936 resultados para Repetitive-element-based PCR assays


Relevância:

30.00% 30.00%

Publicador:

Resumo:

Model updating methods often neglect that in fact all physical structures are damped. Such simplification relies on the structural modelling approach, although it compromises the accuracy of the predictions of the structural dynamic behaviour. In the present work, the authors address the problem of finite element (FE) model updating based on measured frequency response functions (FRFs), considering damping. The proposed procedure is based upon the complex experimental data, which contains information related to the damped FE model parameters and presents the advantage of requiring no prior knowledge about the damping matrix structure or its content, only demanding the definition of the damping type. Numerical simulations are performed in order to establish the applicability of the proposed damped FE model updating technique and its results are discussed in terms of the correlation between the simulated experimental complex FRFs and the ones obtained from the updated FE model.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

Combined tunable WDM converters based on SiC multilayer photonic active filters are analyzed. The operation combines the properties of active long-pass and short-pass wavelength filter sections into a capacitive active band-pass filter. The sensor element is a multilayered heterostructure produced by PE-CVD. The configuration includes two stacked SiC p-i-n structures sandwiched between two transparent contacts. Transfer function characteristics are studied both theoretically and experimentally. Results show that optical bias activated photonic device combines the demultiplexing operation with the simultaneous photodetection and self amplification of an optical signal acting the device as an integrated photonic filter in the visible range. Depending on the wavelength of the external background and irradiation side, the device acts either as a short- or a long-pass band filter or as a band-stop filter. The output waveform presents a nonlinear amplitude-dependent response to the wavelengths of the input channels. A numerical simulation and two building-blocks active circuit is presented and gives insight into the physics of the device.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

Combined tunable WDM converters based on SiC multilayer photonic active filters are analyzed. The operation combines the properties of active long-pass and short-pass wavelength filter sections into a capacitive active band-pass filter. The sensor element is a multilayered heterostructure produced by PE-CVD. The configuration includes two stacked SiC p-i-n structures sandwiched between two transparent contacts. Transfer function characteristics are studied both theoretically and experimentally. Results show that optical bias activated photonic device combines the demultiplexing operation with the simultaneous photodetection and self amplification of an optical signal acting the device as an integrated photonic filter in the visible range. Depending on the wavelength of the external background and irradiation side, the device acts either as a short- or a long-pass band filter or as a band-stop filter. The output waveform presents a nonlinear amplitude-dependent response to the wavelengths of the input channels. A numerical simulation and a two building-blocks active circuit are presented and give insight into the physics of the device. (c) 2013 Elsevier B.V. All rights reserved.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

The structural integrity of multi-component structures is usually determined by the strength and durability of their unions. Adhesive bonding is often chosen over welding, riveting and bolting, due to the reduction of stress concentrations, reduced weight penalty and easy manufacturing, amongst other issues. In the past decades, the Finite Element Method (FEM) has been used for the simulation and strength prediction of bonded structures, by strength of materials or fracture mechanics-based criteria. Cohesive-zone models (CZMs) have already proved to be an effective tool in modelling damage growth, surpassing a few limitations of the aforementioned techniques. Despite this fact, they still suffer from the restriction of damage growth only at predefined growth paths. The eXtended Finite Element Method (XFEM) is a recent improvement of the FEM, developed to allow the growth of discontinuities within bulk solids along an arbitrary path, by enriching degrees of freedom with special displacement functions, thus overcoming the main restriction of CZMs. These two techniques were tested to simulate adhesively bonded single- and double-lap joints. The comparative evaluation of the two methods showed their capabilities and/or limitations for this specific purpose.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

This study is based on a previous experimental work in which embedded cylindrical heaters were applied to a pultrusion machine die, and resultant energetic performance compared with that achieved with the former heating system based on planar resistances. The previous work allowed to conclude that the use of embedded resistances enhances significantly the energetic performance of pultrusion process, leading to 57% decrease of energy consumption. However, the aforementioned study was developed with basis on an existing pultrusion die, which only allowed a single relative position for the heaters. In the present work, new relative positions for the heaters were investigated in order to optimize heat distribution process and energy consumption. Finite Elements Analysis was applied as an efficient tool to identify the best relative position of the heaters into the die, taking into account the usual parameters involved in the process and the control system already tested in the previous study. The analysis was firstly developed with basis on eight cylindrical heaters located in four different location plans. In a second phase, in order to refine the results, a new approach was adopted using sixteen heaters with the same total power. Final results allow to conclude that the correct positioning of the heaters can contribute to about 10% of energy consumption reduction, decreasing the production costs and leading to a better eco-efficiency of pultrusion process.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

Functionally graded composite materials can provide continuously varying properties, which distribution can vary according to a specific location within the composite. More frequently, functionally graded materials consider a through thickness variation law, which can be more or less smoother, possessing however an important characteristic which is the continuous properties variation profiles, which eliminate the abrupt stresses discontinuities found on laminated composites. This study aims to analyze the transient dynamic behavior of sandwich structures, having a metallic core and functionally graded outer layers. To this purpose, the properties of the particulate composite metal-ceramic outer layers, are estimated using Mod-Tanaka scheme and the dynamic analyses considers first order and higher order shear deformation theories implemented though kriging finite element method. The transient dynamic response of these structures is carried out through Bossak-Newmark method. The illustrative cases presented in this work, consider the influence of the shape functions interpolation domain, the properties through-thickness distribution, the influence of considering different materials, aspect ratios and boundary conditions. (C) 2014 Elsevier Ltd. All rights reserved.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

Sandwich structures with soft cores are widely used in applications where a high bending stiffness is required without compromising the global weight of the structure, as well as in situations where good thermal and damping properties are important parameters to observe. As equivalent single layer approaches are not the more adequate to describe realistically the kinematics and the stresses distributions as well as the dynamic behaviour of this type of sandwiches, where shear deformations and the extensibility of the core can be very significant, layerwise models may provide better solutions. Additionally and in connection with this multilayer approach, the selection of different shear deformation theories according to the nature of the material that constitutes the core and the outer skins can predict more accurately the sandwich behaviour. In the present work the authors consider the use of different shear deformation theories to formulate different layerwise models, implemented through kriging-based finite elements. The viscoelastic material behaviour, associated to the sandwich core, is modelled using the complex approach and the dynamic problem is solved in the frequency domain. The outer elastic layers considered in this work may also be made from different nanocomposites. The performance of the models developed is illustrated through a set of test cases. (C) 2015 Elsevier Ltd. All rights reserved.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

We show here a simplified RT-PCR for identification of dengue virus types 1 and 2. Five dengue virus strains, isolated from Brazilian patients, and yellow fever vaccine 17DD as a negative control, were used in this study. C6/36 cells were infected and supernatants were collected after 7 days. The RT-PCR, done in a single reaction vessel, was carried out following a 1/10 dilution of virus in distilled water or in a detergent mixture containing Nonidet P40. The 50 µl assay reaction mixture included 50 pmol of specific primers amplifying a 482 base pair sequence for dengue type 1 and 210 base pair sequence for dengue type 2. In other assays, we used dengue virus consensus primers having maximum sequence similarity to the four serotypes, amplifying a 511 base pair sequence. The reaction mixture also contained 0.1 mM of the four deoxynucleoside triphosphates, 7.5 U of reverse transcriptase, 1U of thermostable Taq DNA polymerase. The mixture was incubated for 5 minutes at 37ºC for reverse transcription followed by 30 cycles of two-step PCR amplification (92ºC for 60 seconds, 53ºC for 60 seconds) with slow temperature increment. The PCR products were subjected to 1.7% agarose gel electrophoresis and visualized by UV light after staining with ethidium bromide solution. Low virus titer around 10 3, 6 TCID50/ml was detected by RT-PCR for dengue type 1. Specific DNA amplification was observed with all the Brazilian dengue strains by using dengue virus consensus primers. As compared to other RT-PCRs, this assay is less laborious, done in a shorter time, and has reduced risk of contamination

Relevância:

30.00% 30.00%

Publicador:

Resumo:

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do Grau de Mestre em Engenharia Biomédica

Relevância:

30.00% 30.00%

Publicador:

Resumo:

We have developed a procedure for the rapid diagnosis of plague that also allows the identification of prominent virulence markers of Y. pestis strains. This procedure is based upon the use of a single polymerase chain reaction with multiple pairs of primers directed at genes present in the three virulence plasmids as well as in the chromosomal pathogenicity island of the bacterium. The technique allowed the discrimination of strains which lacked one or more of the known pathogenic loci, using as template total DNA obtained from bacterial cultures and from simulated blood cultures containing diluted concentration of bacteria. It also proved effective in confirming the disease in a blood culture from a plague suspected patient. As the results are obtained in a few hours this technique will be useful in the methodology of the Plague Control Program.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

The precise microenvironment of Paracoccidioides brasiliensis has not yet been discovered perhaps because the methods used are not sensitive enough. We applied to this purpose the polymerase chain reaction (PCR) using three sets of specific primers corresponding to two P. brasiliensis genes. This fungus as well as several other fungi, were grown and their DNA obtained by mechanical disruption and a phenol chloroform isoamylalcohol-based purification method. The DNA served for a PCR reaction that employed specific primers from two P. brasiliensis genes that codify for antigenic proteins, namely, the 27 kDa and the 43 kDa. The lowest detection range for the 27 kDa gene was 3 pg. The amplification for both genes was positive only with DNA from P. brasiliensis; additionally, the mRNA for the 27 kDa gene was present only in P. brasiliensis, as indicated by the Northern analysis. The standardization of PCR technology permitted the amplification of P. brasiliensis DNA in artificially contaminated soils and in tissues of armadillos naturally infected with the fungus. These results indicate that PCR technology could play an important role in the search for P. brasiliensis’ habitat and could also be used in other ecological studies.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

Differences were detected in the gene expression of strains of E. histolytica using RNA (RAP-PCR) and DNA fingerprinting (RAPD). Analysis of the electrophoretic profiles of the gels revealed some polymorphic markers that could be used in the individual characterization of the strains. The 260 bands generated by using five different primers for RAP-PCR, as well as RAPD, were employed in the construction of dendograms. The dendogram obtained based on the RAPD products permitted the distinction of symptomatic and asymptomatic isolates, as well the correlation between the polymorphism exhibited and the virulence of the strains. The dendogram obtained for the RAP-PCR products did not show a correlation with the virulence of the strains but revealed a high degree of intraspecific transcriptional variability that could be related to other biological features, whether or not these are involved in the pathogenesis of amebiasis.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

More than 70 species of mycobacteria have been defined, and some can cause disease in humans, especially in immunocompromised patients. Species identification in most clinical laboratories is based on phenotypic characteristics and biochemical tests and final results are obtained only after two to four weeks. Quick identification methods, by reducing time for diagnosis, could expedite institution of specific treatment, increasing chances of success. PCR restriction-enzyme analysis (PRA) of the hsp65 gene was used as a rapid method for identification of 103 clinical isolates. Band patterns were interpreted by comparison with published tables and patterns available at an Internet site (http://www.hospvd.ch:8005). Concordant results of PRA and biochemical identification were obtained in 76 out of 83 isolates (91.5%). Results from 20 isolates could not be compared due to inconclusive PRA or biochemical identification. The results of this work showed that PRA could improve identification of mycobacteria in a routine setting because it is accurate, fast, and cheaper than conventional phenotypic identification.

Relevância:

30.00% 30.00%

Publicador:

Resumo:

Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquímica

Relevância:

30.00% 30.00%

Publicador:

Resumo:

A high incidence of cytomegalovirus (CMV) infections is observed in Brazil. These viruses are causatives of significant morbidity and mortality among patients with advanced human immunodeficiency virus (HIV) infection. This work, shows the application of a PCR on determination of CMV load in the buffy coat and plasma. We analyzed the samples of 247 HIV infected patients in order to diagnose CMV infection and disease. We developed a semi-quantitative PCR that amplifies part of the glycoprotein B (gB) gene of CMV. The semi-quantitative PCR was carried out only in positive clinical samples in a qualitative PCR confirmed by a nested-PCR. CD4 lymphocyte count, HIV viral load and CMV disease symptom were correlated with CMV load. CMV genome was detected in the buffy coat of 82 of 237 (34.6%) patients, in 10 of these the CMV load was determined varying between 928 and 332 880 viral copies/mug DNA. None of these 237 patients developed any suggestive manifestation of CMV disease. For the other 10 HIV infected patients selected based on the suspicion of CMV disease, CMV genome was detected in only one case. This patient presented a high CMV load, 8 000 000 copies/mug DNA, and developed a disseminated form of CMV disease including hepatitis and retinitis. Our results were greatly influenced by the impact of the highly active antiretroviral therapy that reduced incidence of CMV viremia and occurrence of CMV disease in the HIV infected patients.