996 resultados para Vetores virais


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O material trata dos conceitos de Variáveis Compostas Homogêneas, mais conhecidas como vetores ou arranjos. Vetores ou arranjos são variáveis capazes de manter uma coleção de valores e, todos os valores desses vetores são dos mesmos tipos de dados e cada valor individual é referenciado por um índice do tipo inteiro. O material, após as explicitações dos conceitos, aborda exemplos práticos da utilização das Variáveis Compostas Homogêneas.

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Esta videoaula explica sobre a dependência linear, ou seja, um tipo de relação entre os vetores de um conjunto. Um conjunto de vetores é linearmente dependente se, e só se, um dos vetores do conjunto for combinação linear dos demais vetores do conjunto. Assim, quando escrevemos um vetor como combinação linear e seus coeficientes são todos nulos, ele é linearmente dependente. Quando não existe relação linear entre os vetores diz-se que o conjunto é linearmente independente

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Parte 1: Introdução às planilhas eletrônicas Parte 2: O ambiente de trabalho do programa BrOffice.org Calc Parte 3: Primeiros passos Parte 4: Trabalhando com vetores e matrizes Parte 5: Construindo gráficos Parte 6: Resolvendo problemas Parte 07: “Travando” uma célula na planilha

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A terapia genética tem se revelado uma ferramenta potente na Medicina, na tentativa de revolucionar o tratamento de várias doenças hereditárias e adquiridas. A introdução de genes em células pretende a expressão estável e prolongada de proteínas com efeitos terapêuticos. O silenciamento de genes, através da terapia genética que faz uso de oligonucleótidos antisense, pequenos RNA de interferência (siRNA) ou ribozimas, visa o decréscimo ou anulação do funcionamento de um gene cuja expressão amplificada, por algum motivo, leva ao desenvolvimento de umapatologia. A internalização de material genético nas células, usualmente, carece de métodos e/ou sistemas de entrega (vectores). Estes podem pertencer a duas categorias, designadamente, métodos virais e métodos não-virais. O primeiro é considerado o mais eficiente, apresentando porém, sérias desvantagens como o risco de carcinogénese. A solução é a utilização de métodos não virais,que podem ser físicos ou químicos. O objectivo principal desta dissertação foi a utilização de dendrímeros para o silenciamento do gene da proteína fluorescente optimizada (EGFP) em células HeLa, previamente modificadas para expressarem esta proteína. Dendrímeros poli(amidoamina) geração 5 (PAMAM G5) modificados com 4 ou 8 moléculas de ácidos gordos de diferentes comprimentos foram complexados com oligonucleótidos antisense. A vantagem que estes apresentam em relação aos dendrímeros nativos é que são capazes de interagir com os lípidos da membrana celular, esperando-se, por isso, uma melhor eficiência de transfecção e efeitos antisense. Isto foi efectivamente verificado, sendo que o nível de silenciamento do gene da EGFP obtido, está directamente relacionado com o aumento da razão NP, o número e o comprimento das cadeias hidrofóbicas. O silencimento de genes tem sofrido grandes avanços, havendo actualmente uma série de ensaios clínicos para a sua utilização no tratamento de doenças como cancros de origem hereditária ou viral, prevendo-se que venha para ficar, juntamente com o silenciamento mediado por siRNA.

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A Wolbachiapertence a um grupo de bactérias intracelulares, transmitidas maternalmente, que se encontram amplamente distribuídas nos artrópodes. Estes endossimbiontes encontram-se normalmente nos tecidos do sistema reprodutor dos artrópodes e têm a capacidade manipular a sua reproduçãode modo a garantir a sua transmissão à descendência e rápida dispersão na população.A capacidade de manipulação reprodutiva da Wolbachia tornou-a o alvo de diversos estudos para uma maior e melhor perceção da sua implicação em processos biológicos e evolutivos e por acreditar-se que esta bactéria é uma promissora ferramenta no controlo de populações de insetos que são pragas agrícolas. Os afídeos são um grupo de insetos associados às plantas que podem ter um efeito devastador nas culturas agrícolas e hortícolas pois não só retiram nutrientes às plantas como podem ser vetores de doenças. Embora durante muito tempo se pensasse que estes insetos não albergavam a Wolbachia estudos recentes mostram que são várias as espécies de afídeos infetados com esta bactéria. O principal objetivo deste trabalho é estudar a prevalência de infeção por Wolbachia assim como a caracterização das suas estirpes em amostras de afídeos dos Arquipélagos da Madeira e dos Açores. Neste estudo foram analisadas 545 amostras de afídeos, 361 provenientes do Arquipélago da Madeira e 184 dos Açores. Utilizando a técnica da “Polymerase Chain Reaction” (reação em cadeia da polimerase) amplificou-se o gene 16S rRNA (RNA ribossomal) e verificou-se que 32 destas amostras encontravam-se infetadas com Wolbachia sendo a maior parte das amostras infetadas provenientes dos Açores. Para determinar a estirpe que infeta estes afídeos utilizou-se a tipagem sequencial multilocus (MLST) com os genesglutamil-tRNA amidotransferase, subunidade B (gatB), citocromo c oxidase, subunidade I (coxA), proteína hipotética conservativa (hcpA) e proteína da divisão celular (ftsZ). A análise filogenética realizada para os diferentes genes mostrou que grande parte das amostras analisadas estão incluídas em dois dos novos supergrupos descobertos para Wolbachia, supergrupo M e N.Foi detetada a presença da mesma estirpe de Wolbachia, supergrupo N, em duas espécies diferentes de afídeos, Neophyllaphis podocarpi e Aphis spiraecola da mesma planta hospedeira, Podocarpus macrophyllus. Esta infeção reforça a ideia de que a Wolbachia não recorre só a transmissão vertical para se difundir na população mas utiliza também a transmissão horizontal. A deteção e caracterização das estirpes de Wolbachia é essencial para um maior entendimento sobre a sua origem e forma de disseminação. Esta informação é importante para desenvolvimento de estratégias de controlo de pestes recorrendo a estes endossimbiontes.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

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American visceral leishmaniasis is a zoonosis caused by Leishmania infantum and transmitted by the bite of the sand flies Lutzomia longipalpis.The main domestic reservoir is the dog, while foxes and opposums are the known wild reservoirs. However, identification of natural infections with L. infantum in rodents appears for need of investigating the participation of these rodents how source of infection of the parasite. In the present work the Leishmania infantum infection was investigated in rodents captured in Rio Grande do Norte, aiming at to offer subsidies to the understanding of the epidemic chains of LVA in the State. Thirteen Galea spixii were distributed in four groups, being G1 the group control with four animals and the others, G2, G3 and G4, with three animals each. Those animals were intraperitoneally inoculated with 107 promastigotas of L. infantum and accompanied for, respectively, 30, 90 and 180 days. Weekly the animals were monitored as for the corporal weight and rectal temperature. At the end of each stipulated period the animals were killed. Blood were used for determination of the parameters biochemical and haematological, PCR, ELISA, microscopic examination and cultivation in NNN medium. Liver, spleen and lymph node were used in Giemsa-stained impression and cultivation in NNN medium. Liver and spleen fragments were still used in PCR and histopathological, respectively. At the same time 79 rodents of the species Rattus rattus, Bolomys lasiurus, Oligoryzomys nigripis, Oryzomys subflavus and Trichomys apereoides were captured in the Municipal districts of Brejinho, Campo Grande, Coronel Ezequiel, Passa e Fica and Vázea for identification of natural infection with L. infantum. Evidence of infection was checked by direct examination of Giemsa-stained impression of liver, spleen and blood and culture of these tissues in NNN medium. Antibodies were researched by ELISA. They were not found differences among the weigh corporal final, rectal temperature and biochemical and haematological parameters of the Galea spixii controls and infected. The rectal temperature of the animals varied from 36OC to 40OC. For the first time values of the haematocrit (33,6% to 42,8%), hemoglobin (10,2 to 14,5g/dl), erythrocyts number (4,67x106 to 6,90x106/mm3), total leukocytes (0,9x103 to 9,2x103/mm3), platelets (49x103 to 509x103/mm3) total proteins (1,56 to 6,06 g/dl), albumin (1,34 to 3,05 g/dl) and globulins (0,20 to 3,01 g/dl) of the Galea spixii were determined. The lymphocytes were the most abundant leucocytes. Infection for L. infantum was diagnosed in two animals euthanasied 180 days after the infection. In one of the animals was also identified antibodies anti-Leishmania. The parasite was not found in none of the five other species of rodents captured. Galea spixii are resistant to the infection for L. infantum and they are not good models for the study for visceral leishmaniose, although they can act as infection sources. More studies are necessary to determine the paper of the rodents in the epidemic chain of transmission of the visceral leishmaniose in the State of Rio Grande do Norte

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Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites

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The State of Paraíba is one of the most dynamic states of Brazil, strategically located in the northeast, is notable for the excellent potential for integration of different transportation modes forming the states of Rio Grande do Norte, Pernambuco and Alagoas. The dynamic that occurs with port activity causes changes in the space where it is installed. And the elements of this space are always more than suffering direct or indirect influences as the flow in the port is expanded. Therefore, this region became subject to the accidental spillage of oil, because it presents a heavy traffic of ships of various sizes that can run aground or collide with oil causing accidental events. The study of geomorphological and sedimentological compositions of seafloor becomes important as more is known about the relationships between these parameters and associated fauna, and can identify their preferred habitats. The database background, acoustically collected along the proposed study area, is a wealth of information, which were duly examined, cataloged and made available. Such information can serve as an important tool, providing a geomorphological survey of the sedimentary area studied, and come to subsidize, in a flexible, future decision making. With the study area Port of Cabedelo, Paraíba - Brazil, this research aimed to evaluate the influence of the tidal surface and background in modeling the seabed, including the acquisition of information about the location of submerged rocky bodies and the depth of these bodies may turn out to be natural traps for the trapping of oil in case of leaks, and obtain the relationship between types of bed and the hydrodynamic conditions present in the region. In this context, for this study were collected bathymetric data (depth) and physical oceanographic (height of water column, water temperature, intensity and direction of currents, waves and turbidity), meteorological (rainfall, air temperature, humidity, winds and barometric pressure) of the access channel to the Port of Cabedelo / PB and its basin evolution (where the cruise ships dock), and includes tools of remote sensing (Landsat 7 ETM +, 2001), so that images and the results are integrated into Geographic Information Systems and used in the elaboration of measures aimed at environmental protection areas under the influence of this scale facilities, serving as a grant to prepare a contingency plan in case of oil spills in the region. The main findings highlight the techniques of using hydroacoustic data acquisition together bathymetric surveys of high and low frequency. From there, five were prepared in bathymetric pattern of Directorate of Hydrography and Navigation - DHN, with the depth in meters, on a scale of 1:2500 (Channel and Basin Evolution of Access to Port of Cabedelo), where there is a large extent possible beachrocks that hinder the movement of vessels in the port area, which can cause collisions, running aground and leaking oil. From the scatter diagram of the vectors of currents, it can be seen as the tidal stream and undergoes a channeling effect caused by the bidirectional effect of the tide (ebb and flood) in the basin of the Port of Cabedelo evolution in NW-direction SE and the highest speed of the currents occurs at low tide. The characterization weather for the period from 28/02 to 04/07/2010 values was within the expected average for the region of study. The multidisciplinary integration of products (digital maps and remote sensing images), proved to be efficient for the characterization of underwater geomorphological study area, reaching the aim to discriminate and enhance submerged structures, previously not visible in the images

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Toxoplasmosis is a zoonosis of worldwide distribution caused by the protozoan Toxoplasma gondii, triggering dangerous complications in immunocompromised patients and pregnant women, as well as having great economic impact for the livestock. So far the control of toxoplasmosis is made primarily by chemotherapy. However, most drugs used routinely have some limitations. In order to control this disease, several research groups, including ours, has been working to develop a medical-veterinary vaccine based on parasite antigens, vectors and protocols of immunization. In this study were implemented and standardized methodologies for amplification and cloning of recombinant immunogens in the system for the development of a prototype vaccine, based on the surface antigens of T. gondii and recombinant adenovirus encoding these antigens. Genes encoding BAG1, GRA2 and SAG1 proteins were amplified. We established a strategy for cloning SAG1, SAG2, SAG3 and TgAMA1- genes in recombinant system. The genes encoding SAG1 and SAG2 were cloned and their sequences showed high similarity with sequences from GenBank. The virtual translation of these proteins showed polymorphisms in the amino acid sequence, which can be correlated with levels of antigenicity. Simultaneously, the adenovirus encoding the SAGs (HAdSAGs) were expanded, purificated and characterizated. Immunization of C57bl/6 mice, using viral supernatant was not enought to elicit immune responses at high levels, being required HAdSAGs titration for future immunizations. Therefore, this study allowed the cloning of the two genes important for the development of a prototype vaccine. Besides, implementations methodologies that permit advancements in the development of a vaccine against toxoplasmosis using adenovirus to express proteins of the parasite

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Control of human visceral leishmaniasis in endemic regions is hampered in part by the lack of knowledge with respect of the role reservoirs and vector. In addition, there is not yet an understanding of how non-symptomatic subclinical infection might influence the maintenance of infection in a particular locality. Of worrisome is the limited accessibility to medical care in places with emerging drug resistance. There is still no available protective vaccine either for humans or other reservoirs. Leishmania species are protozoa that express multiple antigens which are recognized by the vertebrate immune system. Since there is not one immunodominant epitope recognized by most hosts, strategies must be developed to optimize selection of antigens for prevention and immunodiagnosis. For this reason, we generated a cDNA library from the intracellular amastigote form of Leishmania chagasi, the causative agent of South American visceral leishmaniasis. We employed a two-step expression screen of the library to systematically identify T and T-dependent B cell antigens. The first step was aimed at identifying the largest possible number of clones producing an epitope-containing polypeptide with a pool of sera from Brazilians with documented visceral leishmaniasis. After removal of clones encoding heat shock proteins, positive clones underwent a second step screen for their ability to cause proliferation and IFN-γ responses of T cells from immune mice. Six unique clones were selected from the second screen for further analysis. The clones encoded part of the coding sequence of glutamine synthetase, transitional endoplasmic reticulum ATPase, elongation factor 1γ, kinesin K-39, repetitive protein A2, and a hypothetical conserved protein. Humans naturally infected with L. chagasi mounted both cellular and antibody responses to these protein Preparations containing multiple antigens may be optimal for immunodiagnosis and protective vaccines against Leishmania

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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The drug targeting has been the subject of extensive studies in order to develop site-specific treatments that minimize side effects and become more effective anticancer therapy. Despite considerable interest in this class, drugs like antibiotics also have limitations, and have been neglected. Using new pharmaceutical technologies, the use of magnetic vectors appear as promising candidate for drug delivery systems in several studies. Small magnetic particles bound to the drug of interest can be modulated according to the orientation of a magnet outside the body, locating and holding in a specific site. In this work, we propose the use of High Energy Milling (HEM) for synthesis of a magnetic vector with characteristics suitable for biomedical applications by intravenous administration, and for the formation of an oxacillin-carrier complex to obtain a system for treating infections caused by Staphylococcus aureus. The results of the variation of milling time showed that the size and structural properties of the formed material change with increasing milling time, and in 60 hours we found the sample closest to the ideal conditions of the material. The vector-drug system was studied in terms of structural stability and antimicrobial activity after the milling process, which revealed the integrity of the oxacillin molecule and its bactericidal action on cultures of Staphylococcus aureus ATCC

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Gene therapy is based on the transfer of exogenous genetic material into cells or tissues in order to correct, supplement or silencing a particular gene. To achieve this goal, efficient vehicles, viral or non-viral, should be developed. The aim of this work was to produce and evaluate a nanoemulsion system as a possible carrier for no-viral gene therapy able to load a plasmid model (pIRES2-EGFP). The nanoemulsion was produced by the sonication method, after been choose in a pseudo-ternary phase diagram build with 5 % of Captex 355®, 1.2 % of Tween 80®, 0.8 % of Span 80®, 0.16% of stearylamine and water (to 100 %). Measurements of droplet size, polydispersity index (PI), zeta potential, pH and conductivity, were performed to characterize the system. Results showed droplets smaller than 200 nm (PI < 0.2) and zeta potential > 30 mV. The formulation pH was near to 7.0 and conductivity was that expected to oil in water systems (70 to 90 μS/s) A scale up study, the stability of the system and the best sterilization method were also evaluated. We found that the system may be scaled up considering the time of sonication according to the volume produced, filtration was the best sterilization process and nanoemulsions were stable by 180 days at 4 ºC. Once developed, the complexation efficiency of the plasmid (pDNA) by the system was tested by agarose gel electrophoresis retardation assay.. The complexation efficiency increases when stearylamine was incorporated into aqueous phase (from 46 to 115 ng/μL); regarding a contact period (nanoemulsion / pDNA) of at least 2 hours in an ice bath, for complete lipoplex formation. The nanoemulsion showed low toxicity in MRC-5 cells at the usual transfection concentration, 81.49 % of survival was found. So, it can be concluded that a nanoemulsion in which a plasmid model was loaded was achieved. However, further studies concerning transfectation efficiency should be performed to confirm the system as non-viral gene carrier

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This dissertation aims to comprehend how we meet with the other at cinema. For this, it receives the doubt, the uncertainty, the way that the human being is unfinished as stance of observation. It takes into consideration the multiples social vectors, historical, subjective, temporal and cultural which constitute these phenomena. The cinema is used as a cognitive resource in order to incorporate the poetic and the imaginary, making possible not forgetting gestures and knowledge that hatch from sounds, colours, remembrances, images, words, social exchanges, cognitive reciprocity that allow us to think about the culture, the society and the communication. For this, it has as theoretical prerequisites ideas of double (myselfthe other) and effective participation (projection-identification) developed by Edgar Morin, as well as the idea of communication as possibility/impossibility of constructions of bonds with the other through the body, our primary media, concepts developed by Harry Pross and Norval Baitello Jr. The chosen film to analysis is Lavoura Arcaica, it is an emblematic movie in the discussion of the human condition and in the relationship between the man and its body, its desires, with its nature/culture ambivalence. About this film are made reflections about the process of activation of the double (myself-the other) in the searching for the other through cinematographically elements of construction, as: montage, direction, screenplay, actors, scenography, costumes and soundtrack. In conclusion, the cinema, when it allows the effective participation and provides t e experiment of the double, becomes a gateway line in the path to development of the alterity