Asthma and allergies are some of the most common illnesses worldwide that almost everybody will come in contact with. This thesis studied persons with allergic asthma, non-allergic asthma, allergic rhinitis and atopic dermatitis in a population-based sample. At an early stage, these illnesses were regarded as psychosomatic. Over time, as knowledge about asthma/allergy has increased more of a biomedical perspective was taken by the research field. In considering early documentations well as contemporary research, a psychobiosocial perspective was taken in this thesis when conducting the three studies. Thus, as psychological factors may affect the illness and be a result of the illness, it is important to incorporate these factors to better understand asthma and allergy. Study I examined the co- and multimorbidity in asthma/allergy with the environmental intolerances in the form of chemical and building-related intolerance. Study II investigated psychological distress in the four forms of asthma and allergy. Psychological distress was in this study defined as stress, burnout, anxiety, depression and environmental health worries. Study III examined usage of problem and emotion focused coping strategies and perceived social support from the surrounding in high and low asthma/allergy severity. All studies were performed using data from the Västerbotten Environmental Health Study, a questionnaire-based survey with focus on various environmental hypersensitivities and asthma and allergy. The result showed that the co- and multimorbidity with the environmental intolerances in asthma/allergy was larger than what was statistically excepted. Those with allergic asthma and atopic dermatitis experienced more stress, burnout and anxiety than those with non-allergic asthma, allergic rhinitis and non-asthma/allergy. Moreover, the most common way of coping with asthma and allergy was found to be strategies such as avoiding environments that are believed to affect health, and trying to accept the situation, independent of asthma/allergy severity. Finally, in general, those with asthma and allergy reported receiving most support from their partner, other family members and health care, and least support was perceived by those with low asthma/allergy severity.The findings suggest that co- and multimorbidity with environmental intolerances is relatively common in asthma and allergy, and should therefore be included in the clinical anamnesis for this patient group. The elevated level of distress in allergic asthma and atopic dermatitis evokes the question of use of therapies such as mindfulness maybe beneficial in certain afflicted persons. The results on coping and social support provide a foundation for further research regarding informing the asthma/allergy patient and family members about effective coping strategies and the importance of adequate social support. A metaperspective is taken in which interrelations between important variables in the thesis are discussed.

Introdução: A dermatite atópica (DA) é uma doença cutânea inflamatória de caráter crônico, recidivante, em que o prurido intenso e a xerose cutânea são frequentes. A etiopatogenia da DA é multifatorial, envolvendo fatores genéticos, ambientais e imunológicos. Eosinófilos são leucócitos polimorfonucleares multifuncionais que estão implicados na patogênese de diversos processos inflamatórios, incluindo a DA. Além da produção e secreção de diversas proteínas presentes nos grânulos citoplasmáticos, os eosinófilos também apresentam potencial para secretar metaloproteinases, enzimas proteolíticas que degradam vários componentes da matriz extracelular, e estão presentes em diversos processos fisiológicos e patológicos. Objetivo: Avaliar: 1) o perfil fenotípico dos eosinófilos na dermatite atópica do adulto, através da expressão das moléculas CCR3, CD23, CD38, CD69 e CD62L; 2) o perfil funcional, a partir da secreção de metaloproteinases, inibidores teciduais de metaloproteinases e RANTES por eosinófilos purificados. Métodos: Foram incluídos 41 adultos diagnosticados com DA, de acordo com os critérios de Hanifin & Rajka e 45 controles adultos sadios. A gravidade da doença foi mensurada através do escore de gravidade EASI (Eczema Area and Severity Index). Eosinófilos (LIN 1- CCR3+) do sangue periférico foram analisados para os marcadores CCR3, CD38, CD69, CD23 e CD62L através da citometria de fluxo (LSRFortessa, BD Biosciences) a análise foi realizada com o FlowJo 7.5.6 software. Eosinófilos purificados de indivíduos com DA e indivíduos controles foram estimulados com enterotoxina de Staphylococcus aureus B (SEB) e FSL-1 (agonista de receptores Toll-like 2 e 6), e os sobrenadantes foram coletados para dosagem de metaloproteinases (MMPs), inibidores teciduais de metaloproteinases 1 e 2 (TIMP-1 e TIMP-2) e RANTES por ELISA e por Cytometric bead array. Resultados: Indivíduos com DA apresentaram maior frequência de eosinófilos (LIN1- CCR3+), relacionada à gravidade da doença. Observou-se também, que a frequência de CD62L (L-selectina) e de CD23 (receptor de baixa afinidade para IgE) em eosinófilos (LIN1- CCR3+) diminui em pacientes com DA. Os receptores de ativação precoce (CD69) e tardio (CD38) não mostraram diferença estatística entre os grupos analisados. Os níveis séricos de MMPs e de TIMPs foram similares entre os controles e pacientes. Ao analisarmos a secreção de MMPs e de (TIMPs), a partir de eosinófilos purificados de pacientes com dermatite atópica, observamos diminuição dos níveis basais de TIMP-1 e TIMP-2 e de RANTES. Conclusões: Na DA do adulto, o perfil fenotípico e funcional dos eosinófilos mostrou: perfil de ativação da fase aguda, com expressão aumentada de CCR3; potencial de migração elevado, em decorrência da diminuição da expressão de CD62L; falhas no processo de ativação dos eosinófilos via CD23, bem como, no remodelamento tecidual mediado por TIMP-1 e TIMP-2 e na quimotaxia mediada por RANTES / Introduction: Atopic dermatitis (AD) is an inflammatory, chronic and recurrent skin disease characterized by intense pruritus and xerosis. AD has a complex etiopathogenesis, which involves the influence of genetics, environment, and immunological disorders, among others. Eosinophils are multifunctional polymorphonuclear leukocytes that contribute to the pathogenesis of several inflammatory processes, such as AD. In addition to the production and secretion of diverse proteins of the cytoplasmic granules, eosinophils have also the potential to secrete metalloproteinases (MMPs), proteolytic enzymes with a primary role for degrading several extracellular matrix components, present in distinct physiological and pathological processes. Objective: To evaluate:1) the phenotypic profile of eosinophils in adults with atopic dermatitis through the expression of CCR3, CD23, CD38, CD69 and CD62L molecules; 2) the functional profile through secretion of MMPs, tissue inhibitors of metalloproteinases 1 and 2 ( TIMP-1 and TIMP-2) and RANTES by purified eosinophils. Methods: This work enrolled 41 patients with AD, diagnosed according to Hanifin & Rajka\'s criteria) and 45 healthy controls. Severity of the disease was established utilizing EASI (Eczema Area and Severity Index). Eosinophils (Lineage cocktail 1- CCR3+) from peripheral blood were analyzed for CCR3, CD38, CD69, CD23 and CD62L by flow cytometry (LSRFortessa, BD Biosciences), and analysis was performed using the FlowJo 7.5.6 software. Purified eosinophils were stimulated with Staphylococcus aureus enterotoxin B (SEB) FSL-1 (Toll-like receptor 2/6 agonist), and supernatants were collected for MMPs, TIMPs and RANTES secretion, evaluated by ELISA and cytometric bead array (CBA). Results: Patients with AD have a higher frequency of eosinophils (LIN1- CCR3+), related to disease severity. Moreover, the frequency of CD62L (L-selectin) and CD23 (low-affinity receptor for IgE) in (LIN1- CCR3+) eosinophils was reduced in individuals with AD. CD69 and CD38 (early and late activation receptors) did not show significant difference in the studied groups. Serum levels of MMPs and of TIMP-1 and TIMP-2 were similar in healthy controls and AD patients. When analyzing secretion of MMPs and TIMPs by purified eosinophils from AD individuals, we detected a decrease in baseline levels of TIMP-1, TIMP-2, and reduced RANTES-mediated chemotaxis. Conclusions: Eosinophils in AD exhibit an activation profile of acute phase, with enhanced CCR3 expression, high potential for migration due to reduced expression of CD62, defective activation mechanisms via CD23, altered tissue remodeling process mediated by TIMP-1 and TIMP-2 and reduced RANTES-mediated chemotaxis

Introdução: A dermatite atópica (DA) é uma enfermidade cutânea inflamatória de caráter crônico, na qual o prurido é constante, e com marcada xerose. Dermatose que geralmente se inicia na infância, e pode surgir em indivíduos com história pessoal ou familiar de asma, rinite alérgica e/ou DA. A pele com DA apresenta colonização por Staphylococcus aureus (S. aureus) em 80-100% dos casos, sendo responsável pela produção enterotoxinas, capazes de exacerbar a resposta inflamatória na DA. Nesta enfermidade, existem distintos subtipos de células apresentadoras de antígeno ou dendríticas (DC), tanto na pele quanto circulantes. As DC exercem papel relevante na inflamação da DA, em especial um subgrupo de células dendríticas mieloides (mDC), as chamadas células dendríticas inflamatórias epidérmicas (IDEC). Objetivo: Avaliar o fenótipo e a função das mDC (IDEC-like) em células mononucleares do sangue periférico (PBMC) na DA do adulto. Métodos: Foram selecionados 21 pacientes com DA (idades entre18 e 65 anos, sendo 13 homens e oito mulheres) e 21 controles (idades entre 21 e 41 anos, sendo oito homens e 13 mulheres), nos quais foram realizadas as avaliações fenotípica e funcional das mDC (IDEC-like) em PBMC. Para tal, foram analisadas as expressões de: Fc?RI, TNF, IFN-y, IL-10, CD36 e CD83 nas mDC, estimuladas com enterotoxina estafilocócica B (SEB), agonistas de TLR2 (Pam3CSK4), TLR4 (LPS) e de TLR7/8 (CL097) através da citometria de fluxo. Resultados: Os principais achados nos pacientes com DA foram: aumento da frequência de células IDEC-like frente ao estímulo com agonista de TLR2 (Pam3CSK4); aumento da frequência de IFN-y em condição não estimulada, e de IL-10 frente a estímulo com agonista de TLR7/8 (CL097) nesta população de células dendríticas. Conclusão: A caracterização das mDC circulantes na DA evidencia perfil pró-inflamatório em condição não estimulada, impactando na resposta imune adaptativa. O aumento significativo na frequência de células IDEC-like nos pacientes com DA sugere sua participação na perpetuação do processo inflamatório da DA / Introduction: Atopic dermatitis (AD) is an inflammatory skin disease with a chronic course, with constant pruritus and marked xerosis. It usually starts during childhood, and a personal or familial history of skin and/or respiratory allergy may be present. Around 80-100% of the patients show a cutaneous colonization of Staphylococcus aureus (S. aureus), which produces enterotoxins that may exacerbate the inflammatory response in AD. In this disease, there are distinct subtypes of antigen-presenting cells or dendrytic cells (DC), either circulating or present in the affected tissue. DC exert a relevant role in AD inflammation, especially a subgroup of myeloid cells (mDC), known as epidermal inflammmatory dendritic cells (IDEC). Objective: To evaluate the phenotype and function of mDC (IDEC-like) in mononuclear cells of the peripheral blood (PBMC) of adults with AD. Methods: Twenty-one adults with AD (age 18/65; male/female: 13/8) and 21 healthy controls (age 21/41; male/female: 8/13) were selected for the current study. Phenotypical and functional analysis of mDC (IDEC-like) of PBMC were performed, through the expression of Fc?RI ,TNF, IFN-y, IL-10, CD36 and CD83 in mDC, stimulated with enterotoxin B (SEB) and with agonists of TLR2 (Pam3CSK4), TLR4 (LPS) and TLR7/8 (CL097) by flow cytometry. Results: Main findings of AD patients included: elevation of IDEC-like cell frequency with TLR2 (Pam3CSK4) agonist, augmented unstimulated frequency of IFN-y, and of IL-10 with TLR7/8(CL097) agonist of this population of dendritic cells. Conclusion: Characterization of circulating mDC on AD shows proinflammatory profile in unstimulated conditions, therefore causing impact on the adaptive immune response. The significant increase in the frequency of IDEC-like cells in AD patients suggest a role in the maintenance of inflammation in AD

La dermatite atopique, mieux connue sous le nom d’eczéma, est une condition chronique fréquente dont la prise en charge est souvent complexe et difficile à comprendre. Cette étude prospective vise la conception et la validation d'un guide pratique écrit ainsi qu'un questionnaire de connaissances en eczéma dans le but de combler un besoin concernant la disponibilité d'un programme d'éducation thérapeutique accessible à la population souffrant de cette maladie. Les objectifs secondaires sont d'évaluer l'impact de ces outils sur les connaissances en eczéma à court terme ainsi que l'impact du niveau de littératie en santé et des variables sociodémographiques sur ces mêmes connaissances. Un projet pilote a d’abord été conduit auprès d’onze futurs parents d'un cours prénatal de l'Université de Sherbrooke, suivi d'une phase expérimentale auprès de 107 patients âgés entre 18 et 45 ans de la clinique de dermatologie du Centre Hospitalier Universitaire de Sherbrooke (CIUSSS de l’Estrie-CHUS). Les analyses de validité et de fiabilité performées lors du projet pilote ont permis de faire les modifications nécessaires sur le guide pratique écrit ainsi que sur le questionnaire de connaissances afin de les rendre applicables à plus grande échelle. Ces outils modifiés ont par la suite été utilisés dans les études comparatives menées dans la phase expérimentale. Ces dernières ont démontré une amélioration statistiquement significative de la connaissance en eczéma à court terme par une comparaison des scores moyens au questionnaire de connaissances en pré-test et en post-test. Dans l’ensemble, aucune différence de progression de connaissance (delta entre le pré-test et le post-test) entre les différents sous-groupes de chaque variable sociodémographique n’a été démontrée de façon statistiquement significative. De plus, la courbe de progression de connaissances en pré-test et post-test est comparable entre les différents niveaux de littératie en santé. Ces derniers résultats démontrent l’absence d’impact des variables sociodémographiques étudiées ainsi que du niveau de littératie sur les outils à l’étude, ce qui supporte leur applicabilité à grande échelle. Mots clés: dermatite atopique, eczéma, éducation thérapeutique, guide pratique écrit, éducation en santé. / Atopic dermatitis, also known as eczema, is a common chronic disease for which treatments are often complex and difficult to understand. This prospective study aims to develop and validate a written action plan and a knowledge questionnaire about eczema in order to fill an existing gap in literature concerning the availability of a self-management training program that would be accessible to patients suffering from this condition. The secondary end-points aim to evaluate the impact of the written action plan as well as the impact of health literacy and other demographic variables on eczema knowledge over a short period of time. A pilot over 11 subjects was firstly conducted in a prenatal class at the University of Sherbrooke. Validity and reliability analyses were performed and allowed for alterations on the written action plan as well as the knowledge questionnaire so that they could be used on a larger scale. These modified assessment tools were then used on the 107 patients recruited at the dermatology clinic of the Centre Hospitalier Universitaire de Sherbrooke (CIUSSS de l’Estrie-CHUS) for comparative analyses. Results demonstrate a significant increase on the short-term eczema knowledge when comparing mean knowledge scores on the pre-test and post-test. On a general basis, there is no significant difference on the knowledge progression for the many demographic variables analysed (delta between pre-test and post-test mean scores). Also, knowledge progression between the different levels of health literacy is comparable. These results demonstrate a lack of impact of health literacy and other demographic variables on the studied assessment tools, which support their applicability on a larger scale. Keywords : Atopic Dermatitis, Eczema, Self-Management, Written Action Plan, Health Education.

Depuis plusieurs années, la vectorisation de molécules est considérée comme la stratégie la plus prometteuse pour améliorer la pénétration cutanée des principes actifs et pour cibler et contrôler leur libération, augmentant ainsi l'efficacité thérapeutique des traitements tout en limitant leurs effets secondaires. Les nanoparticules polymériques ont fait l'objet de nombreuses études car elles possèdent une très grande stabilité et une capacité supérieure aux autres vecteurs à libérer les principes actifs de façon prolongée. Récemment, le laboratoire de Pharmacie Galénique de Besançon a montré, in vivo, que les nanoparticules polymériques de diamètre inférieure ou égale à 100 nm pénétraient spécifiquement dans la peau inflammée de la souris, alors qu'aucune pénétration n'était observée dans la peau saine. Le premier objectif de cette thèse était de confirmer ces hypothèses dans la peau inflammée d'un autre animal, le porc. Les résultats de notre étude in vivo confirment l'absence de pénétration des nanoparticules polymériques dans la peau saine du porc et montrent une pénétration taille dépendante dans la peau inflammée. Le second objectif poursuivi était de vérifier ces résultats chez l'Homme. Pour cela, une preuve de concept a été mise en place dans le service de Dermatologie du CHRU de Besançon. Les premiers résultats de cette étude clinique semblent confirmer l'absence de pénétration des nanoparticules polymérique de I 00 nm dans la peau des volontaires sains et dans la peau non lésée des patients souffrant de dermatite atopique. A l'inverse, une forte pénétration des nanoparticules est observée au niveau des plaques d'eczéma des patients. Si les résultats de l'étude clinique se confirment, nous prévoyons d'encapsuler un anti­inflammatoire ou un immunosuppresseur pour vérifier l'intérêt thérapeutique de ces vecteurs en médecine humaine dans le traitement de la dermatite atopique. Parallèlement, une évaluation pourrait être réalisée en médecine vétérinaire pour le traitement de cette pathologie fréquente chez le chien, et dont le traitement actuel repose sur l'administration per os de corticoïdes à l'origine de nombreux effets secondaires. / For several years now, nanocarriers have been considered as the most promising strategy to improve skin penetration of active ingredients. Moreover, these carriers are more efficient at targeting and controlling drug release into the skin, which leads to increased treatment efficiency and reduced side effects. Polymeric nanoparticles have been the object of an increasing number of studies due to their good physicochemical stability and prolonged release of active ingredients which is superior to any other carriers. Recently, the Laboratory of Pharmaceutical Engineering at Besançon proved in vivo, that polymeric nanoparticles with a diameter smaller or equivalent to I 00 nm specifically penetrated in inflamed skin of mice whilst no penetration was observed in healthy skin. The first aim ofthis thesis was to confirm this hypothesis on another animal's inflamed skin, in instance the pig. Our results confirm the poor penetration of polymeric nanoparticles in healthy skin ofpigs and show various degree ofpenetration depending on the size of the nanoparticles into the inflamed skin area. The second objective ofthis work was to evaluate the skin penetration of our polymeric nanoparticles in humans. A proof of concept has been developed in the Department of Dermatology at Besancon University Hospital. The first results ofthis clinical trial tend to confim1 the greater penetration ofour carrier, specifically in inflamed skin. In fact, no penetration of polymeric nanoparticles with a size close to 100 nm was observed in healthy skin ofvolunteers or in the non-inflamed skin of patients suffering from atopic dennatitis. Conversely, a high penetration ofthese carriers was observed in the skin lesions of patients with atopic dermatitis. If the results ofthis clinical trial are confirmed, we plan to load an anti-inflammatory or an immunosuppressive drug into the nanoparticles to evaluate the therapeutic value ofthese nanocarriers in human medicine in the treatment ofatopic dermatitis. Meanwhile, a similar study may be undertaken in veterinary medicine for the treatment of atopic dermatitis in dogs which is a common disease whose current treatment is based on the oral administration of corticosteroids and cause many undesirable side effects.

La dermatite atopique (DA) est une dermatose inflammatoire dont la prévalence élevée est en constante augmentation dans les pays industrialisés. La physiopathologie de la DA est complexe et associe des facteurs immunologiques, environnementaux et génétiques. L'ensemble de ces facteurs touchent principalement l'épiderme et sont responsables d'une altération de la fonction barrière, ce qui facilite la pénétration transcutanée des molécules en contact avec la peau et la sensibilisation des individus. Ce travail de thèse vise à mieux caractériser le rôle de l'épiderme dans la physiopathologie de la DA. Il repose sur l'utilisation d'un modèle in vitro d'épiderme reconstruit humain généré à partir de cellules progénitrices de follicules pileux prélevés chez des patients atteints de DA, ainsi que chez des individus sains. Ce travail a tout d'abord montré que les épidermes DA générés à partir de patients non porteurs de mutations pour le gène de la filaggrine (FLG+/+) se comportaient comme des épidermes normaux, (i) tant à l'homéostasie, (ii) que suite à une stimulation par de l'interleukine-1beta (IL-1 β) ou par un allergène majeur de la DA, tel qu'un extrait d'acarien (Dermatophagoïdes farinae). Cependant, de façon très intéressante, les épidermes- DA se sont avérés davantage sensibles à l'apoptose induite par une exposition aux ultraviolets (UV), que les épidermes normaux. A cet égard, nous avons observé une modulation différente de certains gènes impliqués dans l'induction/régulation de l'apoptose par les épidermes provenant de patients DA après exposition aux UV. En outre, ce travail a démontré que des épidermes normaux sont profondément affectés par l'IL-1 β qui induit un phénotype atopique caractéristique associant (i) la production de quantité notable de TSLP (thymic stromal lymphopoietin) et (ii) une altération de la fonction barrière. L'impact de l'IL-1 β sur la biologie de l'épiderme et la sensibilité accrue exprimée par les épidermes-DA à l'apoptose induite par les UV sont donc de nouveaux éléments qui confirment l'importance de l'épiderme dans la physiopathologie de la DA ce qui permettra certainement de nouvelles stratégies thérapeutiques / Atopic dermatitis (AD) is an inflammatory skin disease with a high prevalence constantly increased in developed countries. The AD physiopathology is complex and associated with immunological, environmental and genetic factors. These factors impact mainly the epidermis and are responsible of the impaired barrier function which increases antigen penetration and people sensitization. The aim of this work is to improve the understanding of the epidermis role in AD physiopathology. For this, we used an in vitro reconstructed human epidermis model generated from outer root sheath cells taken off AD patients or normal individuals.First of all, this work has shown that epidermis generated from AD patients (AD- epidermis) with no mutation on filaggrine gene (FLG+/+) behave like those from normal individuals (Normal- epidermis), (i) at the steady state, and (ii) after stimulation with interleukin-1beta (IL-1 β) or a major allergen found in AD, Dermatophagoïdes farinae. However, interestingly, AD-epidermis are more sensitized to ultra-violets radiation-induced (UVR) apoptosis than Normal-epidermis. In this regard, we observed a higher expression of genes involved in the induction / regulation of apoptosis by the epidermis from AD patients after UV exposure. Moreover, this work has demonstrated that normal epidermis are profoundly affected by IL-1 β which induces an AD like phenotype associating (i) production of large amount of TLSP (thymic stromal lymphopoietin) and (ii) alterated barrier function. The impact of IL-1 β on epidermis biology and the increased sensitivity that seems to express AD-epidermis to UVR-induced apoptosis are new evidences that confirm the importance of the epidermis in pathophysiology of AD which certainly will lead to new therapeutic strategies

A prevalência de doenças alérgicas incluindo asma e dermatite atópica aumentou nos últimos anos. A asma é uma doença crônica caracterizada por hiper-responsividade das vias aéreas e limitação variável ao fluxo aéreo, reversível espontaneamente ou com tratamento. A elevada prevalência, mortalidade e custos associados tornam a doença um importante problema de saúde pública que requer atenção. A dermatite atópica (DA) é uma doença inflamatória crônica da pele, caracterizada por prurido intenso, eritema, escoriações, liquenificação na pele entre outras características. A DA causa profundo impacto na vida do indivíduo e da família e geralmente mostra os primeiros sintomas durante a infância. Ambas as doenças podem ser associadas a sensibilização a alérgenos. Nosso grupo mostrou que entre pacientes com rinite e/ou asma alérgicos a barata a tropomiosina da barata Periplaneta americana é um alérgeno principal. Em invertebrados, a tropomiosina induz resposta IgE e reatividade cruzada entre invertebrados incluindo ácaros, barata, camarão e parasitas. No presente estudo, produzimos tropomiosina recombinante de barata (alérgeno Per a 7) com elevado grau de pureza e com quantidades mínimas de endotoxina. Em modelo experimental de asma em camundongos, com sensibilização e desencadeamento com Per a 7 recombinante, houve aumento do número de células totais, e de eosinófilos, neutrófilos e linfócitos no lavado broncoalveolar. Alérgeno Per a 7 recombinante foi também utilizado em ELISA quimérico para investigar a resposta IgE a este alérgeno em 112 pacientes com dermatite atópica. A idade dos pacientes variou de 3 a 67 anos com média de 24,9 (± 15,4) anos, com 75 indivíduos do sexo feminino (67%). Nos 112 pacientes, o SCORAD apresentou média de 43,1 (± 18,1) e foram relatadas asma e rinite em 42 e 85 indivíduos, respectivamente. Níveis de IgE total apresentaram ampla variação, de 14,2 a 63.000 UI/mL, com média geométrica de 2.193 UI/mL. A idade de início dos sintomas e o tempo de doença apresentaram médias, respectivamente, de 9,5 (± 12,9) e 15,4 (± 12,3) anos. Trinta pacientes (26,8%) apresentaram níveis detectáveis de IgE para Per a 7 recombinante, com variação de 2,3 a 3.191 UI/mL. A razão de IgE específica para Per a7/IgE total nestes pacientes variou de 0,03% a 33,8%. Dividindo os pacientes em sensibilizados e não sensibilizados ao alérgeno de barata, observamos que não houve diferença significante entre os grupos com relação à idade de início dos sintomas, tempo de doença, SCORAD, presença de rinite ou asma e níveis de IgE total. Nossos resultados mostraram que o alérgeno Per a 7 recombinante induziu resposta inflamatória com caraterísticas semelhante à observada em humanos, em modelo experimental de asma em camundongos. Frequência menor de reatividade IgE ao alérgeno Per a 7 foi observada entre pacientes com dermatite atópica em nosso meio, quando comparada à observada previamente em pacientes com asma e/ou rinite. Não houve associação entre a presença de sensibilização IgE ao alérgeno Per a 7 e a gravidade da dermatite atópica, presença de asma ou rinite, idade de início e tempo de doença, e níveis de IgE total. Entretanto, a investigação do perfil de sensibilização IgE tem importância ao se considerar o uso de imunoterapia alérgenoespecífica em pacientes com dermatite atópica. / The prevalence of allergic diseases has increased in recent years, including asthma and atopic dermatitis. Asthma is a chronic disease resulting from airway hyperresponsiveness and variable airflow limitation, reversible spontaneously or with treatment. The high prevalence, mortality, and associated costs make the disease an important public health problem which requires attention. Atopic dermatitis (AD) is a chronic inflammatory skin disease, as asthma has high prevalence, and it is characterized by intense itching, erythema, excoriations, lichenification in the skin among other characteristics. The AD causes profound impact on individual and family´s life and usually shows the first symptoms during childhood. Both diseases could be related and associated with sensitization to allergens. In the case of asthma, allergy to cockroach is well known to be related with the disease. In 1999, a Brazilian study in our group has shown that among patients with allergic rhinitis and /or asthma the tropomyosin of the American cockroach Periplaneta americana (Per a 7) is a major allergen. This protein is present in vertebrates and invertebrates, and was related to induction of IgE response and cross-reactivity against among invertebrates, including mites, cockroaches, shrimp and parasites. In the present study, we produced recombinant tropomyosin from cockroach (Per a 7 allergen) with high purity and minimal amounts of endotoxin. In an experimental model of asthma in mice, with sensitization and triggering with recombinant Per a 7, there was an increase in the number of total cells, and eosinophils, neutrophils and lymphocytes in the bronchoalveolar lavage. Per a 7 allergen was also used in chimeric ELISA to investigate IgE response to this allergen in 112 patients with atopic dermatitis. Patients\' ages ranged from 3 to 67 years, mean of 24.9 (± 15.4) years, 75 female subjects (67%). In the 112 patients, a mean of SCORAD presented 43.1 (± 18.1) and asthma and rhinitis were reported in 42 and 85 individuals, respectively. Total IgE levels varied from 14.2 to 63.000 IU / mL, with a geometric mean of 2,193 IU / mL. The age at onset of symptoms and disease time presented averages, respectively, of 9.5 (± 12.9) and 15.4 (± 12.3) years. Thirty patients (26.8%) had detectable IgE recombinant Per a 7 levels, ranging from 2.3 to 3191 IU / mL. The ratio of IgA specific for Per a7 / total IgE in these patients ranged from 0.03% to 33.8%. Dividing the patients into sensitized and not sensitized to the cockroach allergen, we observed that there was no significant difference between the groups regarding the age of onset of symptoms, disease time, SCORAD, presence of rhinitis or asthma, and total IgE levels. Our results showed that recombinant Per a 7 allergen induced an inflammatory response with characteristics similar to that observed in humans in an experimental model of asthma in mice. Minor frequency reactivity of IgE Per a 7 allergen was observed among patients with atopic dermatitis in our group, when compared to that previously observed in patients with asthma and / or rhinitis. There was no association between the presence of IgE sensitization to the Per 7 allergen and the severity of atopic dermatitis, presence of asthma or rhinitis, age at onset and disease time, and total IgE levels. However, the investigation of the IgE sensitization profile is important when considering the use of allergen-specific immunotherapy in patients with atopic dermatitis

Oncogenic microRNAs (miRNAs) are small RNA molecules that inhibit post-translational regulatory mechanisms at the epigenetic level. miRNAs are often deregulated in malignancies and due to their stability are detectable in non-cellular fractions of peripheral blood. In our laboratory, we have performed several studies that have investigated and utilized miRNAs as biomarkers for various hematological tumors (e.g., chronic lymphocytic leukemia, Hodgkin`s lymphoma) and solid tumors (e.g., breast cancer). The aim of these studies was to find the association of miRNAs with pathophysiological and clinical aspects of each disease. Here, we confirmed the importance of particular miRNA or its complex during disease monitoring. Combining clinical, molecular biological and statistical analyses, we were able to find miRNA sets that fulfilled not only a diagnostic role but also a prognostic role beyond expectations. The main focus of this thesis is on the investigation of microRNAs in the diagnosis of a hematological malignancy - primary cutaneous T-cell lymphoma (CTCL). Tumor specificity of some miRNAs has been demonstrated. Their aberrant expression in tissue samples of CTCL patients obtained from skin biopsies, correctly distinguished malignant disease from control samples of benign skin lesions. Here, we...

The thesis deals with design and implementation of a tool for simulating marks of chosen skin diseases into a synthetic fingerprint. The diseases selected to work with are warts and atopic eczema. The marks of diseases are generated into a synthetic fingerprint image created by the SFinGe application. Existing disesase-affected fingerprints from the STRaDe database are analysed in detail. Then, methods for simulating the diseases into a synthetic fingerprint are proposed, implemented, and the results are evaluated.

Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.