995 resultados para Vetores virais recombinantes
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This dissertation presents a new proposal for the Direction of Arrival (DOA) detection problem for more than one signal inciding simultaneously on an antennas array with linear or planar geometry by using intelligent algorithms. The DOA estimator is developed by using techniques of Conventional Beam-forming (CBF), Blind Source Separation (BSS), and the neural estimator MRBF (Modular Structure of Radial Basis Functions). The developed MRBF estimator has its capacity extended due to the interaction with the BSS technique. The BSS makes an estimation of the steering vectors of the multiple plane waves that reach the array in the same frequency, that means, obtains to separate mixed signals without information a priori. The technique developed in this work makes possible to identify the multiple sources directions and to identify and to exclude interference sources
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Nowadays, classifying proteins in structural classes, which concerns the inference of patterns in their 3D conformation, is one of the most important open problems in Molecular Biology. The main reason for this is that the function of a protein is intrinsically related to its spatial conformation. However, such conformations are very difficult to be obtained experimentally in laboratory. Thus, this problem has drawn the attention of many researchers in Bioinformatics. Considering the great difference between the number of protein sequences already known and the number of three-dimensional structures determined experimentally, the demand of automated techniques for structural classification of proteins is very high. In this context, computational tools, especially Machine Learning (ML) techniques, have become essential to deal with this problem. In this work, ML techniques are used in the recognition of protein structural classes: Decision Trees, k-Nearest Neighbor, Naive Bayes, Support Vector Machine and Neural Networks. These methods have been chosen because they represent different paradigms of learning and have been widely used in the Bioinfornmatics literature. Aiming to obtain an improvment in the performance of these techniques (individual classifiers), homogeneous (Bagging and Boosting) and heterogeneous (Voting, Stacking and StackingC) multiclassification systems are used. Moreover, since the protein database used in this work presents the problem of imbalanced classes, artificial techniques for class balance (Undersampling Random, Tomek Links, CNN, NCL and OSS) are used to minimize such a problem. In order to evaluate the ML methods, a cross-validation procedure is applied, where the accuracy of the classifiers is measured using the mean of classification error rate, on independent test sets. These means are compared, two by two, by the hypothesis test aiming to evaluate if there is, statistically, a significant difference between them. With respect to the results obtained with the individual classifiers, Support Vector Machine presented the best accuracy. In terms of the multi-classification systems (homogeneous and heterogeneous), they showed, in general, a superior or similar performance when compared to the one achieved by the individual classifiers used - especially Boosting with Decision Tree and the StackingC with Linear Regression as meta classifier. The Voting method, despite of its simplicity, has shown to be adequate for solving the problem presented in this work. The techniques for class balance, on the other hand, have not produced a significant improvement in the global classification error. Nevertheless, the use of such techniques did improve the classification error for the minority class. In this context, the NCL technique has shown to be more appropriated
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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In vivo production of viral biopesticides is the major source of viral insecticides currently in the marketplace. However, this system presents limitations during production scale-up. For the Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV), the insect used for replication has cannibalistic characteristics, thus production is even more difficult. Insect cells are commonly used for in vitro baculovirus production. Most of these cell lines are derived from Lepidoptera species. The Sf21 cell line is derived from Spodoptera frugiperda caterpillar ovarian tissue, and its clonal isolate Sf9 has been used for biopesticide production due to its ease of growth in suspension cultures. In this work, the in vitro production capabilities of a Brazilian SfMNPV isolate obtained from cornfields was evaluated. Comparison of polyhedra production was carried out using both Sf21 and Sf9 cells, based on volumetric and specific yields. Both cell lines were cultivated in Hyclone medium supplemented with different fetal bovine serum concentrations (2,5 and 5%). The best results were obtained using Sf9 cells supplemented with 5% serum. These results were further confirmed quantitively through kinetic parameter estimation for both cells lines and different serum concentrations. After seven successive passages, this system still presented high specific polyhedra production
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Visceral leishmaniosis caused by Leishmania chagasi, also known as calazar, presented, in the period from 1990 to 2005, tax of incidence in Brazil varying between 1 and 3 cases for 100 000 inhabitants. The Northeast region that up to the year of 2000 contributed with almost 90% of the registered cases is reducing his participation in the current decade, reaching 56% in 2005. Conventional leishmaniasis treatment is costly and it shows high toxicity, demanding more research for alternative treatments, with special interest in development of vaccines and diagnosis kits which include production of recombinant antigens by host cells. Escherichia coli has been the microorganism most studied and used as a host for recombinant protein production. Therefore, the aim of this work was to study the influence of induction on cellular growth and to verify the type of Leishmania chagasi antigens expression (intra or extracellular) during two recombinant E. coli clones (kmp11 and P36) cultivation in rotary incubator (shaker) using three different media (2xTY, TB, FASS+EL). For that, tests were carried out using conditions established in the literature for E. coli (37°C and 200 rpm) and media supplemented with antibiotics to guarantee that only competent cells grows. First, tests were carried out without induction in order to verify the two microorganisms kinetic behavior (growth and substrate consumption) in different media. Next, the induction was carried out through the addition of IPTG (1mM as final concentration), at the first hour of cultivation. It was observed that protein expression were intracellular for all clones and media tested, however the highest level of expression was clearly observed by the electrophoresis band density (intensity) for 2xTY medium and kmp11 protein. Although it contains the lowest substrate concentration, consequently, a reduced cellular concentration when compared to other media, it appeared that this medium and clone combination is the most indicated for recombinant protein production. Therefore, the objective of this work was achieved, since the interested proteins were produced. Consequently, this result motivates new studies for production optimization using different cultivation strategies
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Among the pests that attack corn crop in Brazil, there is Spodoptera frugiperda (JE Smith, 1797) (Lepidoptera: Noctuidae), known as fall armyworm, which is the major corn pest. Due to genetic instability during serial passage of baculoviruses in insect cell culture, the viral bioinseticides in vitro production development is the greatest challenge for mass production of this bioproduct. Successive passages of virus using extracellular viruses (BVs), necessary during viral bioinseticides production scaling up, leads to the appearance of aberrant forms of virus, a process so called as "passage effect ". The main consequence of passage effect is the production of occlusion bodies (OB) decrease, preventing its production using in vitro process. In this study, it was carried out a serial passage of baculovirus Spodoptera frugiperda multiple nucleopolyhedrovirus, isolate 18, using Sf21 cells. A decrease in the production of occlusion bodies from 170 to 92 in the third to fourth passage was observed. A factorial experimental design (22) was employed to verify the influence of two input variables, concentration of the hormone 20 - hydroxyecdysone (CH) and cholesterol (CC) on the values of response variables (volumetric and the specific OB production) of the process, seeking to define the optimum operating ranges trying to reverse or minimize the passage effect. The result indicated a negative influence of the cholesterol addition and positive effect in the hormone supplementation which the optimum range found for the concentrations studied were 8 to 10μg/mL and 5 to 6.5 mg / mL, for cholesterol and hormone concentrations respectively. New experiments were performed with addition of hormone and cholesterol in order to check the influence of these additives on the OB production independently. While the best result obtained from the factorial experiment was 9.4 x 107 OB/mL and 128.4 specific OB/cell, with the addition of only 6μg/mL 20-hydroxyecdysone these concentrations increased to 1.9 x 108 OB/mL and 182.9 OB/cell for volumetric and specific OB production, respectively. This result confirms that the addition of the hormone 20-hydroxyecdysone enhances the SfMNPV in vitro production process performance using Sf21 cells
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O presente artigo apresenta uma análise do Plano Nacional de Saúde publicado em 2004. Este documento expressa um importante período de transição na gestão do SUS, uma vez que foi predecessor do Pacto pela Saúde. A partir de um estudo descritivo com base em procedimentos quantitativos e qualitativos, o objetivo foi compreender as ideias centrais do documento, identificando as conexões existentes entre seus princípios, objetivos e prioridades. O principal resultado do estudo foi a identificação da integralidade das ações, da capacitação dos recursos humanos e mudança do marco regulatório com base numa visão intersetorial como núcleo central do documento. Essas ideias, por sua vez, circulam pelo discurso das diretrizes do plano, fortalecendo os laços do eixo central do texto na reorganização da atenção ambulatorial e na qualificação profissional. Por fim, quando comparadas metas e ações previstas nas diretrizes, observa-se uma tensão entre o que foram denominados vetores da verticalidade e da horizontalidade, deixando em aberto o rumo do lugar social em disputa.
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A aceitação de 11 amostras de aguardentes de cana envelhecidas e não envelhecidas foi avaliada por testes sensoriais afetivos e análises estatísticas uni e multivariada. As aguardentes estudadas compreenderam seis amostras comerciais de diferentes marcas, (sendo três não envelhecidas e três envelhecidas) e ainda outras cinco amostras correspondentes a zero, 12, 24, 36 e 48 meses de envelhecimento em um tonel de carvalho de 200L. As amostras foram avaliadas por 100 provadores consumidores do produto, recrutados por questionário de avaliação quanto à afetividade. Para os testes afetivos foi utilizada escala hedônica não estruturada de 9cm, sendo os dados obtidos avaliados por dois métodos estatísticos distintos: o Mapa de Preferência Interno (MDPREF) e a análise de variância univariada (ANOVA) com comparação de médias pelo teste de Tukey e análise de correlação. As amostras de aguardente envelhecidas por 12, 36 e 48 meses obtiveram maior aceitação, com médias ao redor de 7,0 na escala hedônica. A amostra com menor aceitação foi a correspondente ao tempo zero de envelhecimento (controle). As demais amostras obtiveram aceitação intermediária. A análise por MDPREF gerou em espaço multidimensional (onde as variações com relação aos dados de preferência foram extraídas em eixos ortogonais e para cada dimensão de preferência), coordenadas relativas aos produtos, que foram geradas em função da resposta dos consumidores. Os dados de aceitação de cada provador foram utilizados para o desenvolvimento de vetores individuais de preferência, resultando na construção de um mapa mutidimensional das amostras, em função dos dados de aceitação. No presente estudo o MDPREF foi gerado pelas primeira e segunda dimensões de preferência, as quais explicaram em conjunto 89,83% das variações observadas entre as amostras com relação à aceitação. O MDPREF confirmou os resultados da ANOVA, indicando uma maior preferência dos provadores pelas amostras de aguardentes envelhecidas. Os resultados sugerem também que aguardentes envelhecidas por mais de 24 meses em tonel de carvalho de 200L são preferidas pelos consumidores, em detrimento das comerciais não envelhecidas e mesmo das comerciais envelhecidas, que podem ser adicionadas de aguardente não envelhecida (processo denominado corte) e também ter correção da cor, conforme permite a Legislação Brasileira. O conteúdo de polifenóis totais e a intensidade de cor também foram determinados, e ambos apresentaram correlação linear positiva significativa (p<=0,05) com o aumento do tempo de envelhecimento das amostras.
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INTRODUÇÃO: Populações de Triatoma sordida Stål, 1859 foram investigadas quanto à suscetibilidade à deltametrina. MÉTODOS: Análise por meio de bioensaios por aplicação tópica em 11 populações de T. sordida procedentes dos Estados de Goiás, Mato Grosso e Mato Grosso do Sul. RESULTADOS: As estimativas de DL50 e RR50 demonstraram elevados níveis de suscetibilidade (DL50 < 1 e RR50 < 2). Entretanto, as análises do coeficiente angular da curva dose resposta revelaram que as populações de triatomíneos dos municípios de Firminópolis/GO, Posse/GO, Poxoréu/MT, Douradina/MS e Aparecida do Taboado/MS apresentam maiores probabilidades de evolução de resistência, portanto, mais propícias a tolerar o tratamento com deltametrina. CONCLUSÕES: Detectaram-se pequenas alterações de suscetibilidade e baixos níveis de resistência, porém as alterações temporais de suscetibilidade deverão ser continuamente monitoradas, a fim de nortear adequadamente as ações de controle dos vetores da DC.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico
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The magnetic order of bylayers composed by a ferromagnetic film (F) coupled with an antiferromagnetic film (AF) is studied. Piles of coupled monolayers describe the films and the interfilm coupling is described by an exchange interaction between the magnetic moments at the interface. The F has a cubic anisotropy while the AF has a uniaxial anisotropy. We analyze the effects of an external do magnetic field applied parallel to the interface. We consider the intralayer coupling is strong enough to keep parallel all moments of the monolayer an then they are described by one vector proportional to the magnetization of the layer. The interlayer coupling is represented by an exchange interaction between these vectors. The magnetic energy of the system is the sum of the exchange. Anisotropy and Zeeman energies and the equilibrium configuration is one that gives the absolute minimum of the total energy. The magnetization of the system is calculated and the influence of the external do field combined with the interfilm coupling and the unidirectional anisotropy is studied. Special attention is given to the region near of the transition fields. The torque equation is used to study dynamical behavior of these systems. We consider small oscillations around the equilibrium position and we negleet nonlinear terms to obtain the natural frequencies of the system. The dependence of the frequencies with the external do field and their behavior in the phase transition region is analized
Caracterização da resistência do algodoeiro a Ramularia areola e variabilidade molecular do patógeno
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This research was conducted with the aim to study the genetic and pathogenic structure of Ramularia areola isolates collected in Brazil and to characterize the resistance response in cotton plants to ramularia spot. The genetic variability of 28 isolates of R. areola was studied using RAPD markers. The pathogenicity evaluation was realized by the inoculation of 6 isolates on cotton varieties Guazuncho-2 (Gossypium hirsutum) and VH8-4602 (Gossypium barbadense). The inheritance of disease resistance was studied using an artificially inoculated population of F2 individuals derived from the intercross of Guazuncho-2 (susceptible variety) end VH8-4602 (resistant variety), and also the parents and F1 individuals. Molecular polymorphism between the G. hisutum varieties DeltaOpal (suscetible) and CNPA CO-11612 (resistant) was estimated by 118 SSR and 24 AFLP markers. The parental genotypes Guazuncho-2 and VH8-4602 were selected for mapping, and then Recombinant Inbred Lines (RIL´s) derived from this crossing were evaluated with SSR 12 markers. The analysis of population structure of R. areola revealed that the three subpopulations were genetically simillar (Gst=0.18), and the isolates from Goiás and Minas Gerais were more similar to each other (0,92). This probability can be related to the relatively high gene flow among the three subpopulations (Nm=2.20). The isolates R. areola 9.1, from Minas Gerais State and 8.1 and 8.3 from Goiás State were the most aggressive ones to the susceptible variety Guazuncho-2. The variety VH8-4602 presented high level of resistance to ramularia spot. No differential interaction was observed between the pathogens and the analyzed varieties, and the resistance was classified as horizontal. The quantification of disease by number of necrotic lesions and number of spores in individual plants of F1 and F2 generations from the crossing between the varieties Guazuncho-2 and VH8-4602 presented continuous distribution, suggesting polygenic resistance. The resistance is probabilly recessive, since necrotic lesions and sporulation were observed on F1 plants. The molecular polymorphism between DeltaOpal e CNPA CO-11612 lineages was low (6%), then would be difficult to accomplish molecular mapping of disease resistance using this intercross. With the genotyping of the RIL s it was verified that 25% of the markers segregated in the proportions proposed by Mendel s Law and 75% of the studied markers presented segregation distortion in favor to the parental G. hirsutum. Both the low genetic variability of the pathogen and the number of resistance genes suggest that durable genetic resitance may be achieved
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The development of epidemiological practices in the last years of the nineteenth and early twentieth century was characterized by both an influence of medical geography and the emergence of microbes and vectors of diseases. Both theories were used to explain outbreaks in Rio Grande do Norte specially in Natal. In this process were organized new institutions linked to public health, unhealthy spaces and prescribed hygiene measures. The redefinitions of the spaces were linked to updated elements of Hippocratic medicine such as aerism and emphasis on medical topography. How the physicians of the town were organized in the face of new meanings and fields of expertise in the demarcation of diseases and regulation of their own practices against the illegal medical practitioners? Likewise, the very occurrence of epidemics mobilized people, urban institutions and apparatuses. But how the Hippocratic legacy that leads to the idea of bad air originated by swamps from the eighteenth and nineteenth century has been linked to new microbial assumptions and disease vectors in the early twentieth century? How an invader from Africa, (the mosquito A. gambiae) mobilized transnational efforts to combat malaria and redefined the epidemiological practices? The aim of this work is to understand how epidemiological practices redefine the way we define spaces, practices and disease from both an approach influenced by a relational history of spaces and a theoretical synergy which includes topics in Science Studies, Post Structuralist Geography and some elements of Feminist Studies. Documentary research were surveyed in the reports of the provincial presidents, government posts to the Provincial Assembly, specialized medical articles and theses, and documents from the Rockefeller Foundation and national and international journals. In this regard shall be given to both material and discursive aspects of space-related practical epidemiological that Natal as much (in general) Rio Grande do Norte between bad air and malaria.
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Oral squamous cell carcinoma (OSCC) is the most common malignancy in oral cavity and human papillomavirus (HPV) may have an important role in its development. The aim of this experiment was to investigate the HPV DNA and viral types in 90 cases of OSCC. Moreover, a comparative analysis between the cases of OSSC with and without HPV DNA was performed by using cell cycle markers p21 and pRb in order to detect a possible correlation of these proteins and HPV infection. DNA was extracted from paraffin embedded tissue and amplified by PCR (polymerase chain reaction) with primers PCO3+ e PCO4+ for a fragment of human β-globin gene. After this procedure, PCR for HPV DNA detection was realized using a pair of generic primers GP5+ e GP6+. Immunohistochemical study was performed by streptoavidin-biotin technique and antibodies against p21 and pRb proteins were employed. Eighty-eight cases were positive for human β-globin gene and HPV DNA was found in 26 (29.5%) of then. It could not be detected significant correlation between HPV and age, sex and anatomical sites of the lesion. The most prevalent viral type was HPV 18 (80.8%). Regarding the immunohistochemical analysis, it was detected significant association between HPV presence and pRb immunoexpression (p=0,044), nevertheless, the same was not observed in relation to p21 protein (p =0,416). It can be concluded that the low detection of HPV DNA in OSCC by the present experiment suggests a possible role of the virus in the development and progression in just a subset of this disease
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Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the oral cavity and reach a large number of individuals, has become an important public health problem. Studies have demonstrated changes in pathway components BMP in various types of cancers as prostate, colon, breast, gastric and OSCCs. Is the current knowledge that these proteins may exert pro-tumor effect in more advanced stages of neoplastic development coming to favor progression and invasion tumor. The inhibition of the signaling pathway BMP-2 through its antagonists, have shown positive results of antitumor activity and use of Noggin may be a novel therapeutic target for cancer. Given this evidence and the few studies with BMP-2, Noggin and OSCC, the objective of this research was to evaluate the effect of BMP-2 and its antagonist Noggin on proliferation and migration cell in line of cell cultures of human tongue squamous cell carcinoma (SCC25). The study was divided in three groups, a control group, where SCC25 cells suffered no treatment, a BMP-2 group, in which cells were treated with 100ng/ml of BMP-2 and a group of cells that were treated with 100ng/ml of Noggin. For the proliferation assay and cell cycle were established three time intervals (24, 48 and 72 hours). Proliferative activity was investigated by trypan blue and cell cycle analysis by staining with propidium iodide flow cytometry. The potential for migration / invasion of SCC25 cells was performing by a cell invasion assay using Matrigel in a 48-hour interval. The proliferation curve showed a higher proliferation in cells treated with BMP-2 in 72 hours (p < 0.05), and lower overgrowth and cell viability in Noggin group. Recombinant proteins favored a greater percentage of cells in cell cycle phase Go/G1 with a statistically significant difference in the interval of 24 hours (p < 0.05). BMP- 2 produced a greater invasion of cells studied as well as its antagonist Noggin inhibits invasion of cells (p < 0.05). Thus, these results indicate that BMP-2 promotes malignant phenotype, dues stimulates proliferation and invasion of SCC25 cells and, its antagonist Noggin may be an alternative treatment, due to inhibit the tumor progression