Cerebral vasculopathy in children with sickle cell disease A study of genetic modulators of the disease

Sickle cell disease (SCD) is a genetic disorder with recessive transmission, caused by the mutation HBB:c.20A>T. It originates hemoglobin S that forms polymers inside the erythrocyte, upon deoxygenation, deforming it and ultimately leading to premature hemolysis. The disease presents with high heterogeneity of clinical manifestations, the most devastating of which, ischemic stroke, occurs in 11% of patients until 20 years of age. In this study, we tried to identify genetic modifiers of risk and episodes of stroke by studying 66 children with SCD, grouped according to the degree of cerebral vasculopathy (Stroke, Risk and Control). Association studies were performed between the three phenotypic groups and hematological and biochemical parameters of patients, as well as with 23 polymorphic regions in genes related to vascular cell adhesion (VCAM-1, THBS-1 and CD36), vascular tonus (NOS3 and ET-1) and inflammation (TNF-α and HMOX-1). Relevant data was collected from patient’s medical records. Known genetic modulators of SCD (beta-globin cluster haplotype and HBA and BCL11A genotypes) and putative genetic modifiers of cerebral vasculopathy were characterized. Differences in their distribution among groups were assessed. VCAM-1 rs1409419 allele C and NOS3 rs207044 allele C were associated to stroke events, while VCAM-1 rs1409419 allele T was found to be protective. Alleles 4a and 4b of NOS3 27 bp VNTR appeared to be respectively associated to stroke risk and protection. HMOX-1 longer STRs seemed to predispose to stroke. Higher hemoglobin F levels were found in Control group, as a result of Senegal haplotype or of BCL11A rs11886868 allele T, and higher lactate dehydrogenase levels, marker of hemolysis, were found in Risk group. Molecular mechanisms underlying the modifier functions of the relevant genetic variants are discussed.

Chitosan-based magnetic nanoparticles for osteosarcoma theranostic

Cancer is a well-known disease with a significant impact in society not only due to its incidence, more evident in more developed countries, but also due to the expenses related to medical treat-ments. Cancer research is considered an increasingly logical science with great potential for the development of new treatment options. Advances in nanomedicine have resulted in rapid devel-opment of nanomaterials with considerable potential in cancer diagnostics and treatment. The combination of diagnosis and treatment in a single nano-platform is named theranostic. In this PhD thesis a theranostic system for osteosarcoma was proposed, composed by a magnetic core, a polymeric coating, and a chemotherapeutic drug. The presence of a specific targeting agent, in this case a monoclonal antibody, provides high specificity to the proposed theranostic system. For the core of the proposed theranostic system, stable aqueous suspensions of superparamagnetic iron oxide nanoparticles with an average diameter of 9 nm were produced. Chitosan-based poly-meric nanoparticles with a hydrodynamic diameter around 150 nm were successfully produced. Incorporation of iron oxide nanoparticles into the polymeric ones increased their hydrodynamic diameter to at least 250 nm. A monoclonal antibody specific for a transmembranar protein (car-bonic anhydrase IX) present in solid tumors was developed by hybridoma technology. Functional hybridomas producing the desired monoclonal antibodies were obtained. The proposed theranostic system functionality was evaluated in separated parts of its components. Uncoated and coated iron oxide nanoparticles with chitosan-based polymers generated heat under the application of an external alternating magnetic field. Uncoated iron oxide nanoparticles sta-bilized with oleic acid were able to enhance contrast in magnetic resonance imaging. Drug deliv-ery studies were conducted in chitosan-based polymeric nanoparticles without and with the in-corporation of iron oxide nanoparticles, demonstrating to be an effective drug delivery platform for doxorubicin. The theranostic system proposed in this PhD thesis is very promising for cancer theranostic, demonstrating to be applicable in solid tumors such as osteosarcoma.

Polyhydroxyalkanoates production by photosynthetic mixed cultures

Polyhydroxyalkanoates (PHAs) are natural biologically synthesized polymers that have been the subject of much interest in the last decades due to their biodegradability. Thus far, its microbial production is associated with high operational costs, which increases PHA prices and limits its marketability. To address this situation, this thesis’ work proposes the utilization of photosynthetic mixed cultures (PMC) as a new PHA production system that may lead to a reduction in operational costs. In fact, the operational strategies developed in this work led to the selection of PHA accumulating PMCs that, unlike the traditional mixed microbial cultures, do not require aeration, thus permitting savings in this significant operational cost. In particular, the first PHA accumulating PMC tested in this work was selected under non-aerated illuminated conditions in a feast and famine regime, being obtained a consortium of bacteria and algae, where photosynthetic bacteria accumulated PHA during the feast phase and consumed it for growth during the famine phase, using the oxygen produced by algae. In this symbiotic system, a maximum PHA content of 20% cell dry weight (cdw) was reached, proving for the first time, the capacity of a PMC to accumulate PHA. During adaptation to dark/light alternating conditions, the culture decreased its algae content but maintained its viability, achieving a PHA content of 30% cdw. Also, the PMC was found to be able to utilize different volatile fatty acids for PHA production, accumulating up to 20% cdw of a PHA co-polymer composed of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (HV) monomers. Finally, a new selective approach for the enrichment of PMCs in PHA accumulating bacteria was tested. Instead of imposing a feast and famine regime, a permanent feast regime was used, thus selecting a PMC that was capable of simultaneously growing and accumulating PHA, being attained a maximum PHA content of 60% cdw, the highest value reported for a PMC thus far. The results presented in this thesis prospect the utilization of cheap, VFA-rich fermented wastes as substrates for PHA production, which combined with this new photosynthetic technology opens up the possibility for direct sunlight illumination, leading to a more cost-effective and environmentally sustainable PHA production process.

The perfect paint in modern art conservation: a comparative study of 21st century vinyl emulsions

Contemporary painting places, and will continue to place, several questions about its meaning, its chemical nature, its durability and the best way to preserve it. This research aims at putting together comprehensive data on vinyl based paints, including their components, their properties, their aging behavior and their response to selected cleaning products. In this project degradation mechanisms of vinyl binders and formulations used in the 20th and 21st century were studied. Stability over time of selected vinyl polymers was assessed through natural indoor and artificially aging. The objective was to enhance knowledge and understanding of vinyl emulsion formulations and their performance over time. Overall conservation state of pictorial layers namely, adhesion, cohesion and discoloration of selected case studies from the Portuguese artist Julião Sarmento (b.1948) was correlated with the observed molecular level changes studied in laboratory experiments. Sarmento’s paintings were chosen due to conservation concerns (discoloration) on some of his works from the 90’s. Besides, research was carried out to start increasing the knowledge of what can be expected of PVAc based paints in terms of response to conservation treatments namely, surface cleaning. Artificial aging showed that the most recent formulations which are based on a poly(vinyl acetate), poly(vinyl chloride) and polyethylene terpolymer are less stable when compared to some homopolymer formulations. From the four pigments studied, titanium dioxide rutile and a carbon based black proved to be stabilizers for both types of polymer. The mixture lithopone plus calcium carbonate has showed to have a photocatalytic effect on the binders. The studied paintings showed to be in an overall good state of conservation except for the paintings created in the 90’s with white glue and a mixture of white lithoponeand calcium carbonate. Discoloration of this white paint seems to be irreversible and ongoing and is still a major concern. The disapearance of the plasticizer was the only change detected. The current works created by Sarmento are expected to be more stable as they were painted using the rutile titanium dioxide. Immersion/cleaning tests showed that vinyl based paints can be susceptible to water and organic solvents like ethanol as some evidences point to the removal/diffusion of additives from the paint. The observations made point to the need to further proceed in this research field.

A technical investigation of an oil painting on copper support, including a study on consolidants for treatment

Paper submitted to e-conservation Journal: Maria Leonor Oliveira, Leslie Carlyle, Sara Fragoso, Isabel Pombo Cardoso and João Coroado, “Investigations into paint delamination and consolidation of an oil painting on copper support”.

Bioactive nanocomposite spheres with proved shape memory capability in bone defects

Bioactive glass nanoparticles (BGNPs) promote an apatite surface layer in physiologic conditions that lead to a good interfacial bonding with bone.1 A strategy to induce bioactivity in non-bioactive polymeric biomaterials is to incorporate BGNPs in the polymer matrix. This combination creates a nanocomposite material with increased osteoconductive properties. Chitosan (CHT) is a polymer obtained by deacetylation of chitin and is biodegradable, non-toxic and biocompatible. The combination of CHT and the BGNPs aims at designing biocompatible spheres promoting the formation of a calcium phosphate layer at the nanocomposite surface, thus enhancing the osteoconductivity behaviour of the biomaterial. Shape memory polymers (SMP) are stimuli-responsive materials that offer mechanical and geometrical action triggered by an external stimulus.2 They can be deformed and fixed into a temporary shape which remains stable unless exposed to a proper stimulus that triggers recovery of their original shape. This advanced functionality makes such SMPs suitable to be implanted using minimally invasive surgery procedures. Regarding that, the inclusion of therapeutic molecules becomes attractive.  We propose the synthesis of shape memory bioactive nanocomposite spheres with drug release capability.3   1.  L. L. Hench, Am. Ceram. Soc. Bull., 1993, 72, 93-98. 2.  A. Lendlein and S. Kelch, Angew Chem Int Edit, 2002, 41, 2034-2057. 3.  Ã . J. Leite, S. G. Caridade and J. F. Mano, Journal of Non-Crystalline Solids (in Press)

Produtos têxteis inteligentes incorporando filamentos compósitos com nanotubos de carbono

Programa Doutoral em Engenharia Têxtil.

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.

Prestress losses in NSM-CFRP flexurally strengthened RC beams

The use of prestressed near surface mounted fibre reinforced polymers (NSM-FRP) has been long acknowledged to be a suitable approach to strengthen and retrofit existing reinforced concrete structures. The application of a certain amount of prestress to the FRP prior to its installation provides a number of benefits, mainly related to crack width and deflection requisites at serviceability limit state conditions. After transferring the prestress to a structural element, some of the existing cracks can be closed, decreasing the vulnerability of the element to corrosion and, a certain amount of deflection can be recovered due to the introduced negative curvature. However, these benefits can only be assured if the prestress is properly preserved over time. In this context, three series of reinforced concrete beams, in a total of 10 beams, were strengthened with a prestressed carbon FRP laminate (CFRP) and monitored for about 40 days. The data obtained from these tests is in this paper presented and analysed. The observed losses of strain 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 initial strain was well-preserved over time. Additionally, the highest CFRP strain losses were observed in the first 6 to 12 days after prestress transfer, suggesting that the benefits of prestressed NSM-FRP will not be considerably lost over time.

Evaluation of the bond performance in FRP-brick components re-bonded after initial delamination

The bond behavior between Fiber Reinforced Polymers (FRPs) and masonry substrates has been the subject of many studies during the last years. Recent accelerated aging tests have shown that bond degradation and FRP delamination are likely to occur in FRP-strengthened masonry components under hygrothermal conditions. While an investigation on the possible methods to improve the durability of these systems is necessary, the applicability of different bond repair methods should also be studied. This paper aims at investigating the debonding mechanisms after repairing delaminated FRP-strengthened masonry components. FRP-strengthened brick specimens, after being delaminated, are repaired with two different adhesives: a conventional epoxy resin and a highly flexible polymer. The latter is used as an innovative adhesive in structural applications. The bond behavior in the repaired specimens is investigated by performing single-lap shear bond tests. Digital image correlation (DIC) is used for deeper investigation of the surface deformation and strains development. The effectiveness of the repair methods is discussed and compared with the strengthened specimens.

Moisture-induced degradation of interfacial bond in FRP-strengthened masonry

Externally bonded strengthening of masonry structures using Fiber Reinforced Polymers (FRPs) has been accepted as a promising technique. Although the effectiveness of FRPs in improving the performance of masonry components has been extensively investigated, their long-term performance and durability remain poorly addressed. This paper, tackling one of the aspects related to durability of these systems, presents an experimental investigation on the effect of long-term (one year) water immersion on the performance of GFRP-strengthened bricks. The tests include materials' mechanical tests, as well as pull-off and single-lap shear bond tests, to investigate the changes in material properties and bond behavior with immersion time, respectively. The effect of mechanical surface treatment on the durability of the strengthened system as well as the reversibility of the degradation upon partial drying are also investigated. The experimental results are presented and critically discussed.

Tensile creep of a structural epoxy adhesive: experimental and analytical characterization

Epoxy adhesives are nowadays being extensively used in Civil Engineering applications, mostly in the scope of the rehabilitation of reinforced concrete (RC) structures. In this context, epoxy adhesives are used to provide adequate stress transference from fibre reinforced polymers (FRP) to the surrounding concrete substrate. Most recently, the possibility of using prestressed FRPs bonded with these epoxy adhesives is also being explored in order to maximize the potentialities of this strengthening approach. In this context, the understanding of the long term behaviour of the involved materials becomes essential. Even when non-prestressed FRPs are used a certain amount of stress is permanently applied on the adhesive interface during the serviceability conditions of the strengthened structure, and the creep of the adhesive may cause a continuous variation in the deformational response of the element. In this context, this paper presents a study aiming to experimentally characterize the tensile creep behaviour of an epoxy-based adhesive currently used in the strengthening of concrete structures with carbon FRP (CFRP) systems. To analytically describe the tensile creep behaviour, the modified Burgers model was fitted to the experimental creep curves, and the obtained results revealed that this model is capable of predicting with very good accuracy the long term behaviour of this material up to a sustained stress level of 60% of the adhesive’s tensile strength.

Investigating the durability of FRP-masonry elements immersed in water

Nowadays, there is an increasing interest in using fiber reinforced polymers (FRP) for strengthening masonry elements. It has been observed that these materials, when used for externally bonded reinforcement (EBR), improve the performance of masonry components. However, issues such as durability and long-term performance of strengthened elements are still open. The bond between composite material and masonry substrate is a critical mechanism in EBR strengthening techniques, and therefore its durability and long-term performance should be deeply investigated and characterized. In the present study, the influence of water immersion on the bond performance is investigated by performing single-lap shear bond tests on two sets of GFRP-strengthened specimens immersed in water for six months. Different surface preparation techniques are used for each set of specimens to study their effect on the bond degradation. The specimens are prepared following the wet lay-up procedure. The observations and the obtained results are presented and discussed.

Bond performance in NSM-strengthened masonry brick

The use of Near Surface Mounted (NSM) Fiber Reinforced Polymers (FRPs) for strengthening masonry structures can be a suitable substitute for Externally Bonded Reinforcement (EBR) technique. NSM technique has many advantages such as larger bonded area, better anchorage capacity, higher resistance, higher percentage exploitation of the FRP and reduced installation time. However, information regarding the effectiveness of this strengthening technique for masonry structures is scarce and characterization of the critical mechanisms such as bond behavior is necessary. This paper presents experimental investigation of the bond performance in NSM-strengthened brick specimens. CFRP laminates are used for NSM strengthening of masonry bricks with different bonded lengths. The bond between FRP and masonry substrate is investigated by performing conventional pull-out tests and the experimental results are presented and discussed.

Seismic evaluation of environmentally degraded FRP-strengthened masonry walls

Fiber Reinforced Polymers (FRPs) have been extensively used for externally bonded reinforcement of masonry structures during the last years. Available information shows that FRPs can significantly improve the seismic performance of masonry elements without altering their structural mass. However, the durability and long-term performance of the strengthened elements are not clearly known yet. Recent experimental results show that environmental conditions can lead to degradation of the bond between FRP and masonry and FRP delaminations. But the effect of these local degradation mechanisms on the global structural response is not studied yet. This paper is therefore aimed at numerically investigating the effect of environmental degradation on the global performance of strengthened masonry walls. The nonlinear behavior of masonry walls strengthened with FRP composites is initially simulated with the aim of a FE package. The adopted numerical modeling strategy is verified by comparison of numerical and experimental results. The model, once validated, is used for investigating the effect of materials and bond degradation on the global behavior and failure modes of strengthened walls. The effect of strengthening scheme on the long-term performance of strengthened walls is also investigated. The degradation data are taken from experimental tests previously performed by the authors. The numerical results show that the effect of local material degradation on the global response of strengthened walls depends on the strengthening schemes, and severity of the environmental conditions. Moreover, environmental induced degradations and FRP delaminations can lead to change of expected failure modes in the strengthened elements. These observations, that are usually neglected at the design stage, can be critical in the long-term performance of strengthened structures.