Phage display as a tool for generation of bio-therapeutic agents for breast cancer

Notch is a conserved signalling pathway, which plays a crucial role in a multiple cellular processes such as stem cell self-renewal, cell division, proliferation and apoptosis. In mammalian, four Notch receptors and five ligands are described, where interaction is achieved through their extracellular domains, leading to a transcription activation of different target genes. Increased expression of Notch ligands has been detected in several types of cancer, including breast cancer suggesting that these proteins represent possible therapeutic targets. The goal of this work was to generate quality protein targets and, by phage display technology, select function-blocking antibodies specific for Notch ligands. Phage display is a powerful technique that allows the generation of highly specific antibodies to be used for therapeutics, and it has also proved to be a reliable approach in identifying and validating new cancer-related targets. Also, we aimed at solving the tri-dimensional structure of the Notch ligands alone and in complex with selected antibodies. In this work, the initial phase focused on the optimization of the expression and purification of a human Delta-like 1 ligand mutant construct (hDLL1-DE3), by refolding from E. coli inclusion bodies. To confirm the biological activity of the produced recombinant protein cellular functional studies were performed, revealing that treatment with hDLL1-DE3 protein led to a modulation of Notch target genes. In a second stage of this study, Antibody fragments (Fabs) specific for hDLL1-DE3 were generated by phage display, using the produced protein as target, in which one good Fab candidate was selected to determine the best expression conditions. In parallel, multiple crystallization conditions were tested with hDLL1-DE3, but so far none led to positive results.

Development and modulation of magnetic responsive supported ionic liquid membranes

The work presented in this thesis aims at developing a new separation process based on the application of supported magnetic ionic liquid membranes, SMILMs, using magnetic ionic liquids, MILs. MILs have attracted growing interest due to their ability to change their physicochemical characteristics when exposed to variable magnetic field conditions. The magnetic responsive behavior of MILs is thus expected to contribute for the development of more efficient separation processes, such as supported liquid membranes, where MILs may be used as a selective carrier. Driven by the MILs behavior, these membranes are expected to switch reversibly their permeability and selectivity by in situ and non-invasive adjustment of the conditions (e.g. intensity, direction vector and uniformity) of an external applied magnetic field. The development of these magnetic responsive membrane processes were anticipated by studies, performed along the first stage of this PhD work, aiming at getting a deep knowledge on the influence of magnetic field on MILs properties. The influence of the magnetic field on the molecular dynamics and structural rearrangement of MILs ionic network was assessed through a 1H-NMR technique. Through the 1H-NMR relaxometry analysis it was possible to estimate the self-diffusion profiles of two different model MILs, [Aliquat][FeCl4] and [P66614][FeCl4]. A comparative analysis was established between the behavior of magnetic and non-magnetic ionic liquids, MILs and ILs, to facilitate the perception of the magnetic field impact on MILs properties. In contrast to ILs, MILs show a specific relaxation mechanism, characterized by the magnetic dependence of their self-diffusion coefficients. MILs self-diffusion coefficients increased in the presence of magnetic field whereas ILs self-diffusion was not affected. In order to understand the reasons underlying the magnetic dependence of MILs self-diffusion, studies were performed to investigate the influence of the magnetic field on MILs’ viscosity. It was observed that the MIL´s viscosity decreases with the increase of the magnetic field, explaining the increase of MILs self-diffusion according to the modified Stokes- Einstein equation. Different gas and liquid transport studies were therefore performed aiming to determine the influence of the magnetic behavior of MILs on solute transport through SMILMs. Gas permeation studies were performed using pure CO2 andN2 gas streams and air, using a series of phosphonium cation based MILs, containing different paramagnetic anions. Transport studies were conducted in the presence and absence of magnetic field at a maximum intensity of 1.5T. The results revealed that gas permeability increased in the presence of the magnetic field, however, without affecting the membrane selectivity. The increase of gas permeability through SMILMs was related to the decrease of the MILs viscosity under magnetic field conditions.(...)

Empowerment and intentions to resist future change: the moderating effect organization-based self-esteem

Nowadays, organizations face a constant need for adaptability, increasing the importance of change management. Our study focuses of how empowering leadership influences intentions to resist future changes, mediated by the effects of psychological and structural empowerment. From the responses of the two questionnaires (N1=230; Ntf=113), we found that empowering leadership fosters psychological and structural empowerment. Structural empowerment was the main driver in reducing intentions to resist future change when an employee has high organization-based self-esteem. Our findings add to the literature by examining how we can anticipate and manage change under an empowering context, building on social exchange and uncertainty reduction theories

How to reduce the causes of call center contact by implementing web self-service at Sonae MC?

This project aims to provide feasible solutions to improve customer´s Help Area at Continente Online. The goal is to increase satisfaction and loyalty by reducing the main causes that lead customers to appeal to Call Center or abandon the website. The pursued solution is the implementation of Web Self-Service and the vision taken is focused not only on providing customers basic help tools but also innovate with international best practices to sustain Sonae MC´s present and future market leader position. Customer´s feedback, costs and impact are taken in consideration to find the best fit for the company.

Self-monitoring of freeze–thaw damage using triphasic electric conductive concrete

The effect of freeze–thaw cycles on concrete is of great importance for durability evaluation of concrete structures in cold regions. In this paper, damage accumulation was studied by following the fractional change of impedance (FCI) with number of freeze–thaw cycles (N). The nano-carbon black (NCB), carbon fiber (CF) and steel fiber (SF) were added to plain concrete to produce the triphasic electrical conductive (TEC) and ductile concrete. The effects of NCB, CF and SF on the compressive strength, flexural properties, electrical impedance were investigated. The concrete beams with different dosages of conductive materials were studied for FCI, N and mass loss (ML), the relationship between FCI and N of conductive concrete can be well defined by a first order exponential decay curve. It is noted that this nondestructive and sensitive real-time testing method is meaningful for evaluating of freeze–thaw damage in concrete.

A FEM-based model to study the behaviour of corroded RC beams shear repaired by NSM CFRP rods technique

This paper presents the main features of finite element FE numerical model developed using the computer code FEMIX to predict the near-surface mounted NSM carbon-fiber-reinforced polymer CFRP rods shear repair contribution to corroded reinforced concrete RC beams. In the RC beams shear repaired with NSM technique, the Carbon Fibre Reinforced Polymer (CFRP) rods are placed inside pre-cut grooves onto the concrete cover of the RC beam’s lateral faces and are bonded to the concrete with high epoxy adhesive. Experimental and 3D numerical modelling results are presented in this paper in terms of load-deflection curves, and failure modes for 4 short corroded beams: two corroded beams (A1CL3-B and A1CL3-SB) and two control beams (A1T-B and A1T-SB), the beams noted with B were let repaired in bending only with NSM CFRP rods while the ones noted with SB were repaired in both bending and shear with NSM technique. The corrosion of the tensile steel bars and its effect on the shear capacity of the RC beams was discussed. Results showed that the FE model was able to capture the main aspects of the experimental load-deflection curves of the RC beams, moreover it has presented the experimental failure modes and FE numerical modelling crack patterns and both gave similar results for non-shear repaired beams which failed in diagonal tension mode of failure and for shear-repaired beams which failed due to large flexural crack at the middle of the beams along with the concrete crushing, three dimensional crack patterns were produced for shear-repaired beams in order to investigate the splitting cracks occurred at the middle of the beams and near the support.

HLT Hilotec: self-building manual for sustainable housing

The future of the construction industry will require changes at many levels. One is the ability of companies to adapt to new challenges, converting needs to opportunities and simultaneously contributing to the solving of social and environmental problems. In the coming decades we will see a change in attitude in the industry, with a strong tendency to adopt natural and recycled materials, as well as bet on green technology and social innovation oriented to emerging countries. On the other hand, emerging countries have a high demand for housing construction on a large scale, but the current techniques in the developed countries for building requires a large amount of natural resources and skilled labor. This contextualization brings sustainability problems for the construction sector in emerging countries, often with scarce natural resources and with the construction sector underdeveloped. Through a cooperative action between the construction company Mota-Engil Engineering and the University of Minho in Portugal, a construction technology was developed based on the use of Compressed Earth Blocks as part of a social concept for innovative small houses, favoring the adoption of local and natural materials and with the main premise of being dedicated to self-construction. The HiLoTec project - Development of a Sustainable Self-Construction System for Developing Countries was based on this idea. One of the several results of this project is this construction manual. To Mota-Engil the project was a platform for incubation of knowledge about earth construction and to obtain a constructive solution validated technically and scientifically, suitable to be implemented in the markets where it operates. For the University of Minho the project was an opportunity to strengthen skills in research, laboratory and scientific development, through the development of engineering studies, architecture and sustainability, as well as supporting the doctoral scholarships and dissemination of scientific publications. May the knowledge of this project be of benefit, in the future, for the welfare of those who build a HiLoTec house.

Tensile stress–crack width law for steel fibre reinforced self-compacting concrete obtained from indirect (splitting) tensile tests

In this work, the fracture mode I parameters of steel fibre reinforced self-compacting concrete (SFRSCC) were derived from the numerical simulation of indirect splitting tensile tests. The combined experimental and numerical research allowed a comparison between the stress-crack width (σ - w) relationship acquired straightforwardly from direct tensile tests, and the σ - w response derived from inverse analysis of the splitting tensile tests results. For this purpose a comprehensive nonlinear 3D finite element (FE) modeling strategy was developed. A comparison between the experimental results obtained from splitting tensile tests and the corresponding FE simulations confirmed the good accuracy of the proposed strategy to derive the σ – w for these composites. It is concluded that the post-cracking tensile laws obtained from inverse analysis provided a close relationship with the ones obtained from the experimental uniaxial tensile tests.

Numerical simulation of the punching shear behaviour of self-compacting fibre reinforced flat slabs

This paper presents the numerical simulations of the punching behaviour of centrally loaded steel fibre reinforced self-compacting concrete (SFRSCC) flat slabs. Eight half scaled slabs reinforced with different content of hooked-end steel fibres (0, 60, 75 and 90 kg/m3) and concrete strengths of 50 and 70 MPa were tested and numerically modelled. Moreover, a total of 54 three-point bending tests were carried out to assess the post-cracking flexural tensile strength. All the slabs had a relatively high conventional flexural reinforcement in order to promote the occurrence of punching failure mode. Neither of the slabs had any type of specific shear reinforcement rather than the contribution of the steel fibres. The numerical simulations were performed according to the Reissner-Mindlin theory under the finite element method framework. Regarding the classic formulation of the Reissner-Mindlin theory, in order to simulate the progressive damage induced by cracking, the shell element is discretized into layers, being assumed a plane stress state in each layer. The numerical results are, then, compared with the experimental ones and it is possible to notice that they accurately predict the experimental force-deflection relationship. The type of failure observed experimentally was also predicted in the numerical simulations.

Assessment of the performance of steel fibre reinforced self-compacting concrete in elevated slabs

The reinforcement mechanisms at the cross section level assured by fibres bridging the cracks in steel fibre reinforced self-compacting concrete (SFRSCC) can be significantly amplified at structural level when the SFRSCC is applied in structures with high support redundancy, such is the case of elevated slab systems. To evaluate the potentialities of SFRSCC as the fundamental material of elevated slab systems, a ¼ scale SFRSCC prototype of a residential building was designed, built and tested. The extensive experimental program includes material tests for characterizing the relevant properties of SFRSCC, as well as structural tests for assessing the performance of the prototype at serviceability and ultimate limit conditions. Three distinct approaches where adopted to derive the constitutive laws of the SFRSCC in tension that were used in finite element material nonlinear analysis to evaluate the reliability of these approaches in the prediction of the load carrying capacity of the prototype.

Time-dependent flexural behaviour of cracked steel fibre reinforced self-compacting concrete panels

In the present work are described and discussed the results of an extensive experimental program that aims to study the long-term behaviour of cracked steel fibre reinforced self-compacting concrete, SFRSCC, applied in laminar structures. In a first stage, the influence of the initial crack opening level (wcr = 0.3 and 0.5 mm), applied stress level, fibre orientation/dispersion and distance from the casting point, on the flexural creep behaviour of SFRSCC was investigated. Moreover, in order to evaluate the effects of the creep phenomenon on the residual flexural strength, a series of monotonic tests were also executed. It was found that wcr = 0.5 mm series showed a higher creep coefficient comparing to the series with a lower initial crack opening. Furthermore, the creep performance of the SFRSCC was influenced by the orientation of the extracted prismatic specimens regarding the direction of the concrete flow within the cast panel.

Embedded Through-Section shear strengthening technique using steel and CFRP bars in RC beams of different percentage of existing stirrups

The Embedded Through-Section (ETS) technique is a promising technique for the shear strengthening of existing (RC) elements. According to this technique, holes are drilled through the beam section, and bars of steel or FRP material are introduced into these holes and bonded to the concrete with adhesive materials. An experimental program was carried out with RC T-cross section beams strengthened in shear using the ETS steel bars and ETS CFRP rods. The research is focused on the evaluation of the ETS efficiency on beams with different percentage of existing internal transverse reinforcement (ρsw=0.0%, ρsw=0.1% and ρsw=0.17%). The effectiveness of different ETS strengthening configurations was also investigated. The good bond between the strengthening ETS bars and the surrounding concrete allowed the yield initiation of the ETS steel bars and the attainment of high tensile strains in the ETS CFPR rods, leading to significant increase of shear capacity, whose level was strongly influenced by the inclination of the ETS bars and the percentage of internal transverse reinforcement.

A FEM-based model to study the behaviour of corroded RC beams shear repaired by NSM CFRP rods technique

This paper presents the main features of finite element FE numerical model developed using the computer code FEMIX to predict the near-surface mounted NSM carbon-fiber-reinforced polymer CFRP rods shear repair contribution to corroded reinforced concrete RC beams. In the RC beams shear repaired with NSM technique, the Carbon Fibre Reinforced Polymer (CFRP) rods are placed inside pre-cut grooves onto the concrete cover of the RC beam’s lateral faces and are bonded to the concrete with high epoxy adhesive. Experimental and 3D numerical modelling results are presented in this paper in terms of load-deflection curves, failure modes and slip information of the tensile steel bars for 4 short corroded beams: two corroded beams (A1CL3-B and A1CL3-SB) and two control beams (A1T-B and A1T-SB), the beams noted with B were let repaired in bending only with NSM CFRP rods while the ones noted with SB were repaired in both bending and shear with NSM technique. The corrosion of the tensile steel bars and its effect on the shear capacity of the RC beams was discussed. Results showed that the FE model was able to capture the main aspects of the experimental load-deflection curves of the RC beams, moreover it has presented the experimental failure modes and FE numerical modelling crack patterns and both gave similar results for non-shear repaired beams which failed in diagonal tension mode of failure and for shear-repaired beams which failed due to large flexural crack at the middle of the beams along with the concrete crushing, three dimensional crack patterns were produced for shear-repaired beams in order to investigate the splitting cracks occurred at the middle of the beams and near the support.

Transfer zone of prestressed CFRP reinforcement applied according to NSM technique for strengthening of RC structures

This study presents an experimental program to assess the tensile strain distribution along prestressed carbon fiber reinforced polymer (CFRP) reinforcement flexurally applied on the tensile surface of RC beams according to near surface mounted (NSM) technique. Moreover, the current study aims to propose an analytical formulation, with a design framework, for the prediction of distribution of CFRP tensile strain and bond shear stress and, additionally, the prestress transfer length. After demonstration the good predictive performance of the proposed analytical approach, parametric studies were carried out to analytically evaluate the influence of the main material properties, and CFRP and groove cross section on the distribution of the CFRP tensile strain and bond shear stress, and on the prestress transfer length. The proposed analytical approach can also predict the evolution of the prestress transfer length during the curing time of the adhesive by considering the variation of its elasticity modulus during this period.

Application of acoustic emission technique for bond characterization in FRP-masonry systems

The acoustic emission (AE) technique is used for investigating the interfacial fracture and damage propagation in GFRP-and SRG-strengthened bricks during debonding tests. The bond behavior is investigated through single-lap shear bond tests and the fracture progress during the tests is recorded by means of AE sensors. The fracture progress and active debonding mechanisms are characterized in both specimen types with the aim of AE outputs. Moreover, a clear distinction between the AE outputs of specimens with different failure modes, in both SRG-and GFRP-strengthened specimens, is found which allows characterizing the debonding failure mode based on acoustic emission data.