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.

The use of the boundary element method in the analysis of single lap joints

The most common techniques for stress analysis/strength prediction of adhesive joints involve analytical or numerical methods such as the Finite Element Method (FEM). However, the Boundary Element Method (BEM) is an alternative numerical technique that has been successfully applied for the solution of a wide variety of engineering problems. This work evaluates the applicability of the boundary elem ent code BEASY as a design tool to analyze adhesive joints. The linearity of peak shear and peel stresses with the applied displacement is studied and compared between BEASY and the analytical model of Frostig et al., considering a bonded single-lap joint under tensile loading. The BEM results are also compared with FEM in terms of stress distributions. To evaluate the mesh convergence of BEASY, the influence of the mesh refinement on peak shear and peel stress distributions is assessed. Joint stress predictions are carried out numerically in BEASY and ABAQUS®, and analytically by the models of Volkersen, Goland, and Reissner and Frostig et al. The failure loads for each model are compared with experimental results. The preparation, processing, and mesh creation times are compared for all models. BEASY results presented a good agreement with the conventional methods.

Adherend thickness effect on the tensile fracture toughness of a structural adhesive using an optical data acquisition method

Adhesive bonding is nowadays a serious candidate to replace methods such as fastening or riveting, because of attractive mechanical properties. As a result, adhesives are being increasingly used in industries such as the automotive, aerospace and construction. Thus, it is highly important to predict the strength of bonded joints to assess the feasibility of joining during the fabrication process of components (e.g. due to complex geometries) or for repairing purposes. This work studies the tensile behaviour of adhesive joints between aluminium adherends considering different values of adherend thickness (h) and the double-cantilever beam (DCB) test. The experimental work consists of the definition of the tensile fracture toughness (GIC) for the different joint configurations. A conventional fracture characterization method was used, together with a J-integral approach, that take into account the plasticity effects occurring in the adhesive layer. An optical measurement method is used for the evaluation of crack tip opening and adherends rotation at the crack tip during the test, supported by a Matlab® sub-routine for the automated extraction of these quantities. As output of this work, a comparative evaluation between bonded systems with different values of adherend thickness is carried out and complete fracture data is provided in tension for the subsequent strength prediction of joints with identical conditions.

Preliminary study on the production of functionally graded materials by friction stir processing

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

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

Estimation of the Crustal Bulk Properties Beneath Mainland Portugal from P-Wave Teleseismic Receiver Functions

In this work, we present results from teleseismic P-wave receiver functions (PRFs) obtained in Portugal, Western Iberia. A dense seismic station deployment conducted between 2010 and 2012, in the scope of the WILAS project and covering the entire country, allowed the most spatially extensive probing on the bulk crustal seismic properties of Portugal up to date. The application of the H-κ stacking algorithm to the PRFs enabled us to estimate the crustal thickness (H) and the average crustal ratio of the P- and S-waves velocities V p/V s (κ) for the region. Observations of Moho conversions indicate that this interface is relatively smooth with the crustal thickness ranging between 24 and 34 km, with an average of 30 km. The highest V p/V s values are found on the Mesozoic-Cenozoic crust beneath the western and southern coastal domain of Portugal, whereas the lowest values correspond to Palaeozoic crust underlying the remaining part of the subject area. An average V p/V s is found to be 1.72, ranging 1.63-1.86 across the study area, indicating a predominantly felsic composition. Overall, we systematically observe a decrease of V p/V s with increasing crustal thickness. Taken as a whole, our results indicate a clear distinction between the geological zones of the Variscan Iberian Massif in Portugal, the overall shape of the anomalies conditioned by the shape of the Ibero-Armorican Arc, and associated Late Paleozoic suture zones, and the Meso-Cenozoic basin associated with Atlantic rifting stages. Thickened crust (30-34 km) across the studied region may be inherited from continental collision during the Paleozoic Variscan orogeny. An anomalous crustal thinning to around 28 km is observed beneath the central part of the Central Iberian Zone and the eastern part of South Portuguese Zone.

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.

The development of the N1 and N2 components in auditory oddball paradigms: a systematic review with narrative analysis and suggested normative values

Auditory event-related potentials (AERPs) are widely used in diverse fields of today’s neuroscience, concerning auditory processing, speech perception, language acquisition, neurodevelopment, attention and cognition in normal aging, gender, developmental, neurologic and psychiatric disorders. However, its transposition to clinical practice has remained minimal. Mainly due to scarce literature on normative data across age, wide spectrumof results, variety of auditory stimuli used and to different neuropsychological meanings of AERPs components between authors. One of the most prominent AERP components studied in last decades was N1, which reflects auditory detection and discrimination. Subsequently, N2 indicates attention allocation and phonological analysis. The simultaneous analysis of N1 and N2 elicited by feasible novelty experimental paradigms, such as auditory oddball, seems an objective method to assess central auditory processing. The aim of this systematic review was to bring forward normative values for auditory oddball N1 and N2 components across age. EBSCO, PubMed, Web of Knowledge and Google Scholarwere systematically searched for studies that elicited N1 and/or N2 by auditory oddball paradigm. A total of 2,764 papers were initially identified in the database, of which 19 resulted from hand search and additional references, between 1988 and 2013, last 25 years. A final total of 68 studiesmet the eligibility criteria with a total of 2,406 participants from control groups for N1 (age range 6.6–85 years; mean 34.42) and 1,507 for N2 (age range 9–85 years; mean 36.13). Polynomial regression analysis revealed thatN1latency decreases with aging at Fz and Cz,N1 amplitude at Cz decreases from childhood to adolescence and stabilizes after 30–40 years and at Fz the decrement finishes by 60 years and highly increases after this age. Regarding N2, latency did not covary with age but amplitude showed a significant decrement for both Cz and Fz. Results suggested reliable normative values for Cz and Fz electrode locations; however, changes in brain development and components topography over age should be considered in clinical practice.

Soliton-type and other travelling wave solutions for an improved class of nonlinear sixth-order Boussinesq equations

An improved class of nonlinear bidirectional Boussinesq equations of sixth order using a wave surface elevation formulation is derived. Exact travelling wave solutions for the proposed class of nonlinear evolution equations are deduced. A new exact travelling wave solution is found which is the uniform limit of a geometric series. The ratio of this series is proportional to a classical soliton-type solution of the form of the square of a hyperbolic secant function. This happens for some values of the wave propagation velocity. However, there are other values of this velocity which display this new type of soliton, but the classical soliton structure vanishes in some regions of the domain. Exact solutions of the form of the square of the classical soliton are also deduced. In some cases, we find that the ratio between the amplitude of this wave and the amplitude of the classical soliton is equal to 35/36. It is shown that different families of travelling wave solutions are associated with different values of the parameters introduced in the improved equations.

Utilização de técnicas de análise fotogramétricas em quebra-mares de taludes

Dissertação de Natureza Científica elaborada no Laboratório Nacional de Engenharia Civil (LNEC) para obtenção do grau de mestre em Engenharia Civil na Área de Especialização de Hidráulica no âmbito do protocolo de cooperação entre o ISEL e o LNEC

Local Fractional Variational Iteration Method for Local Fractional Poisson Equations in Two Independent Variables

The local fractional Poisson equations in two independent variables that appear in mathematical physics involving the local fractional derivatives are investigated in this paper. The approximate solutions with the nondifferentiable functions are obtained by using the local fractional variational iteration method.

Simulation of wave action on a moored container carrier inside Sines’ Harbour

The integrated numerical tool SWAMS (Simulation of Wave Action on Moored Ships) is used to simulate the behavior of a moored container carrier inside Sines’ Harbour. Wave, wind, currents, floating ship and moorings interaction is discussed. Several case scenarios are compared differing in the layout of the harbour and wind and wave conditions. The several harbour layouts correspond to proposed alternatives for the future expansion of Sines’ terminal XXI that include the extension of the East breakwater and of the quay. Additionally, the influence of wind on the behavior of the ship moored and the introduction of pre tensioning the mooring lines was analyzed. Hydrodynamic forces acting on the ship are determined using a modified version of the WAMIT model. This modified model utilizes the Haskind relations and the non-linear wave field inside the harbour obtained with finite element numerical model, BOUSS-WMH (Boussinesq Wave Model for Harbors) to get the wave forces on the ship. The time series of the moored ship motions and forces on moorings are obtained using BAS solver. © 2015 Taylor & Francis Group, London.

Road drivers performance appraisal using a MCDA method: a case study in the road transport company

Performance evaluation increasingly assumes a more important role in any organizational environment. In the transport area, the drivers are the company’s image and for this reason it is important to develop and increase their performance and commitment to the company goals. This evaluation can be used to motivate driver to improve their performance and to discover training needs. This work aims to create a performance appraisal evaluation model of the drivers based on the multi-criteria decision aid methodology. The MMASSI (Multicriteria Methodology to Support Selection of Information Systems) methodology was adapted by using a template supporting the evaluation according to the freight transportation company in study. The evaluation process involved all drivers (collaborators being evaluated), their supervisors and the company management. The final output is a ranking of the drivers, based on their performance, for each one of the scenarios used.

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.