Development and application of NMR techniques to study water-in-CO2 microemulsions

Self-assembly is a phenomenon that occurs frequently throughout the universe. In this work, two self-assembling systems were studied: the formation of reverse micelles in isooctane and in supercritical CO2 (scCO2), and the formation of gels in organic solvents. The goal was the physicochemical study of these systems and the development of an NMR methodology to study them. In this work, AOT was used as a model molecule both to comprehensively study a widely researched system water/AOT/isooctane at different water concentrations and to assess its aggregation in supercritical carbon dioxide at different pressures. In order to do so an NMR methodology was devised, in which it was possible to accurately determine hydrodynamic radius of the micelle (in agreement with DLS measurements) using diffusion ordered spectroscopy (DOSY), the micellar stability and its dynamics. This was mostly assessed by 1H NMR relaxation studies, which allowed to determine correlation times and size of correlating water molecules, which are in agreement with the size of the shell that interacts with the micellar layer. The encapsulation of differently-sized carbohydrates was also studied and allowed to understand the dynamics and stability of the aggregates in such conditions. A W/CO2 microemulsion was prepared using AOT and water in scCO2, with ethanol as cosurfactant. The behaviour of the components of the system at different pressures was assessed and it is likely that above 130 bar reverse microemulsions were achieved. The homogeneity of the system was also determined by NMR. The formation of the gel network by two small molecular organogelators in toluene-d8 was studied by DOSY. A methodology using One-shot DOSY to perform the spectra was designed and applied with success. This yielded an understanding about the role of the solvent and gelator in the aggregation process, as an estimation of the time of gelation.

Plasma nitriding of AA2011 alloy and surface characterization by NDT techniques

Al-Cu alloys are widely used in the aerospace and automotive industries due to their high specific strength in some tempered conditions. However, due to poor corrosion and wear resistance, they are often anodized and/or painted. Plasma nitriding has been proposed as an alternative, though the developments in this technique are still in a recent stage for Al alloys. Electrical characterization techniques are well implemented NDTs in the industry because of good accuracy associated with lower cost, compared to other methods. Some, like eddy currents and 4-point probe techniques, are often used in coating inspection. The objective of this study was to perform Al nitriding at low temperatures to minimize the tempering initial condition damage and to assess the feasibility of eddy currents technique as a method for evaluating surface properties. The work developed can be divided in two stages. The first one was the process tuning, done at the Shibaura Institute of Technology, in Tokyo; and the second was the electrical characterization done in Faculdade de Ciências e Tecnologia, UNL. Low temperature nitriding of AA2011 alloy specimens was successfully achieved. Electrical conductivity results show that lift-off measurements by eddy currents testing can be related to surface properties.

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.

Bio-valorization of crude glycerol through polyhydroxyalkanoates production

Due to the prospective partial replacement of fossil fuels by biodiesel, its production has continuously grown in the last decade. The increase in global biodiesel production demands the development of sustainable applications of its main by-product, crude glycerol. In this thesis the feasibility of producing polyhydroxyalkanoates (PHA) by a mixed microbial community using crude glycerol as feedstock was investigated. Several incubation conditions were studied in order to maximize PHA production. The microbial population selected under aerobic dynamic feeding conditions had the ability to consume both major carbon fractions present in the crude, glycerol and methanol. Two biopolymers were stored, poly-3-hydroxybutyrate (PHB) and glucose biopolymer (GB), apparently using glycerol as the only carbon source for their production. The microbial enrichment obtained was able to accumulate up to 47% PHB of cell dry weight with a productivity of 0.24 g HA/L d. The overall PHA yield on total substrate consumed (0.32 g COD HB/g COD crude glycerol) was in the middle range of those reported in literature (0.08–0.58 g COD PHA/g COD real waste). The increase of temperature from 23ºC to 30ºC favored the culture fraction that accumulates glucose biopolymer with a maximum accumulation value of 25% of cell dry weight, which is an interesting value but not the main goal of this thesis. The fact that crude glycerol can be used to produce PHA without any pre-treatment step, makes the overall production process economically more competitive, reducing polymer final cost. This was the first study that demonstrates the valorization of the glycerol fraction present in the crude glycerol into PHA using an aerobic mixed microbial consortium.

ReedCob – An eco-efficient building technology for monolithic walls based on earth and reeds

4th Conference COST ACTION FP1303 – Designing with Bio-based Materials – Challenges and opportiunities. INIA – CSIC, Madrid, 24-25 February 2016. Book of abstracts, T.Troya, J.Galván, D.Jones (Eds.), INIA and IETcc – CSIS, pg. 79-80 (ISBN: 978-91-88349-16-3)

Endovascular techniques for placement of long-term chemotherapy catheters

PURPOSE: To analyze the results from using endovascular techniques to place long-term chemotherapy catheters when advancing the catheter using the external jugular vein is difficult due to obstructions or kinking. METHODS: Between July 1997 and August 2000, 320 long-term chemotherapy catheters were placed, and in 220 cases the external jugular vein was used as the primary venous approach. In 18 of these patients, correct positioning was not achieved and several endovascular techniques were then utilized to overcome these obstacles, including manipulation of a J-wire with a moveable core, venography, and the exchange wire technique. RESULTS: In 94.5% of the patients with difficulties in obtaining the correct positioning, we were able to advance the long-term catheter to the desired position with the assistance of endovascular techniques. CONCLUSIONS: Venography and endovascular guidance techniques are useful for the placement of long-term catheters in the external jugular vein.

The reasons behind the progression in PISA scores: An education production function approach using semi-parametric techniques

Research Masters

Advancing tendon regeneration through the development of bio-stimulating tissue engineering approaches

Tendon tissue engineering (TE) requires tailoring scaffolds designs and properties to the anatomical and functional requirements of tendons located in different regions of the body. Cell sourcing is also of utmost importance as tendon cells are scarce. Recently, we have found that it is possible to direct the tenogenic differentiation of Amniotic fluid and Adipose tissue derived stem cells (hAFSCs and hASCs), and also that there are hASCs subpopulations that might be more prone to tenogenic differentiation. Nevertheless, biochemical stimulation may not be enough to develop functional TE substitutes for a tissue that is known to be highly dependent on mechanical loading.

Toxicity measurement techniques for building materials with wastes

The innovative Horizon 2020 program sponsored by the European Union (EU) aims to promote and develop processes of waste integration in construction materials. However, several potential health hazards caused by building materials have been identified and, there-fore, there is an ongoing need to develop new recycling methods for hazardous wastes and effi-cient barriers in order to prevent toxic releases from the new construction solutions with wastes. This paper presents an overview that focus on two main aspects: the identification of the health risks related to radioactivity and heavy metals present in building materials and identification of these toxic substances in new construction solutions that contain recycled wastes. Different waste materials were selected and distinct methodologies of toxicity evaluation are presented to analyse the potential hazardous, the feasibility of using those wastes and the achievement of op-timal construction solutions involving wastes.

Biologia e controle microbiano de Urbanus acawoios (Williams, 1926) (Lepidoptera: Hesperiidae): I. descrição morfológica e aspectos Bio-ecológicos

Devido aos danos dos ataques de Urbanus acawoios (Williams, 1926) (Lepidoptera: Hesperiidae) em palheteiras (Clitoria racemosa) (Leguminosae) em Manaus, AM e a total ausência de qualquer informação a respeito desta espécie, o presente trabalho foi desenvolvido visando a obtenção de conhecimentos que facilitem a identificação de métodos seguros e eficientes para o seu controle. Os estudos biológicos revelaram que este inseto é multivoltino, tendo duração média de 41,89 ± 0,98 dias por geração. As condições climáticas na cidade de Manaus, a disponibilidade de alimentos, e possivelmente a ausência de inimigos naturais eficientes, parecem ser os fatores principais responsáveis pela ocorrência de erupções populacionais desta espécie. Enquanto o estágio larval consiste de 5 fases (ínstares) e dura em média 18,46 ± 1,28 dias, o da pupa necessita 9,96 ± 1,05, e o adulto 13,47 ± 0,27 dias, sob condições de 30°C e 85% UR. Estudos morfológicos foram incluídos.

Ancient materials and techniques to improve the earthen building durability

A substantial part of the world building heritage has been performed by earthen building. The durability of this existing heritage and mainly of the new buildings built with earth is particularly conditioned by the erosion caused by water action, especially in countries with high levels of rainfall. This research aims to contribute to the increase of knowledge about the ancient building techniques that provide enhanced durability. It is possible to analyse the ancestral practices used to protect the earth material from the water action in order to understand how the old earthen buildings were preserved over the centuries, resisting to harsh weather conditions. Among these techniques are: the incorporation of biopolymers (such as oils or fats from animal or vegetable origin); the addition of some minerals; and the earth stabilization with lime. However, this knowledge seems to be forgotten, probably due to the prejudice related to earthen constructions, which several times are associated with a poor building. This research also focuses on the study of new methods of earth stabilization with lime and biopolymers, adapting the ancient knowledge to improve the durability related to the water action. Therefore, alternative solutions can be obtained to improve the performance of earthen buildings, mainly the resistance of the material in the presence of water, reducing its permeability to water. In addition, with the proposed solutions it is possible to obtain good levels of water vapour permeability, one of the major advantages of the construction with earth.

Implementation and comparison of different switching techniques for shunt active power filters

This paper presents a comparison between three switching techniques that can be used in three-phase four-wire Shunt Active Power Filters (SAPFs). The implemented switching techniques are: Periodic-Sampling (PS), Triangular Carrier Pulse-Width Modulation (TC-PWM) and Space Vector PWM (SVPWM). The comparison between them is made in terms of the compensated currents THD%, implementation complexity, necessary CPU time and SAPF efficiency. To perform this comparison are presented and analyzed several experimental results, obtained with a 20 kVA Shunt Active Power Filter prototype, specially developed for this purpose. The control system of the developed SAPF is based in the p-q Theory with a grid synchronization algorithm p-PLL.

Quantitative image analysis as a tool for Yarrowia lipolytica dimorphic growth evaluation in different culture media

Yarrowia lipolytica, a yeast strain with a huge biotechnological potential, capable to produce metabolites such as γ-decalactone, citric acid, intracellular lipids and enzymes, possesses the ability to change its morphology in response to environmental conditions. In the present study, a quantitative image analysis (QIA) procedure was developed for the identification and quantification of Y. lipolytica W29 and MTLY40-2P strains dimorphic growth, cultivated in batch cultures on hydrophilic (glucose and N-acetylglucosamine (GlcNAc) and hydrophobic (olive oil and castor oil) media. The morphological characterization of yeast cells by QIA techniques revealed that hydrophobic carbon sources, namely castor oil, should be preferred for both strains growth in the yeast single cell morphotype. On the other hand, hydrophilic sugars, namely glucose and GlcNAc caused a dimorphic transition growth towards the hyphae morphotype. Experiments for γ-decalactone production with MTLY40-2P strain in two distinct morphotypes (yeast single cells and hyphae cells) were also performed. The obtained results showed the adequacy of the proposed morphology monitoring tool in relation to each morphotype on the aroma production ability. The present work allowed establishing that QIA techniques can be a valuable tool for the identification of the best culture conditions for industrial processes implementation.

Refinement and variability techniques in model transformation of software requirements

Tese de Doutoramento em Tecnologias e Sistemas de Informação

Rockburst laboratory tests database - Application of data mining techniques

Rockburst is characterized by a violent explosion of a block causing a sudden rupture in the rock and is quite common in deep tunnels. It is critical to understand the phenomenon of rockburst, focusing on the patterns of occurrence so these events can be avoided and/or managed saving costs and possibly lives. The failure mechanism of rockburst needs to be better understood. Laboratory experiments are undergoing at the Laboratory for Geomechanics and Deep Underground Engineering (SKLGDUE) of Beijing and the system is described. A large number of rockburst tests were performed and their information collected, stored in a database and analyzed. Data Mining (DM) techniques were applied to the database in order to develop predictive models for the rockburst maximum stress (σRB) and rockburst risk index (IRB) that need the results of such tests to be determined. With the developed models it is possible to predict these parameters with high accuracy levels using data from the rock mass and specific project.