<p>Hereditary ichthyosis is a collective name for many dry and scaly skin disorders ranging in frequency from common to very rare. The main groups are autosomal recessive lamellar ichthyosis, autosomal dominant epidermolytic hyperkeratosis and ichthyosis vulgaris, and x-linked recessive ichthyosis. Anhidrosis, ectropion and keratodermia are common symptoms, especially in lamellar ichthyosis, which is often caused by mutations in the transglutaminase 1 (TGM1) gene. The aim of this work was to study patients with different types of ichthyosis regarding (i) the patho-aetiology (TGM1 and electron microscopy [EM] analysis), (ii) skin signs and symptoms (clinical score and subjective measure of disease activity), (iii) quality of life (questionnaires DLQI, SF-36 and NHP and face-to-face interviews) and (iv) a search for new ways of topical treatment. Patients from Sweden and Estonia with autosomal recessive congenital ichthyosis (n=83) had a broader clinical spectrum than anticipated, but a majority carried TGM1 mutations. Based on DNA analysis and clinical examinations the patients were classified into three groups, which could be further subdivided after EM analysis. Our studies indicate that patients with ichthyosis have reduced quality of life as reflected by DLQI and by some domains of SF-36, by NHP and the interviews. All the interviewees reported that their skin disease had affected them negatively to varying degrees during their entire lives and that the most problematic period was childhood. All patients with ichthyosis use topical therapy. In a double-blind study creams containing either 5% urea or 20% propylene glycol were found inferior to a cream formulation containing lactic acid 5% and propylene glycol 20% both regarding clinical improvement and thinning of the skin barrier. Improved topical therapy may reduce the need of more toxic, oral drugs. Future studies should elucidate whether this increases the quality of life of ichthyosis patients, especially if combined with more detailed information about the aetiology and inheritance of the diseases.</p>

Hereditary ichthyosis is a collective name for many dry and scaly skin disorders ranging in frequency from common to very rare. The main groups are autosomal recessive lamellar ichthyosis, autosomal dominant epidermolytic hyperkeratosis and ichthyosis vulgaris, and x-linked recessive ichthyosis. Anhidrosis, ectropion and keratodermia are common symptoms, especially in lamellar ichthyosis, which is often caused by mutations in the transglutaminase 1 (TGM1) gene. The aim of this work was to study patients with different types of ichthyosis regarding (i) the patho-aetiology (TGM1 and electron microscopy [EM] analysis), (ii) skin signs and symptoms (clinical score and subjective measure of disease activity), (iii) quality of life (questionnaires DLQI, SF-36 and NHP and face-to-face interviews) and (iv) a search for new ways of topical treatment. Patients from Sweden and Estonia with autosomal recessive congenital ichthyosis (n=83) had a broader clinical spectrum than anticipated, but a majority carried TGM1 mutations. Based on DNA analysis and clinical examinations the patients were classified into three groups, which could be further subdivided after EM analysis. Our studies indicate that patients with ichthyosis have reduced quality of life as reflected by DLQI and by some domains of SF-36, by NHP and the interviews. All the interviewees reported that their skin disease had affected them negatively to varying degrees during their entire lives and that the most problematic period was childhood. All patients with ichthyosis use topical therapy. In a double-blind study creams containing either 5% urea or 20% propylene glycol were found inferior to a cream formulation containing lactic acid 5% and propylene glycol 20% both regarding clinical improvement and thinning of the skin barrier. Improved topical therapy may reduce the need of more toxic, oral drugs. Future studies should elucidate whether this increases the quality of life of ichthyosis patients, especially if combined with more detailed information about the aetiology and inheritance of the diseases.

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No. of bitstreams: 1 Tese - Claudia Dantas Comandolli Wyrepkowski.pdf: 5493553 bytes, checksum: 2facc8046c9c0fe6bc72cf077d977196 (MD5) Previous issue date: 2016-09-24 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / The American Cutaneous Leishmaniasis (ACL) is a protozoonosis caused by parasites of the genus Leishmania, which is transmitted by the bite of the sand fly insect. The LTA, patients can present cutaneous form, with localized skin lesions that may progress to chronic lesions. The treatment of choice is pentavalent antimony and drugs of second choice are pentamidine and amphotericin B, but all have side effects and require parenteral administration. Natural products are presented as an alternative for the treatment of many diseases, and there are studies that demonstrate potential against Leishmania. In the Amazon region, Libidibia ferrea (Mart. Ex Tul) LP Queiroz stands out for its economic and pharmacological importance, due to anti-inflammatory and antimicrobial reported. This study aimed to evaluate the activity of extracts and fractions of L. ferrea against promastigotes and amastigotes of L. (L.) amazonensis and L. (V.) guyanensis also conducting its chemical characterization, and study the effect in topical formulations containing pentamidine, extracts and most promising fractions. And hexane extracts were prepared methanolic the epicarp, leaves and branches of L. ferrea, who underwent in vitro bioassays against promastigotes and amastigotes of L. (L.) amazonensis and L. (V) guyanensis, the counting method direct. The most active extracts were evaluated in vitro for cytotoxic activity in J774 macrophage lineage. The extracts and active fractions were analyzed by TLC assay, the quantized flavonoid and phenolic content, and analyzed by NMR and HPLC. The in vivo assays employed as an animal model hamsters infected with L. (L.) amazonensis. They were prepared and evaluated by topical application on lesions initially hydrating and anhydrous emulsions containing pentamidine isethionate (10%). The results of the bioassays showed that the methanolic extract from epicarp (EpMeOH) showed a higher activity against species of Leishmania, with EC50 15.01 µg/ml against L (L.) amazonensis. This extract had the highest concentration of phenolic (5.14%). A 2-DCM fraction also demonstrated activity with EC50 12.77 µg/mL against L. (L.) amazonensis. No cytotoxic activity was observed in macrophages for both samples. The moisturizing emulsions containing pentamidine were most effective, although viable parasites were detected and all groups after eight days of treatment. These moisturizing emulsions were used as a base for creams with addition of 2.5% of EpMeOH extract and evaluated in two areas with skin lesions - snout and paw. The groups treated with cream EpMeOH extract (2.5%) and placebo showed differences in lesion volume compared to the control group after 20 days of treatment, but showed the presence of parasites, with no statistical difference between the control group. These results suggest that the treatment time and extract concentration to be increased beyond the proportion of use of EpMeOH extract in a more hydrophilic formulation. The hydrogel EpMeOH was then evaluated for lesions on the snout of hamsters as well as cream Fr2. After 40 days of treatment, the groups treated with these topical formulations showed significant differences between the control group and between their respective placebos, as volume, clinical aspect of the lesion, and as the parasitic quantification. Minor inflammatory infiltrate was observed in the hydrogel EpMeOH. This study identified extracts promising in the treatment of experimental cutaneous leishmaniasis and supports pointing new opportunities and innovation serving as a basis for herbal formulations to treat this neglected disease. / A Leishmaniose Tegumentar Americana (LTA) é uma protozoonose causada por parasitas do gênero Leishmania, que são transmitidos ao homem pela picada do inseto flebotomíneo. Na LTA, os pacientes podem apresentar a forma cutânea, com lesões localizadas na pele que podem evoluir para lesões crônicas. O tratamento de primeira escolha são antimoniais pentavalentes e os fármacos de segunda escolha são pentamidina e anfotericina B, porém todos apresentam efeitos colaterais e requerem administração parenteral. Os produtos naturais apresentam-se como uma alternativa para o tratamento de inúmeras enfermidades, e há trabalhos que demonstram potencial contra Leishmania. Na região Amazônica, a espécie Libidibia ferrea (Mart. ex Tul) LP Queiroz destaca-se pela sua importância econômica e farmacológica, devido à atividade anti-inflamatória e antimicrobiana relatada. O presente trabalho teve por objetivo avaliar a atividade de extratos e frações de L. ferrea contra promastigotas e amastigotas de L. (L.) amazonensis e L. (V.) guyanensis, realizando também a sua caracterização química, e estudar o efeito in vivo de formulações tópicas contendo pentamidina, os extratos e frações mais promissoras. Foram preparados extratos hexânicos e metanólicos do epicarpo, folhas e galhos de L. ferrea, os quais foram submetidos a bioensaios in vitro contra promastigotas e amastigotas de L. (L.) amazonensis e L. (V.) guyanensis, pelo método da contagem direta. Os extratos mais ativos foram avaliados in vitro quanto a atividade citotóxica em macrófagos de linhagem J774. Os extratos e frações ativos foram analisados por CCDC, quantificados o teor de fenólicos e flavonoides, e analisados por RMN e CLAE. Os ensaios in vivo empregaram hamsters como modelo animal, infectados com L. (L.) amazonensis. Foram preparadas e avaliadas por aplicação tópica nas lesões, inicialmente, emulsões hidratantes e anidras contendo isetionato de pentamidina (10%). Os resultados dos bioensaios demostraram que o extrato metanólico de epicarpo (EpMeOH) apresentou maior atividade frente as espécies de Leishmania, com EC50 de 15,01 µg/mL contra L (L.) amazonensis. Este extrato apresentou a maior concentração de fenólicos (5,14%). A fração 2-DCM demonstrou também atividade, com EC50 12,77 µg/mL contra L. (L.) amazonensis. Não foi observada atividade citotóxica em macrófagos para ambas as amostras. As emulsões hidratantes contendo pentamidina foram mais eficientes, embora fossem detectados parasitas viáveis em todos os grupos após oito dias de tratamento. Estas emulsões hidratantes foram utilizadas como base para cremes com adição de 2,5% do extrato EpMeOH e avaliados em duas áreas com lesões cutâneas - focinho e pata. Os grupos tratados com creme com extrato EpMeOH (2,5%) e o placebo mostraram diferenças quanto ao volume da lesão quando comparados ao grupo controle após 20 dias de tratamento, mas mostraram a presença de parasitas, sem diferença estatística entre o grupo controle. Estes resultados sugerem que o tempo de tratamento e a concentração do extrato devem ser aumentadas, além da proporção do uso do extrato EpMeOH em uma formulação mais hidrofílica. O hidrogel EpMeOH foi avaliado em lesões no focinho de hamsters, assim como emulsão Fr2. Após 40 dias de tratamento, os grupos tratados com estas formulações tópicas mostraram diferenças estatísticas entre o grupo controle e também entre seus respectivos placebos, quanto ao volume, aspecto clínico da lesão, e quanto à quantificação parasitária. Menor infiltrado inflamatório foi observado no grupo hidrogel EpMeOH. Este estudo permitiu identificar o extratos promissores no tratamento da leishmaniose tegumentar experimental e corrobora apontando novas oportunidades e inovação servindo como base para formulações de fitoterápicos para o tratamento desta doença negligenciada.