Cryopreservation of Cell Sheets of Adipose Stem Cells: Limitations and Successes

Cell Sheets of hASCs (hASCs-CS) have been previously proposed for wound healing applications(1, 2) and despite the concern for production time reduction, the possibility of having these hASCs-CS off-the-shelf is appealing. The goal of this work was to define a cryopreservation methodology allowing to preserve cells viability and the properties CS matrix. hASCs-CS obtained from three different donors were created in UP-cell thermoresponsive dishes(Nunc, Germany) as previously reported(1,2). Different cryopreservation conditions were considered: i)FBS plus DMSO(5% and10%); ii)0.4M of Trehalose plus DMSO (5% and 10%); iii)cryosolution PLL (Akron Biotech, USA); and iv)vitrification. The cryopreservation effect was first assessed for cellular viability by flow cytometry using 7-AAD, and after dissociating the hASCs-CS with collagenase and trypsin-EDTA 0.25%. The expression (RT-PCR) and deposition (western blot and immunocytochemistry) of collagen type I, laminin and fibronectin, and the organization (TEM) of the extracellular matrix was further assessed before and after hASCs-CS cryopreservation to determine a potential effect of the method over matrix composition and integrity. The obtained results confirmed that cell viability is affected by the cryopreservation methodology, as shown before for different CS(3). Interestingly, the matrix properties were not significantly altered and the typical cell sheetâ s easiness of manipulation for transplantation was not lost.

Prestress losses in reinforced concrete beams strengthened with NSM-CFRP laminates

Using prestressed near surface mounted fibre reinforced polymers (NSM-FRP) is nowadays regaining the attention from the scientific community for the strengthening of existing reinforced concrete (RC) structures. The application of prestressed internal FRP bars and externally bonded prestressed FRPs has already been deeply investigated and revealed considerable benefits when compared to the corresponding passive solutions. A certain amount of prestress provides benefits mainly associated to structural integrity and material durability. Immediately after prestress transference, it is possible to close some of the existing cracks, decreasing the susceptibility of the element to corrosion and, a certain amount of deflection can be recovered due to the creation of a negative curvature. However, very few studies have been carried out to properly assess the preservation of prestress over time. In this context, several reinforced concrete beams strengthened with prestressed NSM carbon FRP (CFRP) laminates were prestressed and monitored for about 40 days. The data obtained from these experimental programs is in this paper presented and analysed. The observed prestress losses were later modelled using finite elements analysis and, although this topic is not addressed in this paper, the obtained results revealed considerable precision. The largest strain losses in the CFRP laminate were found to be mainly located in the extremities of the bonded length, while in the central zone most of the applied pre-strain was retained over time. The highest CFRP strain losses were observed in the first 6 to 12 days after prestress transfer, suggesting that the application of prestressed NSM-FRP will be very effective over time.

Design of a case-based reasoner for information security in military organizations

Information security is concerned with the protection of information, which can be stored, processed or transmitted within critical information systems of the organizations, against loss of confidentiality, integrity or availability. Protection measures to prevent these problems result through the implementation of controls at several dimensions: technical, administrative or physical. A vital objective for military organizations is to ensure superiority in contexts of information warfare and competitive intelligence. Therefore, the problem of information security in military organizations has been a topic of intensive work at both national and transnational levels, and extensive conceptual and standardization work is being produced. A current effort is therefore to develop automated decision support systems to assist military decision makers, at different levels in the command chain, to provide suitable control measures that can effectively deal with potential attacks and, at the same time, prevent, detect and contain vulnerabilities targeted at their information systems. The concept and processes of the Case-Based Reasoning (CBR) methodology outstandingly resembles classical military processes and doctrine, in particular the analysis of “lessons learned” and definition of “modes of action”. Therefore, the present paper addresses the modeling and design of a CBR system with two key objectives: to support an effective response in context of information security for military organizations; to allow for scenario planning and analysis for training and auditing processes.

Analysis of polydicyclopentadiene-metal hybrid adhesive joints for automotive exterior body applications

An exterior body panel solution containing a polydicyclopentadiene skin attached to an interior metallic reinforcement through adhesive bonding is being studied to be applied in the MobiCar bonnet. With this solution is expected to achieve lightness, adequate structural integrity and cost-efficiency. However, there is uncertainty regarding to the bonnet adhesiveness since different metallic materials and adhesive types are being considered for its development. Thus, in this paper, several samples are tested through shear loading with the aim of understanding the loading magnitude expected by using polydicyclopentadiene, steel DC04+ZE and aluminum alloy AW5754-H111 as substrates adhesively bonded by an epoxy or a methacrylate. Methacrylate adhesive have shown greater shear strength in all types of adhesive joints. PDCPD joints presented the highest displacements. Surface degradation was considered adequate over abrading once none strength difference was seen between the different surface treatments. Steel treated by cataphoresis has shown the highest joint interface strength.

Aging and Alzheimer's disease: Searching for novel molecular cues

Tese de Doutoramento em Ciências da Saúde

O Esclavagismo dos Tempos Modernos e a Tutela Jurídica das suas Vítimas: os meios jurídicos ao serviço da recuperação psicológica das vítimas de tráfico de seres humanos

Dissertação de mestrado em Direitos Humanos

Construção de um manual de codificação e interpretação de memórias autobiográficas

Dissertação de mestrado integrado em Psicologia

The role of social cognition and motor imagery in gesture processing: a patients' study

Dissertação de mestrado integrado em Psicologia

Analysis and selection of indicators for a sustainability assessment method for school buildings based on SBTool ‐ PT

Nowadays, the sustainability of buildings has an extreme importance. This concept goes towards the European aims of the Program Horizon 2020, which concerns about the reduction of the environmental impacts through such aspects as the energy efficiency and renewable technologies, among others. Sustainability is an extremely broad concept but, in this work, it is intended to include the concept of sustainability in buildings. Within the concept that aims the integration of environmental, social and economic levels towards the preservation of the planet and the integrity of the users, there are, currently, several types of tools of environmental certification that are applicable to the construction industry (LEED, BREEAM, DGNB, SBTool, among others). Within this context, it is highlighted the tool SBTool (Sustainable Building Tool) that is employed in several countries and can be subject to review in institutions of basic education, which are the base for the formation of the critical masses and for the development of a country. The main aim of this research is to select indicators that can be used in a methodology for sustainability assessment (SBTool) of school buildings in Portugal and in Brazil. In order to achieve it, it will also be analyzed other methodologies that already incorporate parameters directly related with the schools environment, such as BREEAM or LEED.

Influence of climate on clinical diagnostic dry eye tests: pilot study

Purpose. To analyze dry eye disease (DED) tests and their consistency in similar nonsymptomatic population samples living in two geographic locations with different climates (Continental vs. Atlantic). Methods. This is a pilot study including 14 nonsymptomatic residents from Valladolid (Continental climate, Spain) and 14 sex-matched and similarly aged residents from Braga (Atlantic climate, Portugal); they were assessed during the same season (spring) of two consecutive years. Phenol red thread test, conjunctival hyperemia, fluorescein tear breakup time, corneal and conjunctival staining, and Schirmer test were evaluated on three different consecutive visits. Reliability was assessed using the intraclass correlation coefficient and weighted kappa (J) coefficient for quantitative and ordinal variables, respectively. Results. Fourteen subjects were recruited in each city with a mean (TSD) age of 63.0 (T1.7) and 59.1 (T0.9) years (p = 0.08) in Valladolid and Braga, respectively. Intraclass correlation coefficient and J values of the tests performed were below 0.69 and 0.61, respectively, for both samples, thus showing moderate to poor reliability. Subsequently, comparisons were made between the results corresponding to the middle and higher outdoor relative humidity (RH) visit in each location as there were no differences in mean temperature (p Q 0.75) despite RH values significantly differing (p e 0.005). Significant (p e 0.05) differences were observed between Valladolid and Braga samples on tear breakup time (middle RH visit, 2.76 T 0.60 vs. 5.26 T 0.64 seconds; higher RH visit, 2.61 T 0.32 vs. 5.78 T 0.88 seconds) and corneal (middle RH, 0.64 T 0.17 vs. 0.14 T 0.10; higher RH, 0.60 T 0.22 vs. 0.0 T 0.0) and conjunctival staining (middle RH, 0.61 T 0.17 vs. 0.14 T 0.08; higher RH, 0.57 T 0.15 vs. 0.18 T 0.09). Conclusions. This pilot study provides initial evidence to support that DED test outcomes assessing the ocular surface integrity and tear stability are climate dependent. Future large-sample studies should support these outcomes also in DED patients. This knowledge is fundamental for multicenter clinical trials. Lack of consistency in diagnostic clinical tests for DED was also corroborated. (Optom Vis Sci 2015;92:e284Ye289)

Inferior frontal gyrus white matter abnormalities in obsessive-compulsive disorder

The aim of the present study is to explore obsessive-compulsive disorder (OCD)-related abnormalities in white matter connectivity in OCD for a core region associated with inhibitory control [i.e. inferior frontal gyrus (IFG)]. Fifteen patients with OCD (11 men) and 15 healthy controls (nine men) underwent diffusion tensor imaging scanning to study four diffusivity indexes of white matter integrity [fractional anisotropy, mean diffusivity (MD), axial diffusivity and radial diffusivity (RD)]. The results showed that persons with OCD manifested significantly lower fractional anisotropy levels in the bilateral IFG as well as its parcellations in the pars opercularis, pars triangularis, and pars orbitalis. Significantly higher levels of MD, RD were evident for the OCD group in the IFG as a whole as well as in the bilateral subregions of the pars triangularis and pars opercularis (for MD and RD), the right side of the pars orbitalis (for RD), and the left side of the pars triangularis and right side pars opercularis (for axial diffusivity). Overall, the results suggest significant alterations in structural connectivity, probably associated with myelination and axonal abnormalities in the IFG of OCD patients.

Reducing seismic vulnerablity: Retrofitting historic buildings

The value of preserving historic buildings is increasingly accepted by society, which not only recognizes built cultural heritage as a part of its identity but is also more cognizant of its economic value. In Europe, for example, tourism accounts for 10 percent of the GDP in the EU and 12 percent of employment.1 Built cultural heritage is a fundamental element of what draws tourists to European destinations. To a great extent, the value of historic buildings rests in the integrity of their components as unique products of the technology of their time and place. Unfortunately, cultural heritage buildings are particularly vulnerable to disasters, for a variety of reasons. They are often damaged or in a state of deterioration; they were built with materials with low resistance; they are heavy; and the connections among their various structural components are frequently insufficient. The main causes of damage are lack of maintenance, water-induced deterioration (from rain or rising damp), soil settlement, and extreme events such as earthquakes. Earthquakes have caused hundreds of thousands of deaths in the last decade, in addition to the tremendous losses in built cultural heritage.

The impact of carbon source in Candida albicans virulence: participation of RLM1 in pathogen host interaction

Dissertação de mestrado em Genética Molecular

Exploring the role of O-Mannosylation in the modulation of E-Cadherin function in Gastric Cancer cell line models

Dissertação de mestrado em Bioquímica Aplicada – Biomedicina

Biological behaviour of thin films consisting of Au nanoparticles dispersed in a TiO2 dielectric matrix

In this work it was studied the possible use of thin films, composed of Au nanoparticles (NPs) embedded in a TiO2 matrix, in biological applications, by evaluating their interaction with a well-known protein, Bovine Serum Albumin (BSA), as well as with microbial cells (Candida albicans). The films were produced by one-step reactive DC magnetron sputtering followed by heat-treatment. The samples revealed a composition of 8.3 at.% of Au and a stoichiometric TiO2 matrix. The annealing promoted grain size increase of the Au NPs from 3 nm (at 300 °C) to 7 nm (at 500 °C) and a progressive crystallization of the TiO2 matrix to anatase. A broad localized surface plasmon resonance (LSPR) absorption band (λ = 580–720 nm) was clearly observed in the sample annealed at 500 °C, being less intense at 300 °C. The biological tests indicated that the BSA adhesion is dependent on surface nanostructure morphology, which in turn depends on the annealing temperature that changed the roughness and wettability of the films. The Au:TiO2 thin films also induced a significant change of the microbial cell membrane integrity, and ultimately the cell viability, which in turn affected the adhesion on its surface. The microstructural changes (structure, grain size and surface morphology) of the Au:TiO2 films promoted by heat-treatment shaped the amount of BSA adhered and affected cell viability.