Introdução. A dengue é uma doença reemergente preocupante para a saúde pública. O seu agente, Aedes aegypti, é a principal espécie responsável pela transmissão do vírus da dengue. Atualmente a Organização Mundial da Saúde (OMS) estima que entre 50 a 100 milhões de pessoas se infectem anualmente em países de todos os continentes com exceção da Europa. Objetivo. Descrever e conhecer a estrutura dos serviços de controle do Aedes aegypti e os aspectos epidemiológicos da dengue no município de Mogi das Cruzes, São Paulo. Métodos. A área de estudo foi o município de Mogi das Cruzes, São Paulo, no período de 2003 a 2007. Foram utilizados dados de bases sucundários da Secretaria de Saúde do município para a descrição quanto a estrutura de vigilância. Os registros de temperatura e pluviosidades foram confrontados por meio de correlação com os focos positivos. Resultados. Observou-se deficiência quanto a estrutura e funcionamento dos serviços de vigilância diante do preconizado pelo PNCD, Ministério da Saúde. Os casos de dengue no município são todos importados, e a maioria são de regiões próximas como o litoral norte e sul. Foi fraca, porém significativa a correlação entre a pluviosidade, temperatura e o foco positivo para o vetor. Observou-se por meio de correlação utilizando o time-lag em mês, que houve correlação significativa no mês em que chove e até três meses após a chuva. Conclusões. Os resultados indicam que o município reúne condições favoráveis à epidemia de dengue e necessita aperfeiçoar o seu sistema de prevenção à doença / Introdution: Dengue is a preoccupying re-emergent illness for public health. The agent, Aedes Aegypti, is the main responsible specie for the transmission of dengue virus. Currently the World Health Organization (OMS) estimate that between 50 until 100 million of people are infected annually in all countries of continents with exception of Europe. Aim. Describe and know the structure of control services of Aedes aegypti and the epidemiologists aspects of dengue in Mogi das Cruzes city, São Paulo. Method. The study area was the Mogi das Cruzes city, Sao Paulo, in the period of 2003 until 2007. It had been used data base of Health Secretary of city to describe as the structure of vigilance. The registers of temperature and rainfalls had been collated by correlation with positive focus. Results. Observed the deficiency for the structure and functionality of vigilance services in the presence of described by PCND, Minister of Health. The all cases of dengue in the city are imported, and the majority is from next regions as north and south coast. It was weak, however the significant correlation between the rainfall, temperature and positive focus for the vector. Observed by correlation mode using the time-lag in month, that had significant correlation in the rain month and until three month after rain. Conclusions. The results indicate that the city congregate favorable conditions to the dengue epidemic and needs to improve their system of illness prevention

A dengue é uma doença infecciosa transmitida pela picada de mosquitos do gênero Aedes. O vírus da dengue (DENV), pertencente ao gênero Flavivirus, família Flaviviridae, é atualmente um importante problema de saúde pública em todo o mundo. São reconhecidos quatro sorotipos antigenicamente distintos (DENV-1, -2, -3 e -4). A infecção por qualquer um dos sorotipos cursa de forma assintomática ou com quadro clínico que varia desde uma febre indiferenciada e autolimitada, passando pela febre clássica da dengue (FD) até quadros graves de febre hemorrágica da dengue (DHF). O diagnóstico clínico é difícil de ser realizado principalmente na faixa etária pediátrica em que os sintomas são muito similares aos de outras infecções febris agudas, ficando a cargo do laboratório o diagnóstico confirmatório. Este estudo descritivo do tipo série de casos teve como objetivo analisar as características clínicas e laboratoriais da dengue em pacientes pediátricos. A população de estudo foi constituída por 110 crianças com idade média de 9,3 ± 3,7 anos recrutadas em uma unidade de saúde no município de Ribeirão Preto, São Paulo. Amostras de sangue, saliva e urina foram coletadas das crianças 1-14 dias após o início do quadro clínico. Os sinais e sintomas mais frequentes foram a febre (105/110, 95%), seguida de cefaleia (66/110, 60%), mialgia (49/110, 45%), vômitos (27/110, 25%) e exantema (16/110, 15%). A infecção pelo vírus da dengue foi confirmada laboratorialmente em 96 crianças, as quais apresentavam sinais e sintomas compatíveis com a classificação de caso suspeito de dengue de acordo com os critérios da Organização Mundial da Saúde. A análise clínica inicial falhou em classificar como caso suspeito de dengue 46% das crianças. A carga viral foi significativamente maior no soro quando comparado àquela observada em saliva e urina, mas ainda assim estas amostras podem ser utilizadas como alternativa para diagnóstico da doença. O sequenciamento nucleotídico da região codificadora da proteína NS5 mostrou a circulação de DENV-3, principalmente durante o ano de 2010, e de DENV-1 e DENV-2, predominantemente no ano de 2011 na população do estudo. Houve predomínio de infecções primárias com quadro clínico leve sem complicações. / Dengue is an infectious disease transmitted by the biting of mosquitoes of Aedes genus. Dengue virus (DENV), belonging to the Flavivirus genus, Flaviviridae family, is an important public health problem worldwide. Four antigenically distinct viruses are recognized (DENV-1, -2, -3, e -4). Infection with any of the virus serotypes causes a spectrum of clinical manifestations ranging from inapparent or mid viral syndrome to classic dengue fever (DF) and severe hemorrhagic disease (DHF). The clinical diagnosis of dengue is difficult, especially in children because the symptoms are very similar to those observed in other febrile illness; thus, the confirmatory diagnosis is carried out by laboratory tests. This descriptive study aimed to analyze the clinical and laboratory features of dengue in pediatric patients. The study population consisted of 110 children; mean age 9.3 ± 3.7 years enrolled in a health center in Ribeirao Preto, Sao Paulo. Samples of blood, saliva and urine were collected from children 1-14 days after the onset of symptoms. The most common signs and symptoms were fever (105/110, 95%), followed by headache (66/110, 60%), myalgia (49/110, 45%), vomiting (27/110, 25%) and rash (16/110, 15%). Dengue virus infection was confirmed by laboratory tests in 96 children whom presented signs and symptoms compatible with the suspected dengue case classification in accordance with the criteria of the World Health Organization. The initial clinical examination failed to classify as a suspected dengue case 46% of children. The viral load in the serum was significantly higher when compared to saliva and urine. Even though, saliva and urine might be used as alternative samples for the diagnosis of the disease. The nucleotide sequencing of a partial region of NS5 protein gene showed the circulation of DENV-3, especially during the year 2010, and DENV-1 and DENV-2, predominantly in the year 2011 in the study population. There was a predominance of primary instead of secondary infections, all of them with self-limiting dengue fever.

Introdução: A dengue é uma doença infecciosa causada por um arbovírus relatada em regiões tropicais e subtropicais. Os maiores números de casos costumam ocorrer nos períodos quentes e chuvosos do ano, que favorecem as condições para o desenvolvimento do vetor da doença, o mosquito Aedes aegypti. Modelos estatísticos podem ser úteis para a compreensão das variações mensais dos números de casos registrados da doença e podem trazer razoáveis predições dos números mensais de casos em um período subsequente a uma série temporal estudada. Métodos: Esta dissertação trata de um estudo ecológico com componente de série temporal. Foram usados registros mensais de casos confirmados de dengue, entre os anos de 1998 a 2008, obtidos do Sistema de Informação de Agravos de Notificação (SINAN), de dois municípios do interior paulista (Ribeirão Preto e Campinas). Modelos estatísticos de séries temporais foram utilizados para descrever o comportamento das séries de dados e predizer o número esperado de casos da doença em um período subsequente. Utilizou-se modelos de Box & Jenkins, ou seja, o modelo SARIMA (Seasonal Autoregressive Integrated Moving Average) com extensões (modelo SARIMAX) que permitem incorporar covariáveis como as temperaturas máxima, média e mínima e a precipitação média a cada mês. Utilizou-se o critério de Akaike (AIC) para a comparação entre diferentes especificações dos modelos. A partir dos modelos ajustados aos dados, foram obtidas predições mensais para o ano de 2009, sendo estas comparadas com os valores observados. Resultados: Sem considerar a inclusão de covariáveis, o melhor modelo para os dados de Campinas foi o SARIMA(2,1,2)(1,1,1)12 , e para os dados de Ribeirão Preto o melhor foi o SARIMA(2,1,3)(1,1,1)12 (modelos com menores valores de AIC). Ao incorporar nos modelos as variáveis climáticas, observou-se um melhor ajuste para o município de Campinas, o modelo SARIMAX(1,1,1)(1,1,1)12 que inclui a precipitação observada nos 2 meses antecedentes às ocorrências mensais de casos de dengue e a temperatura mínima registrada nos 3 meses antecedentes. No município de Ribeirão Preto, o modelo que mostrou-se mais adequado aos dados foi o SARIMAX(1,1,1)(0,1,1)12 que inclui precipitação observada nos 3 meses antecedentes às ocorrências mensais de casos de dengue e a temperatura mínima registrada em um mês anterior. As predições mensais obtidas para o ano de 2009 mostraram-se razoavelmente próximas àquelas observadas, em ambos municípios. Conclusões: Os resultados do presente estudo, em adição a resultados da literatura, evidenciam que os números de componentes autorregressivos e de média móvel adequados ao ajustes de modelos SARIMA variam de uma população a outra, sugerindo diferentes padrões temporais da doença de acordo com características locais. Comparações entre valores preditos e observados sugerem que os modelos SARIMA são ferramentas úteis para predição do número de casos de dengue. Sugere-se que os modelos utilizados nesta dissertação sejam utilizados por serviços municipais de saúde para a vigilância da doença, o que pode trazer benefícios em programas de prevenção e planejamentos de serviços públicos. / Introduction: Dengue is an infection disease which is caused by an arbovirus related to tropical and subtropical region. Most cases usually occur on warm and rainy weathers of the years that facilitate the conditions of the development vector of the disease, the Aedes Aegypti mosquito. Statistical models can be very useful to comprehend the monthly variations of the number of registered cases of the disease, and they might bring reasonable predictions to the monthly number of cases in a subsequent period to a studied time series. Metodology: This dissertation is about ecologic study with a time series component. It was used monthly registry of confirmed dengue cases, between 1998 and 2008 years, obtained by the Information Center of Notification Diseases (ICND), out of two cities in the state of São Paulo (Campinas and Ribeirão Preto). Statistic time series models were used to describe the data series behavior and to predict the average number of disease cases in the subsequent period. We used the Box & Jenkins model, that in, the SARIMA (Seasonal Autoregressive Integrated Moving Average) model, with extensions (SARIMAX model) that allow to incorporate co-variables to the maximum, mean and minimum temperatures and the average monthly precipitation. We used the Akaike criterio (AIC) for comparison between different model specifications. From the fit data models, we obtained monthly predictions to the 2009 year, being compared to the observed data. Results: Without considerating the inclusion of the covariables, the best model for Campinas data was the SARIMA(2,1,2)(1,1,1)12, and for Ribeirão Preto data the best was SARIMA(2,1,3)(1,1,1)12 (smaller AIC values models). As we incorporate the climate variables in the models, we observe that the best fit for Campinas city was the SARIMAX(1,1,1)1,1,1)12 that includes the observed precipitation in 2 months before the monthly dengue cases and the minimum registered temperature in 3 months before. In Ribeirão Preto city, the most adequate model was the SARIMAX(1,1,1)(0,1,1)12 that includes the observed precipitation in 3 months before the monthly dengue case occurrence and the minimum temperature in one month before. The monthly prediction obtained for 2009 year show reasonably close to the observed, in both cities. Conclusion: The results of this study, in addition to the literature results, show that the adequate number of autoregressive and moving average components fitted in SARIMA models, vary between one population and other, suggesting different disease time patterns according to local characteristics. Comparing predict and observed values suggest that the SARIMA models are useful tools to predict the number of dengue cases. We suggest that the models in this dissertation be used in city health services to surveillance the disease, which may bring benefits for prevention programs and public service planning.

La dengue est une maladie virale endémique dans les pays tropicaux. Bénigne dans sa forme classique (FC), elle peut être redoutable lors de complications associées à sa forme sévère (FS). Actuellement, l’établissement du pronostic d’évolution vers une dengue sévère est peu fiable. Afin d’identifier des marqueurs précoces d’évolution vers une forme sévère, une étude de caractérisation différentielle de plasmas FC et FS prélevés précocement, a été effectuée par différentes approches de protéomique. La protéine non-structurale NS1 est une des protéines virale associée à la pathogénicité. Une méthode de profilage des plasma utilisant la technologie SELDI-TOF/MS (Surface Enhanced Laser Desorption Ionization-Time of Flight/Mass Spectrometry) couplée à un anticorps monoclonal anti-NS1 a été utilisée afin d’identifier les ligands de cette protéine. La protéine NS1 a été détectée spécifiquement dans des plasmas de patients dengue. Aucun partenaire de la NS1 n’a pu être identifié. La méthode peut néanmoins être utilisée pour sérotyper les échantillons.Une analyse différentielle en ICPL (Isotope Coded Protein Labelling) des protéines plasmatiques de patients FC ou FS a permis d’identifier trois marqueurs potentiellement liés la sévérité de la maladie qui ont été validés en ELISA. La synthèse du virus mettant en jeu des partenaires cellulaires, le viroprotéome associé à chaque pathologie a été caractérisé par LC-MS/MS. Une méthode de purification du virus de la dengue a préalablement été mise au point sur un modèle in vitro. Cette méthode a ensuite été appliquée sur des pools de plasmas de patients prélevés en phase virémique. Des protéines virales ainsi que des protéines de l’hôte potentiellement associées aux virus ont été identifiées. Après analyse, une « empreinte » mettant en jeux des voies canoniques spécifiques a pu être déterminée pour les FS. Des ELISA ont été réalisés afin de valider le différentiel d’expression sur un choix de protéines.Au-delà de l’identification d’outils susceptibles d’aider les praticiens à poser un pronostic, ces études s’inscrivent dans la compréhension des mécanismes complexes qui sous-tendent le passage d’une FC à une FS. / Dengue is a endemic viral disease in tropical countries. If its classical form (CF) is benign, its severe form (SF) leads however to serious complications. Currently, the prognosis of severe dengue is unreliable. Differential proteomic studies on acute CF and FS plasma specimens were performed in order to identify early markers of progression to severe forms.The non-structural protein 1 (NS1) is a viral protein associated with pathogenicity. A method using SELDI-TOF/MS (Surface Enhanced Laser Desorption Ionization-Time of Flight/Mass Spectrometry) coupled to anti-NS1 monoclonal antibodies was developed in order to profile the proteins interacting with NS1 in plasma. Whereas NS1 protein was specifically detected in acute dengue plasma specimens, no NS1 ligand was identified. This method however allowed for sample serotyping.Plasma proteins of SF and CF patients were analyzed differentially using ICPL (Isotope Coded Protein Labeling). Three markers potentially related to disease severity were identified and validated by ELISA.As cellular partners are involved in virus biosynthesis, the viroproteome associated to each disease was characterized by LC-MS/MS. A method of dengue virus purification was first developed on an in vitro model. This method was then applied to pools of acute plasma of either CF or SF patients. We identified viral proteins as well as host proteins potentially associated with the viral particles. A footprint involving specific canonical pathways were subsequently identified in SF patients. Finally, a set of proteins found differentially expressed was validate by ELISA.Beyond identification of tools allowing assessment of dengue severity prognosis, these results give clues on the complex mechanisms underlying the transition from the CF to the SF.

Au cours des dernières années, de nombreuses épidémies ont emergé ou ré émergé, et sont causées par des arbovirus (arthropod-borne viruses), des virus transmis à des vertébrés par des insectes piqueurs vecteurs. Avec l'augmentation de la densité humaine dans certaines zones géographiques et le réchauffement climatique qui contribuent à l'expansion géographique des vecteurs, les maladies induites par ces virus (arboviroses) ont un impact de plus en plus important sur la santé humaine et l'économie mondiale. Il est donc déterminant d'augmenter nos connaissances sur les systèmes mis en jeux pour garantir la sécurité sanitaire des populations exposées. Les enjeux actuels reposent aussi bien sur la compréhension des virus que sur la compréhension de l'alternance d'hôtes, directement responsables de l'émergence et la dissémination des agents infectieux. Les moustiques sont des vecteurs majeurs des arbovirus comme la dengue (genre Flavivirus) et le Chikungunya (genre Alphavirus). Transmis par les mêmes moustiques Aedes aegypti et Aedes albopictus, le virus de la Dengue (DENV) est responsable de la plus importante arbovirose en zone tropicale, et le virus Chikungunya (CHIKV) est responsable dans le monde entier de centaines de milliers de cas d'infection, et les épidémies récentes ont touché les pays européens. Ainsi, il a été observé que le moustique Ae. albopictus pouvait porter simultanément CHIKV et DENV, et des cas de co-infections humaines ont été observés en Afrique. Toutefois, bien qu'en théorie les deux virus soient capables d'infecter les mêmes cellules chez l'insecte ou l'homme, il n'y a aucune étude détaillée sur les interactions au niveau cellulaire entre CHIKV et DENV lors de la co-infection d'une cellule. C'est pourquoi il est indispensable d'accroitre nos connaissances sur l'interférence éventuelle entre les virus Chikungunya et Dengue pour l'utilisation de voies cellulaires communes chez les insectes vecteurs et l'hôte humain lors de co-infection / Emergence and geographical extension of dengue (DENV), Zika (ZIKV) and chikungunya (CHIKV) viruses increase simultaneous outbreak in an increasing number of countries. To date, no vaccine or cure have yet been developed against these diseases those cause a tremendous impact on human health and in the economy worldwide. During recent simultaneous outbreaks, up to 12% of patients have been diagnosed to be co-infected by CHIKV and DENV. In addition, it was shown that the mosquitoes Aedes albopictus could carry and transmit simultaneously CHIKV and DENV. However, the pathology, as well as the epidemiology of a pathogen, relies on the interactions between several infectious agents present within an organism or a community in the environment. It is crucial to consider to which extent a host infected by a first microorganism is modified and whether its reaction to the infection by a second microorganism is consequently altered. However, there is no extensive report of Alphavirus-Flavivirus or Flavivirus- Flavivirus interactions. Our global objective is to characterize these co-infections in both mosquitoes and humans, at the cell and molecular level. To this aim, we started this project by performing sequential co- infection in different cell lines from Aedes albopictus and Aedes aegypti. We found that the permissiveness and production of DENV is enhanced in presence of CHIKV. On the contrary, there is no effect of DENV pre-infection on subsequent CHIKV co-infection. We generalized the synergistic phenomena and we showed that CHIKV pre-infection also increased the infection by DENV-1, DENV-3 and DENV-4, but also by two others re-emerging Flaviviruses, the Yellow Fever Virus (YFV), and the Zika Virus (ZIKV). Remarkably, we succeeded to establish a mosquito model of co-infection of Aedes aegypti mosquito after by different two feedings at 4 days interval. Using this sequential co-infection, we were able to show that a pre-infection of Aedes aegypti by CHIKV increase the level of DENV-2 RNA in salivary glands compare to mono-infected mosquitos. This phenotype is reminiscent of the phenotype we observed in vitro during successive infections. Altogether, our study paves the way to the characterization of molecular interaction between Flaviviruses and Alphaviruses in mosquito in vitro and in vivo. This study can be crucial for a better understanding of disease and epidemiology during simultaneous outbreaks

La dengue constitue actuellement l'arbovirose la plus répandue sur la planète avec 40% de la population mondiale exposée, soit environ 2,5 milliards de personnes. La maladie est endémique dans tous les continents excepté l'Europe même si les premiers cas de transmission autochtone ont été observés en France métropolitaine et en Croatie en 2010. L'augmentation régulière de son incidence fait actuellement de la dengue un problème de santé publique majeur dans les pays tropicaux et une menace pour d'autres pays du monde. En raison de son mode de transmission, la dengue constitue une maladie étroitement liée à l'environnement. L'urbanisation constitue un facteur important dans l'émergence ou la réémergence d'épidémies de dengue dans les pays tropicaux. Cette étude menée a pour objet d'analyser les liens existant entre le domaine de la santé (situation sanitaire et épidémiologique de la dengue) et de celui de l'environnement (évolution de l'urbanisation) en Guadeloupe continentale entre 2000 et 2008.Pour ce faire, les cas suspects et confirmés de dengue ont été recueillis. L'incidence pour ces deux types de cas a été calculée puis représentée à deux échelles administratives différentes: communes et sections. Les facteurs environnementaux comprenant les variables physiques et socio-démographiques ont été pris en compte pour la définition, l'identification et l'analyse de l'évolution des contextes d'habitation durant la période étudiée. Deux méthodes ont été utilisées: la classification «Raster» consistant en une analyse régulière basée sur des mailles de taille identique et la classification « Vecteur» s'appuyant sur les limites administratives des sections. Ces classifications ont permis de décrire la répartition des différents contextes d'habitation sur le territoire de la Guadeloupe et d'obtenir des indicateurs qui ont été analysés en relation avec les données sanitaires. Il a été mis en évidence que la situation épidémiologique de la dengue dans le département s'est significativement aggravée au cours de la période étudiée. Des épidémies majeures sont apparues en 2001, 2005/2006 et 2007. Le nord Basse-Terre, la région Pointoise ainsi que le sud-est de la Grande-Terre, de la ville du Gosier à Saint-François le long de la côte, sont les régions les plus concernées par l'augmentation d'incidence de la maladie. Concernant les contextes d'habitation, huit contextes distincts ont été identifiés: touristique, naturel, intermédiaire, agricole, résidentiel, rural, périurbain et urbain. Le contexte intermédiaire correspond à des régions de transition entre les autres contextes d'habitation. La différence entre les contextes agricoles et ruraux réside dans la prédominance de surface agricole pour le premier alors que le contexte rural est caractérisé par les espaces cultivés habités. L'évolution surfacique de ces contextes au cours du temps a été analysée en relation avec les incidences. Les résultats montrent que les contextes d'habitation sont statistiquement.associés à l'incidence de la dengue en Guadeloupe. En particulier, les contextes touristiques, résidentiels et ruraux possèdent une association positive élevée avec l'incidence de la dengue. / At present, dengue fever has become the arbovirose the most wide-spread worldwide with 40 % ofthe exposed population, 2,5 billion people. The disease is endemie in all the continents excepted Europe even though the fust cases ofautochthonoustransmissionwere observedinmetropolitanFranceandCroatiain2010.Because oftherecentincrease of its incidence, dengue fever is now a major public health problem in the tropical countries and a threat for the other countries in the worldwide. Due to its mode oftransmission, the disease is closely linked to the environment. Urbanization is an important factor ofthe emergence or the re-emergence of dengue in the tropical countries. This study aims to analyze the links between health and environment domains, which correspond to the epidemiological situation of the dengue and the evolution ofthe urbanization in Guadeloupe.Information about suspect and confirmed cases of dengue were collected. The incidence of suspect and confirmed cases has been estimated, then represented at two different administrative boundaries: communes and sections. Environmental factors including the physical and socio-demographic variables were taken into account to the definition, the identification and the analyze ofevolution of the housing patterns during the studied period. Two methods were used: Il Raster" consisting of a regular analysis based on squares units and "Vector" based on the administrative limits of sections. These classifications allowed to describe the distribution ofthe different housing patterns ofGuadeloupe and to estimate indicators for the analyzing of sanitary data. The results highlight a worsening ofepidemiological situation ofthe dengue over the time in the department. Major outbreaks occurred in 2001,2005/2006 and 2007. The north Basse-Terre, the Pointe-à-Pitre urban area as weIl as the whole coast (frOID the city of Gosier to Saint-François) ofthe southeast ofGrande-Terre were the most affected regions by the increase ofincidence of dengue. Concerning the housing classes, eight different patterns were identified: tourist, natural, intermediate, agricultural, residential, rural, suburban and urban. The intermediate class corresponds to buffer regions between the other patterns. The difference between the agricultural and rural contexts lies in the predominance ofagricultural surface for the first one while the rural context is characterized by the inhabited farmland. The evolution over time of spatial extend ofthese patterns was described in relation with incidence. Results showed that the housing patterns are statically associated with the incidence ofdengue in Guadeloupe. In particular, the touristic, residential and rural patterns have a high positive association with dengue incidence.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2017-07-24T14:50:01Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Tese_Luciana_Holmes_FOLHA_APROVA ÇÃO_Ficha_Catalográfica_25_01.pdf: 3698873 bytes, checksum: f3ad9bfbfbdb382b59bf0c7b2216b2b2 (MD5) / Made available in DSpace on 2017-07-24T14:50:01Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Tese_Luciana_Holmes_FOLHA_APROVA ÇÃO_Ficha_Catalográfica_25_01.pdf: 3698873 bytes, checksum: f3ad9bfbfbdb382b59bf0c7b2216b2b2 (MD5) Previous issue date: 2016-02-29 / A nova classificação da dengue (dengue sem sinais de alarme, dengue com sinais de alarme e dengue grave) da Organização Mundial de Saúde e adotada pelo Ministério da Saúde do Brasil objetiva identificar os pacientes que apresentam um maior risco de complicações. Entre março de 2009 e maio de 2011 formou-se uma coorte prospectiva com suspeitos de dengue em dois hospitais da capital da Paraíba no nordeste do Brasil. De 770 pacientes, 554 preencheram os critérios de inclusão, representados por crianças e adultos, com confirmação laboratorial do diagnóstico de dengue, sendo coletadas informações clínicas, laboratoriais e exame ultrassonográfico. O estudo realizado foi para validar a classificação revisada da Organização Mundial da Saúde, na identificação de casos graves, baseado nos níveis de intervenção clínica (Categoria I, II e III) e nos achados ultrassonográficos. Foram testadas a sensibilidade, especificidade, valores preditivos positivo e negativo da nova classificação com os níveis de intervenção. A sensibilidade e a especificidade da dengue grave para os casos da Categoria III foram de 100% e 91%, respectivamente, com valor preditivo positivo de 15,5% e valor preditivo negativo de 100%. Esses dados demonstram uma maior sensibilidade da classificação revisada na detecção de pacientes com risco de evoluírem para formas graves da doença. A ultrassonografia mostrou ser uma ferramenta útil na identificação de casos suspeitos de dengue e na detecção precoce de sinais de extravasamento plasmático. Os achados ecográficos mais relevantes foram espessamento difuso da parede da vesícula biliar, líquido livre na cavidade abdominal e pélvica, derrame pleural, esplenomegalia e hepatomegalia. O espessamento da parede da vesícula biliar e os derrames cavitários mostraram significância estatística com a nova classificação. / The new classification of Dengue (Dengue without Warning Signs, Dengue with Warning Signs and Severe Dengue) from the World Health Organization and adopted by the Department of Health of Brazil aims to identify patients with a higher risk of complications. From March 2009 to May 2011 formed a prospective cohort with suspected dengue fever in two hospitals in the capital of Paraíba in northeastern Brazil. In 770, 554 patients met the inclusion criteria, represented by children and adults, presenting laboratory confirmation of the diagnosis of dengue, and collected clinical information, laboratory and ultrasound examination. The study was to validate the revised WHO classification to identification of severe cases, based on the clinical intervention (Category I, II e III) and ultrasound findings. Tested the sensitivity, specificity, positive predictive values and negative of new classification with the levels of intervention. The sensitivity and specificity of the Severe Dengue for the cases of category III were 100% and 91%, respectively, with positive predictive value of 15.5% and negative predictive value 100%. These data demonstrate a greater sensitivity of the revised classification in detecting patients with risk of evolve to serious disease. The ultrasonography was a useful tool in the confirmation of suspected cases of dengue fever and on early detection of signs of plasma leakage. Ultrasound findings more important were: thickened gall bladder wall, free fluid in the abdominal and pelvic cavities, pleural effusion, splenomegaly and hepatomegaly. The gallbladder wall thickening and effusions showed statistical significance with the new classification.

A dengue é uma doença infecciosa viral transmitida pela picada de mosquitos do gênero Aedes e é um importante problema de saúde pública mundial. A infecção por qualquer um dos quatro sorotipos (DENV 1-4) pode apresentar diferentes quadros clínicos: pode ser assintomática, causar uma síndrome febril indiferenciada, ou a febre da dengue (DF), a evolução do quadro clínico pode levar a dengue hemorrágica com ou sem choque (DHF/DSS). Um número crescente de casos de infecção por dengue em crianças têm sido observados nos últimos anos. Estudos de prevalência da dengue em regiões endêmicas são importantes para avaliar a incidência da infecção por dengue em crianças. Assim, o objetivo deste estudo é investigar a prevalência de anticorpos anti-dengue IgM e IgG em crianças atendidas no Centro de Saúde Escola Dr. Edgard Aché, localizado na região oeste de Ribeirão Preto-SP. Crianças (n = 271) de 1-15 anos de idade foram recrutadas de março de 2010 até maio de 2011. Depois de um termo de consentimento ter sido assinado pelo responsável, uma amostra de sangue das crianças, participantes deste estudo, foi coletada. As crianças foram classificadas em assintomáticas (n= 174) ou sintomáticas (n = 97), quando estas apresentavam um ou mais de um sintoma sugestivo de dengue, de acordo com critérios da Organização Mundial de Saúde. Anticorpos IgM e IgG anti-dengue foram detectado nas amostras de soro por um ELISA de captura padronizado em nosso laboratório. O ELISA de captura para IgG mostrou uma positividade de 9,23% (25/271) e o ELISA de captura para IgM de 8,49% (23/271). O ELISA de captura para IgG foi positivo em 10,31% (10/97) das crianças sintomáticas e 8,62% (15/174) das crianças assintomáticas, enquanto que o ELISA de captura para IgM foi positivo em 15,46% (15/97) das crianças sintomáticas e 4,6% (8/174) em crianças assintomáticas. Este estudo mostrou a alta prevalência de anticorpos anti-dengue em crianças da região oeste do município de Ribeirão Preto; mostrou também que a infecção pode ser causada de forma assintomática, e que a infecção por este vírus pode ser um grave problema de saúde nesta população, servindo como um alerta às autoridades de saúde. / Dengue is an infectious viral disease transmitted by the biting of mosquitoes of Aedes genus and it is an important public health problem worldwide. Infection with any of the four serotypes (DENV 1-4) may be asymptomatic or causes illness ranging from mild viral syndrome, dengue fever (DF) to dengue hemorrhagic fever (DHF). An increasing number of dengue infection cases in children have been noted in the last years. A dengue surveillance study might be an important tool in endemic region to evaluate the incidence of dengue infection in children. Thus, the aim of this study was to investigate the prevalence of anti-dengue IgM and IgG antibodies in children in the Primary Health Care Center, Dr. Edgard Aché, located in the west region of Ribeirão Preto-SP. Children (n=271) from 1 to 15 years old were recruited during March 2010 until May 2011. After a signed consent by the person responsible for the children to participate in this study, a blood sample was collected. The children were classified in asymptomatic (n=174) or symptomatic (n=97) when they had more than one symptom suggestive of dengue according to the World Health Organization criterions. Anti-dengue IgM and IgG were detected in serum samples by a capture ELISA standardized in our laboratory. IgG capture ELISA was positive in 9,23% (25/271) and IgM capture ELISA in 8,49% (23/271) of the children. IgG capture ELISA was positive in 10,31% (10/97) of the symptomatic children and 8,62% (15/174) in asymptomatic children; while IgM capture ELISA was positive in 15,46% (15/97) of symptomatic children and 4,6% (8/174) in asymptomatic children. This study showed the high prevalence of anti-dengue antibodies in children in the west region of Ribeirão Preto. In addition, a high prevalence of dengue-infected children without symptoms was observed. The present survey demonstrated that dengue virus infection might be a problem in children\'s from Ribeirão Preto city and serve as an alert to the health authorities.

El virus Dengue (VDEN) es el responsable de más de 50-100 millones de casos anualmente en el mundo. La infección del dengue es causada por cuatro serotipos (VDEN-1, VDEN-2, VDEN-3 y VDEN-4) y el espectro de la enfermedad varía desde una fiebre indiferenciada, fiebre hemorrágica por dengue (FHD), síndrome de shock por dengue, y muerte. Información epidemiológica liga el desarrollo de FHD con una infección secundaria y además sugiere que ciertas cepas son más virulentas que otras. En 2001, aparecieron los primeros casos de FHD asociados con VDEN-2 en la costa del Perú. En el 2010 ocurrió un brote en Iquitos con casos severos convirtiéndose en el más largo en la historia de la región. Sin embargo estudios orientados en determinar el origen distribución y diversidad genética de cepas peruanas de VDEN-2 durante los diez últimos años no han sido realizados. Para atender este vacío de conocimiento en la epidemiología del VDEN-2 en Perú, extractos de ARN de 30 aislamientos virales en células C6/36 (Aedes albopictus) fueron procesados por RT-PCR. Secuencias del gen de la envoltura (E) fueron determinadas y comparadas con muestras globales de VDEN-2. El análisis filogenético reveló la circulación de dos genotipos en Perú: Americano (hasta el 2000) y Americano/Asiático (2000- 2010). Adicionalmente se identificaron cepas variantes del genotipo Americano/Asiático distribuidos en dos clados principales (1 y 2) que ingresaron al Perú por la costa norte (Ecuador) y por la selva (Brasil o Bolivia). Con el aparente incremento de la virulencia relacionada a cepas Americano/Asiático del clado 2, nuestros resultados soportan la necesidad de un continuo monitoreo de cepas emergentes de nuevos variantes o genotipos de dengue que podrían estar asociados a casos severos de la enfermedad. Palabras claves: Arbovirus, Dengue 2, Americano/Asiático y relación filogenética.

<p>Our ability to convert basic knowledge into robust anti-viral therapeutics requires discovery of novel host-virus interactions as well as an informed anti-viral discovery pipeline. We used a genome-scale RNAi-based screen followed by a chemical screen of FDA-approved therapeutics to identify scores of novel dengue virus (DENV) human host dependency factors (HDF) and identified more than 20 potential anti-Zika virus (ZIKV) therapeutics. </p><p>Two genes in particular, TTC35 and TMEM111, strongly inhibited DENV infection and, based on comparisons with published literature, implicated a larger protein, the ER Membrane Protein Complex (EMC), as a pan-flavivirus HDF. The EMC is a poorly characterized multiprotein complex that may function in ER-associated protein biogenesis and/or lipid metabolism. Based on our screen data, we hypothesized that the EMC is an uncharacterized HDF that functions through a common mechanism to promote replication of flaviviruses. We report that DENV, ZIKV, and yellow fever virus (YFV) infections were impressively inhibited, while West Nile Virus (WNV) infection was unchanged, in cell lines engineered to lack EMC subunit 4 (EMC4). Furthermore, targeted depletion of EMC subunits in live mosquitos significantly reduced DENV-2 propagation in vivo. In addition, the accumulation of DENV proteins shortly after infection in EMC4 knockout cells was significantly reduced, suggesting that the EMC promotes viral protein biogenesis. </p><p>We interrogated a library of FDA-approved drugs for their ability to block infection of human HuH-7 cells by a newly isolated ZIKV strain. Selected compounds were further validated for inhibition of ZIKV infection in human cervical, placental, and neural stem cell lines, as well as primary human amnion cells. Established anti-flaviviral drugs (e.g., bortezomib and mycophenolic acid) and others that had no previously known antiviral activity (e.g., daptomycin) were identified as inhibitors of ZIKV infection. Several drugs reduced ZIKV infection across multiple cell types.</p><p>We propose that the EMC may be exploited as a novel therapeutic target for multiple flaviviruses in the future. Also we identified drugs that could be tested in clinical studies of ZIKV infection and provides a resource of small molecules to study ZIKV pathogenesis.</p> / Dissertation