Heparinized nanohydroxyapatite/collagen granules for controlled release of vancomycin

The purpose of this study was to develop a bone substitute material capable of preventing or treating osteomyelitis through a sustainable release of vancomycin and simultaneously inducing bone regeneration. Porous heparinized nanohydroxyapatite (nanoHA)/collagen granules were characterized using scanning electron microscopy, micro-computed tomography and attenuated total reflectance Fourier transform infrared spectroscopy. After vancomycin adsorption onto the granules, its releasing profile was studied by UV molecular absorption spectroscopy. The heparinized granules presented a more sustainable release over time, in comparison with nonheparinized nanoHA and nanoHA/collagen granules. Vancomycin was released for 360 h and proved to be bioactive until 216 h. Staphylococcus aureus adhesion was higher on granules containing collagen, guiding the bacteria to the material with antibiotic, improving their eradication. Moreover, cytotoxicity of the released vancomycin was assessed using osteoblast cultures, and after 14 days of culture in the presence of vancomycin, cells were able to remain viable, increasing their metabolic activity and colonizing the granules, as observed by scanning electron microscopy and confocal laser scanning microscopy. These findings suggest that heparinized nanoHA/collagen granules are a promising material to improve the treatment of osteomyelitis, as they are capable of releasing vancomycin, eliminating the bacteria, and presented morphological and chemical characteristics to induce bone regeneration.

Competitive paths for methanol decomposition on ruthenium: a DFT study

Methanol decomposition is one of the key reactions in direct methanol fuel cell (DMFC) state-of-the-art technology, research, and development. However, its mechanism still presents many uncertainties, which, if answered, would permit us to refine the manufacture of DMFCs. The mechanism of methanol decomposition on ruthenium surfaces was investigated using density functional theory and a periodic supercell approach. The possible pathways, involving either initial C−H, C−O or O−H scission, were defined from experimental evidence regarding the methanol decomposition on ruthenium and other metallic surfaces. The study yielded the O−H scission pathway as having both the most favorable energetics and kinetics. The computational data, which present a remarkable closeness with the experimental results, also indicate methanol adsorption, the starting point in all possible pathways, to be of weak nature, implying a considerable rate of methanol desorption from the ruthenium, compromising the reaction.

Estudo eletroquímico do dano oxidativo promovido por RNS e PAH e o efeito protetor de antioxidantes no ADN

No dia-a-dia, os organismos vivos estão sujeitos a vários tipos de agressões de origem endógena e exógena. A produção endógena exagerada de contaminantes que ocorre nos processos metabólicos dos seres vivos está intimamente associada ao aparecimento e desenvolvimento de várias patologias. Por outro lado, e devido às atividades antropogénicas, muitos contaminantes de origem ambiental e alimentar entram por via exógena no organismo dos seres vivos provocando igualmente danos a nível celular. De modo a protegerem-se dos efeitos pejorativos provocados por estes compostos, os organismos vivos desenvolveram mecanismos complexos de defesa antioxidante. Este trabalho consistiu no estudo eletroquímico do dano oxidativo induzido por contaminantes (PAH (hidrocarbonetos aromáticos policíclicos), H2O2, NO• e HClO) e do efeito protetor, ao dano oxidativo, promovido por antioxidantes no material baseado no ADN recorrendo à utilização de um biossensor de bases púricas, adenina-EPC (elétrodo pasta de carbono) e dA20-EPC, utilizando a voltametria de onda quadrada (VOQ) como técnica de deteção. A aplicação da eletroquímica apresenta várias vantagens para a quantificação da capacidade antioxidante total (CAT) pois, permite a redução da quantidade de reagentes e amostra em análise, elimina a etapa de remoção de cor (a cor é um interferente nos métodos óticos) e não requer equipamentos dispendiosos. Foram seguidas diferentes abordagens para a construção dos biossensores. A primeira consistiu na construção de um adenina-EPC em três etapas: i) condicionamento do EPC, ii) eletrodeposição da adenina no EPC e iii) leitura do sinal eletroquímico. Assim, foram otimizados diversos parâmetros: concentração de adenina (150,0 mg/L), potencial de condicionamento (Ec) (+ 1,80 V), potencial de deposição (Ed) (+ 0,40 V), tempo de condicionamento (tc) (180 s) e tempo de deposição (td) (240 s). Foi aplicado o adenina-EPC no estudo do dano oxidativo provocado por PAH (benzo (g,h,i) perileno) e constatou-se que era necessário transformar o benzo (g,h,i) perileno num radical para se possível observar danos oxidativos induzidos no biossensor. A nova estratégia consistiu na construção de um dA20-EPC, através da adsorção física de uma gota de dA20 na superfície do EPC, com posterior secagem e leitura do sinal eletroquímico. Neste procedimento foi otimizada a concentração de dA20 (100,0 mg/L). O dano oxidativo provocado pelo H2O2, NO• e HClO foi estudado sobre o dA20-EPC e verificou-se que os três contaminantes induziam dano oxidativo no dA20-EPC. Confirmou-se a capacidade do ácido ascórbico (AA) em proteger o dA20-EPC do dano oxidativo induzido por H2O2 e NO•. O biossensor desenvolvido (dA20-EPC) foi aplicado na avaliação da CAT de diferentes amostras reais (café, sumo de laranja e água aromatizada de laranja) usando-se como contaminantes o H2O2 e NO•. Todas as amostras analisadas apresentaram ter capacidade antioxidante. Quando se usou o dA20-EPC na presença de H2O2, verificou-se que as amostras de café apresentam valores mais elevados de CAT (1130-1488 mg AAE/L) do que as amostras de bebidas (110 mg AAE/L em água aromatizada e 775 mg AAE/L em sumo). Os valores de CAT obtidos para amostras de sumo e água aromatizada na presença de NO• indicam que a amostra de sumo possui maior teor de CAT (526 mg AAE/L) conforme era esperado, do que a amostra de água aromatizada (172 mg AAE/L).

Heavy metals removal in dual media filters

Dissertação para obtenção do Grau de Mestre em Engenharia do Ambiente – Perfil de Engenharia Sanitária

Utilização de sensores de baixo custo na medição de monóxido de carbono no ar ambiente

Nos últimos anos tem-se assistido à introdução de novos dispositivos de medição da poluição do ar baseados na utilização de sensores de baixo custo. A utilização menos complexa destes sistemas, possibilita a obtenção de dados com elevada resolução temporal e espacial, abrindo novas oportunidades para diferentes metodologias de estudos de monitorização da poluição do ar. Apesar de apresentarem capacidades analíticas distantes dos métodos de referência, a utilização destes sensores tem sido sugerida e incentivada pela União Europeia no âmbito das medições indicativas previstas na Diretiva 2008/50/CE, com uma incerteza expandida máxima de 25%. O trabalho desenvolvido no âmbito da disciplina de Projeto consistiu na escolha, caracterização e utilização em medições reais de um sensor de qualidade do ar, integrado num equipamento protótipo desenvolvido com esse fim, visando obtenção uma estimativa da incerteza de medição associada à utilização deste dispositivo através da aplicação da metodologia de demonstração de equivalência de métodos de medição de qualidade do ar definida pela União Europeia. A pesquisa bibliográfica realizada permitiu constatar que o monóxido de carbono é neste momento o parâmetro de qualidade do ar que permite ser medido de forma mais exata através da utilização de sensores, nomeadamente o sensor eletroquímico da marca Alphasense, modelo COB4, amplamente utilizado em projetos de desenvolvimento neste cotexto de monitorização ambiental. O sensor foi integrado num sistema de medição com o objetivo de poder ser utlizado em condições de autonomia de fornecimento de energia elétrica, aquisição interna dos dados, tendo em consideração ser o mais pequeno possível e de baixo custo. Foi utlizado um sistema baseado na placa Arduino Uno com gravação de dados em cartão de memória SD, baterias e painel solar, permitindo para além do registo das tensões elétricas do sensor, a obtenção dos valores de temperatura, humidade relativa e pressão atmosférica, com um custo global a rondar os 300 euros. Numa primeira fase foram executados um conjunto de testes laboratoriais que permitiram a determinação de várias características de desempenho em dois sensores iguais: tempo de resposta, a equação modelo do sensor, avaliação da repetibilidade, desvio de curto e longo termo, interferência da temperatura e histerese. Os resultados demonstraram um comportamento dos sensores muito linear, com um tempo de resposta inferior a um minuto e com uma equação modelo do sensor dependente da variação da temperatura. A estimativa da incerteza expandida laboratorial ficou, para ambos os sensores, abaixo dos 10%. Após a realização de duas campanhas reais de medição de CO em que os valores foram muito baixos, foi realizada uma campanha de quinze dias num parque de estacionamento subterrâneo que permitiu a obtenção de concentrações suficientemente elevadas e a comparação dos resultados dos sensores com o método de referência em toda a gama de medição (0 a 12 mol.mol-1). Os valores de concentração obtidos pelos dois sensores demonstraram uma excelente correlação com o método de referência (r2≥0,998), obtendo-se resultados para a estimativa da incerteza expandida de campo inferiores aos obtidos para a incerteza laboratorial, cumprindo o objetivo de qualidade de dados definido para as medições indicativas de incerteza expandida máxima de 25%. Os resultados observados durante o trabalho realizado permitiram confirmar o bom desempenho que este tipo de sensor pode ter no âmbito de medições de poluição do ar com um caracter mais indicativo.

Construção de um sensor eletroquímico molecularmente impresso para monitorização do cancro da mama

O Cancro da mama é uma doença cuja incidência tem vindo a aumentar de ano para ano e além disso é responsável por um grande número de mortes em todo mundo. De modo a combater esta doença têm sido propostos e utilizados biomarcadores tumorais que permitem o diagnóstico precoce, o acompanhamento do tratamento e/ou a orientação do tipo tratamento a adotar. Atualmente, os biomarcadores circulantes no sangue periférico recomendados pela Associação Americana de Oncologia Clinica (ASCO) para monitorizar os pacientes durante o tratamento são o cancer antigen 15-3 (CA 15-3), o cancer antigen 27.29 (CA 27.29) e o cancer embryobic antigen (CEA). Neste trabalho foi desenvolvido um sensor eletroquímico (voltamétrico) para monitorizar o cancro da mama através da análise do biomarcador CA 15-3. Inicialmente realizou-se o estudo da adsorção da proteína na superfície do elétrodo para compreender o comportamento do sensor para diferentes concentrações. De seguida, estudaram-se três polímeros (poliaminofenol, polifenol e polifenilenodiamina) e selecionou-se o poliaminofenol como o polímero a utilizar, pois possuía a melhor percentagem de alteração de sinal. Após a seleção do polímero, este foi depositado na superfície do elétrodo por eletropolimerização, formando um filme polimérico molecularmente impresso (MIP) à volta da proteína (molde). Posteriormente, foram analisados cinco solventes (água, mistura de dodecil sulfato de sódio e ácido acético, ácido oxálico, guanidina e proteinase K) e o ácido oxálico revelou ser mais eficaz na extração da proteína. Por último, procedeu-se à caraterização do sensor e analisou-se a resposta analítica para diferentes concentrações de CA 15-3 revelando diferenças claras entre o NIP (polímero não impresso) e o MIP.

Study of the valorisation of the solid by-products obtained in the co-pyrolysis of different wastes

Dissertação para obtenção do grau de doutor em Energia e Bioenergia

Role of surfactants in filtration and fouling of colloidal silica

The EM3E Master is an Education Programme supported by the European Commission, the European Membrane Society (EMS), the European Membrane House (EMH), and a large international network of industrial companies, research centres and universities

Gas gap heat switch for CryoFree project

Cryogen-free superconducting magnet systems have become popular during the last two decades for the simple reason that with the use of liquid helium is rather cumbersome and is a scarce resource. Some available CFMS uses a mechanical cryocooler as cold source of the superconductor magnet. However, the cooling of the sample holder is still made through an open circuit of helium. A thermal management of a completely cryogen-free system is possible to be implemented by using a controlled gas gap heat switch (GGHS) between the cryocooler and the variable temperature insert (VTI). This way it would eliminate the helium open circuit. Heat switches are devices that allow to toggle between two distinct thermal states (ON and OFF state). Several cryogenic applications need good thermal contact and a good thermal insulation at different stages of operation. A versatile GGHS was designed and built with a 100 mm gap and tested with helium as exchange gas. An analytic thermal model was developed and a good agreement with the experimental data was obtained. The device was tested on a crycooler at 4 to 80 K ranges. A 285 mW/K thermal conductance was measured at ON state and 0.09 mW/K at OFF. 3000 ON/OFF thermal conductance ratio was obtained at 4 K with helium.

Earthen plasters: the potential of the clayey soils of barrocal region in Algarve

Due to their high adsorption capacity of water vapor, earthen plasters can act as a moisture buffer, contributing to balance the relative humidity of the indoor environment of buildings. As a consequence of this capacity earthen plasters may also contribute to the perception of thermal comfort, since a high relative humidity increases the thermal conductivity of air and restricts skin evaporation, increasing the discomfort associated with the perception of heat or cold. Simultaneously, earthen plasters may also contribute to the indoor air quality. In one hand, by mitigating health problems of the respiratory system associated with indoor environment with high relative humidity, in which increases the risk of development of microorganisms usually responsible for infections, allergies or asthma. In the other hand, by mitigating the probability of inflammation of the respiratory system airways associated to exceedingly dry indoor environments. Therefore it also becomes expectable that earthen plasters may contribute for reducing the needs for air conditioning and mechanical ventilation in buildings and, thereby, also allowing the reduction of the associated energy consumption. The «Barrocal» region, located in the sedimentary basin of Algarve, South Portugal, presents geomorphological characteristics that promote the occurrence of soils with a clay mineralogy dominated by illite, which is a clay mineral characterized by a high adsorption capacity of water vapor and low expansibility. This fact turns expectable that these soils have a high potential for interior plastering. In order to evaluate this potential four mortars were formulated with an increasing content of clayey soil extracted from a selected clay quarry from «Barrocal» region. The results from the preliminary characterization campaign confirmed the reduced linear shrinkage of these mortars, as well as their high adsorption-desorption capacity, that is positively correlated with the content of clayey soil present in mortar formulation. However, the mechanical tests showed that the mechanical resistance of these mortars should be improved, for instance through the addition of natural fibers for reinforcement, which will be investigated in future research. This research contributed to increase certainty regarding the potential of clayey soils of the «Barrocal» sub-region of Algarve to produce mortars suitable for eco-efficient interior plastering.

Storage of water molecules into biomimetic heterostructures: the role of roughness

The development of devices based on heterostructured thin films of biomolecules conveys a huge contribution on biomedical field. However, to achieve high efficiency of these devices, the storage of water molecules into these heterostructures, in order to maintain the biological molecules hydrated, is mandatory. Such hydrated environment may be achieved with lipids molecules which have the ability to rearrange spontaneously into vesicles creating a stable barrier between two aqueous compartments. Yet it is necessary to find conditions that lead to the immobilization of whole vesicles on the heterostructures. In this work, the conditions that govern the deposition of open and closed liposomes of 1.2-dipalmitoyl-sn-Glycero-3-[Phospho-rac-(1-glycerol)] (sodium Salt) (DPPG) onto polyelectrolytes cushions prepared by the layer-by-layer (LbL) method were analyzed. Electronic transitions of DPPG molecules as well as absorption coefficients were obtained by vacuum ultraviolet spectroscopy, while the elemental composition of the heterostructures was characterized by x-ray photoelectron spectroscopy (XPS). The presence of water molecules in the films was inferred by XPS and infrared spectroscopy. Quartz crystal microbalance (QCM) data analysis allowed to conclude that, in certain cases, the DPPG adsorbed amount is dependent of the bilayers number already adsorbed. Moreover, the adsorption kinetics curves of both adsorbed amount and surface roughness allowed to determine the kinetics parameters that are related with adsorption processes namely, electrostatic forces, liposomes diffusion and lipids re-organization on surface. Scaling exponents attained from atomic force microscopy images statistical analysis demonstrate that DPPG vesicles adsorption mechanism is ruled by the diffusion Villain model confirming that adsorption is governed by electrostatic forces. The power spectral density treatment enabled a thorough description of the accessible surface of the samples as well as of its inner structural properties. These outcomes proved that surface roughness influences the adsorption of DPPG liposomes onto surfaces covered by a polyelectrolyte layer. Thus, low roughness was shown to induce liposome rupture creating a lipid bilayer while high roughness allows the adsorption of whole liposomes. In addition, the fraction of open liposomes calculated from the normalized maximum adsorbed amounts decreases with the cushion roughness increase, allowing us to conclude that the surface roughness is a crucial variable that governs the adsorption of open or whole liposomes. This conclusion is fundamental for the development of well-designed sensors based on functional biomolecules incorporated in liposomes. Indeed, LbL films composed of polyelectrolytes and liposomes with and without melanin encapsulated were successfully applied to sensors of olive oil.

Application of superconducting bulks and stacks of tapes in electrical machines

The present dissertation focuses on the research of the recent approach of innovative high-temperature superconducting stacked tapes in electrical ma-chines applications, taking into account their potential benefits as an alternative for the massive superconducting bulks, mainly related with geometric and me-chanical flexibility. This work was developed in collaboration with Institut de Ciència de Ma-terials de Barcelona (ICMAB), and is related with evaluation of electrical and magnetic properties of the mentioned superconducting materials, namely: analysis of magnetization of a bulk sample through simulations carried out in the finite elements COMSOL software; measurement of superconducting tape resistivity at liquid nitrogen and room temperatures; and, finally, development and testing of a frequency controlled superconducting motor with rotor built by superconducting tapes. In the superconducting state, results showed a critical current density of 140.3 MA/m2 (or current of 51.15 A) on the tape and a 1 N∙m developed motor torque, independent from the rotor position angle, typical in hysteresis motors.

Optimal design and operation of compact simulated moving bed processes for enantioseparations

Simulated moving bed (SMB) chromatography is attracting more and more attention since it is a powerful technique for complex separation tasks. Nowadays, more than 60% of preparative SMB units are installed in the pharmaceutical and in the food in- dustry [SDI, Preparative and Process Liquid Chromatography: The Future of Process Separations, International Strategic Directions, Los Angeles, USA, 2002. http://www. strategicdirections.com]. Chromatography is the method of choice in these ¯elds, be- cause often pharmaceuticals and ¯ne-chemicals have physico-chemical properties which di®er little from those of the by-products, and they may be thermally instable. In these cases, standard separation techniques as distillation and extraction are not applicable. The noteworthiness of preparative chromatography, particulary SMB process, as a sep- aration and puri¯cation process in the above mentioned industries has been increasing, due to its °exibility, energy e±ciency and higher product purity performance. Consequently, a new SMB paradigm is requested by the large number of potential small- scale applications of the SMB technology, which exploits the °exibility and versatility of the technology. In this new SMB paradigm, a number of possibilities for improving SMB performance through variation of parameters during a switching interval, are pushing the trend toward the use of units with smaller number of columns because less stationary phase is used and the setup is more economical. This is especially important for the phar- maceutical industry, where SMBs are seen as multipurpose units that can be applied to di®erent separations in all stages of the drug-development cycle. In order to reduce the experimental e®ort and accordingly the coast associated with the development of separation processes, simulation models are intensively used. One impor- tant aspect in this context refers to the determination of the adsorption isotherms in SMB chromatography, where separations are usually carried out under strongly nonlinear conditions in order to achieve higher productivities. The accurate determination of the competitive adsorption equilibrium of the enantiomeric species is thus of fundamental importance to allow computer-assisted optimization or process scale-up. Two major SMB operating problems are apparent at production scale: the assessment of product quality and the maintenance of long-term stable and controlled operation. Constraints regarding product purity, dictated by pharmaceutical and food regulatory organizations, have drastically increased the demand for product quality control. The strict imposed regulations are increasing the need for developing optically pure drugs.(...)

Biomimetic mineralization: encapsulation in calcium carbonate shells

Calcium carbonate biomineralization is a self-assembly process that has been studied to be applied in the biomedical field to encapsulate biomolecules. Advantages of engineering mineral capsules include improved drug loading efficiencies and protection against external environment. However, common production methods result in heterogeneous capsules and subject biomolecules to heat and vibration which cause irreversible damage. To overcome these issues, a microfluidic device was designed, manufactured and tested in terms of selectivity for water and oil to produce a W/O/W emulsion. During the development of this work there was one critical challenge: the selective functionalization in closed microfluidic channels. Wet chemical oxidation of PDMS with 1M NaOH, confirmed by FTIR, followed by adsorption of polyelectrolytes - PDADMAC/PSS - confirmed by UV-Vis and AFM results, render the surface of PDMS hydrophilic. UV-Vis spectroscopy also confirmed that this modification did not affect PDMS optical properties, making possible to monitor fluids and droplets. More important, with this approach PDMS remains hydrophilic over time. However, due to equipment constrains selectivity in microchannels was not achieved. Therefore, emulsion studies took place with conventional methods. Several systems were tried, with promising results achieved with CaCO3 in-situ precipitation, without the use of polymers or magnesium. This mineral stabilizes oil droplets in water, but not in air due to incomplete capsule formation.

Earthen plasters based on illitic soils from Barrocal region of Algarve: contributions for building performance and sustainability

Clayish earth-based mortars can be considered eco-efficient products for indoor plastering since they can contribute to improve important aspects of building performance and sustainability. Apart from being products with low embodied energy when compared to other types of mortars used for interior plastering, mainly due to the use raw clay as natural binder, earth-based plasters may give a significant contribution for health and comfort of inhabitants. Due to high hygroscopicity of clay minerals, earth-based mortars present a high adsorption and desorption capacity, particularly when compared to other type of mortars for interior plastering. This capacity allows earth-based plasters to act as a moisture buffer, balancing the relative humidity of the indoor environment and, simultaneously, acting as a passive removal material, improving air quality. Therefore, earth-based plasters may also passively promote the energy efficiency of buildings, since they may contribute to decreasing the needs of mechanical ventilation and air conditioning. This study is part of an ongoing research regarding earth-based plasters and focuses on mortars specifically formulated with soils extracted from Portuguese ‘Barrocal’ region, in Algarve sedimentary basin. This region presents high potential for interior plastering due to regional geomorphology, that promote the occurrence of illitic soils characterized by a high adsorption capacity and low expansibility. More specifically, this study aims to assess how clayish earth and sand ratio of mortars formulation can influence the physical and mechanical properties of plasters. For this assessment four mortars were formulated with different volumetric proportions of clayish earth and siliceous sand. The results from the physical and mechanical characterization confirmed the significantly low linear shrinkage of all the four mortars, as well as their extraordinary adsorption-desorption capacity. These results presented a positive correlation with mortars´ clayish earth content and are consistent with the mineralogical analysis, that confirmed illite as the prevalent clay mineral in the clayish earth used for this study. Regarding mechanical resistance, although the promising results of the adhesion test, the flexural and compressive strength results suggest that the mechanical resistance of these mortars should be slightly improved. Considering the present results the mortars mechanical resistance improvement may be achieved through the formulation of mortars with higher clayish earth content, or alternatively, through the addition of natural fibers to mortars formulation, very common in this type of mortars. Both those options will be investigated in future research.