated the desensitizing treatment in the progression of the erosive lesions and in the dentin hypersensitivity (DH). 90 specimens of root dentin were immersed in 6% citric acid (initial erosion) and they were observed through the Confocal Laser Scanning Microscopy (CLM) (initial evaluation). The half their buccal surface was covered (control area). The specimens were randomly divided into 6 groups (n=15): G1(no treatment); G2(Oxagel); G3(Desensibilize Nano P); G4(MI Paste); G5(experimental paste); G6(970-nm diode laser-0.7W/10Hz, 70mJ). After that, they were cycled through erosive challenge (0.3% citric acid, 9-days). The treatments were applied every 3 days. The evaluations of the wear profile, roughness, step and the volume loss of treated area were carried out by the CLM at different times of the erosive challenge. After control area exposure, the final analysis was performed on the 9th day. The permeability test was performed at the end of the experiment (optical microscopy). For superficial analysis, the treatments were compared by ANOVA and the time factor was analyzed by the Friedman and Tukeys Tests (&alpha;=5%). For the morphological and dentin permeability analysis, ANOVA and Tukey\'s Test (&alpha;= 5%) were performed. For the clinical study, 20 patients were selected with erosive cervical lesions in two different quadrants and they were randomly divided into two groups: control group (placebo agent) and treated group (Desensibilize Nano P). Four treatment sessions (7 days interval) and 3 post-treatment follow-up sessions (1, 3 and 6 months) were performed. For the progression analysis of the lesions, replicas were obtained by molding them at each session and evaluated by the MCL. The DH evaluation was performed through the application of air jet in the lesion and by the numerical rating scale. The analysis of variance, the Friedman, Kruskall-Wallis and Tukeys Tests was performed (&alpha;=5%). For DH analysis, data were submitted to the non-parametric Wilcoxons Test (&alpha;=5%). In vitro evaluation of the wear: significant difference was observed in all response variables; G3 showed the lowest values for step and volume loss. Morphological analysis: G5 showed the lowest number and area of exposed tubules during the cycling. Dentin permeability: G3 showed the highest values and G5 exhibited the lowest values; the other groups were similar to each other and to G3 and G5. Clinical evaluation of the wear: there was 18 19 no significant difference between the groups. Regarding lesion dimension, there was no difference, but comparing the initial and final values, there was a similar increase in both groups. DH: there was no significant difference between the groups, only between the initial and final. In vitro, Desensililize Nano P was able to control the erosive process and the experimental paste showed positive results on dentin tubule occlusion and dentin permeability. Clinically, Desensililize Nano P was not able to control the progression of erosive lesions and demonstrated positive results for the treatment of DH.

Made available in DSpace on 2018-08-01T23:26:23Z (GMT). No. of bitstreams: 1 tese_9826_DISSERTAÇÃO FINAL.pdf: 516782 bytes, checksum: f8a8e0ac8e3f39b30abbc544c45a8cf3 (MD5) Previous issue date: 2016-04-12 / Não existem estudos conclusivos que determinem a exata relação entre a recessão gengival e a hipersensibilidade dentinária cervical. Diante da prevalência do problema de hipersensibilidade dentinária cervical associado com a recessão gengival, justifica-se a importância do conhecimento da relação destas condições clinicas. O objetivo deste estudo é avaliar pacientes com recessão gengival vestibular e sua associação com a hipersensibilidade dentinária cervical em estudo clínico em 61 pacientes atendidos na Clínica Odontológica do Curso de Odontologia da UFES. Eles foram selecionados quanto à presença de recessão gengival e após identificada a recessão o paciente foi submetido a avaliação com o intuito de identificar a relação com hipersensibilidade da dentinária cervical. Foram anotados de todos os pacientes dados referentes à idade, sexo, e o tipo de dente. No exame clínico foram avaliados os seguintes parâmetros: determinação da presença ou não do sangramento, presença visível de placa, altura da recessão gengival e hipersensibilidade dentinária cervical. O grau de sensibilidade foi classificado utilizando estímulo térmico (Endo-Ice Spray MAQUIRA®- PR/ Brasil), segundo UCHIDA em grau 0,1,2,3 sendo 0 (sem desconforto significativo),1 (desconforto, mas sem dor considerável), 2 (dor aguda durante a aplicação do estímulo), 3 (dor aguda durante e após a aplicação do estímulo). A associação entre os dentes que apresentam recessão gengival e a presença de hipersensibilidade dentinária cervical apresentou significância estatística com p<0,001, sendo a soma do grau sensibilidade em 69,5%, com índice 3 em 39,6% seguido de índice 2 com 29,9%. A recessão gengival e hipersensibilidade dentinária cervical são mais comuns do lado esquerdo do que o lado direito da arcada dentária com mais recessão gengival do lado esquerdo 56.4% (87 dentes) do que do lado direito 43,6% (67 dentes), o sangramento à sondagem e o índice de placa visível não apresentaram significância estatística ao nível de 5% (p = 0,227 e p = 0,687). Conclui-se que houve relação entre hipersensibilidade dentinária cervical e recessão gengival. / The gingival recession is commonly associated to cervical dentinal hypersensitivity. However, there are no conclusive studies which determine such relationship. The prevalence between these two clinical conditions is usually originated in cervical dentinal hypersensitivity, associated with gingival recession and it justifies the importance of understanding the relationship of these clinical conditions. The objective of this clinical study is to evaluate patients with vestibular gingival recession and the relationship of such condition with the cervical dentin hypersensitivity. Sixtyone patients were submitted to treatment at the Dental Clinic of the UFES School of Dentistry with gingival recession. Subjects were examined in search for the presence of gingival recession and evaluated in order to identify the relationship of each lesion with the cervical dentinal hypersensitivity. Age, gender and teeth type were taken into consideration. The following parameters were recorded through clinical examination: presence or absence of bleeding, visible presence of plaque/biofilm, gingival recession and cervical dentin hypersensitivity. The sensitivity factor (UCHIDA) was applied for sensitivity measurement. The association between the teeth with gingival recession and the presence of cervical dentin hypersensitivity showed statistical significance p<0.001, where the sum of the sensitivity factors were: 69.5% with index 3 in 39.6% of subjects, followed by index 2 featuring 29.9. It was observed, within the studied group, that gingival recession and cervical dentine hypersensitivity are more common on the left side than on the right side of the dentin with arch gingival recession on the left 56.4% (87 teeth) than the right side of 43.6% (67 teeth), bleeding on probing and visible biofilm index were not statistically significant at 5% (p=0.227 and p=0.687). It was concluded that there is a relationship between cervical dentin hypersensitivity and gingival recession.

Made available in DSpace on 2017-07-10T17:40:44Z (GMT). No. of bitstreams: 1 Darci_Fontoura.pdf: 1899459 bytes, checksum: 8c49d8ad8328dec589d5f17122d1bf18 (MD5) Previous issue date: 2013-08-16 / In recent years farmers have gradually increased the use of chemical pesticides to mitigate the losses from diseases in corn. An alternative to reduce the use of such products may be the application of resistance inducer, taking advantage of latent genetic resistance, causing less negative impact on the environment and human health. Although, there may be an energetic expense with this induction which could hold up this form of control. With the objective of verifying the resistance induction in off-season (safrinha) corn by acibenzolar-S-methyl (ASM) and phosphorylated mannanoligosaccharide (MOS), experiments were conducted with four corn hybrids (30F90H, 30K64H, 30K73H and P4042H) in two consecutive years (2011 and 2012) in off-season environment in the municipality of Toledo - PR. In addition to the resistance inducers, a fungicide (azoxystrobin + ciproconazole) was applied, as well as water as a check. A randomized block design, with a triple factorial scheme, plus an additional treatment and three replications was used. The factors were: (A) hybrid in four levels (B) product in three levels and, (C) dose in three levels. The additional treatment was carried out with water spray, representing the level zero of the dose factor. The application of the products happened in phenological stages V8 and VT. There was observed an incidence of diverse plant pathogens in the two consecutive years, on the same hybrids, in the same location and under the same manageable conditions, probably due to diverse climatic conditions from one year to another, although hypersensitive response occurred in both years under MOS treatment. In the 2011 off-season, there was an incidence of Exserohilum turcicum, a necrotrophic pathogen, and no damaged kernels were found, while in the 2012 off-season, there was an incidence of Puccinia polysora, a biotrophic pathogen, plus significant percentage of damaged kernels. None of the applied products affected the traits plant height, ear placement, content of foliar nitrogen and percentage of damaged kernels. There was significant effect of resistance inducers on the hypersensitivity response and root lodging, without reducing the productivity compared to the water treatment. MOS increased hypersensitivity response severity and ASM reduced root lodging. The fungicide treatment (azoxystrobin + ciproconazole) showed better control of foliar diseases (P. polysora and E. turcicum) and higher yield in both years, and in 2011 it reduced stalk lodging and increased the mass of thousand kernels / Nos últimos anos os produtores tem aumentado gradativamente o uso de defensivos químicos para amenizar os prejuízos com doenças da cultura do milho. Uma alternativa visando diminuir o uso de tais produtos poderá ser a aplicação de indutores de resistência, tirando proveito da resistência genética latente, gerando menor impacto negativo sobre o meio ambiente e a saúde humana. Entretanto, poderá haver elevado dispêndio de energia nessa indução que torne inviável essa forma de controle. Objetivando verificar a indução de resistência em milho safrinha pelos indutores acibenzolar-S-metil (ASM) e mananoligossacarídeo fosforilado (MOS), foram conduzidos experimentos com quatro híbridos de milho (30F90H, 30K64H, 30K73H e P4042H), em dois anos consecutivos (2011 e 2012), na safrinha no município de Toledo PR. Além dos indutores de resistência, realizou-se também tratamento com fungicida (azoxistrobina + ciproconazol), além de água como testemunha. Foi utilizado delineamento experimental de blocos casualizados com esquema fatorial triplo e tratamento adicional, em três repetições. Os fatores testados foram: (A) híbrido em quatro níveis, (B) produto em três níveis e, (C) dose em três níveis. O tratamento adicional foi realizado com pulverização de água, representando o nível zero do fator dose. As aplicações dos produtos aconteceram nos estádios fenológicos V8 e VT. Observou-se incidência de patógenos distintos nos dois anos consecutivos sobre os mesmos híbridos, no mesmo local e nas mesmas condições de manejo, provavelmente em função da condição climática distinta de um ano para o outro, embora a resposta de hipersensibilidade tenha acontecido em ambos os anos no tratamento com MOS. Na safrinha de 2011 houve incidência de Exserohilum turcicum, um patógeno necrotrófico, e não foram encontrados grãos ardidos, enquanto que na safrinha de 2012 houve incidência de Puccinia polysora, um patógeno biotrófico, além de significativo percentual de grãos ardidos. Nenhum dos produtos aplicados influenciou as características altura de planta, altura de inserção de espiga, teor de nitrogênio foliar e percentual de grãos ardidos. Houve efeito dos indutores de resistência quanto à resposta de hipersensibilidade e acamamento em função da raiz, sem redução da produtividade em relação ao tratamento com água. MOS aumentou a severidade da resposta de hipersensibilidade e acibenzolar-S-metil reduziu o acamamento em função da raiz. O tratamento com fungicida (azoxistrobina + ciproconazol) apresentou o melhor controle das doenças foliares (P. polysora e E. turcicum) e a maior produtividade em ambos os anos, sendo que em 2011 ainda reduziu o acamamento de colmo e aumentou a massa de mil grãos

Yang, Fan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 111-132). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.iii / ABSTRACT --- p.iv / TABLE OF CONTENTS --- p.viii / LIST OF FIGURES --- p.xii / LIST OF TABLES --- p.xv / LIST OF ABBREVIATIONS --- p.xvi / Chapter Chatper 1. --- General Introduction --- p.1 / Chapter Chapter 2. --- Literature Review --- p.5 / Chapter 2.1 --- Facts on food allergy --- p.5 / Chapter 2.1.1 --- Food allergy and its prevalence --- p.5 / Chapter 2.1.2 --- Pathogenesis of food allergy --- p.6 / Chapter 2.1.3 --- Clineal disorders caused and diagnosis of food allergy --- p.8 / Chapter 2.2 --- Allergenicity assessment of genetically engineered food --- p.13 / Chapter 2.2.1 --- The structural and sequence homology of proteins as a criterion for food allergenicity assessment --- p.14 / Chapter 2.2.2 --- Digestion stability as a criterion for food allergenicity assessment --- p.15 / Chapter 2.2.3 --- Animal models for Food Allergenicity Assessment --- p.21 / Chapter 2.3 --- The importance of rice and its nutritional facts --- p.27 / Chapter 2.3.1 --- The importance of rice --- p.27 / Chapter 2.3.2 --- Rice nutritional facts and its relationship with malnutrition --- p.28 / Chapter 2.4 --- Food allergenicity research in rice --- p.30 / Chapter 2.5 --- Glutelin overexpression transgenic rice GT3 --- p.33 / Chapter 2.6 --- Recent and future perspectives for treatment of food allergy --- p.36 / Chapter Chapter 3. --- Materials and Methods --- p.39 / Chapter 3.1 --- Rice Seed Protein Extraction --- p.39 / Chapter 3.1.1 --- Rice varieties for protein extraction --- p.39 / Chapter 3.1.2 --- Protein extraction from rice seeds --- p.39 / Chapter 3.1.3 --- Fractionation of major rice seed storage proteins --- p.40 / Chapter 3.1.4. --- Protein quantification --- p.41 / Chapter 3.1.5 --- Tricine SDS-PAGE --- p.42 / Chapter 3.2 --- Simulated Gastric Digestibility Assay --- p.43 / Chapter 3.2.1 --- Assay System --- p.43 / Chapter 3.2.2 --- Preparation of Simulated Gastric Fluid --- p.43 / Chapter 3.2.3 --- Assay Procedures --- p.44 / Chapter 3.2.4 --- Results Interpretation --- p.44 / Chapter 3.3 --- Construction of Mouse Models --- p.45 / Chapter 3.3.1 --- Mouse strain and reagents used --- p.45 / Chapter 3.3.2 --- Mouse Model I --- p.46 / Chapter 3.3.3 --- Mouse Model II --- p.50 / Chapter 3.3.4 --- Mouse Model III --- p.51 / Chapter 3.4 --- Bioinformatic Analysis of Glutelin Sequence --- p.52 / Chapter 3.5 --- Epitope Mapping of Glutelin --- p.55 / Chapter 3.5.1 --- Bioinformatic Analysis --- p.55 / Chapter 3.5.2 --- Direct and Competitive ELISA --- p.56 / Chapter 3.5.3 --- Western Blot Analysis --- p.57 / Chapter 3.5.4 --- IgE-binding assay --- p.58 / Chapter Chapter 4. --- Results and Discussion --- p.60 / Chapter 4.1 --- Rice Seed Protein Extraction --- p.60 / Chapter 4.1.1 --- Rice Protein Extraction --- p.60 / Chapter 4.1.2 --- Extraction of rice major seed storage protein fractions --- p.62 / Chapter 4.2 --- Simulated Gastric Digestibility Assay --- p.64 / Chapter 4.2.1 --- Pepsin Digestibility of total protein from GT3 and WT rice seeds --- p.64 / Chapter 4.2.2 --- Pepsin Digestibility of major storage protein fractions in GT3 and WT rice --- p.68 / Chapter 4.2.3 --- Summary of Pepsin Digestibility Assay --- p.74 / Chapter 4.3 --- Mouse Model I --- p.75 / Chapter 4.3.1 --- Protein-specific IgE levels --- p.75 / Chapter 4.3.2 --- Protein-specific IgG1 and IgG2a levels --- p.77 / Chapter 4.3.3 --- Allergic Response Test --- p.79 / Chapter 4.3.4 --- Summary from Mouse Model I --- p.81 / Chapter 4.4 --- Mouse Model II --- p.83 / Chapter 4.4.1 --- Proteins specific IgE levels --- p.84 / Chapter 4.4.2 --- Proteins specific IgG1 and IgG2a levels --- p.85 / Chapter 4.4.3 --- Allergic Response Test --- p.87 / Chapter 4.4.4 --- Summary from Mouse Model II --- p.88 / Chapter 4.5 --- Mouse Model III --- p.90 / Chapter 4.5.1 --- Protein-specific IgE levels --- p.90 / Chapter 4.5.2 --- Proteins specific IgG1 and IgG2a levels --- p.91 / Chapter 4.5.3 --- Allergic Response Test --- p.93 / Chapter 4.5.4 --- Summary from Mouse Model III --- p.93 / Chapter 4.6 --- Potential allergenicity of rice glutelin by bioinformatics and epitope mapping --- p.94 / Chapter 4.6.1 --- Bioinformatic analysis --- p.94 / Chapter 4.6.2 --- ELISA analysis of synthesized epitopes --- p.97 / Chapter 4.6.3 --- Western Blot Analysis --- p.99 / Chapter 4.6.4 --- IgE-binding assay --- p.103 / Chapter Chapter 5. --- Conclusion and Future Perspectives --- p.109 / References --- p.111

Accumulating evidence has indicated that microbial infection could intensify allergic responses. Previous findings demonstrated that eosinophil activation could be elicited by bacterial and viral conserved molecular pattern through TLR. Recently, two cytoplasmic pattern recognition receptors, NLR protein NOD1 and NOD2, have been discovered and the important roles in innate immunity have been elucidated. Eosinophils alone have little responses upon the stimulation with ligands of NOD1 and NOD2. Since airway eosinophils increase in more numbers of asthmatic patients compared to control subjects, we investigated the co-culture system of eosinophils and human bronchial epithelial cells to illustrate the potential immunopathological roles of NOD1 and NOD2 in asthma processes. In the co-culture system, NOD1 ligand gamma-D-Glu-mDAP (iE-DAP) and NOD2 ligand muramyl dipeptide (MDP) could upregulate cell surface expression of CD1 8 and ICAM-1 on eosinophils and ICAM-1 and vascular cell adhesion molecule-1 (VCAM-1) on bronchial epithelial cells, as well as induce chemokines CCL2 and CXCL8 release. These findings therefore imply the direct interaction and activation between the two cells upon NOD1 and NOD2 ligand stimulation. / Allergic diseases are prevalent and their incidences have been increasing worldwide. Eosinophils are the principal effector cells for the late phase response in allergic inflammation. The infiltration of eosinophils together with other inflammatory cells at the local inflammatory sites is the major characteristic in allergic inflammation. However, the detailed innnunopathological responses and mechanisms of the activation of eosinophils in allergic inflammation are not well defined. In the present study, we investigated and attempted to elucidate the mechanisms of eosinophil activation induced by various stimuli, including thymic stromal lymphopoietin (TSLP), the novel interleukin (IL)-12 family cytokine IL-27, and ligands of nucleotide-binding oligomerization domain (NOD) like receptor (NLR) protein NOD1 and NOD2 upon interaction with bronchial epithelial cells. / In conclusion, the above findings demonstrated that eosinophils could be potently activated by diverse stimuli and regulated by multiple intracellular regulatory mechanisms. The elucidation of eosinophil activation may offer new therapeutic stategies and clues for the treatment of allergic diseases. / Recently, the novel IL-12 family member IL-27 was found to regulate immune responses, exerting either stimulation or suppression effects. We found that eosinophils constitutively expressed IL-27 receptor heterodimer, gp130 and WSX-1. IL-27 could prolong eosinophil survival by reducing apoptosis, modulate the expression of adhesion molecules to facilitate eosinophil adhesion and accumulation, and induce the release of proinflammatory cytokines IL-6, tumor necrosis factor (TNF)-aalpha IL-1beta and chemokines CCL2, CXCL8 and CXCL1. The stimulatory effects of IL-27 on eosinophils could not be abrogated by IL-25, hematopoietic cytokine granulocyte-macrophage colony stimulating factor (GM-CSF) and toll-like receptor (TLR) 4 ligand lipopolysaccharide (LPS). These findings were different from the effects of IL-27 and LPS on monocytes. Intracellular signaling mechanistic studies showed that IL-27-mediated eosinophil activation was differentially regulated by MAPKs and NF-kappaB. Based on the above results, IL-27 could play crucial roles in allergic diseases by the activation of eosinophils via differential intracellular signaling cascades. However, IL-27 has been shown to suppress allergic diseases in mouse models. According to our findings of its activating effects on human eosinophils, IL-27 may play pleiotropic roles in human allergic responses. / TSLP is a novel IL-7-like cytokine highly expressed by bronchial epithelial cells and skin keratinocytes in allergic diseases. TSLP acts as a master switch for allergic inflammation through the activation of dendritic cells and mast cells for initiating inflammatory Th2 responses. To elucidate the immunological cascades of epithelium/keatinocyte-eosinophil mediated allergic inflammation, we examined the modulating effects of TSLP on human eosinophils. We observed that human eosinophils constitutively expressed TSLP receptor complex comprising TSLP-binding chain TSLPR and IL-7Ralpha chain. TSLP could significantly delay eosinophil apoptosis, up-regulate the cell surface expression of adhesion molecule CD18 and intercellular adhesion molecule-1 (ICAM-1) but down-regulate L-selectin, enhance eosinophil adhesion to fibronectin, and induce the release of inflammatory cytokine 1L-6 and chemokines CXCL8, CXCL1 and CCL2. All these effects were concentration-dependent and TSLP-specific. TSLP regulated the above effects through the activation of extracellular signal-regulated protein kinase (ERK), p38 mitogen activated protein kinase (MAPK) and nuclear factor (NF)-kappaB signaling pathway, but not signal transducer and activator of transcription (STAT)-5 and STAT-3 which were usually activated in other effector cells upon TSLP stimulation. Collectively, the above findings elucidated the pro-allergic mechanisms of TSLP via the activation of distinct intracellular signaling pathways in eosinophils. / Hu, Shuiqing. / Adviser: Wong Chin Kwok. / Source: Dissertation Abstracts International, Volume: 72-04, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 176-216). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.