A comprehensive high-throughput FTIR spectroscopy-based method for evaluating the transfection event: estimating the transfection efficiency and extracting associated metabolic responses

Reporter genes are routinely used in every laboratory for molecular and cellular biology for studying heterologous gene expression and general cellular biological mechanisms, such as transfection processes. Although well characterized and broadly implemented, reporter genes present serious limitations, either by involving time-consuming procedures or by presenting possible side effects on the expression of the heterologous gene or even in the general cellular metabolism. Fourier transform mid-infrared (FT-MIR) spectroscopy was evaluated to simultaneously analyze in a rapid (minutes) and high-throughput mode (using 96-wells microplates), the transfection efficiency, and the effect of the transfection process on the host cell biochemical composition and metabolism. Semi-adherent HEK and adherent AGS cell lines, transfected with the plasmid pVAX-GFP using Lipofectamine, were used as model systems. Good partial least squares (PLS) models were built to estimate the transfection efficiency, either considering each cell line independently (R 2 ≥ 0.92; RMSECV ≤ 2 %) or simultaneously considering both cell lines (R 2 = 0.90; RMSECV = 2 %). Additionally, the effect of the transfection process on the HEK cell biochemical and metabolic features could be evaluated directly from the FT-IR spectra. Due to the high sensitivity of the technique, it was also possible to discriminate the effect of the transfection process from the transfection reagent on KEK cells, e.g., by the analysis of spectral biomarkers and biochemical and metabolic features. The present results are far beyond what any reporter gene assay or other specific probe can offer for these purposes.

Slaughterhouses fungal burden assessment: a contribution for the pursuit of a better assessment strategy

In slaughterhouses, the biological risk is present not only from the direct or indirect contact with animal matter, but also from the exposure to bioaerosols. Fungal contamination was already reported from the floors and walls of slaughterhouses. This study intends to assess fungal contamination by cultural and molecular methods in poultry, swine/bovine and large animal slaughterhouses. Air samples were collected through an impaction method, while surface samples were collected by the swabbing method and subjected to further macro- and micro-scopic observations. In addition, we collected air samples using the impinger method in order to perform real-time quantitative PCR (qPCR) amplification of genes from specific fungal species, namely A. flavus, A. fumigatus and A. ochraceus complexes. Poultry and swine/bovine slaughterhouses presented each two sampling sites that surpass the guideline of 150 CFU/m3. Scopulariopsis candida was the most frequently isolated (59.5%) in poultry slaughterhouse air; Cladosporium sp. (45.7%) in the swine/bovine slaughterhouse; and Penicillium sp. (80.8%) in the large animal slaughterhouse. Molecular tools successfully amplified DNA from the A. fumigatus complex in six sampling sites where the presence of this fungal species was not identified by conventional methods. This study besides suggesting the indicators that are representative of harmful fungal contamination, also indicates a strategy as a protocol to ensure a proper characterization of fungal occupational exposure.

Toroid inductor development for a SIC DC-DC converter up to 150kW, based on finite element method

Dissertação para a obtenção do grau de Mestre em Engenharia Electrotécnica Ramo de Energia

Otimização de placas compósitas laminadas utilizando redes neuronais e enxames de partículas

Trabalho Final de mestrado para obtenção do grau de Mestre em engenharia Mecância

Multiobjective optimization for node adaptation in the analysis of composite plates using a meshless collocation method

The bending of simply supported composite plates is analyzed using a direct collocation meshless numerical method. In order to optimize node distribution the Direct MultiSearch (DMS) for multi-objective optimization method is applied. In addition, the method optimizes the shape parameter in radial basis functions. The optimization algorithm was able to find good solutions for a large variety of nodes distribution.

Multiobjective optimization of viscoelastic laminated sandwich structures using the Direct MultiSearch method

A multiobjective approach for optimization of passive damping for vibration reduction in sandwich structures is presented in this paper. Constrained optimization is conducted for maximization of modal loss factors and minimization of weight of sandwich beams and plates with elastic laminated constraining layers and a viscoelastic core, with layer thickness and material and laminate layer ply orientation angles as design variables. The problem is solved using the Direct MultiSearch (DMS) solver for derivative-free multiobjective optimization and solutions are compared with alternative ones obtained using genetic algorithms.

Análise do comportamento sísmico da barragem de Luzzone: desenvolvimento de um programa de EF3D utilizando uma formulação em deslocamentos e pressões

Trabalho Final de Mestrado elaborado no Laboratório Nacional de Engenharia Civil (LNEC) para a obtenção do grau de Mestre em Engenharia Civil pelo Instituto Superior de Engenharia de Lisboa no âmbito do protocolo entre o ISEL e o LNEC

Modelação do comportamento dinâmico de sistemas barragem-fundação-albufeira: análise sísmica de uma barragem de abóbadas múltiplas

Trabalho Final de Mestrado elaborado no Laboratório Nacional de Engenharia Civil (LNEC) para a obtenção do grau de Mestre em Engenharia Civil pelo Instituto Superior de Engenharia de Lisboa no âmbito do protocolo de cooperação entre o ISEL e o LNEC

Parallel GPU architecture for hyperspectral unmixing based on augmented Lagrangian method

Hyperspectral imaging has become one of the main topics in remote sensing applications, which comprise hundreds of spectral bands at different (almost contiguous) wavelength channels over the same area generating large data volumes comprising several GBs per flight. This high spectral resolution can be used for object detection and for discriminate between different objects based on their spectral characteristics. One of the main problems involved in hyperspectral analysis is the presence of mixed pixels, which arise when the spacial resolution of the sensor is not able to separate spectrally distinct materials. Spectral unmixing is one of the most important task for hyperspectral data exploitation. However, the unmixing algorithms can be computationally very expensive, and even high power consuming, which compromises the use in applications under on-board constraints. In recent years, graphics processing units (GPUs) have evolved into highly parallel and programmable systems. Specifically, several hyperspectral imaging algorithms have shown to be able to benefit from this hardware taking advantage of the extremely high floating-point processing performance, compact size, huge memory bandwidth, and relatively low cost of these units, which make them appealing for onboard data processing. In this paper, we propose a parallel implementation of an augmented Lagragian based method for unsupervised hyperspectral linear unmixing on GPUs using CUDA. The method called simplex identification via split augmented Lagrangian (SISAL) aims to identify the endmembers of a scene, i.e., is able to unmix hyperspectral data sets in which the pure pixel assumption is violated. The efficient implementation of SISAL method presented in this work exploits the GPU architecture at low level, using shared memory and coalesced accesses to memory.

Parallel hyperspectral compressive sensing method on GPU

Remote hyperspectral sensors collect large amounts of data per flight usually with low spatial resolution. It is known that the bandwidth connection between the satellite/airborne platform and the ground station is reduced, thus a compression onboard method is desirable to reduce the amount of data to be transmitted. This paper presents a parallel implementation of an compressive sensing method, called parallel hyperspectral coded aperture (P-HYCA), for graphics processing units (GPU) using the compute unified device architecture (CUDA). This method takes into account two main properties of hyperspectral dataset, namely the high correlation existing among the spectral bands and the generally low number of endmembers needed to explain the data, which largely reduces the number of measurements necessary to correctly reconstruct the original data. Experimental results conducted using synthetic and real hyperspectral datasets on two different GPU architectures by NVIDIA: GeForce GTX 590 and GeForce GTX TITAN, reveal that the use of GPUs can provide real-time compressive sensing performance. The achieved speedup is up to 20 times when compared with the processing time of HYCA running on one core of the Intel i7-2600 CPU (3.4GHz), with 16 Gbyte memory.

Parallel hyperspectral unmixing method via split augmented lagrangian on GPU

One of the main problems of hyperspectral data analysis is the presence of mixed pixels due to the low spatial resolution of such images. Linear spectral unmixing aims at inferring pure spectral signatures and their fractions at each pixel of the scene. The huge data volumes acquired by hyperspectral sensors put stringent requirements on processing and unmixing methods. This letter proposes an efficient implementation of the method called simplex identification via split augmented Lagrangian (SISAL) which exploits the graphics processing unit (GPU) architecture at low level using Compute Unified Device Architecture. SISAL aims to identify the endmembers of a scene, i.e., is able to unmix hyperspectral data sets in which the pure pixel assumption is violated. The proposed implementation is performed in a pixel-by-pixel fashion using coalesced accesses to memory and exploiting shared memory to store temporary data. Furthermore, the kernels have been optimized to minimize the threads divergence, therefore achieving high GPU occupancy. The experimental results obtained for the simulated and real hyperspectral data sets reveal speedups up to 49 times, which demonstrates that the GPU implementation can significantly accelerate the method's execution over big data sets while maintaining the methods accuracy.

Parallel method for sparse semisupervised hyperspectral unmixing

Parallel hyperspectral unmixing problem is considered in this paper. A semisupervised approach is developed under the linear mixture model, where the abundance's physical constraints are taken into account. The proposed approach relies on the increasing availability of spectral libraries of materials measured on the ground instead of resorting to endmember extraction methods. Since Libraries are potentially very large and hyperspectral datasets are of high dimensionality a parallel implementation in a pixel-by-pixel fashion is derived to properly exploits the graphics processing units (GPU) architecture at low level, thus taking full advantage of the computational power of GPUs. Experimental results obtained for real hyperspectral datasets reveal significant speedup factors, up to 164 times, with regards to optimized serial implementation.

Parallel hyperspectral unmixing method on GPU

Many Hyperspectral imagery applications require a response in real time or near-real time. To meet this requirement this paper proposes a parallel unmixing method developed for graphics processing units (GPU). This method is based on the vertex component analysis (VCA), which is a geometrical based method highly parallelizable. VCA is a very fast and accurate method that extracts endmember signatures from large hyperspectral datasets without the use of any a priori knowledge about the constituent spectra. Experimental results obtained for simulated and real hyperspectral datasets reveal considerable acceleration factors, up to 24 times.

Vertex component analysis: a geometric-based approach to unmix hyperspectral data

Hyperspectral remote sensing exploits the electromagnetic scattering patterns of the different materials at specific wavelengths [2, 3]. Hyperspectral sensors have been developed to sample the scattered portion of the electromagnetic spectrum extending from the visible region through the near-infrared and mid-infrared, in hundreds of narrow contiguous bands [4, 5]. The number and variety of potential civilian and military applications of hyperspectral remote sensing is enormous [6, 7]. Very often, the resolution cell corresponding to a single pixel in an image contains several substances (endmembers) [4]. In this situation, the scattered energy is a mixing of the endmember spectra. A challenging task underlying many hyperspectral imagery applications is then decomposing a mixed pixel into a collection of reflectance spectra, called endmember signatures, and the corresponding abundance fractions [8–10]. Depending on the mixing scales at each pixel, the observed mixture is either linear or nonlinear [11, 12]. Linear mixing model holds approximately when the mixing scale is macroscopic [13] and there is negligible interaction among distinct endmembers [3, 14]. If, however, the mixing scale is microscopic (or intimate mixtures) [15, 16] and the incident solar radiation is scattered by the scene through multiple bounces involving several endmembers [17], the linear model is no longer accurate. Linear spectral unmixing has been intensively researched in the last years [9, 10, 12, 18–21]. It considers that a mixed pixel is a linear combination of endmember signatures weighted by the correspondent abundance fractions. Under this model, and assuming that the number of substances and their reflectance spectra are known, hyperspectral unmixing is a linear problem for which many solutions have been proposed (e.g., maximum likelihood estimation [8], spectral signature matching [22], spectral angle mapper [23], subspace projection methods [24,25], and constrained least squares [26]). In most cases, the number of substances and their reflectances are not known and, then, hyperspectral unmixing falls into the class of blind source separation problems [27]. Independent component analysis (ICA) has recently been proposed as a tool to blindly unmix hyperspectral data [28–31]. ICA is based on the assumption of mutually independent sources (abundance fractions), which is not the case of hyperspectral data, since the sum of abundance fractions is constant, implying statistical dependence among them. This dependence compromises ICA applicability to hyperspectral images as shown in Refs. [21, 32]. In fact, ICA finds the endmember signatures by multiplying the spectral vectors with an unmixing matrix, which minimizes the mutual information among sources. If sources are independent, ICA provides the correct unmixing, since the minimum of the mutual information is obtained only when sources are independent. This is no longer true for dependent abundance fractions. Nevertheless, some endmembers may be approximately unmixed. These aspects are addressed in Ref. [33]. Under the linear mixing model, the observations from a scene are in a simplex whose vertices correspond to the endmembers. Several approaches [34–36] have exploited this geometric feature of hyperspectral mixtures [35]. Minimum volume transform (MVT) algorithm [36] determines the simplex of minimum volume containing the data. The method presented in Ref. [37] is also of MVT type but, by introducing the notion of bundles, it takes into account the endmember variability usually present in hyperspectral mixtures. The MVT type approaches are complex from the computational point of view. Usually, these algorithms find in the first place the convex hull defined by the observed data and then fit a minimum volume simplex to it. For example, the gift wrapping algorithm [38] computes the convex hull of n data points in a d-dimensional space with a computational complexity of O(nbd=2cþ1), where bxc is the highest integer lower or equal than x and n is the number of samples. The complexity of the method presented in Ref. [37] is even higher, since the temperature of the simulated annealing algorithm used shall follow a log( ) law [39] to assure convergence (in probability) to the desired solution. Aiming at a lower computational complexity, some algorithms such as the pixel purity index (PPI) [35] and the N-FINDR [40] still find the minimum volume simplex containing the data cloud, but they assume the presence of at least one pure pixel of each endmember in the data. This is a strong requisite that may not hold in some data sets. In any case, these algorithms find the set of most pure pixels in the data. PPI algorithm uses the minimum noise fraction (MNF) [41] as a preprocessing step to reduce dimensionality and to improve the signal-to-noise ratio (SNR). The algorithm then projects every spectral vector onto skewers (large number of random vectors) [35, 42,43]. The points corresponding to extremes, for each skewer direction, are stored. A cumulative account records the number of times each pixel (i.e., a given spectral vector) is found to be an extreme. The pixels with the highest scores are the purest ones. N-FINDR algorithm [40] is based on the fact that in p spectral dimensions, the p-volume defined by a simplex formed by the purest pixels is larger than any other volume defined by any other combination of pixels. This algorithm finds the set of pixels defining the largest volume by inflating a simplex inside the data. ORA SIS [44, 45] is a hyperspectral framework developed by the U.S. Naval Research Laboratory consisting of several algorithms organized in six modules: exemplar selector, adaptative learner, demixer, knowledge base or spectral library, and spatial postrocessor. The first step consists in flat-fielding the spectra. Next, the exemplar selection module is used to select spectral vectors that best represent the smaller convex cone containing the data. The other pixels are rejected when the spectral angle distance (SAD) is less than a given thresh old. The procedure finds the basis for a subspace of a lower dimension using a modified Gram–Schmidt orthogonalizati on. The selected vectors are then projected onto this subspace and a simplex is found by an MV T pro cess. ORA SIS is oriented to real-time target detection from uncrewed air vehicles using hyperspectral data [46]. In this chapter we develop a new algorithm to unmix linear mixtures of endmember spectra. First, the algorithm determines the number of endmembers and the signal subspace using a newly developed concept [47, 48]. Second, the algorithm extracts the most pure pixels present in the data. Unlike other methods, this algorithm is completely automatic and unsupervised. To estimate the number of endmembers and the signal subspace in hyperspectral linear mixtures, the proposed scheme begins by estimating sign al and noise correlation matrices. The latter is based on multiple regression theory. The signal subspace is then identified by selectin g the set of signal eigenvalue s that best represents the data, in the least-square sense [48,49 ], we note, however, that VCA works with projected and with unprojected data. The extraction of the end members exploits two facts: (1) the endmembers are the vertices of a simplex and (2) the affine transformation of a simplex is also a simplex. As PPI and N-FIND R algorithms, VCA also assumes the presence of pure pixels in the data. The algorithm iteratively projects data on to a direction orthogonal to the subspace spanned by the endmembers already determined. The new end member signature corresponds to the extreme of the projection. The algorithm iterates until all end members are exhausted. VCA performs much better than PPI and better than or comparable to N-FI NDR; yet it has a computational complexity between on e and two orders of magnitude lower than N-FINDR. The chapter is structure d as follows. Section 19.2 describes the fundamentals of the proposed method. Section 19.3 and Section 19.4 evaluate the proposed algorithm using simulated and real data, respectively. Section 19.5 presents some concluding remarks.