Characterization and surface reconstruction of objects in tomographic images of composite materials

Dissertação para obtenção do Grau de Mestre em Engenharia Informática

Gold nanoparticles for plasmid delivery

Gene therapy presents an ideal strategy for the treatment of genetic as well as acquired diseases, such as cancer and typically involves the insertion of a functioning gene into cells to correct a cellular dysfunction or to provide a new cellular function. Gene delivery vectors are based in two models: viral and non-viral. Viral vectors have high transfection efficiency but their major barrier is immunogenicity. Since the non-viral vectors have no immunogenicity, these have been widely studied. Gold nanoparticles have been proposed as optimal delivery systems of genetic material, due their small size, high surface-to-volume ratio and the ability to be functionalized with multiple molecules. In the present work, an AuNP-based formulation was developed to deliver a plasmid in a colorectal cancer cell line, containing as reporter gene the gene encoding to EGFP. The delivery system resulted from the functionalization of 14 nm AuNP with a PEG layer (4300114 PEG chains/AuNP), which increases stability and biocompatibility of AuNPs; quaternary ammonium groups which provide positive charges that allow electrostatic binding of plasmid, which is considered the therapeutic agent to be transported into cells. The system developed was characterized by UV-vis spectroscopy, DLS, TEM and by electrophoretic mobility, yielding a formulation with 113.5 nm.Transfection efficiency of the formulation developed was evaluated through PCR and through EGFP expression by fluorescence microscopy and fluorescence spectroscopy. The internalization was observed 3h post transfection; however a low level of EGFP expression was achieved. After 24h of incubation, EGFP expression increases just 3 times compared to non-transfected cells. The commercial system (Lipofectamine) expressed EGFP 5 times more than the system developed AuNP@PEG@R4N+@pEGFP. This difference could be related to lower translocation to the nucleus.

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.

Antioxidant extraction process for Andean Oca by a photodiode array detector using response surface methodology

Application of Experimental Design techniques has proven to be essential in various research fields, due to its statistical capability of processing the effect of interactions among independent variables, known as factors, in a system’s response. Advantages of this methodology can be summarized in more resource and time efficient experimentations while providing more accurate results. This research emphasizes the quantification of 4 antioxidants extraction, at two different concentration, prepared according to an experimental procedure and measured by a Photodiode Array Detector. Experimental planning was made following a Central Composite Design, which is a type of DoE that allows to consider the quadratic component in Response Surfaces, a component that includes pure curvature studies on the model produced. This work was executed with the intention of analyzing responses, peak areas obtained from chromatograms plotted by the Detector’s system, and comprehending if the factors considered – acquired from an extensive literary review – produced the expected effect in response. Completion of this work will allow to take conclusions regarding what factors should be considered for the optimization studies of antioxidants extraction in a Oca (Oxalis tuberosa) matrix.

Fabrication and characterization of transparent conductive oxide surface sensors for electrocorticography – TrECoG

Understanding how the brain works has been one of the greatest goals of mankind. This desire fuels the scientific community to pursue novel techniques able to acquire the complex information produced by the brain at any given moment. The Electrocorticography (ECoG) is one of those techniques. By placing conductive electrodes over the dura, or directly over the cortex, and measuring the electric potential variation, one can acquire information regarding the activation of those areas. In this work, transparent ECoGs, (TrECoGs) are fabricated through thin film deposition of the Transparent Conductive Oxides (TCOs) Indium-Zinc-Oxide (IZO) and Gallium-Zinc-Oxide (GZO). Five distinct devices have been fabricated via shadow masking and photolithography. The data acquired and presented in this work validates the TrECoGs fabricated as efficient devices for recording brain activity. The best results were obtained for the GZO- based TrECoG, which presented an average impedance of 36 kΩ at 1 kHz for 500 μm diameter electrodes, a transmittance close to 90% for the visible spectrum and a clear capability to detect brain signal variations. The IZO based devices also presented high transmittance levels (90%), but with higher impedances, which ranged from 40 kΩ to 100 kΩ.

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.

Assessment of NDVI, land surface temperature and precipitation anomalies for drought monitoring in Bayankhongor province, Mongolia

During the last decade Mongolia’s region was characterized by a rapid increase of both severity and frequency of drought events, leading to pasture reduction. Drought monitoring and assessment plays an important role in the region’s early warning systems as a way to mitigate the negative impacts in social, economic and environmental sectors. Nowadays it is possible to access information related to the hydrologic cycle through remote sensing, which provides a continuous monitoring of variables over very large areas where the weather stations are sparse. The present thesis aimed to explore the possibility of using NDVI as a potential drought indicator by studying anomaly patterns and correlations with other two climate variables, LST and precipitation. The study covered the growing season (March to September) of a fifteen year period, between 2000 and 2014, for Bayankhongor province in southwest Mongolia. The datasets used were MODIS NDVI, LST and TRMM Precipitation, which processing and analysis was supported by QGIS software and Python programming language. Monthly anomaly correlations between NDVI-LST and NDVI-Precipitation were generated as well as temporal correlations for the growing season for known drought years (2001, 2002 and 2009). The results show that the three variables follow a seasonal pattern expected for a northern hemisphere region, with occurrence of the rainy season in the summer months. The values of both NDVI and precipitation are remarkably low while LST values are high, which is explained by the region’s climate and ecosystems. The NDVI average, generally, reached higher values with high precipitation values and low LST values. The year of 2001 was the driest year of the time-series, while 2003 was the wet year with healthier vegetation. Monthly correlations registered weak results with low significance, with exception of NDVI-LST and NDVI-Precipitation correlations for June, July and August of 2002. The temporal correlations for the growing season also revealed weak results. The overall relationship between the variables anomalies showed weak correlation results with low significance, which suggests that an accurate answer for predicting drought using the relation between NDVI, LST and Precipitation cannot be given. Additional research should take place in order to achieve more conclusive results. However the NDVI anomaly images show that NDVI is a suitable drought index for Bayankhongor province.