Cutaneous melanoma is associated with significant morbidity and mortality representing the most significant cutaneous malignancy. As it is known that early diagnosis and treatment are the most efficient approaches to cure cutaneous melanoma, an improved understanding of the molecular pathogenesis of melanoma and exploration of more reliable molecular biomarkers are particularly essential. Two different types of molecular biomarker for melanoma have been investigated in this thesis. microRNAs (miRNAs) are single-stranded RNA molecules of 20-23 nucleotides in length that are found in both animal and plant cells. miRNAs are involved in the RNA interference (RNAi) machinery to regulate gene expression posttranscriptionally. miRNAs have important roles in cancer: by controlling the expression level of their target genes they can affect cell signalling pathways and have been shown to have both prognostic and therapeutic potential. Importantly for melanoma research, reproducible miRNA expression profiles from formalin-fixed paraffin-embedded (FFPE) tissues can be obtained that are comparable to those from fresh-frozen samples. The aims of the miRNA project were: first, to identify a melanoma-specific miRNA expression profile; secondly, to investigate roles of some of the melanoma-specific miRNAs identified in melanomagenesis. Using miRNA microarray on FFPE samples, I obtained a melanoma-specific miRNA expression profile. 9 of these differentially expressed miRNAs between benign naevi and melanomas (7 downregulated, 2 upregulated in malignancies) were verified by qRT-PCR and the functions of four of these miRNAs were studied. Ectopic overexpression of miR- 200c and miR-205 in A375 melanoma cells inhibited colony forming ability in methylcellulose, an in vitro surrogate assay for tumourigenicity. Moreover, elevation of miR-200c resulted in increased expression levels of E-cadherin through negative regulation of the zinc finger E-box-binding homeobox 2 (ZEB2) gene. Ectopic overexpression of miR-211 in A375 melanoma cells repressed both colony formation in methylcellulose and migratory ability in matrigel, an in vitro surrogate assay for invasiveness. These findings indicate that miR-200c, miR-205 and miR-211 act as tumour suppressors in melanomagenesis. The second biomarker investigated, mutated BRAF, has been seen in 50-70% of spontaneous cutaneous melanoma. The commonest mutation in melanoma is a glutamic acid for valine substitution at position 600 (V600E). Oncogenic BRAF controls many aspects of melanoma cell biology. The aim of this part of the work was: firstly, to study BRAF V600E mutation status in our melanoma tissue microarray (TMA) panel; secondly, to correlate this mutation to various clinicopathological features and evaluate its prognostic value through statistical analyses. BRAF V600E mutations were seen in 20% of the primary and 69% of the metastatic melanomas, respectively. More BRAF V600E mutations were seen in males relative to females. The mutation was also related to cell pigmentation, but not to age, ulceration or solar elastosis. Melanoma patients with the BRAF V600E mutation relapse earlier than patients without this mutation. However, no significant association between the BRAF V600E mutation and overall survival and melanoma specific survival was found.

Les lymphomes T cutanés constituent un ensemble hétérogène de lymphomes non-hodkinniens. Les lymphomes T cutanés sont définis par une prolifération clonale de lymphocytes T malins et de cellules NK (natural killers) de localisation cutanée. Dans l'Union Européenne, l'indication du bexarotène par voie orale est le traitement des manifestations cutanées des lymphomes cutanés T épidermotropes (LCT), au stade avancé (ou dès un stade précoce aux États Unis). L'objectif de ce travail a été d'étudier les effets indésirables du bexarotène chez les patients atteints de lymphome cutané. Nous nous sommes plus particulièrement intéressés aux interactions du bexarotène avec le système de la coagulation et avec le métabolisme lipidique.Dans la première partie de nos travaux, nous avons étudié l'origine de la coagulopathie induite par le bexarotène. Nous avons montré que le bexarotène inhibe les facteurs IX et X de la coagulation, ce qui provoque le prolongement du temps de coagulation ; ces effets pourraient être à l'origine de la coagulopathie observée chez les patients traités au bexarotène.Dans la deuxième partie, nous avons montré que le bexarotène interagit également avec le métabolisme lipidique par l'activation du CETP et par l'inhibition de la lipoprotéine lipase, ce qui provoque une chlolésterémie et une triglyéridémie.Enfin, nous avons présenté les résultats préliminaires de l'essai clinique organisé au niveau de la région ouest de la France sur les effets indésirables du bexarotène.Les résultats de ce travail mettent en évidence l'importance de la structure chimique du bexarotène (effets de charges) dans la neutralisation des facteurs de coagulation d'une part et dans le mécanisme d'interaction avec le métabolisme lipidique (CETP et lipoprotéine lipase) d'autre part. La compréhension des mécanismes moléculaires conduisant à une coagulopathie et à la dyslipidémie représente un enjeu majeur pour prévenir la toxicité du bexarotène et pour permettre une meilleure prise en charge du patient. / The cutaneous T cell lymphomas are a heterogeneous group of non-lymphoma hodkinniens. The cutaneous T cell lymphomas are defined by a clonal proliferation of malignant T cells and NK (natural killer) cells from skin location. In the European Union, the indication of oral bexarotene is the treatment of cutaneous manifestations of cutaneous T-cell lymphomas epidermotropic (LCT) to advanced (or at an early stage in the United States). The objective of this work was to study the adverse effects of bexarotene in patients with cutaneous lymphoma. We are particularly interested in the interactions of bexarotene with the coagulation system and lipid metabolism.In the first part of our work, we studied the origin of the coagulopathy induced by bexarotene. We have shown that bexarotene inhibited factors IX and X of the coagulation causing the extension of clotting time and coagulopathy that was observed in bexarotene-treated patients. In a second part, we showed that Bexarotene induces in vitro cholesteryl ester transfer protein activity, and suppress lipoprotein lipase activity in human plasma.Finally, we presented the preliminary results of the clinical trial conducted at the western region of France on the adverse effects of bexarotene.The results of this study highlight the importance of the chemical structure of bexarotene (charge effects) in the neutralization of coagulation factors mechanisms on the one hand and in its interaction with lipid metabolism (CETP and lipoprotein lipase) on the other hand. Understanding the molecular mechanisms leading to coagulopathy and dyslipidemia is a major issue to prevent the toxicity of bexarotene and to elicit better management of the bexarotene-treated patient.

Master of Science / Department of Kinesiology / Brett J. Wong / Beetroot juice (BRJ) has been shown to increase NO-dependent dilation through both NOS-dependent and NOS-independent pathways. We hypothesized BRJ supplementation would augment reflex cutaneous active vasodilation. Subjects were equipped with two microdialysis fibers on the forearm and randomly assigned as control (Ringer’s) or NOS inhibition (20mM L-NAME). Whole-body heating was achieved via water-perfused suits to raise core temperature (Tc; ingestible telemetric pill) 0.8°C. Maximal cutaneous vasodilation was reached by administering 54mM SNP and local heating to 43°C. Skin blood flow was measured via laser-Doppler flowmetry and mean arterial pressure determined; cutaneous vascular conductance (CVC) was calculated and expressed as %CVCmax. Subjects underwent heat stress pre- and post-nitrate supplementation (3 days of BRJ: 5mM, 0.45g nitrates per day). BRJ increased the plateau CVC at control (pre: 57 ± 3 vs. post: 80 ± 5 %CVCmax) and L-NAME (pre: 36 ± 3 vs. post: 52 ± 6 %CVCmax; p<0.05 for all conditions) sites. The %NO contribution increased from pre- to post-BRJ (pre: 44 ± 5 %CVCmax vs. post: 64 ± 6 %CVCmax; p<0.05). These data suggest that BRJ augments the NOS-dependent and NOS-independent component of reflex cutaneous vasodilation.

Master of Science / Department of Kinesiology / B.J. Wong / Recent data suggests nNOS mediates the NO-component of reflex cutaneous vasodilation with passive heat stress. Our hypothesis was nNOS, but not eNOS, inhibition would attenuate reflex cutaneous vasodilation during dynamic exercise. Protocol 1: subjects performed a VO[subscript]2 peak test on a supine cycle ergometer. Protocol 2: with experimental arm at heart level subjects cycled in supine posture at 60% VO[subscript]2 peak to raise core temperature (Tc) 0.8-1.0°C (35-45 min). In protocol 2 subjects were equipped with 4 microdialysis fibers on the forearm and each randomly assigned as: 1) lactated Ringer’s (control); 2) 5mM NPLA (nNOS inhibition); 3) 10mM L-NIO (eNOS inhibition); and 4) 20mM L-NAME (non- selective NOS inhibition). At the end of protocol 2 all sites were locally heated to 43°C and infused with SNP to elicit maximal dilation. Mean arterial pressure (MAP), skin blood flow via laser- Doppler flowmetry (LDF), and Tc via ingestible telemetric pill were measured; cutaneous vascular conductance (CVC) was calculated as LDF/MAP and normalized to maximum. In protocol 2 there was no significant difference between control (62±5 %CVCmax) and NPLA (61±6 %CVCmax). L-NIO (38±4 %CVCmax) and L-NAME (41±7 %CVCmax) significantly attenuated CVC compared to control and NPLA (p<0.001 all conditions). There was no difference between L-NIO and L- NAME. We conclude eNOS, not nNOS, contributes to reflex cutaneous vasodilation during dynamic exercise.

Orientador: Renée Laufer Amorim / Banca: Noeme Sousa Rocha / Banca: Antonio Carlos Alessi / Resumo: Os mastócitos são células provenientes do precursor CD34 da medula óssea, o mastocitoma (MCT) pertence ao grupo das neoplasias de células redondas e possui etiologia desconhecida, é a neoplasia cutânea mais comum em cães, sem predileção por raça, sexo ou idade. Seu comportamento biológico é bastante agressivo e de alta frequência. A graduação histopatológia proposta Patnaik et al., (1984), é o critério mais utilizado para a graduação histopatológica da neoplasia sendo o grau 1 uma neoplasia bem diferenciada, o grau 2 moderadamente diferenciada e o grau 3 pouco diferenciada ou anaplásica. O presente trabalho teve como objetivos a avaliação do ínidice proliferativo e apoptotico dos MCT em lâmina de Microarranjo de tecido (TMA) utilizando a técnica de imunoistoquiímica e TUNEL. Para tal utilizou-se 166 casos de MCT compondo um arranjo em matriz de amostra tecidual, ou tissue microarray (TMA) com 190 amostras, sendo avaliado o padrão de marcação da proteína Kit, a proliferação celular com o anticorpo primário Ki67 e apoptose com o anticorpo primário caspase-3 clivada e pela técnica da Terminal Deoxynucleotidyl Transferase Mediated dUTP Nick end Labeling Assey (TUNEL). Quando os casos foram avaliados quanto à imunoexpressão de c-KIT, os tumores (únicos e múltiplos) com expressão citoplasmática da proteína KIT tiveram uma taxa proliferativa maior e uma menor taxa apoptótica quando comparados aos com expressão membranosa. Os MCTs grau 3 tiveram um maior índice proliferativo (imunoexpressão de Ki67) quando comparados aos tumores de graus 1 e 2. Não foi observada correlação entre a imunomarcação do anticorpo primário caspase-3 clivada e a técnica de TUNEL, sendo que a técnica de TUNEL se mostrou mais eficaz na avaliação da apoptose que a imunomarcação para caspase-3 clivada / Abstract: Mast cells are from precursor CD34 cells bone marrow, the mast cell tumor (MCT) belongs to the group of round cell malignancies and has unknown etiology, is the most common skin cancer in dogs without distinction of race, sex or age. Its biological behavior is very aggressive and high frequency. The histopathological grading proposal from Patnaik et al., (1984), is the most commonly used criterion for the histopathological grade of the tumor is a tumor grade 1 are well-differentiated, grade 2 are moderately differentiated and grade 3 are poorly differentiated or anaplastic. This study aimed to evaluate the proliferative and apoptotic index of MCT in blade tissue microarray (TMA) using the TUNEL technique and immunohistochemistry. To this end we used 166 cases of MCT composing an tissue microarray (TMA) with 190 samples, and evaluated the pattern of protein labeling kit, cell proliferation, with the primary antibody Ki67 and apoptosis with primary antibody and cleaved caspase-3 by the technique of terminal deoxynucleotidyl transferase dUTP Nick End Labeling Mediated Assey (TUNEL). When the cases were evaluated for immunoexpression of c-KIT, tumors (single and multiple) with cytoplasmic KIT protein expression had a higher proliferative rate and a lower apoptotic rate when compared to those with membranous expression. Grade 3 MCTs had a greater proliferative index (Ki67 immunostaining) when compared with tumor grades 1 and 2. No correlation was observed between the immunostaining of primary antibody caspase-3 cleavage and TUNEL technique, and TUNEL technique was more effective in assessing apoptosis that immunostaining for cleaved caspase-3 / Mestre

A leishmaniose é uma protozoonose que apresenta diferentes manifestações clínicas, associadas a fatores relacionados ao parasita, a resposta imune do hospedeiro e epidemiologia da doença. Esses fatores podem influenciar no desfecho da doença e na efetividade da terapia antileishmania que até hoje se baseia no uso de fármacos de alto custo, alta toxicidade e que podem causar diversos efeitos colaterais. A pentamidina é um fármaco de segunda escolha no tratamento da leishmaniose utilizada em casos específicos de Leishmaniose Tegumentar nas regiões da América do Sul. Por causa dessas limitações, novos fármacos ou novos regimes terapêuticos vêm se mostrando cada vez mais necessários no tratamento contra as leishmanioses devido ao surgimento de falhas terapêutica e resistência aos fármacos existentes. Linhagens de L. amazonensis resistentes a PMD superexpressoras ou não do gene PRP1 tiveram o fenótipo de resistência revertido após a associação de pentamidina com verapamil. Deste modo, propomos verificar o efeito da associação de pentamidina com verapamil, anfotericina B e miltefosina contra parasitos de L. (L.) amazonensis suscetíveis (La M2269) e resistentes à pentamidina (La PRP1 e La PEN 0,5). O efeito das interações entre a pentamidina e os demais fármacos foi avaliado pelo método modificado de isobolograma e a natureza da interação foi classificada através da análise dos valores de Concentração Inibitória Fracionada (CIF). As associações de pentamidina com o verapamil, anfotericina B e miltefosina contra promastigotas de La M2269 foram classificadas como indiferente a partir da análise dos valores de CIF. Interações indiferentes também foram observadas contra promastigotas de La PRP1 e La PEN 0,5 quando tratadas com as mesmas associações. Macrófagos foram infectados com parasitos de La M2269 e tratados com diferentes concentrações de PMD. Nossos resultados sugerem que altas concentrações de PMD (17,5uM) foram tóxicas contra macrófagos e apresentaram baixa atividade quando utilizada contra amastigotas de La M2269 em até 72 horas de incubação. Observamos que ao prorrogar a incubação para 120 horas, concentrações inferiores a 1uM de PMD reduziram a taxa de infecção para aproximadamente 30%. Já ao analisarmos o efeito das combinações de pentamidina com o verapamil observamos baixa atividades desses fármacos quando combinado, indicando possivelmente uma interação indiferente/ antagônica. A associação de pentamidina com anfotericina B contra amastigotas de La M2269 foi avaliada pelo método de isobolograma e a natureza da interação foi classificada como indiferente por apresentar um valor de CIF = 0,65. / Leishmaniasis is a protozoan disease that presents different clinical manifestations, associated to factors related to the parasite, the host immune response and the epidemiology of the disease. These factors may influence the outcome of the disease and the effectiveness of antileishmania therapy, which is still based on the use of high-cost, high toxicity drugs that can cause several side effects. Pentamidine is a second-choice drug in the treatment of leishmaniasis used in specific cases of cutaneous leishmaniasis in the South American regions. Because of these limitations, new drugs or new therapeutic regimens are proving increasingly necessary in the treatment of leishmaniasis due to the emergence of therapeutic failures and resistance to existing drugs. L. (L.) amazonensis strains resistant to PMD overexpressors or not of the PRP1 gene had the resistance phenotype reverted after the association of pentamidine with verapamil. Thus, we propose to verify the effect of the association of pentamidine with verapamil, amphotericin B and miltefosine against parasites of L. (L.) amazonensis susceptible (La M2269) and resistant to pentamidine (La PRP1 and La PEN 0,5). The effect of the interactions between pentamidine and other drugs was evaluated by the modified isobologram method and the nature of the interaction was classified through the analysis of the Fractional Inhibitory Concentration (FIC) values. The associations of pentamidine with verapamil, amphotericin B and miltefosine against La M2269 promastigotes were classified as indifferent from the analysis of FIC values. Indifferent interactions were also observed against La PRP1 and La PEN 0.5 promastigotes when treated with the same associations. Macrophages were infected with parasites of La M2269 and treated with different concentrations of PMD. Our results suggest that high concentrations of PMD (17.5uM) were toxic against macrophages and showed low activity when used against La M2269 amastigotes within 72 hours of incubation. We observed that by extending the incubation to 120 hours, concentrations below 1uM PMD reduced the infection rate to approximately 30%. When analyzing the effect of the combinations of the pentamidine and the verapamil we observed low activities of these drugs when combined, possibly indicating an indifferent / antagonistic interaction. The association of pentamidine with amphotericin B against amastigotes of La M2269 was evaluated by the isobologram method and the nature of the interaction was classified as indifferent because it presented a FIC = 0.65.

Orientador: Marcos Antônio Correa / Resumo: A busca por ativos naturais que apresentem mais de uma função em uma única formulação é pautada em um novo conceito de praticidade e economia, estando em acordo com a visão crítica do consumidor moderno. Neste contexto, a presença dos compostos fenólicos em plantas tem sido estudada por estes apresentarem diversas atividades farmacológicas e cosméticas. Os ácidos hidroxibenzoicos fazem parte desse grupo de compostos orgânicos, sendo o ácido gálico um de seus representantes, conhecido principalmente por sua atividade antioxidante. Um grande número de formulações tópicas contendo antioxidantes tem sido lançado nos últimos anos. O objetivo deste trabalho foi estudar a eficácia do ácido gálico para uso cosmético. Para escolha dos solventes utilizados no estudo levou-se em consideração o desenvolvimento sustentável da pesquisa, ou seja a utilização de solventes verdes. Para a avaliação da atividade antioxidante foram utilizados dois métodos diferentes o DPPH e o ABTS. Também foi realizada a avaliação das atividades despigmentantes, antimicrobiana, citotóxica, bem como estudos da formulação desenvolvida, como: estabilidade, liberação, permeação e retenção, através de experimentos in vitro. Os resultados demonstram que o ácido gálico é um potente antioxidante, suas ações despigmentante e antimicrobiana também foram comprovadas. Pelos estudos de citotoxidade in vitro pode-se constatar que a porcentagem de ácido gálico utilizada na emulsão desenvolvida é segura. A validação da metodol... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The search for natural assets that present more than one function in a single formulation is based on a new concept of practicality and economy, being in agreement with the critical view of the modern consumer. In this context, the presence of phenolic compounds in plants has been studied because they present several pharmacological and cosmetic activities. Hydroxybenzoic acids are part of this group of organic compounds, and gallic acid is one of its representatives, known mainly for its antioxidant activity. A large number of topical formulations containing antioxidants have been launched in recent years. The objective of this work was to study the efficacy of gallic acid for cosmetic use. To choose the solvents used in the study took into consideration the sustainable development of the research, ie the use of green solvents. For the evaluation of the antioxidant activity, two different methods were used: DPPH and ABTS. The evaluation of depigmenting activities, antimicrobial, cytotoxic, as well as studies of the developed formulation, such as: stability, release, permeation and retention, were carried out through in vitro experiments. The results demonstrate that gallic acid is a potent antioxidant, its depigmenting and antimicrobial actions have also been proven. By in vitro cytotoxicity studies it can be verified that the percentage of gallic acid used in the developed emulsion is safe. The validation of the methodology by HPLC ensured all parameters necessary for the s... (Complete abstract click electronic access below) / Mestre

A espécie Leishmania (Viannia) shawi foi caracterizada recentemente pelo grupo de Lainson. Estudos recentes indicam o importante papel médico epidemiológico deste parasito no Brasil. Portanto, os objetivos do presente estudo foram caracterizar o modelo experimental murino desta infecção, purificar antígenos protéicos e avaliar seus graus de proteção após desafio. Para caracterizar o modelo murino de infecção, camundongos das linhagens BALB/c e C57BL/6 foram infectados na pata com formas promastigotas e os achados histopatológicos e imunológicos foram avaliados durante a evolução da infecção. Para os estudos de imunização foram utilizados 10 diferentes antígenos: três secretados/excretados pelas formas promastigotas de L. (V.) shawi, dois intracelulares solúveis das formas amastigotas (AgAma) e promastigotas (AgPro), e cinco frações protéicas purificadas a partir do antígeno intracelular solúvel das formas promastigotas. Estes antígenos foram utilizados para imunizar camundongos da linhagem BALB/c duas vezes, subcutaneamente no dorso. Após uma semana da última imunização, os animias foram desafiados com formas promastigotas. O desenvolvimento das lesões nos animais foram acompanhadas por seis ou oito semanas pós desafio (PD), quando os animais foram sacrificados para análise da carga parasitária e dos aspectos relacionados às respostas imune celular e humoral. Camundongos da linhagem BALB/c foram altamente susceptíveis à infecção, uma vez que as mudanças histopatológicas e da imunidade humoral foram mais pronunciadas nos camundongos BALB/c que em C57BL/6. Os antígenos secretados/excretados de baixa massa molecular induziram alta taxa de proteção em comparação aos animais não imunizados, já os antígenos secretados/excretados de média massa molecular protegeram intermediariamente os animais, possivelmente pela alta expressão de IFN-g e IL-4 nos linfócitos T CD8+. AgAma e AgPro tiveram uma resposta antagônica nos animais, pois o AgAma suprimiu a produção de IFN-g e IL-12, contudo houve maior produção de TGF-b, facilitando o aumento do parasitismo na pele e em linfonodos. A despeito da detecção de TGF-b nos animais imunizados com AgPro, houve um balanço entre a produção de citocinas, com a participação de IL-12 e IFN-g, que levou a um controle do parasitismo em pele. Através da purificação do AgPro foi visto que os antígenos F1 e F5 protegeram os animais da infecção na pele após desafio, e ainda F1 também protegeu os linfonodos destes animais. Os antígenos F3 e F4 levaram a exacerbação das lesões de pele. A identificação, por espectrometria de massa, do antígeno F1 revelou a presença de 67 componentes, sendo que a maioria deles não possui identificação. Ainda, o antígeno F1 protegeu duradouramente os animais associado à estimulação de linfócitos T CD8+ de memória, contudo a presença de baixos números de parasitos pode ser o reflexo da alta produção de IL-10. Estes dados indicam que o antígeno F1 pode representar um importante candidato vacinal contra a Leishmaniose Tegumentar Americana / Leishmania (Viannia) shawi specie was recently characterized by Lainson group. Currently, studies indicate important medical and epidemiological role of this parasite in Brazil. Therefore, the aims of this study were to characterize the experimental murine model of this infection, purify proteic antigens and evaluate their protection degrees after challenge. To characterize the murine model of infection, BALB/c and C57BL/6 mice were infected in the footpad with promastigote forms, and the histopathological and immunological findings were evaluated during the evolution of infection. For the immunization studies, 10 different antigens were used, as follow: three released/excreted by promastigote forms of L. (V.) shawi; two intracellular soluble antigens from amastigote (AgAma) and promastigote forms (AgPro), and 5 proteic fractions purified from soluble intracellular antigens from promastigote forms. These antigens have been used to immunize BALB/c mice twice, subcutaneously in the rump. After 1 week of last immunization, the animals were challenged. The lesion developments in animals were followed during either six or eight weeks post-challenge (PC), when the animals were sacrificed to evaluate the parasite load and aspects of cellular and humoral immune responses. BALB/c mice were the most susceptible to L. (V.) shawi infection, since the histopathological and humoral changes were higher in BALB/c than C57BL/6 mice. Secreted/excreted antigens of low molecular mass induced high protection rate compared to non-immunized mice, already mice immunized with secreted/released antigens of medium molecular mass showed mild protection, possibly caused by high expression of IFN-g and IL-4 by CD8+ T lymphocytes. AgAma and AgPro showed antagonic response in animals, since AgAma suppressed the IFN-g and IL-12 production, however high level of TGF-b has been detected, allowing the increasing of parasitism in the skin and lymph nodes. In spite of the detection of TGF-b in AgPro-immunized mice, there was a balance in the cytokines production, with the participation of IFN-g and IL-12, leading to parasite control in skin. Through the purification of AgPro, it was observed a protective effect of F1 and F5 antigens in the skin after challenge; in addition, F1 also protected the lymph nodes of BALB/c mice. Both F3 and F4 antigens exacerbated the skin infection. The identification, by mass spectrometry, revealed that F1 was composed by 67 components, and the majority has not been identified till now. Moreover, F1 induced long-lasting immunity in BALB/c mice, associated to generation of memory CD8+ T lymphocytes, however low parasitism could be the reflect of high production of IL-10. These data indicate which F1 antigen could be an important vaccine candidate against American Tegumentar Leishmaniasis

O lúpus eritematoso foi pouco estudado na infância, principalmente nas formas cutânea crônica, subaguda e bolhosa que são infreqüentes nessa faixa etária. Este trabalho tem por objetivo estudar as diversas formas de lúpus eritematoso em crianças. Para tanto, realizamos um estudo retrospectivo por meio da análise de prontuários de pacientes com lúpus eritematoso que teve início até os 16 anos de idade e que foram acompanhados na Divisão de Dermatologia do Hospital das Clínicas da Universidade de São Paulo entre 1991 e 2006. Os pacientes incluídos tiveram o diagnóstico feito pelo quadro clínico e confirmado pela histologia cutânea de uma lesão específica. Foram observados 48 pacientes: 33 com lesões discóides, dois com lúpus profundo, quatro com lúpus eritematoso cutâneo subagudo, quatro com lúpus eritematoso cutâneo agudo sem outras lesões específicas e cinco com lúpus eritematoso bolhoso. Analisadas as características clínicas, laboratoriais, histológicas e de imunofluorescência direta. Dezenove casos preencheram critérios para lúpus eritematoso sistêmico. Houve predomínio discreto no sexo feminino. Casos familiares de lúpus eritematoso estavam presentes em 10% dos pacientes. Vinte e quatro por cento das crianças com lesões discóides preencheram critérios para lúpus eritematoso sistêmico, sendo que 75% apresentavam lesões generalizadas. No lúpus eritematoso sistêmico, as manifestações mais freqüentes foram: cutânea, articular e renal. A glomerulonefrite proliferativa difusa foi a forma mais freqüente de nefrite. Foram observadas alterações hematológicas no limite inferior ou abaixo das taxas relatadas na literatura. Acometimento neurológico ocorreu em 68%. Acometimento cardíaco, pulmonar e do sistema digestivo foram pouco freqüentes. O FAN estava positivo em 95% dos casos e o anticorpo anticardiolipina estava presente em 21%. Dois dos quatro pacientes com lúpus eritematoso cutâneo subagudo preencheram critérios para lúpus eritematoso sistêmico e apresentaram convulsões e nefrite, sendo que um necessitou de transplante renal. Todos referiam fotossensibilidade e o anticorpo anti-Ro estava presente em três dos quatro casos. Todos os pacientes com lúpus eritematoso sistêmico bolhoso preencheram critérios para lúpus eritematoso sistêmico com acometimento renal e baixos níveis de complemento. Nenhum dos dois pacientes com lúpus profundo apresentou critérios para lúpus eritematoso sistêmico. Ambos apresentavam quadro disseminado com histologia típica. O quadro histológico nos demais casos também foi característico e a IgM foi o imunodepósito mais freqüente à imunofluorescência direta. A coloração de PAS (ácido periódico/reagente de Schiff) mostrou espessamento da membrana basal em 37 dos 40 casos estudados. Concluímos que o lúpus eritematoso da infância é semelhante ao do adulto, porém apresenta algumas peculiaridades como menor predomínio no sexo feminino, alta taxa de história familiar de lúpus eritematoso e maior associação com lúpus eritematoso sistêmico nas crianças com lesões discóides. Crianças com lúpus eritematoso cutâneo subagudo e critérios para lúpus eritematoso sistêmico tiveram acometimento sistêmico grave. O acometimento renal foi freqüente nas crianças com lúpus eritematoso sistêmico bolhoso / Lupus erythematosus was not well studied in childhood, especially in its chronic cutaneous, subacute and bullous forms that are uncommon at this age group. The present work aims to study the various forms of childhood lupus erythematosus. We retrospectively studied through medical records patients with lupus erythematosus whose disease had started until 16 years of age who were followed at Dermatology Division of Hospital das Clínicas of University of São Paulo between 1991 and 2006. The patients included had the diagnosis established on clinical grounds and confirmed by histological examination of a specific lesion. We found 48 patients: 33 with discoid lesions, two with lupus profundus, four with subacute cutaneous lupus erythematosus, four with acute cutaneous lupus erythematosus without other specific lesion and five with bullous lupus erythematosus. We analyzed the clinical, laboratorial, histological and direct immunofluorescence findings. Nineteen cases fulfilled the criteria for systemic lupus erythematosus. There was a slight female predominance. Familiar history of lupus erythematosus was present in 10% of cases. Twenty four percent of children with discoid lesions fulfilled the criteria for systemic lupus erythematosus and 75% showed disseminated lesions. The most frequent clinical manifestations in systemic lupus erythematosus were cutaneous, articular and renal. Diffuse proliferative glomerulonephritis was the most frequent form of nephritis. Hematological involvement was bellow or in the bottom of normal limits reported in the literature. Neuropsychiatric disease occurred in 68% of patients. Cardiac, pulmonary and gastrointestinal involvement was not frequent. ANA was positive in 95% of cases and. anticardiolipin antibodies were present in 21%. Two of four subacute cutaneous lupus erythematosus patients fulfilled the criteria for systemic lupus erythematosus and had convulsions and nephritis; one of them was submitted to renal transplantation. All of them referred photosensitivity and anti-Ro was present in three of four cases. All patients with bullous lupus erythematosus fulfilled the criteria for systemic lupus erythematosus and presented renal involvement and low complement levels. None of the two patients with lupus profundus fulfilled the criteria for systemic lupus erythematosus. Both had disseminated lesions and typical histology. Histological examination was also characteristic in the other cases and IgM was the most frequent immunodeposit at direct immunofluorescence. PAS staining showed thickening of basement membrane in 37 of 40 cases studied. To conclude, childhood lupus erythematosus is similar to adult lupus erythematosus, however shows some peculiarities as lower female predominance, high proportion of familiar history of lupus erythematosus and greater association with systemic lupus erythematosus in children with discoid lesions. Children with subacute cutaneous lupus erythematosus and criteria for systemic lupus erythematosus had severe systemic involvement. Renal disease was frequent in children with bullous systemic lupus erythematosus

Tem-se atribuído ao flebotomíneo o papel de transmissor das leishmanioses. Com relação a leishmaniose tegumentar americana, 23 espécies parecem estar incriminadas na transmissão. Contudo, somente em Psychodopygus trapidoi, Ps.ylephiletor, Ps. intermedius, Ps. welcomei, Ps. flaviscutellatus, Ps. panamensis, Ps. olmecus, Lutzomyia gomezi e Pinzomyia pessoai, foi possível o isolamento da Leishmania. O caráter zoonótico desta doença denota uma situação ainda complexa devido aos escassos conhecimentos sobre os hospedeiros, vetores e epidemiologia das diversas Leishmania. A presente revisão confirma que esta função é desempenhada por dípteros da subfamília Phlebotominae. / The function of transmitting Leishmaniasis is generally attributed to phlebotomine sandfly. In relation to American Cutaneous Leishmaniasis, 23 species appears to be involved in the transmission process. However, Leishmania organisms could be isolated from only Psychodopygus trapidoi, Ps. ylephiletor, Ps. intermedius, Ps. welcomei, Ps. flaviscutellatus, Ps. panamensis, Ps. olmecus, Lutzomyia gomezi and Pintomyia pessoai. The zoonotic charater of this disease is yet a complex situation due to lack of adequate knowledge about host, vector and epidemiology of various Leishmania. The present revision confirms that the function is perfomed by diptera of sub-family, Phlebotominae.