Modulation of the secretome of hBMSCs by tailoring the macromolecular gradient in hydrogels to generate tissue-to-tissue interfaces

Tissue-to-tissue interfaces are commonly present in all tissues exhibiting structural, biological and chemical gradients serving a wide range of physiological functions. These interfaces are responsible for mediation of load transfer between two adjacent tissues. They are also important structures in sustaining the cellular communications to retain tissueâ s functional integration and homeostasis. [1] All cells have the capacity to sense and respond to physical and chemical stimulus and when cultured in three-dimensional (3D) environments they tend to perform their function better than in two-dimensional (2D) environments. Spatial and temporal 3D gradient hydrogels better resemble the natural environment of cells in mimicking their extracellular matrix. [2] In this study we hypothesize that differential functional properties can be engineered by modulation of macromolecule gradients in a cell seeded threedimensional hydrogel system. Specifically, differential paracrine secretory profiles can be engineered using human Bone Marrow Stem Cells (hBMSCâ s). Hence, the specific objectives of this study are to: assemble the macromolecular gradient hydrogels to evaluate the suitablity for hBMSCâ s encapsulation by cellular viability and biofunctionality by assessing the paracrine secretion of hBMSCâ s over time. The gradient hydrogels solutions were prepared by blend of macromolecules in one solution such as hyaluronic (HA) acid and collagen (Col) at different ratios. The gradient hydrogels were fabricated into cylindrical silicon moulds with higher ratio solutions assembled at the bottom of the mould and adding the two solutions consecutively on top of each other. The labelling of the macromolecules was performed to confirm the gradient through fluorescence microscopy. Additionally, AFM was conducted to assess the gradient hydrogels stiffness. Gradient hydrogels characterization was performed by HA and Col degradation assay, degree of crosslinking and stability. hBMSCâ s at P3 were encapsulated into each batch solution at 106 cells/ml solution and gradient hydrogels were produced as previously described. The hBMSCâ s were observed under confocal microscopy to assess viability by Live/Dead® staining. Cellular behaviour concerning proliferation and matrix deposition was also performed. Secretory cytokine measurement for pro-inflammatory and angiogenesis factors was carried out using ELISA. At genomic level, qPCR was carried out. The 3D gradient hydrogels platform made of different macromolecules showed to be a suitable environment for hBMSCâ s. The hBMSCâ s gradient hydrogels supported high cell survival and exhibited biofunctionality. Besides, the 3D gradient hydrogels demonstrated differentially secretion of pro-inflammatory and angiogenic factors by the encapsulated hBMSCâ s. References: 1. Mikos, AG. et al., Engineering complex tissues. Tissue Engineering 12,3307, 2006 2. Phillips, JE. et al., Proc Natl Acad Sci USA, 26:12170-5, 2008

Integration of geometric consistency contributory factors in three-leg junctions collision prediction models of Portuguese two-lane national highways

This paper aims at developing a collision prediction model for three-leg junctions located in national roads (NR) in Northern Portugal. The focus is to identify factors that contribute for collision type crashes in those locations, mainly factors related to road geometric consistency, since literature is scarce on those, and to research the impact of three modeling methods: generalized estimating equations, random-effects negative binomial models and random-parameters negative binomial models, on the factors of those models. The database used included data published between 2008 and 2010 of 177 three-leg junctions. It was split in three groups of contributing factors which were tested sequentially for each of the adopted models: at first only traffic, then, traffic and the geometric characteristics of the junctions within their area of influence; and, lastly, factors which show the difference between the geometric characteristics of the segments boarding the junctionsâ area of influence and the segment included in that area were added. The choice of the best modeling technique was supported by the result of a cross validation made to ascertain the best model for the three sets of researched contributing factors. The models fitted with random-parameters negative binomial models had the best performance in the process. In the best models obtained for every modeling technique, the characteristics of the road environment, including proxy measures for the geometric consistency, along with traffic volume, contribute significantly to the number of collisions. Both the variables concerning junctions and the various national highway segments in their area of influence, as well as variations from those characteristics concerning roadway segments which border the already mentioned area of influence have proven their relevance and, therefore, there is a rightful need to incorporate the effect of geometric consistency in the three-leg junctions safety studies.

Strategic planning of electricity systems: Integrating renewable energies

Tese de Doutoramento - Programa Doutoral em Engenharia Industrial e Sistemas (PDEIS)

Bimodal information increases spontaneous interpersonal synchronization of goal directed upper limb movements

When interacting with each other, people often synchronize spontaneously their movements, e.g. during pendulum swinging, chair rocking[5], walking [4][7], and when executing periodic forearm movements[3].Although the spatiotemporal information that establishes the coupling, leading to synchronization, might be provided by several perceptual systems, the systematic study of different sensory modalities contribution is widely neglected. Considering a) differences in the sensory dominance on the spatial and temporal dimension[5] , b) different cue combination and integration strategies [1][2], and c) that sensory information might provide different aspects of the same event, synchronization should be moderated by the type of sensory modality. Here, 9 naïve participants placed a bottle periodically between two target zones, 40 times, in 12 conditions while sitting in front of a confederate executing the same task. The participant could a) see and hear, b) see , c) hear the confederate, d) or audiovisual information about the movements of the confederate was absent. The couple started in 3 different relative positions (i.e., in-phase, anti-phase, out of phase). A retro-reflective marker was attached to the top of the bottles. Bottle displacement was captured by a motion capture system. We analyzed the variability of the continuous relative phase reflecting the degree of synchronization. Results indicate the emergence of spontaneous synchronization, an increase with bimodal information, and an influence of the initial phase relation on the particular synchronization pattern. Results have theoretical implication for studying cue combination in interpersonal coordination and are consistent with coupled oscillator models.

Temporal analysis of CHAVE collection

Lecture Notes in Computer Science, 9309

Reconstructing transcriptional regulatory networks using data integration and text mining

Transcriptional Regulatory Networks (TRNs) are powerful tool for representing several interactions that occur within a cell. Recent studies have provided information to help researchers in the tasks of building and understanding these networks. One of the major sources of information to build TRNs is biomedical literature. However, due to the rapidly increasing number of scientific papers, it is quite difficult to analyse the large amount of papers that have been published about this subject. This fact has heightened the importance of Biomedical Text Mining approaches in this task. Also, owing to the lack of adequate standards, as the number of databases increases, several inconsistencies concerning gene and protein names and identifiers are common. In this work, we developed an integrated approach for the reconstruction of TRNs that retrieve the relevant information from important biological databases and insert it into a unique repository, named KREN. Also, we applied text mining techniques over this integrated repository to build TRNs. However, was necessary to create a dictionary of names and synonyms associated with these entities and also develop an approach that retrieves all the abstracts from the related scientific papers stored on PubMed, in order to create a corpora of data about genes. Furthermore, these tasks were integrated into @Note, a software system that allows to use some methods from the Biomedical Text Mining field, including an algorithms for Named Entity Recognition (NER), extraction of all relevant terms from publication abstracts, extraction relationships between biological entities (genes, proteins and transcription factors). And finally, extended this tool to allow the reconstruction Transcriptional Regulatory Networks through using scientific literature.

Spatio-temporal modelling of environmental data

Programa Doutoral em Matemática e Aplicações.

Spatial-temporal modellization of the NO2 concentration data through geostatistical tools

The nitrogen dioxide is a primary pollutant, regarded for the estimation of the air quality index, whose excessive presence may cause significant environmental and health problems. In the current work, we suggest characterizing the evolution of NO2 levels, by using geostatisti- cal approaches that deal with both the space and time coordinates. To develop our proposal, a first exploratory analysis was carried out on daily values of the target variable, daily measured in Portugal from 2004 to 2012, which led to identify three influential covariates (type of site, environment and month of measurement). In a second step, appropriate geostatistical tools were applied to model the trend and the space-time variability, thus enabling us to use the kriging techniques for prediction, without requiring data from a dense monitoring network. This method- ology has valuable applications, as it can provide accurate assessment of the nitrogen dioxide concentrations at sites where either data have been lost or there is no monitoring station nearby.

Método de desagregação temporal para a previsão da taxa de desemprego em Portugal

Dissertação de mestrado em Estatística

Low f-number microlenses for integration on optical microsystems

This paper presents microlenses (MLs) with low f-number made of AZ4562 photoresist for integration on optical microsystems. The fabrication process was based on the thermal reflow and rehydration. Large series of MLs were fabricated with a width of 35 μm, a thickness of 5 μm, and spaced apart by 3 μm. The MLs were fabricated directly on the surface of a die with type n+/p-substrate junction photodiode fabricated in a standard CMOS process. The measured focal length was 49 μm with a tolerance of ±2 μm (maximum error of ±4%), resulting in a numerical aperture of 33.6 × 10-2 (±1.3 × 10-2). The measurements also revealed an f-number of 1.4.

Finite integration methods for isospectral flows

In this paper we consider the approximate computation of isospectral flows based on finite integration methods( FIM) with radial basis functions( RBF) interpolation,a new algorithm is developed. Our method ensures the symmetry of the solutions. Numerical experiments demonstrate that the solutions have higher accuracy by our algorithm than by the second order Runge- Kutta( RK2) method.

Europa: uma questão profana ou sagrada?: estudo sobre as relações entre o poder político e as religiões no âmbito do processo de integração europeia

Dissertação de mestrado em Direito da União Europeia

Integration methods in dynamic analysis

"Series title: Springerbriefs in applied sciences and technology, ISSN 2191-530X"

Discurso mediático e sociedade: repensar a Análise Crítica do Discurso

A análise do discurso jornalístico e do seu enraizamento social tem conhecido avanços significativos nas últimas duas décadas, especialmente devido ao surgimento e desenvolvimento da Análise Crítica do Discurso. No entanto, há três aspectos importantes que merecem mais investigação: o plano temporal na análise do discurso, as estratégias discursivas dos atores sociais, e os efeitos extra e supra-textual do discurso mediatizado. Em primeiro lugar, a compreensão da biografia dos assuntos públicos exige uma análise longitudinal dos textos mediatizados e dos seus contextos sociais, mas a maioria das formas de análise do discurso jornalístico não tem em conta a sequência temporal dos textos e as suas implicações. Em segundo lugar, como a representação mediática das questões sociais é, em grande medida, função da construção discursiva de eventos, problemas e posições por diferentes atores sociais, as estratégias discursivas que eles empregam numa variedade de arenas e canais ‘antes’ e ‘depois’ dos textos jornalísticos precisam de ser examinados. Em terceiro lugar, o facto de que muitos dos modos de operação do discurso são extra- ou supra-textuais requer que se tenha em consideração vários processos sociais ‘fora’ do texto. Este trabalho tem como objetivo produzir um contributo teórico e metodológico para a integração destas questões em análise do discurso, propondo um quadro analítico que combina uma dimensão textual com uma contextual.

Emigrantes portugueses low cost: as perspetivas de ontem e de hoje

Dissertação de mestrado em Sociologia (área de especialização em Organizações e Trabalho)