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No description available.

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.

Dois experimentos foram conduzidos com o objetivo de avaliar a utilização de dietas com níveis crescentes de glicerina de biodiesel apenas na fase pré-inicial (1 a 7 dias) e apenas na fase final (35 a 42 dias de idade) de frangos de corte, respectivamente. As variáveis analisadas incluíram o desempenho, o rendimento de carcaça e cortes, a retenção de umidade em pintos aos 7 dias, a umidade de cama, a incidência de pododermatite, os níveis plasmáticos de colesterol e triglicerídeos e o custo da ração referente ao ganho de peso. Os tratamentos consistiram de dietas formuladas com 0%, 5% ou 10% de glicerina, sendo isonutritivas e formuladas à base de milho, farelo de soja e óleo de milho. A glicerina usada continha 83,6% de glicerol, 11,2% de água, 1,83% de sódio e 397 mg/kg de metanol, com energia metabolizável estimada de 3258 kcal/kg. No Experimento I, as dietas com glicerina foram fornecidas apenas na fase pré-inicial ou durante todo o ciclo de criação com 1610 aves em um delineamento inteiramente casualizado com 5 tratamentos e 7 repetições. Na primeira semana, as dietas com glicerina promoveram melhora significativa no ganho de peso e na conversão alimentar, sem afetar o consumo de ração e viabilidade. Essa melhora não foi mantida após os 14 dias de idade. Não houve efeito de tratamentos sobre o ganho de peso, consumo de ração e conversão alimentar. Entretanto, houve redução significativa na viabilidade aos 35 dias de idade das aves com 10% de glicerina por todo ciclo, mas não aos 42 dias; ao final do experimento, o índice de eficiência produtiva desse tratamento sofreu redução significativa. A umidade da cama das aves do tratamento com 10% de glicerina por todo ciclo de criação foi mais alta que dos demais a partir dos 21 dias, entretanto a incidência de pododermatite foi mais severa tanto para 5% como para 10% de glicerina por todo ciclo. Os níveis e tempos de fornecimento de glicerina não afetaram o rendimento de carcaça e de cortes ou a gordura abdominal. No experimento II, as dietas experimentais foram fornecidas a 819 frangos apenas na semana final de criação em um experimento inteiramente casualizado com 3 tratamentos e 7 repetições. O desempenho e o custo de ração por kg de ganho de peso não foram afetados pelos tratamentos. Nos Experimentos I e II, as rações contendo glicerina durante todo o ciclo ou apenas na última semana não resultaram em alteração nas concentrações plasmáticas de triglicerídeos e colesterol dos frangos. Dietas contendo 5% ou 10% de glicerina na fase pré-inicial ou 5% de glicerina continuamente não afetam a produtividade de frangos de corte, enquanto o nível de 10% durante todo o ciclo foi prejudicial. Esse mesmo nível de glicerina pode ser usado na dieta dos 35 aos 42 dias de idade sem afetar as aves. / Two experiments were conducted to evaluate the use of diets with increasing levels of biodiesel glycerin only in pre-starter (1-7 days) and only in the final phase (35-42 days of age) of broilers, respectively. The variables analyzed included the performance, carcass and parts yield, moisture retention in chicks at 7 days, litter moisture, incidence of foot pad dermatitis, plasma levels of cholesterol and triglycerides and diet cost related to weight gain. Treatments were isonutritive diets based on corn, soybean meal and corn oil containing 0, 5 and 10% glycerin. The glycerin used contained 83.6% glycerol, 11.2% water, 1.83% sodium and 397 mg/kg methanol, with estimated metabolizable energy value of 3,258 kcal/kg. In the first experiment, diets with glycerin were provided only in the pre-starter or for the entire growth period with 1610 birds in a completely randomized design with 5 treatments and 7 replications. In the first week, diets with glycerin resulted in significant improvement in weight gain and feed conversion without affecting feed intake and viability. This improvement was not sustained after 14 days of age. There was no effect of treatments on weight gain, feed intake and feed conversion. However, there was a significant reduction in viability at 35 days of age for the birds with 10% glycerin throughout the cycle, but not at 42 days; at the end of the experiment, the productivity index of this treatment was significantly decreased. Litter moisture for the treatment with 10% glycerin throughout the cycle was higher than the for the others from the 21 days, however the incidence of foot pad dermatitis was more severe for both 5% and 10% glycerin treatments for the entire period. The levels and periods of glycerin feeding did not affect carcass and parts yield and abdominal fat. In experiment II, the experimental diets were provided to 819 chickens only in the final week of growth in a completely randomized design with 3 treatments and 7 replications. The performance and feed cost per kg of weight gain were not affected by treatments. In Experiments I and II, the diets containing glycerin throughout the cycle or just in the last week resulted in no change in plasma concentrations of triglycerides and cholesterol of chickens. Diets containing 5% or 10% glycerin in the pre-starter phase or 5% glycerin continually did not affect the productivity of broilers, while the 10% level throughout the cycle was detrimental. This same level of glycerin can be used in the diet from 35 to 42 days without affecting the birds.

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

<p>Atopic dermatitis (AD) is a common pruritic skin disease in people and domestic animals that can be severely debilitating and stressful to the patient and the caregiver. The diagnosis of AD requires time consuming and expensive procedures, and treatment is often lifelong at considerable cost. Alterations in the lipid composition of the epidermis are a hallmark of the disease, and these may represent changes caused by the inflammation and defects in the lipid barrier. Liquid chromatography tandem mass spectrometry (LC-MS/MS) and, more recently, untargeted profiling using high-resolution time-of-flight instruments have been used to quantify the lipid composition in skin and other tissues, but these approaches are highly demanding in sample preparation and instrument time. In addition, these methods either detect only a limited number of lipids at the time or the identification of detected mass-to-charge ratio (m/z) is problematic when untargeted profiling is used. New lipidomic approaches that generate lipid profiles in a faster and more efficient manner can lead to a better understanding of these lipid changes. </p><p>The mass spectrometry analytical strategy used in this study, multiple reaction monitoring (MRM)-profiling, rapidly identifies discriminant lipids of the epidermis by flow injection. MRM-profiling is a small molecule accelerated discovery workflow performed in two parts using a triple quadrupole mass spectrometer with electrospray ionization as the ion source. Briefly, the first step consists of discovery experiments based on neutral loss and precursor ion scans to detect lipids in pooled samples by targeting class-specific chemical motifs such as polar heads of phospholipids or sphingoid bases of ceramides. The second step of the MRM-profiling is the screening of individual samples for the transitions detected in the discovery phase. </p><p>We first developed the experimental approach of the MRM-profiling methodology using epidermal samples of mice with AD-like inflammatory skin disease (chronic proliferative dermatitis, cpdm). Subsequently, we investigated lipid changes as the disease in mice progressed from minimal to severe. In order to select the most relevant ions, we utilized a two-tiered filter/wrapper feature-selection strategy. First, we built linear models linking the presence of every lipid monitored to disease stage information. The top 10 lipids, ranked based on η2 effect size, were used to build a predictive elastic-net (E-net) regression model linking the lipid ions detected by MRM-profiling with disease progression. The developed model accurately identified disease stages based on the variations in relative amounts of lipid ions corresponding to phosphatidylcholines, cholesterol esters, and glycerolipids-containing and eicosapentaenoic acid fatty acyl residues. Finally, we investigated the lipid profile of the epidermis in dogs with canine AD using the previously developed methodology. Epidermis from client owned patients and healthy controls were collected. Patients were sampled from affected and unaffected skin avoiding areas with secondary infections and the canine atopic dermatitis extent and severity index (CADESI-4) was recorded. The monitored lipids substantially separated the samples of healthy dogs from atopic dogs and distinguished the affected from the unaffected skin of patients. Samples were grouped into two cohorts for low-score and high-score CADESI-4, the first principal component was able to differentiate the control group from the low and high-score group. Differences in the lipid composition associated with low and high score CADESI-4 were significantly different only after separating the samples by sex of the dogs, demonstrating sexual dimorphism in the lipid changes associated with disease. The compositional data was feature extracted using the CADESI-4 to build linear models that identified oleic acid-containing triacylglycerides, long-chain acylcarnitines and sphingolipids as highly predictive lipids and were subsequently used to construct a predictive E-net regression. The lipid fingerprint obtained from the MRM-profiling was highly correlated (R2=0.89) with the classification of the standardized CADESI-4 score. </p><p>This research showed that changes in the lipid composition of the epidermis can be detected by MRM-profiling in atopic dogs even when the skin looks clinically healthy and that sex is a modifying factor in the lipid profile of canine atopic dermatitis (CAD). We expect that this research leads to a better understanding of the lipid changes in the epidermis during the onset of AD and as the chronic inflammatory process develops. The high prediction rate given by the lipid biomarkers for disease progression identified here by the machine learning strategy provides a potential molecular assessment tool for the diagnosis and monitoring of atopic dermatitis and the patient response to treatment.</p><div><br></div>

Introdução: A dermatite atópica (DA) é uma doença cutânea inflamatória crônica que cursa em surtos. Possui manifestação clínica variável, mas o prurido e a xerose são características frequentes, e pode estar associada a outras manifestações extra-cutâneas de atopia. Pacientes com DA apresentam maior risco de infecções por bactérias e vírus, destacando-se a erupção variceliforme de Kaposi, causada por herpes simples. A DA mostra-se como exemplo de dermatose com comprometimento da barreira cutânea, aliado a disfunção imunológica. São descritas alterações das proteínas da barreira cutânea na DA (filagrina e claudinas ), relacionadas ao maior risco de infecção . Objetivo: Avaliar a expressão de proteínas relacionadas à barreira cutânea como a filagrina, e as claudinas -1 e -4 na pele de pacientes adultos com dermatite atópica, acompanhados no Departamento de Dermatologia da Faculdade de Medicina da Universidade de São Paulo. Métodos: 32 indivíduos com diagnóstico de DA (estabelecido pelos critérios de Hanifin & Rajka) e 23 controles (indivíduos sem DA), maiores de 18 anos, foram submetidos a biópsias cutâneas. Os indivíduos com DA foram biopsiados em dois pontos, tanto na pele lesada, quanto na pele não-lesada. O material obtido foi analisado por imuno-histoquímica, através de marcadores específicos para filagrina, claudina-1 e claudina-4. As lâminas foram digitalizadas pelo Panoramic Scan - 3DHistech - Hungria, e as imagens analisadas pelo software Image Pro Plus 4,5, quanto à intensidade da expressão do marcador. A espessura média da epiderme do local estudado foi também avaliada. O grupo com DA foi também analisado quanto à gravidade da doença (EASI), níveis séricos de IgE e grau de eosinofilia. Resultados: Houve redução da expressão da filagrina na pele de doentes de DA em relação aos controles, tanto na pele com lesão quanto na pele sem lesão. Demonstrou-se correlação inversa na expressão da filagrina, tanto com relação à gravidade da doença quanto à espessura da epiderme. A análise das claudinas -1 e -4 demonstrou redução de ambas na pele dos doentes de DA, mas não houve correlação com a gravidade, espessura da epiderme, níveis de IgE sérica ou eosinofilia. Conclusão: No adulto com dermatite atópica, existe redução da expressão das proteínas relacionadas à barreira cutânea, como a filagrina e as claudinas -1 e -4. A redução da expressão da filagrina relacionou-se inversamente com a gravidade da doença, e com a espessura da epiderme, sugerindo cronicidade das lesões. Houve redução da expressão das claudinas -1 e -4, sem relação com a gravidade da doença, espessura da epiderme, eosinofilia ou com os níveis séricos de IgE / Introduction: Atopic dermatitis (AD) is a chronic, inflammatory dermatosis with ocasional flares. Its clinical features are variable, but pruritus and xerosis are frequent, and the disease may be associated to extracutaneous atopy. Patients with AD have increased risk for bacterial or viral infection, with emphasis on eczema herpeticum due to herpes simplex. AD is an example of a compromised skin barrier, allied to na imune dysfunction. There are reports on efective proteins of the skin barrier (filaggrin and claudins), related to increased risk for infection. Objectives: To evaluate the expression of proteins related to the skin barrier, such filaggrin and claudins-1 and-4 in the skin of adults with AD, followed at the Department of Dermatology, University of Sao Paulo Medical School. Methods: 32 individuals diagnosed as AD, according to Hanifin & Rajka\'s criteria, and 23 non-atopic controls, above the age of 18, were biopsied. Individuals with AD were biopsied in two different sites (lesional and nonlesional skin). The specimens were analyzed by immunohistochemistry through specific markers for filaggrin, claudins 1 and 4. The slides were scanned utilizing Panoramic Scan - 3DHistech - Hungary, and images analyzed by Image Pro Plus 4,5 for the intensity of each marker. The mean epidermal thickness was also evaluated. AD patients were also analyzed for disease severity (EASI), circulating IgE levels and eosinophilia. Results: In lesional and nonlesional skin of AD patients there was a reduced expression of filaggrin, when compared to nonatopic controls. There was an inverse correlation of filaggrin expression with disease severity and epidermal thickness. In the skin of AD individuals, there was reduced expression of claudins 1-and-4, which did not correlate with disease severity, epidermal thickness or eosinophilia. Conclusion: In adults with AD, there is reduced expression of skin barrier proteins, such as filaggrin, claudins 1 and 4. The reduction of filaggrin expression had an inverse correlation with disease severity and epidermal thickness, suggesting disease chronicity. There was reduction of claudins 1 and 4, with no relation with disease severity, epidermal thickness, circulating IgE levels or eosinophilia

Abstract Bullous pemphigoid (BP) is the most common autoimmune blistering disease. It mostly affects elderly patients and is characterized by intense pruritus and blistering or bullae. Treatment options include topical and systemic corticosteroids, other immunosuppressive drugs and doxycycline. Disease course may be chronic and relapses are common. The incidence of BP has been reported to have increased in the last few decades, but the reason for this trend is not known. The aim of this thesis was to study the risk factors of BP. Firstly, the influence of the use of dipeptidyl peptidase (DPP-4) inhibitors was analyzed as a risk factor, and then those of other oral diabetes medications. This study also aimed to determine whether drugs used for psychiatric and neurologic conditions are risk factors for BP. Finally, previously diagnosed dermatitis herpetiformis (DH) and celiac disease (CD) were examined as potential risk factors for subsequent BP. For this retrospective, matched case-control study, patient data were obtained from the Finnish Care Register for Health Care database, and data on reimbursed drugs from the Social Insurance Institution of Finland. In the present study, prior use of DPP-4 inhibitors was found to increase the risk of BP twofold and in particular, vildagliptin increased the risk tenfold. The mean time between the initiation of vildagliptin and diagnosis of BP was 449 days. Metformin and other conventional diabetes drugs were not risk factors for BP. Several drugs used for neurological and psychiatric diseases were associated with an elevated risk for BP, but no pharmacological or chemical properties of these drugs emerged as candidates to explain the increased risk. A prior diagnosis of DH increased the risk of BP 22-fold and a diagnosis of CD doubled it. Dapsone had been used in the two years before BP diagnosis by 44% of patients whose BP was preceded by DH. The mean time between the diagnoses of DH and BP was 3.3 years. This study confirms the view that DPP-4 inhibitors increase the risk for BP. No such association was found with other classes of diabetes drugs and therefore their use can be continued following a diagnosis of BP. Doctors treating patients with DH should be aware of the association between DH and BP, and be particularly vigilant if a DH patient’s skin symptoms change or become unresponsive to a gluten-free diet and/or dapsone. / Tiivistelmä Rakkulainen pemfigoidi (pemfigoidi) on yleisin ihon autoimmuunirakkulatauti. Pemfigoidi on pääasiassa ikääntyneiden sairaus, ja sen tyypillisiä oireita ovat kova kutina ja rakkulat iholla. Pemfigoidin hoitoon käytetään paikallisia ja systeemisiä kortikosteroideja, muita immunosuppressiivisia lääkkeitä sekä doksisykliiniä. Taudinkulku on usein krooninen ja uusiutumiset ovat yleisiä. Rakkulaisen pemfigoidin ilmaantuvuuden on raportoitu lisääntyneen, mutta syitä tähän muutokseen ei täysin ymmärretä. Tämän tutkimuksen tavoite oli tutkia pemfigoidin riskitekijöitä Suomessa. Retrospektiivisessä tapaus-verrokkitutkimuksessa käytettiin aineistona Terveyden ja hyvinvoinnin laitoksen hoitoilmoitusrekisteristä poimittuja pemfigoidipotilaita (N=3397) ja verrokkeina ihon tyvisolusyöpäpotilaita (N=12941). Tiedot korvatuista lääkeostoista saatiin Kelan lääkekorvausrekisteristä. Tutkimuksessa todettiin DPP-4:n salpaajien kaksinkertaistavan pemfigoidin riskin ja DPP-4:n salpaaja vildagliptiini lisäsi riskiä jopa kymmenkertaiseksi. Vildagliptiinin aloituksen ja pemfigoidin toteamisen välillä kului keskimäärin 449 vuorokautta. Metformiini ja muut tutkitut suun kautta otettavat diabeteslääkkeet eivät lisänneet pemfigoidin riskiä. Useiden psykiatrisiin ja neurologisiin sairauksiin käytettävien lääkkeiden todettiin lisäävän pemfigoidin riskiä. Pemfigoidin on kuvattu voivan puhjeta ihokeliakian jälkeen, mutta laajempia tutkimuksia näiden sairauksien yhteydestä ei oltu aiemmin tehty. Tämän vuoksi samassa potilasaineistossa tutkittiin ihokeliakiaa ja keliakiaa pemfigoidin riskitekijöinä. Edeltävä ihokeliakia lisäsi pemfigoidin toteamisen riskiä selvästi, jopa 22-kertaiseksi ja keliakia kaksikertaiseksi. Huomattava osa potilaista oli ostanut ihokeliakian hoitoon käytettävää dapsonia edeltävän 2 vuoden aikana ennen pemfigoidin toteamista, mikä voi kertoa ihokeliakian oireiden aktiivisuudesta. Tämä tutkimus vahvistaa näkemystä siitä, että DPP-4:n salpaajat ovat pemfigoidin riskitekijä. Muut tutkitut diabeteslääkkeet eivät lisänneet riskiä ja voidaan ajatella, että ne eivät edelleen hankaloita aiemmin todetun pemfigoidin oireita. Koska ihokeliakian todettiin olevan pemfigoidin riskitekijä, tulee näitä potilaita hoitavan lääkärin muistaa pemfigoidin mahdollisuus, jos ihokeliakian oireet muuttuvat tai hoitovaste menetetään.

O crescente aumento da resistência microbiana aos antibióticos disponíveis impulsiona a busca por novas substâncias, com características superiores às correntemente usadas. Neste sentido, este trabalho propôs investigar as propriedades terapêuticas do xilitol visando a sua aplicação no controle da dermatite atópica, patologia que acomete a pele e que é agravada pela presença da bactéria Staphylococcus aureus. No presente estudo foram executados ensaios in vitro de atividade antimicrobiana do xilitol e verificado se este composto atua na aderência bacteriana, sobre a bactéria Staphylococcus aureus ATCC 25923. Foi avaliada a ação do xilitol produzido tanto pela via química quanto pela biotecnológica, sendo este último obtido a partir de fermentação do hidrolisado hemicelulósico da palha de trigo, nas concentrações de 1, 5 e 10 % (p/ v). A seguir, foram executados testes de toxicidade dérmica aguda com doses repetidas com xilitol nas concentrações de 5 e 10 % (p/ p), nas formas farmacêuticas de creme e de gel, em coelhos albinos da raça Nova Zelândia e testes de fototoxicidade, na concentração de 10 % (p/ p), nas formas farmacêuticas de creme e de gel, utilizando cobaias albinas da raça Durkin-Hartley. Em relação aos ensaios in vitro, observou-se que o xilitol, nas concentrações testadas, não impediu o crescimento bacteriano, mas inibiu a aderência da bactéria a uma superfície de prova, evidenciando ser este o provável mecanismo de ação desta substância sobre as bactérias. Em testes toxicológicos realizados, todas as formulações contendo xilitol foram classificadas como não irritantes quando foram avaliadas quanto à toxicidade dérmica aguda com doses repetidas. Entretanto, nos testes da fototoxicidade, as formulações testadas apresentaram certa fototoxicidade, sugerindo ser a formulação a base de creme mais fototóxica do que aquela a base de gel. Estes resultados evidenciam a aplicabilidade do xilitol no controle da dermatite atópica, como princípio ativo de formulações seguras, observando que a aplicação deve ser realizada com o uso de protetor solar. Este estudo buscou contribuir de maneira expressiva na elucidação do mecanismo de ação do xilitol e verificar os cuidados que devem ser considerados quando realizada sua aplicação pela via dérmica. / Since the microorganisms\' resistance to antibiotics increases, it is imperative to look for different substances that can combat these pathogens growth, with greater advantages. The propose of this work was to study therapeutic properties of xylitol in order to control atopic dermatitis, a dermal disease characterized by the presence of Staphylococcus aureus bacterium. Xylitol is a substance that can be obtained by chemical and biotechnological means, being the latter a relevant alternative that produces high-value compounds obtained by fermentation of lignocelullosic hydrolysates. In the present study in vitro assays were performed in order to check if wxylitol (obtained by chemical and biotechnological means) has antimicrobial activity at 1, 5 and 10 % (w/ v) concentrations. Other assays were also performed to verify if xylitol properties, at the same concentrations, hinder the adherence of Staphylococcus aureus ATCC 25923 bacterium. Xylitol was produced by biotechnological means using wheat straw hemicellulosic hydrolysate. Afterwards, in vivo assays were performed to investigate if xylitol is safe for skin application. Acute dermal toxicity tests with repeated doses were done with New Zealand rabbits using concentrations of 5 and 10 % (w/ w) xylitol, in either cream or gel. Phototoxicity assays were also performed with Durkin-Hartley guinea pigs, using only 10 % (w/ w) xylitol, in cream and gel forms. It was observed that xylitol does not have antimicrobial properties on S. aureus at all tested concentrations, but this compound has the capability of inhibiting this bacterium adherence on a surface, at all tested concentrations. In relation to toxicity assays, formulations contaning xylitol are nonirritative. However, xylitol has phototoxicity properties, mainly when cream is the base. Xylitol is an adequate alternative to be applied for atopic dermatitis control, and its application on the skin should be done with sunscreen. This study aimed to clarify xylitol action mechanism and to check the cares that should be taken when xylitol is applied on skin.