M.Tech. (Homoeopathy) / Atopic dermatitis (atopic eczema) is a chronic, relapsing, allergic inflammatory skin disease (Hauk, 2008). The prevalence of atopic dermatitis has increased significantly over the past few decades, with highest rates of 45 – 64% occurring amongst preschool children (Butler, 2009), and 40% amongst older children and adults (Manjra, 2005). This increase in prevalence is attributed to environmental factors such as microbial exposure and poor nutrition, which can all lead to atopic dermatitis (Schnopp, 2006). The quality of life of patients suffering from atopic dermatitis and their family members are significantly affected (Manjra, 2005). Atopic dermatitis is characterized by active skin lesions that are red, flaky, dry and itchy and in children commonly occurs in the flexural areas of the body (Fölster-Hols et al., 2007, Schnopp, 2006). Conventional treatment potions for atopic dermatitis are associated with adverse effects in children (Kalicharan et al., 2005). Homoeopathic remedies may offer an alternative option for this condition. This study aimed to assess the effect of a homoeopathic complex consisting of Graphites 6cH, Histaminum 9cH, Psorinum 6cH and Sulphur 6cH, on atopic dermatitis in children. All the participants of the study received the homoeopathic complex. The atopic dermatitis was evaluated using the SCORAD index (Scoring of Atopic Dermatitis) (Appendix F) and the Children’s Dermatology Life Quality Index (CDLQI) (Appendix E). Thirty four participants who met the inclusion and exclusion criteria were recruited to participate in this pre-test – post-test single group study by means of advertisements (Appendix A) placed in and around primary schools in the Gauteng area (with relevant permission given) and in the local newspaper. Participants were also recruited via word of mouth. Once participants were accepted into the study they were allocated into the treatment group which received the homoeopathic complex containing Graphites 6cH, Histaminum 9cH, Psorinum 6cH and Sulphur 6cH. The study was completed over a four week period. The percentage of the area affected, the intensity of the symptoms, the pruritus and the loss of sleep as well as the quality of life of the participants suffering from atopic dermatitis were aspects of the condition evaluated on a weekly basis. The results for the CDLQI showed improvements in the participant’s perception of itching/ pain of the affected area, as well as their quality of sleep. These improvements were shown to have occurred gradually over the study period. There were however no statistically significant changes noted in the mental and emotional quality of life of the participants.

Indiana University-Purdue University Indianapolis (IUPUI) / Atopic dermatitis (AD) is a pruritic, chronic inflammatory skin disease with a lifetime prevalence of 10-20% in children and 1-3% in adults, worldwide. In the past three decades, prevalence of the disease has increased by two to three-fold in industrialized countries, with higher incidences in urban regions compared to rural regions. Mice with an activating mutation in STAT6, known as STAT6VT, constitutively express STAT6 in T-cells. Our preliminary data suggests significant differences between the STAT6VT transgenic mice from WT littermate controls treated with SLS. These findings correlate with evidence that there are abnormalities in the barrier function between these mice

Dermatological conditions affect between 15 to 25% of canines. Canine Atopic Dermatitis (CAD) is one of the most prevalent of these conditions and is a chronic inflammatory disorder with a wide array of causes and triggers and no known cure. Symptoms of this condition can be persistent and often lead to significant discomfort, leading many owners to seek preventative care in an attempt to avoid expensive veterinary care and discomfort in their dogs. Sebaceous Adenitis (SA) is another skin condition, which while less common, is also a disorder of the inflammatory pathways and affects keratin production in the coats of dogs. It has more persistent symptoms and requires rigorous grooming care to prevent its onset, and as a result is the cause for euthanasia in many of the breeds which it affects. Similar to CAD, owners of dogs with SA seek preventative care to avoid flares of the conditions that are often coupled with secondary infections that perpetuate the skin’s poor condition. Due to the overwhelming variety and misleading advertising that currently exists in the dog food market, one key element to preventative treatment that is confusing to many owners is manipulation of the diet. Research has demonstrated that certain dietary components may be important. These include increased protein composition, supplementation of vitamins A, D, and E, essential fatty acid supplements, and the use of probiotics and prebiotics. Increasing the proportion of dietary protein to total energy intake allows for greater consumption of amino acids, which can be utilized to renew epidermal cells, including hair follicles. It is speculated that Vitamin A supplementation may slow down the rate of hyperkeratinization seen in SA as it attenuates epithelial cell turnover. Supplementation with vitamin E has been found to improve pruritus in CAD and acts as an anti-inflammatory in areas which are affected by a loss of sebum, as seen in SA. It may also act to improve antibody responses in hypersensitivity reactions, which are exaggerated immunologic responses to allergens, seen in CAD. It is known that an increase in dietary vitamin D enhances immunomodulatory roles that lead to its ability to reduce lesions in both CAD and SA. Essential fatty acids (FA), of which two are omega-3 and omega-6 FA, are useful supplements for dogs with CAD, as they may affect mediators of inflammation specific to inflammatory skin disorders. Additionally, supplementation with essential FA may replenish a factor lacking in an impaired skin barrier. The breakdown in the epithelial barrier leads to increased cutaneous permeability and decreased membrane fluidity. It is important to balance the more pro-inflammatory effects of omega-6 FA with the anti-inflammatory effects of omega-3 FA. Two balanced sources of these essential fatty acids that often appear in the literature are evening primrose and borage oils, which supply gamma linoleic acid (GLA), a less inflammatory omega-6 FA. Probiotics/prebiotics aid in repairing possible issues observed in the gut-skin axis seen in AD though the strains which provide benefits are not consistent in the literature. In summary, there are promising developments on the treatment of CAD and SA through diet, although extensive research is needed to validate these studies and clarify the results prior to usage by the general public.

No description available.

Introduction: Staphylococcus aureus skin infection is an almost ubiquitous feature of patients with atopic dermatitis (AD). TLR1, 2 and 6 are important in immune sensing of these bacteria. The aim of this study was to determine whether defects in TLR1, 2 and/or 6 expression/function may explain the propensity to infection in humans and the NC eczema mouse model. Methods: Fibroblast cell lines from severe AD, nonatopic controls, and mouse embryonic fibroblasts (MEFs) from the NC, MSM/Ms wild-type strain and a 3T3 control strain TLR1, 2 and 6 expression were measured by qPCR and FACS. IL-6, TNF-α, TSLP and IL-33 production was measured by qPCR and ELISA at baseline and after stimulation with LPS, HKSA and a live strain of Staphylococcus aureus that produced only SEB. Results: No differences were found in either TLR expression or function in human fibroblasts derived from patients and controls. The MSM/Ms MEFs expressed significantly more TLR1 and 2, as well as exhibited high inflammatory profile after stimulation comparing with 3T3 and the NC MEFs. Live Staphylococcus aureus, but not HKSA, LPS or SEB, was a potent stimulus for the Th2-inducing cytokines (TSLP and IL-33), and induce cell death. Cytokines levels were found to be similar in AD and NC MEFs when compared to healthy controls. Conclusion: Eczema in both the human and NC mouse is not associated with abnormalities in fibroblasts TLR1, 2, and 6. Live, but not killed Staphylococcus aureus or its enterotoxin, is a potent inducer of TSLP and IL-33 in both species.

Over the past four years, a previously unrecognised family of innate effector cells has been identified. Their comprehensive functional capabilities range from lymphoid organogenesis, tissue remodelling, wound healing, immune protection and homeostasis to contribution to inflammation and allergic responses. Here we investigate the presence and function of type 2 innate lymphoid cells (ILC2) in the skin. We show that human ILC2 are resident in human skin and express RORA and GATA3, and skin homing receptors. ILC2 further infiltrate the skin after allergen challenge, where they produce the type 2 cytokines IL-5 and IL-13. Skin-derived ILC2 express the IL-33 receptor ST2, which is up-regulated during activation. Signalling via IL-33 induces type 2 cytokine and amphiregulin expression, and increases ILC2 migration. Atopic dermatitis (AD) is a chronic inflammatory skin disorder. Current evidence suggests that both skin barrier dysfunction and immune system abnormalities, particularly those of a type 2 phenotype, contribute to disease pathogenesis. We demonstrated that ILC2 are enriched in lesional skin biopsies from atopic patients and show higher expression of cytokine receptors, reflecting an activated phenotype. Down-regulation of E-cadherin is characteristic of filaggrin insufficiency, a cardinal feature of AD. Interestingly, E-cadherin binding to KLRG1 on human ILC2 dramatically inhibits IL-5 and IL-13 production. ILC2 may contribute to increases in type 2 cytokine production in the absence of the inhibitory E-cadherin ligation through this novel mechanism of barrier sensing. CRTH2, a receptor for prostaglandin D₂ (PGD₂), is expressed by human ILC2. However, the function of CRTH2 in these cells is unclear. We sought to determine the role of PGD₂ and CRTH2 in human ILC2 and compare it with that of the established ILC2 activators IL-25 and IL-33. PGD₂ binding to CRTH2 induced ILC2 migration and production of type 2 cytokines IL-4, IL-5, IL-13 and release of other pro-inflammatory cytokines IL-3, IL-8, IL-9, IL-21, GM-CSF, and CSF-1 in a dose-dependent manner. ILC2 activation through CRTH2 also upregulated the expression of IL-33 and IL-25 receptor subunits (ST2 and IL-17RA) suggesting a synergistic role. The effects of PGD₂ on ILC2 could be mimicked by the supernatant from activated human mast cells and inhibited by a CRTH2 antagonist. Therefore, PGD₂ can be considered as an important and potent activator of ILC2 through CRTH2 mediating strong inflammatory responses. Cell surface interaction mechanisms that activate ILC2 function are unknown. We observed the expression of NKp30 on ILC2 ex vivo and after culture. Using quantitative PCR we confirmed that ILCs express NKp30c splice variant, an immune-modulatory isoform. Incubation of ILC2 with the NKp30 ligand B7H6 and tumour cell lines expressing this protein induced production of type 2 cytokines. This interaction can be inhibited by NKp30 blocking antibodies. We further established that activation of NKp30 induces the canonical pathway of NFƙB signalling. Overall the work in thesis shows for the first time that ILC2 are resident in human skin and infiltrate rapidly after allergen challenge and in AD lesional skin. We have defined cytokine and lipid mediators that contribute to migration and activation of ILC2 and shown that KLRG1 and NKp30 act as inhibitory and activatory receptors respectively. The work defines novel pathways for barrier sensing and cutaneous inflammation, and identifies potential new targets for therapeutic intervention.

Vaccinia virus (VACV) is primarily known as a vaccine against its relative variola virus, the causative agent of smallpox. In the seventies of the 20th century, the vaccination campaign with VACV led to eradication of smallpox. Consequently, vaccination of the general population was stopped. Currently, the vaccination was reintroduced, namely among army and healthcare professionals. However, vaccination with VACV is accompanied with a high incidence of vaccination-related complications, namely among immunocompromised individuals. One of the complications is eczema vaccinatum, occuring in patients with atopic dermatitis. The laboratory of Dr. Melkova has focused on development of a model of eczema vaccinatum in mice Nc/Nga and on studies of pathogenesis of this complication. The goal of my diploma thesis is to contribute to characterization of imunopathogenesis of eczema vaccinatum in mice Nc/Nga infected either with VACV strain Western Reserve (WR) or with a recombinant VACV with the integrated cDNA for IRF-3 (Interferon Regulatory Factor 3; WR-IRF3). IRF-3 regulates the expression of interferon type I in response to viral infection. This recombinant virus has been constructed in the laboratory of Dr. Melková. The objective of my work was to verify the expression of the integrated cDNA for IRF-3 and to...

Orientador: Luiz Henrique de Araújo Machado / Resumo: Dermatite atópica (DA) canina é uma enfermidade alergoinflamatória de caráter crônico, da pele e orelha externa, geralmente de cães adultos jovens, induzida por distintos alérgenos. A afecção tem etiopatologia complexa e ainda não totalmente elucidada, dificilmente atingindo cura. Porém, há hoje, várias opções medicamentosas para o seu controle. O oclacitinib faz parte da recente remessa de medicamentos para tratamento e controle do prurido dos cães atópicos, com poucos efeitos colaterais. É um fármaco inibidor de janus quinase, com rápida ação e que age na ligação das citocinas aos receptores JAKs. Este trabalho teve como intuito avaliar a resposta inflamatória da pele de animais atópicos, antes e 30 dias após o tratamento com Oclacitinib comercial (Apoquel®), através da mensuração das interlucinas IL-1, IL-4, IL-6, IL-10, IL-17, IL-31, TNF-, IFN- e filagrina, pela técnica de imuno-histoquímica, além da resposta clinica ao tratamento pela avaliação do CADESI-4 e escore do prurido. Os 10 cães utilizados no estudo são provenientes do atendimento dermatológico do HV da UNESP-Botucatu e clínica particular de Sorocaba-SP. Apenas os escores do CADESI-4, do prurido e da IL-1, citocina inflamatória chave da imunidade inata e na patogênese da DA, apresentaram redução na análise estatística. Concluise que a IL-1 pode ter um papel importante na farmacocinética do oclacitinib e maiores estudos são necessários para confirmar a ação do oclacitinib sobre a produção e ligação da IL-1 aos ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Canine atopic dermatitis (DA) is a chronic inflammatory alergo disease of the skin and external ear, usually of young adult dogs, induced by distinct allergens. The disease has complex and still not fully elucidated etiopatology, hardly reaching cure. However, there are today several drug options for your control. Oclacitinib is part of the recent consignment of medications for treatment and control of the itching of atopic dogs, with few side effects. It is an inhibitor drug of Janus Kinase, with rapid action and acting in the binding of cytokines to jaks receptors. The objective of this work was to evaluate the inflammatory response of the skin of atopic animals before and 30 days after treatment with commercial oclacitinib (Apoquel®), through the measurement of interlukines il-1, IL-4, IL-6, IL-10, IL-17, IL-31, TNF-α, IFN-γ and Filagrin, by immunohistochemistry technique, in addition to clinical response to treatment by evaluation of Cadesi-4 and pruritus score. The 10 dogs used in the study are from the dermatological treatment of HV of UNESP-Botucatu and private Clinic of Sorocaba-SP. Only the scores of Cadesi4, pruritus and IL-1, the key inflammatory cytokine of innate immunity and the pathogenesis of DA, showed a reduction in the statistical analysis. It is concluded that IL-1 may play an important role in the pharmacokinetics of oclacitinib and further studies are needed to confirm the action of oclacitinib on the production and binding of IL-1 to cutaneous receptors... (Complete abstract click electronic access below) / Mestre

Dermatite atópica (DA) é uma dermatose inflamatória, de caráter crônico e recidivante, com alta prevalência mundial, caracterizada por eczema localizado ou generalizado, xerose cutânea e prurido intenso, mais comum em crianças, porém podendo acometer adultos, muitas vezes prejudicando de forma considerável a qualidade de vida dos pacientes e seus familiares. O diagnóstico de alergia inclui testes in vivo (testes cutâneos de hipersensibilidade imediata ou prick test) e testes in vitro com mensuração de anticorpos IgE no sangue. Está disponível método para detecção de alergia baseado no princípio de diagnóstico por componentes, o ImmunoCAP Immunosorbent Allergen CHIP (ImmunoCAP-ISAC) que marca a transição para o diagnóstico molecular de alergia. No presente estudo, tivemos por objetivos determinar o perfil de resposta IgE a alérgenos purificados, naturais ou recombinantes, em pacientes adultos com DA e comparar o valor do diagnóstico de alergia por componentes com métodos diagnósticos existentes. Foram selecionados 39 pacientes adultos com DA dentre aqueles Atendidos nos Ambulatórios de Alergia e Dermatologia do HCFMRP-USP. A idade dos pacientes variou de 14-66 anos (média ± DP 34,3 ± 2.1 anos), 64% mulheres. O tempo médio de doença foi de 16 anos. SCORAD médio foi de 36,6 (2-90). Média geométrica (MG) de IgE total foi 1,963 kU/L (24-63,000 kU/L). Alérgenos de ácaros foram dominantes, com sensibilização a Der p1 e a Der f1, Der p2 e Der f2 em 82% e 85% dos pacientes, com MG 27,6; 50; 39,2; 45,4 ISU-E respectivamente, seguidos de gato (38%), cachorro (36%) e pólen de gramíneas (36%). IgE para alérgenos de baratas, fungos, pólen de árvores, látex e veneno de insetos foi encontrada em menos de 20% dos pacientes em baixos níveis. Sensibilização para castanhas (33%) e camarão (31%) foram os alérgenos mais prevalentes entre os alimentos, enquanto a reatividade IgE para leite e ovo esteve presente em 10% ou menos dos pacientes, com baixos níveis de anticorpos IgE na maioria dos casos. Apenas 6 pacientes (15,4%) apresentaram IgE para o panalérgeno tropomiosina, e 3/39 (7,7%) foram negativos para todos os 112 componentes de alérgenos testados no ImmunoCAP-ISAC, com níveis de IgE total de 24, 38,7 e 156 kU/L. Concluímos que o perfil de sensibilização IgE entre os pacientes adultos com dermatite atópica difere daquele entre os pacientes com alergiarespiratória, apresentando menos sensibilização para baratas e para o panalérgeno tropomiosina, e difere ainda do perfil presente em crianças com DA, com predomínio de sensibilização IgE a castanhas e camarão, e baixa taxa de sensibilização a alérgenos de leite de vaca e ovo. / Atopic dermatitis (AD) is an inflammatory, chronic, relapsing dermatosis with a high global prevalence characterized by localized or generalized eczema, cutaneous xerosis and intense pruritus, more common in children, but it can affect adults, often causing considerable damage to the quality of life of patients and their families. The diagnosis of allergy includes in vivo tests (skin tests of immediate hypersensitivity or prick test) and in vitro tests with measurement of IgE antibodies in the blood. An allergy detection method based on the principle of component diagnosis is available, ImmunoCAP Immunosorbent Allergen CHIP (ImmunoCAP-ISAC), and marks the transition to the molecular diagnosis of allergy. In the present study, we aimed to determine the IgE response profile to purified, natural or recombinant allergens in adult patients with AD and to compare the value of component allergy diagnosis with existing diagnostic methods. Thirty-nine adult patients with AD were selected from among those attending the Outpatient Clinic of Allergy and Dermatology at HCFMRP-USP. The age of the patients ranged from 14-66 years (mean ± SD 34,3 ± 2.1 years), 64% women. The mean duration of illness was 16 years. SCORAD mean was 36,6 (2- 90). Geometric mean (GM) of total IgE was 1,963 kU / L (24-63,000 kU / L). Mite allergens were dominant, with sensitization to Der p1 and Der f1, Der p2 and Der f2 in 82% and 85% of the patients, with GM 27,6; 50; 39.2; 45.4 ISU-E respectively, followed by cat (38%), dog (36%) and grass pollen (36%). IgE for cockroach, fungus, tree pollen, latex and insect venom allergens was found in less than 20% of patients at low levels. Sensitization to nuts (33%) and shrimp (31%) were the most prevalent allergens among foods, while IgE reactivity to milk and egg was present in 10% or less of the patients, with low levels of IgE antibodies in most cases. Only 6 patients (15.4%) presented IgE for the pan-allergen tropomyosin, and 3/39 (7.7%) were negative for all 112 allergen components tested on ImmunoCAP-ISAC, with total IgE levels of 24; 38,7 and 156 kU / L. We concluded that the IgE sensitization profile among adult patients with atopic dermatitis differs from that among patients with respiratory allergy, presenting less sensitization to cockroaches and to the pan-allergen tropomyosin, andalso differs from the profile present in children with AD, with a predominance of IgE sensitization to nuts and shrimp, and low sensitization rate to cow\'s milk and egg allergens.

Background: Staphylococcus aureus is an important trigger of flares in atopic dermatitis. The exact mechanisms by which S. aureus induces inflammatory responses and cell death in the skin epithelium is unclear. The aim of this thesis was to elucidate the cellular and molecular mechanisms by which S. aureus induces it's pathogenic effects on keratinocyte and fibroblast cell lines. Methods: Human keratinocytes (HEKa), and mouse embryonic fibroblasts (MEF) from the NC/Nga dermatitis prone mouse strain were used to investigate the induction of Th2-promoting cytokines (IL-33 and TSLP) and cell death by S. aureus. Cytokine levels were measured by ELISA and cytotoxicity by flow cytometry. Results: Live, but not killed S. aureus or other staphylococcal species, induced release of Th2-promoting cytokines (IL-33 and TSLP) and necrosis in both human and mouse cell lines. Cytokines were not induced by TLR2 ligands, and anti-TLR2 antibodies did not inhibit release, suggesting that the TLR2 pathway was not involved. By contrast, the release of cytokines was induced by a secreted, heat-labile factor/s and could be blocked by protease and PAR2 inhibitors, suggesting that the protease-PAR2 pathway was critical. NC/Nga mouse fibroblasts that lacked soluble IL-33 (sST2) receptor were more sensitive to the effects of S. aureus than control MEF. Conclusions: S. aureus is unique amongst staphylococcal species in it's ability to induce an inflammatory response and cytotoxicity in human keratinocytes and mouse fibroblasts. The protease-PAR2 pathway is critical to this bioactivity. Development of specific inhibitors of this pathway may provide novel therapies for treating S. aureus -induced eczema flares.