Development of human central nervous system 3D in vitro models for preclinical research

Neurological disorders are a major concern in modern societies, with increasing prevalence mainly related with the higher life expectancy. Most of the current available therapeutic options can only control and ameliorate the patients’ symptoms, often be-coming refractory over time. Therapeutic breakthroughs and advances have been hampered by the lack of accurate central nervous system (CNS) models. The develop-ment of these models allows the study of the disease onset/progression mechanisms and the preclinical evaluation of novel therapeutics. This has traditionally relied on genetically engineered animal models that often diverge considerably from the human phenotype (developmentally, anatomically and physiologically) and 2D in vitro cell models, which fail to recapitulate the characteristics of the target tissue (cell-cell and cell-matrix interactions, cell polarity). The in vitro recapitulation of CNS phenotypic and functional features requires the implementation of advanced culture strategies that enable to mimic the in vivo struc-tural and molecular complexity. Models based on differentiation of human neural stem cells (hNSC) in 3D cultures have great potential as complementary tools in preclinical research, bridging the gap between human clinical studies and animal models. This thesis aimed at the development of novel human 3D in vitro CNS models by integrat-ing agitation-based culture systems and a wide array of characterization tools. Neural differentiation of hNSC as 3D neurospheres was explored in Chapter 2. Here, it was demonstrated that human midbrain-derived neural progenitor cells from fetal origin (hmNPC) can generate complex tissue-like structures containing functional dopaminergic neurons, as well as astrocytes and oligodendrocytes. Chapter 3 focused on the development of cellular characterization assays for cell aggregates based on light-sheet fluorescence imaging systems, which resulted in increased spatial resolu-tion both for fixed samples or live imaging. The applicability of the developed human 3D cell model for preclinical research was explored in Chapter 4, evaluating the poten-tial of a viral vector candidate for gene therapy. The efficacy and safety of helper-dependent CAV-2 (hd-CAV-2) for gene delivery in human neurons was evaluated, demonstrating increased neuronal tropism, efficient transgene expression and minimal toxicity. The potential of human 3D in vitro CNS models to mimic brain functions was further addressed in Chapter 5. Exploring the use of 13C-labeled substrates and Nucle-ar Magnetic Resonance (NMR) spectroscopy tools, neural metabolic signatures were evaluated showing lineage-specific metabolic specialization and establishment of neu-ron-astrocytic shuttles upon differentiation. Chapter 6 focused on transferring the knowledge and strategies described in the previous chapters for the implementation of a scalable and robust process for the 3D differentiation of hNSC derived from human induced pluripotent stem cells (hiPSC). Here, software-controlled perfusion stirred-tank bioreactors were used as technological system to sustain cell aggregation and dif-ferentiation. The work developed in this thesis provides practical and versatile new in vitro ap-proaches to model the human brain. Furthermore, the culture strategies described herein can be further extended to other sources of neural phenotypes, including pa-tient-derived hiPSC. The combination of this 3D culture strategy with the implemented characterization methods represents a powerful complementary tool applicable in the drug discovery, toxicology and disease modeling.

A behavioral perspective on the effects of using performance measurement systems in the companies: Evidence from a case study

The effects of PMSs on the people’s behaviour represent a high degree of relevance in the context of an organization performance and success. Thus, motivational and behavioural consequences of performance measurements are far from being totally understood (Franco-Santos et al., 2012). This work project (WP) purposes going further regarding the consequences/effects on people’s behaviour of using PMSs in organizations. The researcher conducted 11 interviews to managers during a nine-month internship as a controller in a Portuguese multi-national company. The evidence from this WP suggests that the way how managers understand a PMS determines a lot the way how they behave. Data also supports that PMSs influences in several ways motivation, perceptions, participation and job-related stress of managers.

Access of WetAMD and DME patients to treatment in public hospitals

Degeneration (WetAMD) and Diabetic Macular Edema (DME) patients’ access to treatment in public hospitals, by identifying bottlenecks and stress points that prevent timely and adequate care to patients who suffer from a degenerative disease, and consequently for whom the lack of access to treatment can have disastrous consequences. Considering the specificity and degenerative traits of these conditions, the long queues for specialty appointments in public hospitals are a significant threat to patients’ health, as the disease may be misdiagnosed and or progress significantly, causing unnecessary permanent and non-reversible loss in visual acuity. Therefore optimizing the patient journey will increase patients’ access to adequate treatment, and prevent avoidable progress of a degenerative condition which causes permanent and non-reversible blindness. Following the investigation which supports this thesis, the patient journey was broken down into its different phases, so that key issues could be identified, and referred back to the main stress points highlighted during the interviews with physicians and administrators. Finally results were scrutinized and systematized, and a set of action points was proposed, considering what may cause major impact and is actually feasible to implement.

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.

Risk assessment in a research laboratory during sol–gel synthesis of nano-TiO2

The occupational risks in the nanotechnology research laboratories are an important topic since a great number of researchers are involved in this area. The risk assessment performed by both qualitative and quantitative methods is a necessary step for the management of the occupational risks. Risk assessment could be performed by qualitative methods that gather consensus in the scientific community. It is also possible to use quantitative methods, based in different technics and metrics, as indicative exposure limits are been settled by several institutions. While performing the risk assessment, the information on the materials used is very important and, if it is not updated, it could create a bias in the assessment results. The exposure to TiO2 nanoparticles risk was assessed in a research laboratory using a quantitative exposure method and qualitative risk assessment methods. It was found the results from direct-reading Condensation Particle Counter (CPC) equipment and the CB Nanotool seem to be related and aligned, while the results obtained from the use of the Stoffenmanager Nano seem to indicate a higher risk level.

Hand gesture recognition system based in computer vision and machine learning

"Lecture notes in computational vision and biomechanics series, ISSN 2212-9391, vol. 19"

Information systems assessment in pathologic anatomy service

Information technologies changed the way of how the health organizations work, contributing to their effectiveness, efficiency and sustainability. Hospital Information Systems (HIS) are emerging on all of health institutions, helping health professionals and patients. However, HIS are not always implemented and used in the best way, leading to low levels of benefits and acceptance by users of these systems. In order to mitigate this problem, it is essential to take measures able to ensure if the HIS and their interfaces are designed in a simple and interactive way. With this in mind, a study to measure the user satisfaction and their opinion was made. It was applied the Technology Acceptance Model (TAM) on a HIS implemented on various hospital centers (AIDA), being used the Pathologic Anatomy Service. The study identified weakness and strengths features of AIDA and it pointed some solutions to improve the medical record.

Mercury and methylmercury concentration assessment in children's hair from Manaus, Amazonas state, Brazil

The present study evaluated Hg and MeHg content in hair samples of 201 children 2 to 7 years old, living in six neighborhoods of the city of Manaus, Amazonas State, Brazil. In general, the total Hg and MeHg median ranges in hair were similar (0.91 to 1.71 mg kg-1) except for the São Jose neighborhood, which was lower (0.16 mg kg-1). De spite the fact that the Manaus population consumes fish as part of the normal dietary intake, the Hg hair levels were below the level for an adult population not exposed to mercury (2.0 mg kg-1). These data were compared to demographic, socioeconomic information and eating habits of the families that took part in the study. The results were also compared to other published data from the Amazon region, other regions of Brazil and other countries. Future studies to set Hg and MeHg levels in hair of children in Brazil should take into account and assess the diversity of the country, mainly in terms of eating habits, socio-economic and cultural aspects.

Racionalização e redução de custos das TIC na Administração Pública: o caso da PSP

Dissertação de mestrado integrado em Engenharia e Gestão de Sistemas de Informação

Business intelligence no suporte às autarquias locais e à sua gestão orçamental

Dissertação de mestrado integrado em Engenharia e Gestão de Sistemas de Informação

Hand gesture recognition system based in computer vision and machine learning: Applications on human-machine interaction

Tese de Doutoramento em Engenharia de Eletrónica e de Computadores

Listening to the public: enacting power : citizen access, Standing and influence in public participation discourses

Public participation in environmental governance is typically associated with citizen access to power despite many closures and limitations having been identified in participatory processes. This article proposes an analytical framework to analyse discursive practices involved in public consultation processes. Critical Discourse Analysis is used to examine and appraise citizens’ access, standing and influence. We apply that framework to a ‘notice and comment’ process on a hydroelectric power plan in Portugal and show that it was discursively managed to justify the decision of constructing 10 large dams and to reject critical or alternative views. Citizens’ access, standing and influence were constrained through diverse discursive practices which (re)produced very unequal power relationsbetween policy proponents and participating individuals. More generally, the article illustrates the potential of Critical Discourse Analysis to assess voice(s) in policy processes. Focusing on argumentative, interactional and rhetorical levels, and how they are interwoven in public consultation discourses, the proposed framework is conceivably applicable in other studies.

Hormonal and cardiovascular reflex assessment in a female patient with pure autonomic failure

We report the case of a 72-year-old female with pure autonomic failure, a rare entity, whose diagnosis of autonomic dysfunction was determined with a series of complementary tests. For approximately 2 years, the patient has been experiencing dizziness and a tendency to fall, a significant weight loss, generalized weakness, dysphagia, intestinal constipation, blurred vision, dry mouth, and changes in her voice. She underwent clinical assessment and laboratory tests (biochemical tests, chest X-ray, digestive endoscopy, colonoscopy, chest computed tomography, abdomen and pelvis computed tomography, abdominal ultrasound, and ambulatory blood pressure monitoring). Measurements of catecholamine and plasmatic renin activity were performed at rest and after physical exercise. Finally the patient underwent physiological and pharmacological autonomic tests that better diagnosed dysautonomia.

Avaliação de projetos de eficiência energética na iluminação pública

Dissertação de mestrado em Engenharia Industrial

Impact of nutritional counseling in reducing serum cholesterol in public health service patients

OBJECTIVE: To assess the impact of nutritional attention on the lipid profile and nutritional status of hypercholesterolemic patients attended in health centers of Belo Horizonte. METHODS: Using nutritional attendance patient record cards from two health units, the evolution of the lipid profile and the nutritional state (BMI) was monitored of 96 hypercholesterolemic patients who received diet. The patients were appraised at the following moments: initial (1st consultation), after 3 months (2nd consultation) and last consultation (variable for each patient). RESULTS: On the first attendance, 44,4% of the patients presented not only high total cholesterol and LDL-c, but also hypertriglyceridemia and 70.3% were overweight or obese, but most patients (75.6%) presented adequate HDL-c levels. There was significant reduction in the BMI, total cholesterol, LDL-c values (p < 0.01) and also in the triglyceride levels (p < 0.05) in the first three months, without alteration in the HDL-c levels. A significant reduction (p < 0.01) was observed in the frequency of individuals with high cholesterol (from 89.6% down to 47.9%), high and very high LDL-c (from 82.6% down to 45.7%), as well as high and very high triglyceride (from 43.6% down to 16.7%). The observed reduction in frequency of the low HDL-c was statistically meaningless. CONCLUSION: This study evidences the effect of the nutritional attention on lipid profile in hypercholesterolemic patients, reinforcing the need for a multiprofessional team to attend them at the public health services.