983 resultados para PCR detection
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This article describes an effective microchip protocol based on electrophoretic-separation and electrochemical detection for highly sensitive and rapid measurements of nitrate ester explosives, including ethylene glycol dinitrate (EGDN), pentaerythritol tetranitrate (PETN), propylene glycol dinitrate (PGDN) and glyceryl trinitrate (nitroglycerin, NG). Factors influencing the separation and detection processes were examined and optimized. Under the optimal separation conditions obtained using a 15 mM borate buffer (pH 9.2) containing 20 mM SDS, and applying a separation voltage of 1500 V, the four nitrate ester explosives were separated within less than 3 min. The glassy-carbon amperometric detector (operated at -0.9 V vs. Ag/AgCl) offers convenient cathodic detection down to the picogram level, with detection limits of 0.5 ppm and 0.3 ppm for PGDN and for NG, respectively, along with good repeatability (RSD of 1.8-2.3%; n = 6) and linearity (over the 10-60 ppm range). Such effective microchip operation offers great promise for field screening of nitrate ester explosives and for supporting various counter-terrorism surveillance activities.
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A simple and easy approach to produce polymeric microchips with integrated copper electrodes for capacitively coupled contactless conductivity detection (CD) is described. Copper electrodes were fabricated using a printed circuit board (PCB) as an inexpensive thin-layer of metal. The electrode layout was first drawn and laser printed on a wax paper sheet. The toner layer deposited on the paper sheet was thermally transferred to the PCB surface working as a mask for wet chemical etching of the copper layer. After the etching step, the toner was removed with an acetonitrile-dampened cotton. A poly(ethylene terephthalate) (PET) film coated with a thin thermo-sensitive adhesive layer was used to laminate the PCB plate providing an insulator layer of the electrodes to perform CID measurements. Electrophoresis microchannels were fabricated in poly(dimethylsiloxane) (PDMS) by soft lithography and reversibly sealed against the PET film. These hybrid PDMS/PET chips exhibited a stable electroosmotic mobility of 4.25 +/- 0.04 x 10(-4) V cm(-2) s(-1), at pH 6.1, over fifty runs. Efficiencies ranging from 1127 to 1690 theoretical plates were obtained for inorganic cations.
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Aims: To investigate the expression of sboA and ituD genes among strains of Bacillus spp. at different pH and temperature. Methods and Results: Different Bacillus strains from the Amazon basin and Bacillus subtilis ATCC 19659 were investigated for the production of subtilosin A and iturin A by qRT-PCR, analysing sboA and ituD gene expression under different culture conditions. Amazonian strains presented a general gene expression level lower than B. subtilis ATCC 19659 for sboA. In contrast, when analysing the expression of ituD gene, the strains from the Amazon, particularly P40 and P45B, exhibited higher levels of expression. Changes in pH (6 and 8) and temperature (37 and 42 degrees C) caused a decrease in sboA expression, but increased ituD expression among strains from Amazonian environment. Conclusions: Temperature and pH have an important influence on the expression of genes sboA (subtilosin A) and ituD (iturin A) among Bacillus spp. The strains P40 and P45B can be useful for the production of antimicrobial peptide iturin A. Significance and Impact of the Study: Monitoring the expression of essential biosynthetic genes by qRT-PCR is a valuable tool for optimization of the production of antimicrobial peptides.
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This paper proposes a three-stage offline approach to detect, identify, and correct series and shunt branch parameter errors. In Stage 1 the branches suspected of having parameter errors are identified through an Identification Index (II). The II of a branch is the ratio between the number of measurements adjacent to that branch, whose normalized residuals are higher than a specified threshold value, and the total number of measurements adjacent to that branch. Using several measurement snapshots, in Stage 2 the suspicious parameters are estimated, in a simultaneous multiple-state-and-parameter estimation, via an augmented state and parameter estimator which increases the V - theta state vector for the inclusion of suspicious parameters. Stage 3 enables the validation of the estimation obtained in Stage 2, and is performed via a conventional weighted least squares estimator. Several simulation results (with IEEE bus systems) have demonstrated the reliability of the proposed approach to deal with single and multiple parameter errors in adjacent and non-adjacent branches, as well as in parallel transmission lines with series compensation. Finally the proposed approach is confirmed on tests performed on the Hydro-Quebec TransEnergie network.
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The taxonomy of the N(2)-fixing bacteria belonging to the genus Bradyrhizobium is still poorly refined, mainly due to conflicting results obtained by the analysis of the phenotypic and genotypic properties. This paper presents an application of a method aiming at the identification of possible new clusters within a Brazilian collection of 119 Bradryrhizobium strains showing phenotypic characteristics of B. japonicum and B. elkanii. The stability was studied as a function of the number of restriction enzymes used in the RFLP-PCR analysis of three ribosomal regions with three restriction enzymes per region. The method proposed here uses Clustering algorithms with distances calculated by average-linkage clustering. Introducing perturbations using sub-sampling techniques makes the stability analysis. The method showed efficacy in the grouping of the species B. japonicum and B. elkanii. Furthermore, two new clusters were clearly defined, indicating possible new species, and sub-clusters within each detected cluster. (C) 2008 Elsevier B.V. All rights reserved.
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The main purpose of this paper is to present architecture of automated system that allows monitoring and tracking in real time (online) the possible occurrence of faults and electromagnetic transients observed in primary power distribution networks. Through the interconnection of this automated system to the utility operation center, it will be possible to provide an efficient tool that will assist in decisionmaking by the Operation Center. In short, the desired purpose aims to have all tools necessary to identify, almost instantaneously, the occurrence of faults and transient disturbances in the primary power distribution system, as well as to determine its respective origin and probable location. The compilations of results from the application of this automated system show that the developed techniques provide accurate results, identifying and locating several occurrences of faults observed in the distribution system.
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In this study, the innovation approach is used to estimate the measurement total error associated with power system state estimation. This is required because the power system equations are very much correlated with each other and as a consequence part of the measurements errors is masked. For that purpose an index, innovation index (II), which provides the quantity of new information a measurement contains is proposed. A critical measurement is the limit case of a measurement with low II, it has a zero II index and its error is totally masked. In other words, that measurement does not bring any innovation for the gross error test. Using the II of a measurement, the masked gross error by the state estimation is recovered; then the total gross error of that measurement is composed. Instead of the classical normalised measurement residual amplitude, the corresponding normalised composed measurement residual amplitude is used in the gross error detection and identification test, but with m degrees of freedom. The gross error processing turns out to be very simple to implement, requiring only few adaptations to the existing state estimation software. The IEEE-14 bus system is used to validate the proposed gross error detection and identification test.
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On-line leak detection is a main concern for the safe operation of pipelines. Acoustic and mass balance are the most important and extensively applied technologies in field problems. The objective of this work is to compare these leak detection methods with respect to a given reference situation, i.e., the same pipeline and monitoring signals acquired at the inlet and outlet ends. Experimental tests were conducted in a 749 m long laboratory pipeline transporting water as the working fluid. The instrumentation included pressure transducers and electromagnetic flowmeters. Leaks were simulated by opening solenoid valves placed at known positions and previously calibrated to produce known average leak flow rates. Results have clearly shown the limitations and advantages of each method. It is also quite clear that acoustics and mass balance technologies are, in fact, complementary. In general, an acoustic leak detection system sends out an alarm more rapidly and locates the leak more precisely, provided that the rupture of the pipeline occurs abruptly enough. On the other hand, a mass balance leak detection method is capable of quantifying the leak flow rate very accurately and of detecting progressive leaks.
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Leakage reduction in water supply systems and distribution networks has been an increasingly important issue in the water industry since leaks and ruptures result in major physical and economic losses. Hydraulic transient solvers can be used in the system operational diagnosis, namely for leak detection purposes, due to their capability to describe the dynamic behaviour of the systems and to provide substantial amounts of data. In this research work, the association of hydraulic transient analysis with an optimisation model, through inverse transient analysis (ITA), has been used for leak detection and its location in an experimental facility containing PVC pipes. Observed transient pressure data have been used for testing ITA. A key factor for the success of the leak detection technique used is the accurate calibration of the transient solver, namely adequate boundary conditions and the description of energy dissipation effects since PVC pipes are characterised by a viscoelastic mechanical response. Results have shown that leaks were located with an accuracy between 4-15% of the total length of the pipeline, depending on the discretisation of the system model.
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Hemotropic mycoplasmas, epicellular erythrocytic bacterial parasites lacking a cell wall, are the causative agents of infectious anemia in numerous mammalian species. The presence of hemotropic mycoplasmas in blood samples of neotropical and exotic wild canids and felids from Brazilian zoos were recorded using molecular techniques. Blood samples were collected from 146 Brazilian wild felids, 19 exotic felids, 3 European wolves (Canis lupus), and from 97 Brazilian wild canids from zoos in the Brazilian states of Sao Paulo and Mato Grosso and the Federal District. Using conventional polymerase chain reaction (PCR), this work found 22 (13%) wild felids positive to Candidatus Mycoplasma haemominutum [4 jaguars (Panthera onca); 3 pumas (Puma concolor); 10 ocelots (Leopardus pardalis); 2 jaguarondis (Puma yagouaroundi); and 3 little spotted cats (Leopardus tigrinus)]. Only one little spotted cat (Leopardus tigrinus) was positive to Mycoplasma haemofelis, and none was positive to Candidatus Mycoplasma turicensis. Two bush dogs (Speothos venaticus) were positive for a Mycoplasma sp. closely related to Candidatus Mycoplasma haematoparvum, and two European wolves were positive for a Mycoplasma sp. closely related to candidatus Mycoplasma haemominutum. This is the first study regarding the molecular detection of hemotropic mycoplasmas in wild canids.
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In this paper, a framework for detection of human skin in digital images is proposed. This framework is composed of a training phase and a detection phase. A skin class model is learned during the training phase by processing several training images in a hybrid and incremental fuzzy learning scheme. This scheme combines unsupervised-and supervised-learning: unsupervised, by fuzzy clustering, to obtain clusters of color groups from training images; and supervised to select groups that represent skin color. At the end of the training phase, aggregation operators are used to provide combinations of selected groups into a skin model. In the detection phase, the learned skin model is used to detect human skin in an efficient way. Experimental results show robust and accurate human skin detection performed by the proposed framework.
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Acoustic resonances are observed in high-pressure discharge lamps operated with ac input modulated power frequencies in the kilohertz range. This paper describes an optical resonance detection method for high-intensity discharge lamps using computer-controlled cameras and image processing software. Experimental results showing acoustic resonances in high-pressure sodium lamps are presented.
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Sigma phase is a deleterious one which can be formed in duplex stainless steels during heat treatment or welding. Aiming to accompany this transformation, ferrite and sigma percentage and hardness were measured on samples of a UNS S31803 duplex stainless steel submitted to heat treatment. These results were compared to measurements obtained from ultrasound and eddy current techniques, i.e., velocity and impedance, respectively. Additionally, backscattered signals produced by wave propagation were acquired during ultrasonic inspection as well as magnetic Barkhausen noise during magnetic inspection. Both signal types were processed via a combination of detrended-fluctuation analysis (DFA) and principal component analysis (PCA). The techniques used were proven to be sensitive to changes in samples related to sigma phase formation due to heat treatment. Furthermore, there is an advantage using these methods since they are nondestructive. (C) 2010 Elsevier B.V. All rights reserved.
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This paper reports the use of a non-destructive, continuous magnetic Barkhausen noise (CMBN) technique to investigate the size and thickness of volumetric defects, in a 1070 steel. The magnetic behavior of the used probe was analyzed by numerical simulation, using the finite element method (FEM). Results indicated that the presence of a ferrite coil core in the probe favors MBN emissions. The samples were scanned with different speeds and probe configurations to determine the effect of the flaw on the CMBN signal amplitude. A moving smooth window, based on a second-order statistical moment, was used for analyzing the time signal. The results show the technique`s good repeatability, and high capacity for detection of this type of defect. (C) 2009 Elsevier Ltd. All rights reserved.
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Alpha prime formation leads to material embrittlement and deterioration of corrosion resistance. In the present study, the mechanical and corrosion behavior of super duplex stainless steel UNS S32520 aged at 475 degrees C from 0.5 h to 1,032 h was evaluated using microhardness measurements, Charpy impact tests, electrochemical impedance spectroscopy, and cyclic polarization curves. The sensibility of these tests to the effects of alpha prime phase was investigated. The microhardness test showed a gradual increase in hardness with aging time, whereas the impact tests revealed losses of about 80% in the energy absorption capacity for the material aged for 12 h in comparison with the solution-annealed samples. The most responsive analysis was the impact test, which indirectly revealed the presence of this deleterious phase in samples aged for 0.5 h. The electrochemical impedance spectroscopy and polarization tests were not highly sensitive to the alpha prime phase unless these are present in large amounts in the stainless steel.