The American Visceral Leishmaniasis (AVL) is endemic in tropical and subtropical regions of Latin America. In Brazil, in recent years has been expanding and building up. Historically, the state of Ceara has been presented in several cities a high prevalence of the disease, however, the city of Fortaleza, escaping this epidemiological pattern always reported low incidence of the disease, however, from 2005, showed an increase in the number of cases and spatial distribution, showing an expanding epidemic of the disease. The aim of this study was to evaluate the spatial and temporal distribution of disease incidence in the period 1995 to 2008. We analyzed the cases of AVL reported in SINAN and SIM, considering only patients in Fortaleza, who met the criteria adopted by the Ministry of Health for confirmation of diagnosis and the site of infection. The cases were georeferenced and analyzed based on the indicators: incidence and mortality and the following variables: dependent - confirmed cases of the disease and independent - age, age group, sex, place of residence, clinical manifestation, clinical course and type of diagnosis. We used spatial analysis techniques to analyze the cases. During the study period were recorded human cases of AVL 1267, of which 50% were younger than 4 years and 74 deaths. The year 2007 was presented the highest incidence of the disease. The highest incidence of cases were in males (64.3%). The average age of cases of AVL increased in epidemic period. The disease is clinically expressed differently in the epidemic phase, compared with the endemic phase. The risk of infection was higher in males than in females. Mortality was high only in the age group above 60 years. There was a significant increase in disease incidence in the elderly. Diagnostic tests showed high sensitivity and specificity. Over 70% of cases were confirmed through laboratory tests. We observed a decrease in cure rate and an increase in cases of co-infection with HIV. In the epidemic phase, spatial diffusion of disease was the kind of contagion. The diffusion process was linked to environmental factors and socioeconomic factors. We identified areas of high risk for the disease occurrence based on spatial analysis techniques. The study revealed the establishment of a new epidemiological pattern for kala-azar in Fortaleza, mainly characterized by the appearance of an emerging problem: the increase of cases of co-infection with HIV. Also revealed that environmental factors and socioeconomic factors were crucial in the processes of urbanization and expansion of the disease in the city. / A Leishmaniose Visceral Americana (LVA) à uma doenÃa endÃmica nas regiÃes tropicais e subtropicais da America Latina. No Brasil, nos Ãltimos anos, vem se expandindo e urbanizando-se. Historicamente, o Estado do Cearà tem apresentado, em diversos municÃpios, altas prevalÃncias da enfermidade; entretanto, a cidade de Fortaleza, fugindo desse padrÃo epidemiolÃgico sempre registrou baixa ocorrÃncia da doenÃa, no entanto, a partir de 2005, apresentou incremento significativo no nÃmero de casos e na distribuiÃÃo espacial, evidenciando uma expansÃo epidÃmica da doenÃa. O objetivo deste estudo foi avaliar a distribuiÃÃo espacial e temporal da incidÃncia da doenÃa, no perÃodo de 1995 a 2008. Foram analisados os casos de LVA notificados no SINAN e no SIM, considerando-se apenas os pacientes residentes em Fortaleza, que preenchiam os critÃrios adotados pelo MinistÃrio da SaÃde para confirmaÃÃo diagnÃstica e do local da infecÃÃo. Os casos foram georreferenciados e analisados com base nos indicadores: taxa de incidÃncia e letalidade e nas seguintes variÃveis: dependente â casos confirmados da doenÃa e independentes â sexo,faixa etÃria, grupo etÃrio, local de residÃncia, manifestaÃÃo clÃnica, evoluÃÃo clÃnica, tipo de diagnÃstico, tipo de tratament e escolaridade. Foram utilizadas tÃcnicas de anÃlise espacial para analisar os casos. No perÃodo do estudo foram registrados 1.267 casos humanos de LVA, dos quais 50% eram menores de 4 anos, e 74 Ãbitos. O ano de 2007 foi o que apresentou a maior taxa de incidÃncia da doenÃa. A maior ocorrÃncia de casos foi em indivÃduos do sexo masculino (64,3%). Houve mudanÃa de perfil da idade e da forma de expressÃo clÃnica da doenÃa na fase epidÃmica, comparada com a fase endÃmica. O risco de adoecer no sexo masculino foi maior do que no sexo feminino. A letalidade foi alta apenas na faixa etÃria acima dos 60 anos. Houve um crescimento significativo na incidÃncia da doenÃa em pessoas idosas. Os exames diagnÃsticos apresentaram alta sensibilidade e especificidade. Mais de 70% dos casos foram confirmados por critÃrio laboratorial. Foi observada uma queda na taxa de cura e um aumento dos casos de co-infecÃÃo por HIV. Na fase epidÃmica, a difusÃo espacial da doenÃa foi do tipo por contÃgio. O processo de difusÃo esteve associado aos fatores ambientais e socioeconÃmicos. Foram identificadas as Ãreas de alto risco para a ocorrÃncia da doenÃa com base nas tÃcnicas de anÃlise espacial. O estudo revelou o estabelecimento de um novo padrÃo epidemiolÃgico para o calazar em Fortaleza, caracterizado principalmente pelo surgimento de um problema emergente: o incremento de casos de co-infecÃÃo por HIV. Revelou, ainda, que os fatores ambientais e socioeconÃmicos foram determinantes nos processos de urbanizaÃÃo e expansÃo da doenÃa na cidade.

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Translational control has a direct impact on cancer development and progression. Quantitative and qualitative changes of cap-dependent translation initiation contribute to neoplastic transformation and progression. However, the idea that “alternative” mechanisms of translation initiation, such as IRES-dependent translation, can be involved in the tumorigenesis is emerging. Because the relevance of this kind of translation initiation in cancer progression is not so well clarified, the purpose of my work was to study the impact of IRES-dependent mRNA translation on tumourigenesis and cancer progression with particular regard for breast cancer. The data obtained clarify the function of cap-independent translation in cancer. Particularly they suggested that the deregulation of IRES-dependent translation can be considered a sort of pro-oncogenic stimulus characterized by the inhibition of the expression of some proteins that block cell growth and proliferation and by the over expression of other proteins that contributed to cell survival. In addition, under stress condition, such as a hypoxia, in immortalized epithelial cell lines, changes in cap-independent translation are associated with an induction of expression of stem cell markers, and with the selection of a sub group of cells that have an increased ability to self-renewing and therefore in the acquisition of a more aggressive phenotype.

In cycling cells positive stimuli like nutrient, growth factors and mitogens increase ribosome biogenesis rate and protein synthesis to ensure both growth and proliferation. In contrast, under stress situation, proliferating cells negatively modulate ribosome production to reduce protein synthesis and block cell cycle progression. The main strategy used by cycling cell to coordinate cell proliferation and ribosome biogenesis is to share regulatory elements, which participate directly in ribosome production and in cell cycle regulation. In fact, there is evidence that stimulation or inhibition of cell proliferation exerts direct effect on activity of the RNA polymerases controlling the ribosome biogenesis, while several alterations in normal ribosome biogenesis cause changes of the expression and the activity of the tumor suppressor p53, the main effector of cell cycle progression inhibition. The available data on the cross-talk between ribosome biogenesis and cell proliferation have been until now obtained in experimental model in which changes in ribosome biogenesis were obtained either by reducing the activity of the RNA polymerase I or by down-regulating the expression of the ribosomal proteins. The molecular pathways involved in the relationship between the effect of the inhibition of RNA polymerase III (Pol III) activity and cell cycle progression have been not yet investigated. In eukaryotes, RNA Polymerase III is responsible for transcription of factors involved both in ribosome assembly (5S rRNA) and rRNA processing (RNAse P and MRP).Thus, the aim of this study is characterize the effects of the down-regulation of RNA Polymerase III activity, or the specific depletion of 5S rRNA. The results that will be obtained might lead to a deeper understanding of the molecular pathway that controls the coordination between ribosome biogenesis and cell cycle, and might give useful information about the possibility to target RNA Polymerase III for cancer treatment.

Background: Circulating tumor cells (CTCs) and circulating free plasma DNA (FPDNA) have been proposed as biomarkers predictive of outcome and response to therapy in solid tumors. We investigated the multiple associations of the presence of CTC and the levels of FPDNA with the outcome and/or the response to chemotherapy in patients with localized breast cancer (LBC), metastatic breast cancer (MBC) and advanced ovarian cancer (AOC). Experimental Design: Blood samples were collected before (baseline), during and after therapy in 40 LBC and 50 AOC patients treated with neo-adjuvant chemotherapy. In 20 MBC patients blood was sampled at baseline and every each cycle of adjuvant chemotherapy. Real time PCR was applied to quantify FPDNA using the Quantifiler Human Quantification kit and CTCs through the detection of tumor-cell specific mRNA levels with or without epithelial enrichment. Results: At baseline CTCs were detected in 90% MBC, 42.5% LBC and 33% AOC patients respectively. The presence of baseline CTC was significantly associated with shorter overall survival (OS) in MBC and AOC patients, and shorter progression free survival (PFS) in LBC patients. Presence of CTCs at the end of neo-adjuvant chemotherapy was detected in 42% LBC and 18% AOC patients and was associated with shorter PFS and OS only in LBC. Increased FPDNA levels at baseline were found in 65% MBC, 17.5% LBC and 76% AOC patients but never related to OS. Baseline FPDNA high levels were associated with shorter PFS only in LBC patients. High FPDNA levels after neo-adjuvant chemotherapy were detected in 57% LBC and 48% AOC patients. Increased FPDNA after neo-adjuvant was associated with response to therapy and shorter PFS in AOC patients. Conclusions: Detection of CTCs may represent a prognostic and predictive biomarker in LBC, MBC and AOC. Quantification of FPDNA could be useful for monitoring response to therapy in AOC patients.

E2F-1 is a transcription factor that plays a key role in cell-cycle control at G1/S check-point level by regulating the timely expression of many target genes whose products are required for S phase entry and progression. In mammalian cells, E2F-1 is negatively regulated by hypo-phosphorylated Retinoblastoma protein (pRb) whereas it is protected against degradation by its binding to Mouse Double Minute 2 protein (MDM2). In this study we experimented a drug combination in order to obtain a strong down-regulation of E2F-1 by acting on two different mechanisms of E2F-1 regulation mentioned above. This was achieved by combining drugs inhibiting the phosphorylation of pRb with drugs inactivating the MDM2 binding capability. The mechanism of action of these drugs in down-regulating E2F-1 level and activity is p53 independent. As expected, when combined, these drugs strongly inhibits E2F-1 and hinder cell proliferation in p53-/- and p53-mutated cells by blocking them in G1 phase of cell cycle, suggesting that E2F-1 down-regulation may represent a valid chemotherapeutic approach to inhibit proliferation in tumors independently of p53 status.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Leishmania braziliensis is the main causative agent of American cutaneous leishmaniasis in Brazil. Altough L. braziliensis infection is self-limited it is distinguished by its latency and chronicity and can persist in mice and patients even after spontaneous clinical cure or treatment. Little is known about the persistence feature in L. braziliensisâ infection in humans and mice. Thereby, the aim of this study was to characterize the parasiteâs persistence in the murine model. The parasite load, lesion thickness and pattern of cytokines and chemokines involved in persistence were evaluated through BALB/c mice infection with a L. braziliensis strain and observation for 90 days after infection. The results showed that parasites gradually disappear in the footpad, while the parasite load was sustained in the draining lymph node until 90 days post-infection (p.i). In the footpad, TNF-α expression was higher at 30 days p.i, followed by the decrease in parasite load and lesion thickness in the site. IL-10 and TGF-β were more expressed at first and decreased after 30 days of infection. A higher concentration of TNF-α and IFN-γ was observed at 15 and 30 days post-infection in the draining lymph node, while IL-4 was significantly increased at 15 days post-infection, and IL-10 and TGF-β were the predominant cytokines after 90 days of infection. CCL2, CCL3, CXCL1 e CXCL10 were expressed in the footpad, with a peak at 30 days p.i. and reduction at 60 days. In the draining lymph node, CCL2 and CXCL10 presented a low expression at first, increasing to a peak at 45 days p.i., while CCL3 was more expressed at 30 days p.i., the same period of the maximum lesion thickness. In our work, we observed that Leishmania persists in draining lymph nodes, while CCL2, CXCL10, IL-10 and TGF- are significantly expressed/produced. These results indicate that these citokynes act synergistically, which can determinate parasite persistence in the draining lymph node. / A infecÃÃo por Leishmania braziliensis se distingue por sua latÃncia e cronicidade, podendo persistir em camundongos e nos pacientes apÃs a cura clÃnica espontÃnea ou apÃs o tratamento. A persistÃncia da infecÃÃo por L. braziliensis ainda tem sido pouco estudada, seja em humanos, seja em modelos experimentais. O objetivo deste trabalho, portanto, foi caracterizar a persistÃncia da infecÃÃo por L. braziliensis utilizando o modelo murino. Foram avaliados em diferentes tempos a espessura da lesÃo, a carga parasitÃria e o padrÃo de citocinas e quimiocinas envolvidas no processo. Para isso, camundongos BALB/c foram infectados com uma cepa de L. braziliensis e acompanhados por 90 dias. Os resultados mostraram que os parasitos desapareceram gradativamente da pata, enquanto nos linfonodos a carga parasitÃria se manteve, mesmo 90 dias pÃs-infecÃÃo (p.i.). Houve pouca expressÃo de IFN-γ nas patas durante os perÃodos analisados, enquanto a expressÃo de TNF-α apresentou-se alta aos 30 dias de infecÃÃo. IL-10 e TGF-β apresentaram maior expressÃo inicialmente e regrediram a partir de 30 dias p.i. Quanto Ãs quimiocinas, observou-se expressÃo de CCL2, CCL3, CXCL1 e CXCL10 nas patas; aos 30 dias todas as quimiocinas apresentaram mÃxima expressÃo, e diminuÃram no 60o dia p.i. AtravÃs de detecÃÃo por ELISA no linfonodo de drenagem, observou-se um pico de concentraÃÃo de TNF-α aos 15 dias e de IFN-γ aos 30 dias pÃs-infecÃÃo, o que precedeu a reduÃÃo na espessura da lesÃo e diminuiÃÃo da carga parasitÃria na pata. IL-4 foi predominante aos 15 dias p.i., tendo sua concentraÃÃo muito diminuÃda aos 30 dias; a partir desse perÃodo a citocina foi pouco detectada. JÃ a concentraÃÃo de IL-10 e TGF-β manteve-se alta e estÃvel, a partir dos 30 dias p.i. Nos linfonodos, a expressÃo de CCL2 e CXCL10 aumentou gradativamente, atingindo pico aos 45 dias p.i., enquanto a expressÃo de CCL3 apresentou pico aos 30 dias p.i., regredindo depois. Em conclusÃo, os parasitos persistem no linfonodo de drenagem em perÃodos crÃnicos de infecÃÃo, quando CCL2, CXCL10, IL-10 e TGF- continuam sendo expressos/produzidos em quantidade. O quadro observado sugere um equilÃbrio entre essas citocinas, o que pode ser determinante para a persistÃncia do parasito no linfonodo.