Navegando por Palavras-chave "Aedes Aegypti"
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- ItemAcesso aberto (Open Access)Avaliação dos impactos à saúde e da evolução temporal da dengue nos Municípios de Ribeirão Preto e São Paulo(Universidade Federal de São Paulo (UNIFESP), 2018-09-21) Gabriel, Ana Flavia Barbosa [UNIFESP]; Miraglia, Simone Georges El Khouri [UNIFESP]; Guimarães, Marcelo de Paiva; http://lattes.cnpq.br/5035762125459415; http://lattes.cnpq.br/6423311971848669; http://lattes.cnpq.br/4538424488673789; Universidade Federal de São Paulo (UNIFESP)Objectives: To evaluate the health impacts of the chemical control applied in the fight against dengue, as well as to analyze the relationship between the occurrence of cases of the disease and precipitation. In addition to evaluate the temporal evolution of dengue cases in the municipalities of Ribeirão Preto and São Paulo, in order to observe incidence patterns to fit predictive models for one year out of the sample. Methods: The objectives were divided into scientific article and the data analysis was specific to each article. In first article a bibliographic review was carried out to build a network of impacts, which facilitated the visualization of the possible direct and indirect impacts from the application of insecticides in the fight against dengue. The methodology of Health Impact Assessment was applied for a characterization of the one of the impacts identified in the network, being quantitative and qualitative analysis were conducted. In the second article we used the Spearman’s Correlation, with the concept of time lag, to analyze the association between dengue occurrence and precipitation in the municipally of Ribeirão Preto. In the third article, time series tools were used, more specifically, Seasonal Integrated Autoregressive Integrated and Moving Average methods to predict dengue cases for one year out of the sample. Results: In the first article it was possible to have a broad and systematic view of the economic, environmental and health impacts caused by chemical control. In addition, there was an increase in dengue cases after an insecticide application, demonstrating an inefficiency of this type of control. The second article showed that precipitation contributes to the generation of new cases of dengue one to five months after its occurrence. In the third article, from the evaluation of the temporal evolution it was possible to adjust SARIMA models, which forecast for one year out of the sample. Conclusion: We concluded with this study that understanding the possible impacts of chemical control on health, as well as the temporal evolution of dengue and the factors that may influence its transmission dynamics, is relevant for public awareness and for rethinking public policies in the attempt to eliminate dengue as a burden on public health.
- ItemSomente MetadadadosConstrução De Biblioteca De Inibidores Proteicos Em Sistema Phage Display Para Seleção De Inibidores Específicos Para Proteases De Larvas De Mosquito Aedes Aegypti(Universidade Federal de São Paulo (UNIFESP), 2017-10-26) Manzato, Veronica De Moraes [UNIFESP]; Tanaka, Aparecida Sadae [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)The mosquito Aedes aegypti is well adapted to urban environment and the major vector of several diseases as dengue, yellow fever, zika and chikungunya flaviviruses. It has been considered a global public health problem because, in the last years, the outbreaks of every 3-5 years of dengue have spread with increased virulence. There is no treatment for these diseases and vaccine has low efficacy, as a result, the prophylaxis remains on vector control but the presence of resistant strains of inseticides and larvicidal in use makes the control a difficult and still unsolved problem. Knowing that the larval stage feeds constantly, we intend to interfere with the larvae development by the inhibition of the digestion proteins. A neutrophil elastase and chymotrypsin inhibitor found in Triatoma infestans eggs named TiPI (Triatoma infestans Pacifastin Inhibitor) had the reactive site (P2-P2’) randomly mutated by the phage display technique which allowed their selection against digestive enzymes of the 4th instar larvae midgut. The selected mutants DNA analyzes evidenced group of 11% possible trypsin inhibitors with Arg or Lys at P1 position, 18.5% possible neutrophil elastase inhibitors with Ala, Leu or Val at P1 and curiously, the major group represented by 47% of the mutants with Gln at P1 and 88.8% with Gln at any mutated position. Fusion phages of ten selected mutants were produced, among them 4 possible trypsin inhibitor, mutants 73 (QRLQ), 154 (LKQL), 161 (KRKQ) and 189 (QRHL), 2 possible chymotrypsin inhibitor 29 (ALAR) and 143 (LLQC), and the 4 most frequently mutants, 4 (SQWH), 5 (QSAM), 6 (HQSL) and 22 (AQVF) and their interactions with proteins of the 4th instar larvae midgut and commercial enzymes were evaluated by Surface Plasmon Resonance. The inconclusive results conduct us to protein expression by Pichia pastoris yeast. The yeast supernatant culture containing recombinant mutants was used in the serine 18 protease inhibition assays. Among the trypsin inhibitors, mutant 73.21 showed high inhibitory activity to trypsin, and the mutant 29.14 inhibits neutrophil elastase. Mutants 4.27, 5.26 and 6.27 presented inhibitory activity for subtilisin A. The results confirm that the TiPI1 phage display library allowed the selection of specific mutants against digestive enzymes of larvae of Ae. aegypti. Our results suggested that these molecules can be use in the development of specific synthetic inhibitors with larvicidal activity for Ae. aegypti.
- ItemAcesso aberto (Open Access)Determinação da estrutura tridimensional de um inibidor de serinoproteases do tipo Kazal do mosquito aedes aegypti, vetor da dengue(Universidade Federal de São Paulo (UNIFESP), 2018-06-28) Torquato, Ricardo Jose Soares [UNIFESP]; Tanaka, Aparecida Sadae [UNIFESP]; Pereira, Pedro José Barbosa; http://lattes.cnpq.br/1884820479531031; http://lattes.cnpq.br/1168789309568199; http://lattes.cnpq.br/3995278540776284; Universidade Federal de São Paulo (UNIFESP)The mosquito Ae. aegypti is the main vector of arboviruses for humans in Brazil and it is responsible for outbreaks of dengue, chikungunya, yellow fever, and recently it has been involved in the high number of humans infected with zika virus. The hematophageal activity of Ae. aegypti allowed it to develop several strategies to control the hemostasis of vertebrate host’s. In 2010, our group expressed and characterized a serinepeptidase inhibitor belonging to Kazal family called rAaTI (Aedes aegypti Trypsin Inhibitor). The purified rAaTI was able to inhibit trypsin and affect coagulation time. With this data, the present work had as general objective the determination of three-dimensional structure of AaTI to understand its role in prolonging coagulation time. The rAaTI was purified in amount and an inhibitor preparation containing 20 mg/mL presenting high degree of homogeneity was used in crystallization experiments. The rAaTI crystallized in 100 mM sodium acetate, pH 5.5 containing 25% PEG 3350, 5% PEG 400 and 3% dioxane. Some crystals of rAaTI were dipped in sodium iodide solution with different concentrations (soaking) for 10 s to 1 min, and then they were analyzed (diffracted) on the MX2 line at the National Synchrotron Light Laboratory (LNLS), Campinas. The three-dimensional structure of rAaTI was determined at high resolution of 1.4 Å. The unit cell of the crystal had an unusual low solvent content of approximately 22-23% (Torquato et al., 2017), accommodating two molecules of rAaTI. rAaTI has previously been described as inhibitor of serine protease, bovine trypsin (Ki = 0.15 nM) and human plasmin (Ki = 3.8 nM), and with anticoagulant activity, prolongating the coagulation time, including time of thrombin (TT) (Watanabe et al., 2010). In an attempt to clarify where rAaTI binds to the surface of thrombin, some experiments were performed using surface plasmon resonance (SPR) technology. Thrombin was immobilized on the surface of the CM5TM GE sensorchip. Different concentrations of rAaTI were applied to the surface containing immobilized thrombin and it was obtained a dissociation constant (KD) of 3.68 μM. Watanabe et al. showed a value of KD = 320 nM (Watanabe et al., 2011). The dissociation constant calculated by SPR is approximately ten times higher, which can be explained by partial blocking of thrombin exosites I and II during the enzyme immobilization procedure. The SPR experiments showed inhibition of the binding of rAaTI to thrombin in the presence of heparin, reinforcing the suggestion that AaTI can interacted to exosite II of thrombin.