Produção e avaliação de nanoemulsão catiônica como possível vetor não-viral para pIRES2-EGFP

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

Uso da internet em ambiente acadêmico :estudo com docentes em institruição de educação tecnológica no Brasi

The last decade, characterized by the vertiginous growth of the computers worldwide net, brought radical changes in the use of the information and communication. Internet s use at business world has been largely studied; however, few are the researches about the academic use of this technology, mainly if we take into consideration institutions of technologic education. In this context, this research made an analysis of internet s use in a technologic education institution in Brazil, analyzing, in particular, the Centro Federal de Educação Tecnológica do Rio Grande do Norte CEFET/RN for that standard use of this Information Technology (IT) tools and, at the same time, studying the determinant factors of this use. To reach the considered objectives, a survey research was effected, be given data collected daily through the research s questionnaire application to 150 teachers who answered a set of closed and scaled questions. The quantitative data were qualitatively analyzed, arriving a some significant results related to the standard use and the factors that influenced in the use of these Internet technologies, like: the age scale, the exposition s to the computer level, the area of academic graduation, the area of knowledge where acts and the title, exert significant influence in the academic use of Internet between the professors

Dinâmica da Plasticidade Sináptica em neurônios do hipocampo durante ciclos de sono: um estudo computacional

Several research lines show that sleep favors memory consolidation and learning. It has been proposed that the cognitive role of sleep is derived from a global scaling of synaptic weights, able to homeostatically restore the ability to learn new things, erasing memories overnight. This phenomenon is typical of slow-wave sleep (SWS) and characterized by non-Hebbian mechanisms, i.e., mechanisms independent of synchronous neuronal activity. Another view holds that sleep also triggers the specific enhancement of synaptic connections, carrying out the embossing of certain mnemonic traces within a lattice of synaptic weights rescaled each night. Such an embossing is understood as the combination of Hebbian and non-Hebbian mechanisms, capable of increasing and decreasing respectively the synaptic weights in complementary circuits, leading to selective memory improvement and a restructuring of synaptic configuration (SC) that can be crucial for the generation of new behaviors ( insights ). The empirical findings indicate that initiation of Hebbian plasticity during sleep occurs in the transition of the SWS to the stage of rapid eye movement (REM), possibly due to the significant differences between the firing rates regimes of the stages and the up-regulation of factors involved in longterm synaptic plasticity. In this study the theories of homeostasis and embossing were compared using an artificial neural network (ANN) fed with action potentials recorded in the hippocampus of rats during the sleep-wake cycle. In the simulation in which the ANN did not apply the long-term plasticity mechanisms during sleep (SWS-transition REM), the synaptic weights distribution was re-scaled inexorably, for its mean value proportional to the input firing rate, erasing the synaptic weights pattern that had been established initially. In contrast, when the long-term plasticity is modeled during the transition SWSREM, an increase of synaptic weights were observed in the range of initial/low values, redistributing effectively the weights in a way to reinforce a subset of synapses over time. The results suggest that a positive regulation coming from the long-term plasticity can completely change the role of sleep: its absence leads to forgetting; its presence leads to a positive mnemonic change

Estratégias de Produção in vitro de Bioinseticida Viral: influências do Isolado, da Cinética e do Modo de operação

Societal concerns about environmental sustainability has lead to the development of ecologically-friendly alternatives to chemical insecticides for crop protection. One such alternative is biological pest control. In particular, baculoviruses are well suited as insect biopesticides due to their narrow host specificity and relative ease of propagation. In Brazil, the baculovirus Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) is the main biological control agent employed for the soybean pest, Anticarsia gemmatalis. This baculovirus biopesticide is currently produced using caterpillars, but increasing market demand for the product has encouraged the development of an in vitro manufacturing process, which can be scaled up to much higher virus productivities. In this study, three wild-type AgMNPV isolates (AgMNPV-2D, AgMNPV-MP2 and AgMNPV-MP5) and a recombinant form (vAgEGT-LacZ) were characterised in terms of occlusion body (OB) production and infection kinetics, to enable future optimisation of the in vitro production process. These viruses were propagated using a Spodoptera frugiperda (IPLB-SF21) insect cell line grown in shaker-flask batch cultures. Among the virus isolates tested, AgMNPV-MP5 was found to be the best producer, yielding (5.3±0.85)x108 OB/mL after 8 days post infection. The characterisation of vAgEGT-LacZ propagation in suspension cell cultures has not been previously reported in the literature; hence it became the main focus for this thesis. In particular, it was carried out a study on the effect of the multiplicity of infection (MOI) on OB production. Five successive batches were performed getting a final production (8.9±1.42)x1014 occlusion bodies, considering that production is related for a bioreactor with final volume of 10m3. A low MOI associated with a fed-batch process for vAgEGT-LacZ production was found to support a 3-fold higher OB yield when compared to the default batch process (1.8x107 and 5.3x107 OB/mL, respectively). This yield is competitive with regards to the production process.

Aplicação do Sensoriamento Remoto e do Sistema de Informações Geográficas na Detecção de Manchas de Óleo na Região do Pólo de Exploração de Guamaré, R.N.

Objective to establish a methodology for the oil spill monitoring on the sea surface, located at the Submerged Exploration Area of the Polo Region of Guamaré, in the State of Rio Grande do Norte, using orbital images of Synthetic Aperture Radar (SAR integrated with meteoceanographycs products. This methodology was applied in the following stages: (1) the creation of a base map of the Exploration Area; (2) the processing of NOAA/AVHRR and ERS-2 images for generation of meteoceanographycs products; (3) the processing of RADARSAT-1 images for monitoring of oil spills; (4) the integration of RADARSAT-1 images with NOAA/AVHRR and ERS-2 image products; and (5) the structuring of a data base. The Integration of RADARSAT-1 image of the Potiguar Basin of day 21.05.99 with the base map of the Exploration Area of the Polo Region of Guamaré for the identification of the probable sources of the oil spots, was used successfully in the detention of the probable spot of oil detected next to the exit to the submarine emissary in the Exploration Area of the Polo Region of Guamaré. To support the integration of RADARSAT-1 images with NOAA/AVHRR and ERS-2 image products, a methodology was developed for the classification of oil spills identified by RADARSAT-1 images. For this, the following algorithms of classification not supervised were tested: K-means, Fuzzy k-means and Isodata. These algorithms are part of the PCI Geomatics software, which was used for the filtering of RADARSAT-1 images. For validation of the results, the oil spills submitted to the unsupervised classification were compared to the results of the Semivariogram Textural Classifier (STC). The mentioned classifier was developed especially for oil spill classification purposes and requires PCI software for the whole processing of RADARSAT-1 images. After all, the results of the classifications were analyzed through Visual Analysis; Calculation of Proportionality of Largeness and Analysis Statistics. Amongst the three algorithms of classifications tested, it was noted that there were no significant alterations in relation to the spills classified with the STC, in all of the analyses taken into consideration. Therefore, considering all the procedures, it has been shown that the described methodology can be successfully applied using the unsupervised classifiers tested, resulting in a decrease of time in the identification and classification processing of oil spills, if compared with the utilization of the STC classifier