Mechanisms underlying intracellular delivery

AuNPs are versatile systems used for different biomedical application including imaging, drug and gene delivery. These systems support the intracellular transport of active molecules, a step that is considered one of the crucial problems in drug delivery. Nevertheless, in order to design optimal multifunctional AuNPs for specific and efficient nanomedicine applications, the mechanism by which AuNPs interact with living cells must be fully understand. The main goal of this work consisted in the assessment of the cellular uptake mechanism of 14 nm spherical AuNPs by A549 cells, through fluorescent spectroscopy and microscopy, in combination with quantitative analysis by ICP-MS. TAMRA labeled AuNPs were characterized by UV-visible and fluorescent spectroscopy and the final hydrodynamic diameter of 22.5 ± 0.33 nm was obtained by DLS. Regarding the cellular uptake studies, the AuNPs presented a fast cellular uptake kinetics reaching a saturation point after 6 hours of incubation in A549 cells. Further investigation concerning the internalization mechanism of this AuNPs was evaluated using specific inhibitors for different endocytic pathways. Optimal inhibition was achieved using chlorpromazine, inhibitor of clathrin-mediated endocytosis, resulting in a 23.5 % inhibition of AuNPs after 1 hour of incubation. This preliminary result obtained by fluorescent spectroscopy suggests that these AuNPs were predominantly uptake by clathrin-mediated endocytosis, meaning that other endocytic pathways must be involved in the cellular uptake of this AuNPs. In what cell viability is concern, the prepared AuNPs and the endocytic inhibitors revealed no significant effect on the cell viability in A549 cell line.

What do broadband consumers want? Design and execution of a questionnaire on demand for bundled services on the Portuguese telecomunications market

The report addresses the question of what are the preferences of broadband consumers on the Portuguese telecommunication market. A triple play bundle is being investigated. The discrete choice analysis, adopted in the study, base on 110 responses, mainly from NOVA students. The data for the analysis was collected via manually designed on-line survey. The results show that the price attribute is relatively the most important one while the television attribute is being overlooked in the decision making process. Main effects examined in the research are robust. In addition, "extras" components are being tested in terms of users' preferences.

The role of guarantees in pension plan design: Pension saving decisions

This study centers on the assessment of psychological value of guarantees in pension products and the behavior biases associated with choice. When a guarantee on a product increases from 99% to 99,5% less than half of respondents show willingness to pay in contrast with 73% when going from 99,5% to 100%. Out of 105 respondents, 55 show that their choices concerning pension products are inconsistent with classic utility theory. Financial background proves insignificant thus pointing to behavioral biases. As individuals make choices that leave them worse-off, we argue that pension plan design would highly benefit from public policy interventions.

Design and implementation of an assurance map at Sonae retail business

The present Working Project aims at studying the topic of assurance mapping in a specific organizational context of a Portuguese retail company. For this purpose, an assurance map framework was designed to support the decision making process of stakeholders, through the delivery of comfort concerning risks, operations and control. In the end, the framework was successfully implemented for the process sourcing of goods in two business units of the company. Although, further implementation of the framework proved not to be feasible during the project’s timespan, it is expected to occur in the near future.

Design of bio-inspired ionic liquids for protein stabilisation

Ionic Liquids (ILs) consist in organic salts that are liquid at/or near room temperature. Since ILs are entirely composed of ions, the formation of ion pairs is expected to be one essential feature for describing solvation in ILs. In recent years, protein - ionic liquid (P-IL) interactions have been the subject of intensive studies mainly because of their capability to promote folding/unfolding of proteins. However, the ion pairs and their lifetimes in ILs in P-IL thematic is dismissed, since the action of ILs is therefore the result of a subtle equilibrium between anion-cation interaction, ion-solvent and ion-protein interaction. The work developed in this thesis innovates in this thematic, once the design of ILs for protein stabilisation was bio-inspired in the high concentration of organic charged metabolites found in cell milieu. Although this perception is overlooked, those combined concentrations have been estimated to be ~300 mM among the macromolecules at concentrations exceeding 300 g/L (macromolecular crowding) and transient ion-pair can naturally occur with a potential specific biological role. Hence the main objective of this work is to develop new bio-ILs with a detectable ion-pair and understand its effects on protein structure and stability, under crowding environment, using advanced NMR techniques and calorimetric techniques. The choline-glutamate ([Ch][Glu]) IL was synthesized and characterized. The ion-pair was detected in water solutions using mainly the selective NOE NMR technique. Through the same technique, it was possible to detect a similar ion-pair promotion under synthetic and natural crowding environments. Using NMR spectroscopy (protein diffusion, HSQC experiments, and hydrogen-deuterium exchange) and differential scanning calorimetry (DSC), the model protein GB1 (production and purification in isotopic enrichment media) it was studied in the presence of [Ch][Glu] under macromolecular crowding conditions (PEG, BSA, lysozyme). Under dilute condition, it is possible to assert that the [Ch][Glu] induces a preferential hydration by weak and non-specific interactions, which leads to a significant stabilisation. On the other hand, under crowding environment, the [Ch][Glu] ion pair is promoted, destabilising the protein by favourable weak hydrophobic interactions , which disrupt the hydration layer of the protein. However, this capability can mitigates the effect of protein crowders. Overall, this work explored the ion-pair existence and its consequences on proteins in conditions similar to cell milieu. In this way, the charged metabolites found in cell can be understood as key for protein stabilisation.

Frequency-domain receiver design for doubly-selective channels

This work is devoted to the broadband wireless transmission techniques, which are serious candidates to be implemented in future broadband wireless and cellular systems, aiming at providing high and reliable data transmission and concomitantly high mobility. In order to cope with doubly-selective channels, receiver structures based on OFDM and SC-FDE block transmission techniques, are proposed, which allow cost-effective implementations, using FFT-based signal processing. The first subject to be addressed is the impact of the number of multipath components, and the diversity order, on the asymptotic performance of OFDM and SC-FDE, in uncoded and for different channel coding schemes. The obtained results show that the number of relevant separable multipath components is a key element that influences the performance of OFDM and SC-FDE schemes. Then, the improved estimation and detection performance of OFDM-based broadcasting systems, is introduced employing SFN (Single Frequency Network) operation. An initial coarse channel is obtained with resort to low-power training sequences estimation, and an iterative receiver with joint detection and channel estimation is presented. The achieved results have shown very good performance, close to that with perfect channel estimation. The next topic is related to SFN systems, devoting special attention to time-distortion effects inherent to these networks. Typically, the SFN broadcast wireless systems employ OFDM schemes to cope with severely time-dispersive channels. However, frequency errors, due to CFO, compromises the orthogonality between subcarriers. As an alternative approach, the possibility of using SC-FDE schemes (characterized by reduced envelope fluctuations and higher robustness to carrier frequency errors) is evaluated, and a technique, employing joint CFO estimation and compensation over the severe time-distortion effects, is proposed. Finally, broadband mobile wireless systems, in which the relative motion between the transmitter and receiver induces Doppler shift which is different or each propagation path, is considered, depending on the angle of incidence of that path in relation to the direction of travel. This represents a severe impairment in wireless digital communications systems, since that multipath propagation combined with the Doppler effects, lead to drastic and unpredictable fluctuations of the envelope of the received signal, severely affecting the detection performance. The channel variations due this effect are very difficult to estimate and compensate. In this work we propose a set of SC-FDE iterative receivers implementing efficient estimation and tracking techniques. The performance results show that the proposed receivers have very good performance, even in the presence of significant Doppler spread between the different groups of multipath components.

The mixed impact of design thinking on business collaborations

The present work project investigates the implications of design thinking on an interbusiness collaborative project, unveiling both the benefits and fragilities that derive from the use of such methodology within the mentioned context. The study is organized in two parts. The first one starts with a literature review on the two relevant frames of reference – design thinking and business collaboration. It proceeds to the creation of a conceptual framework to assess the potential value of the design-based approach. The second part focuses on the practical application of to the newly developed instrument to the inTRAIN project – R&D for railway interiors.

The Mechanism of Centriole Inactivation in Starfish Oocytes

The centrosome is the major organizing center in a cell, composed by two centrioles, one mother and one daughter, and surrounded by a pericentriolar material, which nucleates microtubules. Centriole duplication and segregation is tightly coupled to cell cycle, which guarantees that centriole number is maintained over generations. During the somatic cell cycle, a pair of centrioles duplicates, after which each daughter cell receives a pair, forming a closed cycle. However, during fertilization, if both cells were to contribute with their pair of centrioles, gamete fusion would result in the double of the normal centriole number.(...)

Design formula for the flexural strengthening of RC beams using prestressed CFRP reinforcement

Applying a certain prestress level to the carbon fiber reinforced polymer (CFRP) reinforcement according to either externally bonded reinforcing (EBR) or near surface mounted (NSM) techniques can mobilize the strengthening potentialities of this high tensile strength composite material. For the prediction of the flexural behavior of reinforced concrete (RC) structures strengthened with prestressed EBR or NSM CFRPs, however, simplified analytical and design formulations still need to be developed as a guidance for engineers to design this type of strengthened structures by hand calculation without any programming help. Hence, the current work aims to briefly explain a developed simplified analytical approach, with a design framework, to predict the flexural behavior of RC beams flexurally strengthened with either prestressed EBR or NSM CFRP reinforcements. Moreover, an upper limit for the prestress level is proposed in order to optimize the ductility performance of the NSM prestressing technique. The good predictive performance of the analytical approaches was appraised by simulating the results of experimental programs composed of RC beams strengthened with prestressed NSM CFRP reinforcements.

Affordable prefabricated modular houses using cement and polymer based materials and advanced design tools

By taking advantage of the appropriate use of cement and polymer based materials and advanced computational tools, a pre-fabricated affordable house was built in a modular system. Modular system refers to the complete structure that is built-up by assembling pre-fabricated sandwich panels composed of steel fibre reinforced self-compacting concrete (SFRSCC) outer layers that are connected by innovative glass fibre reinforced polymer (GFRP) connectors, resulting in a panel with adequate structural, acoustic, and thermal insulation properties. The modular house was prepared for a typical family of six members, but its living area can be easily increased by assembling other pre-fabricated elements. The speed of construction and the cost of the constructive elements make these houses competitive when compared to traditional solutions. In this paper the relevant research subjacent to this project (LEGOUSE) is briefly described, as well as the construction process of the built real scale prototype.

Assessment of the effectiveness of steel fibre reinforcement for the punching resistance of flat slabs by experimental research and design approach

The present paper deals with the experimental assessment of the effectiveness of steel fibre reinforcement in terms of punching resistance of centrically loaded flat slabs, and to the development of an analytical model capable of predicting the punching behaviour of this type of structures. For this purpose, eight slabs of 2550 x 2550 x 150 mm3 dimensions were tested up to failure, by investigating the influence of the content of steel fibres (0, 60, 75 and 90 kg/m3) and concrete strength class (50 and 70 MPa). Two reference slabs without fibre reinforcement, one for each concrete strength class, and one slab for each fibre content and each strength class compose the experimental program. All slabs were flexurally reinforced with a grid of ribbed steel bars in a percentage to assure punching failure mode for the reference slabs. Hooked ends steel fibres provided the unique shear reinforcement. The results have revealed that steel fibres are very effective in converting brittle punching failure into ductile flexural failure, by increasing both the ultimate load and deflection, as long as adequate fibre reinforcement is assured. An analytical model was developed based on the most recent concepts proposed by the fib Mode Code 2010 for predicting the punching resistance of flat slabs and for the characterization of the behaviour of fibre reinforced concrete. The most refined version of this model was capable of predicting the punching resistance of the tested slabs with excellent accuracy and coefficient of variation of about 5%.

An investigation of safety design practices of metal machines

BACKGROUND: Machinery safety issues are a challenge facing manufacturers who are supposed to create and provide products in a better and faster way. In spite of their construction and technological advance, they still contribute to many potential hazards for operators and those nearby. OBJECTIVE: The aim of this study is to investigate safety aspects of metal machinery offered for sale on Internet market according to compliance with minimum and fundamental requirements. METHODS: The study was carried out with the application of a checklist prepared on the basis of Directive 2006/42/EC and Directive 2009/104/EC and regulations enforcing them into Polish law. RESULTS: On the basis of the study it was possible to reveal the safety aspects that were not met in practice. It appeared that in the case of minimum requirements the most relevant problems concerned information, signal and control elements, technology and machinery operations, whereas as far as fundamental aspects are concerned it was hard to assure safe work process. CONCLUSIONS: In spite of the fact that more and more legal acts binding in the Member Countries of the European Union are being introduced to alleviate the phenomenon, these regulations are often not fulfilled.

Masonry components

Masonry is a non-homogeneous material, composed of units and mortar, which can be of different types, with distinct mechanical properties. The design of both masonry units and mortar is based on the role of the walls in the building. Load-bearing walls relate to structural elements that bear mainly vertical loads, but can serve also to resist to horizontal loads. When a structural masonry building is submitted to in-plane and out-of-plane loadings induced by an earthquake for example, the masonry walls are the structural elements that ensure the global stability of the building. This means that the walls should have adequate mechanical properties that enable them to resist to different combinations of compressive, shear and tensile stresses.The boundary conditions influence the resisting mechanisms of the structural walls under in-plane loading and in a buildings the connection at the intersection walls are of paramount importance for the out-of-plane resisting mechanism. However, it is well established that the masonry mechanical properties are also relevant for the global mechanical performance of the structural masonry walls. Masonry units for load-bearing walls are usually laid so that their perforations are vertically oriented, whereas for partition walls, brick units with horizontal perforation are mostly adopted.

Design of a case-based reasoner for information security in military organizations

Information security is concerned with the protection of information, which can be stored, processed or transmitted within critical information systems of the organizations, against loss of confidentiality, integrity or availability. Protection measures to prevent these problems result through the implementation of controls at several dimensions: technical, administrative or physical. A vital objective for military organizations is to ensure superiority in contexts of information warfare and competitive intelligence. Therefore, the problem of information security in military organizations has been a topic of intensive work at both national and transnational levels, and extensive conceptual and standardization work is being produced. A current effort is therefore to develop automated decision support systems to assist military decision makers, at different levels in the command chain, to provide suitable control measures that can effectively deal with potential attacks and, at the same time, prevent, detect and contain vulnerabilities targeted at their information systems. The concept and processes of the Case-Based Reasoning (CBR) methodology outstandingly resembles classical military processes and doctrine, in particular the analysis of “lessons learned” and definition of “modes of action”. Therefore, the present paper addresses the modeling and design of a CBR system with two key objectives: to support an effective response in context of information security for military organizations; to allow for scenario planning and analysis for training and auditing processes.

Assessing student individual performance within PBL teams: findings from the implementation of a new mechanism

This paper reports on the experience of the implementation of a new mechanism to assess individual student contribution within project work, where students work in teams to solve a large-scale open-ended interdisciplinary project. The study takes place at the University of Minho, with first year engineering students, enrolled in the Industrial Management and Engineering (Integrated Masters) degree. The aim of this paper is to describe the main principles and procedures underlying the assessment mechanism created and also provide some feedback from its first implementation, based on the students, lecturers and tutors perceptions. For data collection, a survey was sent to all course lecturers and tutors involved in the assessment process. Students also contributed with suggestions, both on a workshop held at the end of the project and also by answering a survey on the overall satisfaction with PBL experience. Findings show a positive level of acceptance of the new mechanism by the students and also by the lecturers and tutors. The study identified the need to clarify the criteria used by the lecturers and the exact role of the tutor, as well as the need for further improvement of its features and procedures. Some recommendations are also issued regarding technical aspects related to some of the steps of the procedures, as well as the need for greater support on the adjustment and final setting of the individual grades.