Active bionanoconjugates of laccase and gold nanoparticles: kinetic and structural studies

Thesis for the master degree in Structural and Functional Biochemistry

Desulfovibrio gigas ferredoxin II: redox structural modulation of the [3Fe–4S] cluster

J Biol Inorg Chem (2006) 11: 307–315 DOI 10.1007/s00775-005-0077-2

Structural characterization of the urea-unfolded state of Colicin Immunity Protein Im7 and Im9

Dissertação para obtenção do Grau de Mestre em Bioquímica Estrutural e Funcional

Structural investigation of the Bacillus subtilis morphogenic factor RodZ

A thesis to obtain a Master degree in Structural and Functional Biochemistry

Assessment by doppler ultrasound of entheseal lesions in spondyloarthritis : a longitudial study to determine structural damage and disease activity lesions

RESUMO: Enthesitis is the hallmark of spondyloarthritis (SpA), and is observed in all subtypes. Wide information on SpA abnormalities, including synovitis, tendinitis and enthesitis, can be efficiently perceived by Doppler ultrasound. Furthermore, several studies on imaging of enthesis showed that imaging techniques are better than clinical examination to detect enthesis alterations; and vascularized enthesitis detected by Doppler ultrasound appears to be a valuable diagnostic tool to confirm SpA diagnosis. However, data published until now concerning entheseal elementary alterations that characterize SpA enthesitis (enthesis inflammatory activity) or enthesopathy (permanent structural changes) reflect rather the authors’ empiric opinion than a methodological validation process. In this sense it seems crucial to identify elementary entheseal lesions associated with activity or damage, in order to improve monitoring and treatment response in SpA patients. The development of better assessment tools is today a challenge and a need in SpA. The first study of this thesis focused on the analysis of the reliability of inter-lector and inter-ultrasonography equipment of Madrid sonography enthesitis index (MASEI). Fundamental data for the remaining unrolling project validity. In the second and third studies we concerned about two entheseal elemental lesions: erosions and bursa. In literature erosions represent a permanent structural damage, being useful for monitoring joint injury, disease activity and therapeutic response in many rheumatic diseases; and to date, this concept has been mostly applied in rheumatoid arthritis (RA). Unquestionably, erosion is a tissue-related damage and a structural change. However, the hypothesis that we decided to test was if erosions represent a permanent structural change that can only grow and worsen over time, as occurs in RA, or a transitory alteration. A longitudinal study of early SpA patients was undertaken, and the Achilles enthesis was used as a model. Our results strongly suggested that previously detected erosions could disappear during the course of the disease, being consistent with the dynamic behavior of erosion over time. Based on these striking results it seems reasonable to suggest that the new-bone formation process in SpA could be associated with the resolution of cortical entheseal erosion over time. These results could also be in agreement with the apparent failure of anti-tumor necrosis factor (TNF) therapies to control bone proliferation in SpA; and with the relation of TNF-α, Dickkopf-related protein 1 (Dkk-1) and the regulatory molecule of the Wnt signaling pathway in the bone proliferation in SpA. In the same model, we then proceeded to study the enthesis bursa. Interestingly, the Outcome Measures in Rheumatology Clinical Trials (OMERACT) enthesopathy definition does not include bursa as an elementary entheseal lesion. Nonetheless, bursa was included in 46% of the enthesis studies in a recently systematic literature review, being in agreement with the concept of “synovio-entheseal complex” that includes the link between enthesitis and osteitis in SpA. It has been clarified in recent data that there is not only a close functional integration of the enthesis with the neighboring bone, but also a connection between enthesitis and synovitis. Therefore, we tried to assess the prevalence and relevance of the bursa-synovial lesion in SpA. Our findings showed a significant increase of Achilles bursa presence and thickness in SpA patients compared to controls (healthy/mechanical controls and RA controls). These results raise awareness to the need to improve the enthesopathy ultrasonographic definition. In the final work of this thesis, we have explored new perspectives, not previously reported, about construct validity of enthesis ultrasound as a possible activity outcome in SpA. We performed a longitudinal Achilles enthesis ultrasound study in patients with early SpA. Achilles ultrasound examinations were performed at baseline, six- and twelve-month time periods and compared with clinical outcome measures collected at basal visit. Our results showed that basal erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) are higher in patients with Doppler signal in enthesis, and even that higher basal ESR, CRP and Ankylosing Spondylitis Disease Activity Score (ASDAS) predicted a higher Doppler signal (an ultrasound alteration accepted as representative of inflammation) six months later. Patients with very high disease activity assessed by ASDAS (>3.5) at baseline had significantly higher Achilles total ultrasound score verified at the same time; and ASDAS <1.3 predicted no Doppler signal at six and twelve months. This seems to represent a connection between classical biomarkers and clinical outcomes associated with SpA activity and Doppler signal, not only at the same time, but also for the following months. Remarkably, patients with inactive disease (ASDAS < 1.3) at baseline had no Doppler signal at six and twelve months. These findings reinforce the potential use of ultrasound related techniques for disease progression assessment and prognosis purposes. Intriguingly, Ankylosing Spondylitis Disease Activity Index (BASDAI) didn’t show significant differences between different cut-offs concerning ultrasound lesions or Doppler signal, while verified with ASDAS. These results seem to indicate that ASDAS reflects better than BASDAI what happens in the enthesis. The work herein discussed clearly shows the potential utility of ultrasound in enthesis assessment in SpA patients, and can be important for the development of ultrasound activity and structural damage scores for diagnosis and monitoring purposes. Therefore, local promotion of this technique constitutes a medical intervention that is worth being tested in SpA patients for diagnosis, monitoring and prognosis purposes.

Psychological safety, authentic leadership and social networks : A psycho-structural approach to the study of groups

Tese de Doutoramento em Psicologia na área de especialização de Psicologia das Organizações apresentada ao ISPA - Instituto Universitário

Usefulness of zebrafish model to assess glomerular and tubular alterations induced by tenofovir

Tenofovir (TFV) is one of the most used antiretroviral drugs. However, it is associated with tubular damage with mitochondria as a possible target. Tubulopathy precedes glomerular dysfunction, thus classic markers of renal function like the glomerular filtration rate (GFR) do not detect early TFV damage. Prediction and management of drug induced renal injury (DIRI) rely on the mechanisms of the drug insult and in optimal animal models to explore it. Zebrafish (Danio rerio) offers unique advantages for assessing DIRI, since the pronephros is structurally very similar to its human counterpart and is fully developed at 3.5 days postfertilization. The main aim of the present work was to evaluate the effects of TFV, as well as its pro-drug, tenofovir disoproxil fumarate (TDF), on the GFR and in mitochondria morphology in tubular cells of zebrafish larvae. Lethality curves were performed to understand the relationship between drug concentration and lethality. LC10 was selected to explore the renal function using the FITC-inulin assay and to analyze the mitochondrial toxicity by electron microscopy on larvae exposed to TDF, TFV, paracetamol and gentamicin (positive controls) or water (negative control). Lethality curves showed that gentamicin was the most lethal drug, followed by TDF, TFV and paracetamol. Gentamicin and paracetamol decreased the GFR, but no differences were found for either TDF or TFV, when compared to controls (%FITC Control = 33±8; %FITC TDF = 35±10; %FITC TFV = 30±10; %FITC Gentamicin = 46±17; %FITC Paracetamol = 83±14). Tubular mitochondria from treated larvae were notably different from non-treated larvae, showing swelling, irregular shapes, decreased mitochondria network, cristae disruption and loss of matrix granules. These results are in agreement with the effects of these drugs in humans and thus, demonstrate that zebrafish larvae can be a good model to assess the functional and structural damage associated with DIRI.

Heart hypoplasia in an animal model of congenital diaphragmatic hernia

PURPOSE: In previous papers, we described a new experimental model of congenital diaphragmatic hernia in rabbits, and we also reported noninvasive therapeutic strategies for prevention of the functional and structural immaturity of the lungs associated with this defect. In addition to lung hypoplasia, pulmonary hypertension, biochemical, and structural immaturity of the lungs, the hemodynamics of infants and animals with congenital diaphragmatic hernia are markedly altered. Hence, cardiac hypoplasia has been implicated as a possible cause of death in patients with congenital diaphragmatic hernia, and it is hypothesized to be a probable consequence of fetal mediastinal compression by the herniated viscera. Cardiac hypoplasia has also been reported in lamb and rat models of congenital diaphragmatic hernia. The purpose of the present experiment was to verify the occurrence of heart hypoplasia in our new model of surgically produced congenital diaphragmatic hernia in fetal rabbits. METHODS: Twelve pregnant New Zealand rabbits underwent surgery on gestational day 24 or 25 (normal full gestational time - 31 to 32 days) to create left-sided diaphragmatic hernias in 1 or 2 fetuses per each doe. On gestational day 30, all does again underwent surgery, and the delivered fetuses were weighed and divided into 2 groups: control (non-surgically treated fetuses) (n = 12) and congenital diaphragmatic hernia (n = 9). The hearts were collected, weighed, and submitted for histologic and histomorphometric studies. RESULTS: During necropsy, it was noted that in all congenital diaphragmatic hernia fetuses, the left lobe of the liver herniated throughout the surgically created defect and occupied the left side of the thorax, with the deviation of the heart to the right side, compressing the left lung; consequently, this lung was smaller than the right one. The body weights of the animals were not altered by congenital diaphragmatic hernia, but heart weights were decreased in comparison to control fetuses. The histomorphometric analysis demonstrated that congenital diaphragmatic hernia promoted a significant decrease in the ventricular wall thickness and an increase in the interventricular septum thickness. CONCLUSION: Heart hypoplasia occurs in a rabbit experimental model of congenital diaphragmatic hernia. This model may be utilized for investigations in therapeutic strategies that aim towards the prevention or the treatment of heart hypoplasia caused by congenital diaphragmatic hernia.

Electro-Optical/Infrared sensor turret integration on an aircraft - structural im-pact on LOCKHEED MARTIN C-130H and design methodology

The growing need to patrol and survey large maritime and terrestrial areas increased the need to integrate external sensors on aircraft in order to accomplish those patrols at increasingly higher altitudes, longer range and not depending upon vehicle type. The main focus of this work is to elaborate a practical, simple, effective and efficient methodology for the aircraft modification procedure resulting from the integration of an Elec-tro-Optical/Infra-Red (EO/IR) turret through a support structure. The importance of the devel-opment of a good methodology relies on the correct management of project variables as time, available resources and project complexity. The key is to deliver a proper tool for a project de-sign team that will be used to create a solution that fulfils all technical, non-technical and certi-fication requirements present in this field of transportation. The created methodology is inde-pendent of two main inputs: sensor model and aircraft model definition, and therefore it is in-tended to deliver the results for different projects besides the one that was presented in this work as a case study. This particular case study presents the development of a structure support for FLIR STAR SAPHIRE III turret integration on the front lower fuselage bulkhead (radome) of the LOCKHEED MARTIN C-130 H. Development of the case study focuses on the study of local structural analysis through the use of Finite Element Method (FEM). Development of this Dissertation resulted in a cooperation between Faculty of Science and Technology - Universidade Nova de Lisboa and the company OGMA - Indústria Aeronáutica de Portugal

Experimental and numerical approaches for structural assessment in new footbridge designs (SFRSCC–GFPR hybrid structure)

Within the civil engineering field, the use of the Finite Element Method has acquired a significant importance, since numerical simulations have been employed in a broad field, which encloses the design, analysis and prediction of the structural behaviour of constructions and infrastructures. Nevertheless, these mathematical simulations can only be useful if all the mechanical properties of the materials, boundary conditions and damages are properly modelled. Therefore, it is required not only experimental data (static and/or dynamic tests) to provide references parameters, but also robust calibration methods able to model damage or other special structural conditions. The present paper addresses the model calibration of a footbridge bridge tested with static loads and ambient vibrations. Damage assessment was also carried out based on a hybrid numerical procedure, which combines discrete damage functions with sets of piecewise linear damage functions. Results from the model calibration shows that the model reproduces with good accuracy the experimental behaviour of the bridge.

Structural assessment of a masonry vault in Portugal

This paper reports on a structural safety assessment and performance evaluation of the upper choir of the Santa Maria de Belém Church in the Jerónimos monastery, Lisbon, one of the most important cultural heritage buildings in Portugal. The possibility of adding a new 20 t organ to the upper choir and its effects on the church structure's response are presented. A refined and a simplified finite-element model is developed to investigate the structure's performance under self-weight and seismic actions. A sensitivity analysis is performed to investigate the effect of masonry mechanical properties and rib cross-sections on the structural response, given the difficulty in accurately obtaining this information. The results show that the safety level of the structure is acceptable, even in the case of adding a heavy new organ.

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.

Structural analysis of the masonry envelope of Ica Cathedral, Peru

Dissertação de mestrado em Structural Analysis of Monuments and Historical Constructions

Structural analysis of the timber structure of Ica Cathedral, Peru

Dissertação de mestrado em Structural Analysis of Monuments and Historical Constructions

An innovative framework for probabilistic-based structural assessment with an application to existing reinforced concrete structures

A novel framework for probabilistic-based structural assessment of existing structures, which combines model identification and reliability assessment procedures, considering in an objective way different sources of uncertainty, is presented in this paper. A short description of structural assessment applications, provided in literature, is initially given. Then, the developed model identification procedure, supported in a robust optimization algorithm, is presented. Special attention is given to both experimental and numerical errors, to be considered in this algorithm convergence criterion. An updated numerical model is obtained from this process. The reliability assessment procedure, which considers a probabilistic model for the structure in analysis, is then introduced, incorporating the results of the model identification procedure. The developed model is then updated, as new data is acquired, through a Bayesian inference algorithm, explicitly addressing statistical uncertainty. Finally, the developed framework is validated with a set of reinforced concrete beams, which were loaded up to failure in laboratory.