Crystallographic studies on membrane and cytoplasmic enzymes

Dissertation presented to obtain the Ph.D degree in Biochemistry, Structural Biochemistry

Studies of the assembly of the Streptococcus pneumoniae capsular polysaccharide

Dissertation presented to obtain the Ph.D degree in Biology.

Understanding the microbial ecology and ecophysiology of enhanced biological phosphorus removal processes through metabolic modelling and experimental studies

Dissertation to obtain the degree of Master in Chemical and Biochemical Engineering

Controlling gene expression in enterobacteriaceae: studies on sRNAs and strategies for synthetic biology

Dissertation presented to obtain the Master Degree in Molecular, Genetics and Biomedicine

Functional and structural studies of a mini-ferritin protein

Dissertação para obtenção do Grau de Mestre em Bioquímica

Studies towards modified chitooligosaccharides: a new approach to NAG-NAM moiety

Dissertação para obtenção do Grau de Mestre em Bioorgânica

Upgrade of an experimental volumetric unit for gas adsorption equilibrium studies

In this work, a volumetric unit previously assembled by the research group was upgraded. This unit revamping was necessary due to the malfunction of the solenoid valves employed in the original experimental setup, which were not sealing the gas properly leading to erroneous adsorption equilibrium measurements. Therefore, the solenoid valves were substituted by manual ball valves. After the volumetric unit improvement its operation was validated. For this purpose, the adsorption equilibrium of carbon dioxide (CO2) at 323K and 0 - 20 bar was measured on two different activated carbon samples, in the of extrudates (ANG6) and of a honeycomb monolith (ACHM). The adsorption equilibrium results were compared with data previously measured by the research group, using a high-pressure microbalance from Rubotherm GmbH (Germany) – gravimetric. The results obtained using both apparatuses are coincident thus validating the good operation of the volumetric unit upgraded in this work. Furthermore, the adsorption equilibrium of CO2 at 303K and 0 - 10 bar on Metal-Organic Frameworks (MOFs) Cu-BTC and Fe-BTC was also studied. The CO2 adsorption equilibrium results for both MOFs were compared with the literature results showing good agreement, which confirms the good quality of the experimental results obtained in the new volumetric unit. Cu-BTC sample showed significantly higher CO2 adsorption capacity when compared with the Fe-BTC sample. The revamping of the volumetric unit included a new valve configuration in order to allow testing an alternative method for the measurement of adsorption equilibrium. This new method was employed to measure the adsorption equilibrium of CO2 on ANG6 and ACHM at 303, 323 and 353K within 0-10 bar. The good quality of the obtained experimental data was testified by comparison with data previously obtained by the research group in a gravimetric apparatus.

Double-walled PLLA/PLGA particles for the sustained release of Meloxicam: preparation, characterisation and contolled release studies

There were two main objectives in this thesis investigation, first, the production, characterisation, in vitro degradation and release studies of double walled microspheres for drug release control. The second one, and the most challenging, was the production of double walled nanospheres, also for drug control delivery. The spheres were produced using two polymers, the Poly(L-lactide)Acid, PLLA, and the Poly(L-lactide-co-glycolic)Acid, PLGA.Afterwards, a model drug, Meloxicam, which is an antiinflammatory drug, was encapsulated into the particles. Micro and nanospheres were produced by the solvent extraction/evaporation method, where perfect spherical particles were obtained. By varying the polymers PLLA/PLGA mass ratio, different core and shell composition, as well as several shell and core thickness were observed. In the particles with a PLLA/PLGA mass ratio 1:1, the shell is composed by PLLA and the core by PLGA. It was also verified that the Meloxicam has a tendency to be distributed in the PLGA layer. Micro and nanoparticles were characterised in morphology, size, polymer cristalinity properties and drug distribution. Particles degradation studies was performed, where the particles in a PVA solution of pH 7,4 where placed in an incubator, during approximately 40 days, at 120rpm, and 37ºC, simulating, as much as possible, the human body environment. From these studies, the conclusion was that particles containing a PLGA shell and a PLLA core degrade more rapidly, due to the fact that PLLA is more hydrophobic than the PLGA. Concerning the drug release controlled results, done also for 40 and 50 days, they showed that the microspheres containing a shell of PLLA release more slowly than when the shell is composed of PLGA. This result was predictable, since the drug is solubilised in the PLGA polymer and so, in that case, the PLLA shell works like a barrier between the drug and the outer medium. Another positive aspect presented by this study is the lower initial burst effect, obtained when using double walled particles, which is one of the advantages of the same. In a second part of this investigation, the production of the nanospheres was the main goal, since it was not yet accomplished by other authors or investigators. After several studies, referring to the speed, time and type of agitation, as well as, the concentration and volume of the first aqueous solution of poly-vinyl-alcohol (PVA) during the process of solvent extraction/evaporation it was possible to obtain double walled nanospheres.(...)

Structural and functional studies of a high activity NiFeSe Hydrogenase

Theawareness that fossil fuels exist in limited quantities has stimulated research into energy production from renewable sources. Future energy sources! should! be! plentiful! with! negligible! impact! on! the! environment.! Hydrogen!has!the!potential!to!satisfy!these!requirements.!Nevertheless,!current! methods! of! H2! production! rely! on! nonOrenewable! resources.! Biological! H2! production! from! sunlight! or! biomass! is! an! appealing! alternative! to! current! production!methods.!!(...)

Politicization of non-political events

Spatial analysis and social network analysis typically take into consideration social processes in specific contexts of geographical or network space. The research in political science increasingly strives to model heterogeneity and spatial dependence. To better understand and geographically model the relationship between “non-political” events, streaming data from social networks, and political climate was the primary objective of the current study. Geographic information systems (GIS) are useful tools in the organization and analysis of streaming data from social networks. In this study, geographical and statistical analysis were combined in order to define the temporal and spatial nature of the data eminating from the popular social network Twitter during the 2014 FIFA World Cup. The study spans the entire globe because Twitter’s geotagging function, the fundamental data that makes this study possible, is not limited to a geographic area. By examining the public reactions to an inherenlty non-political event, this study serves to illuminate broader questions about social behavior and spatial dependence. From a practical perspective, the analyses demonstrate how the discussion of political topics fluсtuate according to football matches. Tableau and Rapidminer, in addition to a set basic statistical methods, were applied to find patterns in the social behavior in space and time in different geographic regions. It was found some insight into the relationship between an ostensibly non-political event – the World Cup - and public opinion transmitted by social media. The methodology could serve as a prototype for future studies and guide policy makers in governmental and non-governmental organizations in gauging the public opinion in certain geographic locations.

Information provision improvement with a geofencing event-bases system

Nowadays there is a big percentage of the population, specially young users, which are smartphone users and there is a lot of information to be provided within the applications, information provision should be done carefully and should be accurate, otherwise an overload of information will be produced, and the user will discard the app which is providing the information. Mobile devices are becoming smarter and provide many ways to filter information. However, there are alternatives to improve information provision from the side of the application. Some examples are, taking into account the local time, considering the battery level before doing an action and checking the user location to send personalized information attached to that location. SmartCampus and SmartCities are becoming a reality and they have more and more data integrated every day. With all this amount of data it is crucial to decide when and where is the user going to receive a notification with new information. Geofencing is a technique which allows applications to deliver information in a more useful way, in the right time and in the right place. It consists of geofences, physical regions delimited by boundaries, and devices that are eligible to receive the information assigned to the geofence. When devices cross one of these geofences an alert is pushed to the mobile device with the information.

Crystallographic studies of proteins involved in the mRNA localization mechanisms in Drosophila melanogaster and amidation of the peptidoglycan residues in Staphylococcus aureus

Part of the work described in this chapter, was the subject of the following publication: D. Vieira, T. a. Figueiredo, A. Verma, R. G. Sobral, A. M. Ludovice, H. de Lencastre, and J. Trincao, “Purification, crystallization and preliminary X-ray diffraction analysis of GatD, a glutamine amidotransferase-like protein from Staphylococcus aureus peptidoglycan,” Acta Crystallogr. Sect. F Struct. Biol. Commun., vol. 70, no. 5, pp. 1–4, Apr. 2014.

Foresight exercises as a tool for decision-making: the example of two case studies in health

Based on the report for the unit “Foresight Methods Analysis” of the PhD programme on Technology Assessment at the Universidade Nova de Lisboa, under the supervision of Prof. Dr. António B. Moniz

Experimental and modeling studies on reverse electrodialysis for sustainable power generation

A potentially renewable and sustainable source of energy is the chemical energy associated with solvation of salts. Mixing of two aqueous streams with different saline concentrations is spontaneous and releases energy. The global theoretically obtainable power from salinity gradient energy due to World’s rivers discharge into the oceans has been estimated to be within the range of 1.4-2.6 TW. Reverse electrodialysis (RED) is one of the emerging, membrane-based, technologies for harvesting the salinity gradient energy. A common RED stack is composed by alternately-arranged cation- and anion-exchange membranes, stacked between two electrodes. The compartments between the membranes are alternately fed with concentrated (e.g., sea water) and dilute (e.g., river water) saline solutions. Migration of the respective counter-ions through the membranes leads to ionic current between the electrodes, where an appropriate redox pair converts the chemical salinity gradient energy into electrical energy. Given the importance of the need for new sources of energy for power generation, the present study aims at better understanding and solving current challenges, associated with the RED stack design, fluid dynamics, ionic mass transfer and long-term RED stack performance with natural saline solutions as feedwaters. Chronopotentiometry was used to determinate diffusion boundary layer (DBL) thickness from diffusion relaxation data and the flow entrance effects on mass transfer were found to avail a power generation increase in RED stacks. Increasing the linear flow velocity also leads to a decrease of DBL thickness but on the cost of a higher pressure drop. Pressure drop inside RED stacks was successfully simulated by the developed mathematical model, in which contribution of several pressure drops, that until now have not been considered, was included. The effect of each pressure drop on the RED stack performance was identified and rationalized and guidelines for planning and/or optimization of RED stacks were derived. The design of new profiled membranes, with a chevron corrugation structure, was proposed using computational fluid dynamics (CFD) modeling. The performance of the suggested corrugation geometry was compared with the already existing ones, as well as with the use of conductive and non-conductive spacers. According to the estimations, use of chevron structures grants the highest net power density values, at the best compromise between the mass transfer coefficient and the pressure drop values. Finally, long-term experiments with natural waters were performed, during which fouling was experienced. For the first time, 2D fluorescence spectroscopy was used to monitor RED stack performance, with a dedicated focus on following fouling on ion-exchange membrane surfaces. To extract relevant information from fluorescence spectra, parallel factor analysis (PARAFAC) was performed. Moreover, the information obtained was then used to predict net power density, stack electric resistance and pressure drop by multivariate statistical models based on projection to latent structures (PLS) modeling. The use in such models of 2D fluorescence data, containing hidden, but extractable by PARAFAC, information about fouling on membrane surfaces, considerably improved the models fitting to the experimental data.

How much cultural events are "worth" for host communities: Case of Faz Música Lisboa

This paper studies the economic and social impact of Faz Música Lisboa on the segment of society that feels from any direct or indirect consequence due to its existence. A qualitative research based on surveys and interviews is made to retract a list of the benefits and costs that each stakeholder perceives. Relying on the quantifiable variables, it is performed a cost-benefit analysis to measure how much the event is “worth” for the community. I conclude this is a viable project, as it brings a positive net benefit to the society, value that could increase with a higher institutional support.