Mechanisms of chloroperoxidase-catalyzed enantioselective reactions as probed by site-directed mutagenesis and isotopic labeling

Chloroperoxidase (CPO) is a heme-containing glycoprotein secreted by the marine fungus Caldariomyces fumago. Chloroperoxidase contains one ferriprotoporphyrin IX prosthetic group per molecule and catalyzes a variety of reactions, such as halogenation, peroxidation and epoxidation. The versatile catalytic activities of CPO coupled with the increasing demands for chiral synthesis have attracted an escalating interest in understanding the mechanistic and structural properties of this enzyme. In order to better understand the mechanisms of CPO-catalyzed enantioselective reactions and to fine-tune the catalytic properties of chloroperoxidase, asparagine 74 (N74) located in the narrow substrate access channel of CPO was replaced by a bulky, nonpolar valine and a polar glutamine using site-directed mutagenesis. The CPO N74 mutants displayed significantly enhanced activity toward nonpolar substrates compared to wild-type CPO as a result of changes in space and polarity of the heme distal environment. More interestingly, N74 mutants showed dramatically decreased chlorination and catalase activity but significantly enhanced epoxidation activity as a consequence of improved kinetic perfection introduced by the mutation as reflected by the favorable changes in k cat and kcat/KM of these reactions. It is also noted that the N74V mutant is capable of decomposing cyanide, the most notorious poison for many hemoproteins, as judged by the unique binding behavior of N74V with potassium cyanide. Histidine 105 (H105) was replaced by a nonpolar amino acid alanine using site-directed mutagenesis. The CPO H105 mutant (H105A) displayed dramatically decreased chlorination and catalase activity possibly because of the decreased polarity in the heme distal environment and loss of the hydrogen bonds between histidine 105 and glutamic acid 183. However, significantly increased enantioselectivity was observed for the epoxidation of bulky styrene derivatives. Furthermore, my study provides strong evidence for the proposed histidine/cysteine ligand switch in chloroperoxidase, providing experimental support for the structure of the 420-nm absorption maximum for a number of carbon monoxide complexes of heme-thiolate proteins. For the NMR study, [dCPO(heme)] was produced using 90% deuterated growth medium with excess heme precursors and [dCPO(Phe)] was grown in the same highly deuterated medium that had been supplemented with excess natural phenylalanine. To make complete heme proton assignments, NMR spectroscopy has been performed for high-resolution structural characterization of [dCPO(heme)] and [dCPO(Phe)] to achieve unambiguous and complete heme proton assignments, which also allows important amino acids close to the heme active center to be determined.

Development of a new client -server architecture for context aware mobile computing

This dissertation studies the context-aware application with its proposed algorithms at client side. The required context-aware infrastructure is discussed in depth to illustrate that such an infrastructure collects the mobile user’s context information, registers service providers, derives mobile user’s current context, distributes user context among context-aware applications, and provides tailored services. The approach proposed tries to strike a balance between the context server and mobile devices. The context acquisition is centralized at the server to ensure the reusability of context information among mobile devices, while context reasoning remains at the application level. Hence, a centralized context acquisition and distributed context reasoning are viewed as a better solution overall. The context-aware search application is designed and implemented at the server side. A new algorithm is proposed to take into consideration the user context profiles. By promoting feedback on the dynamics of the system, any prior user selection is now saved for further analysis such that it may contribute to help the results of a subsequent search. On the basis of these developments at the server side, various solutions are consequently provided at the client side. A proxy software-based component is set up for the purpose of data collection. This research endorses the belief that the proxy at the client side should contain the context reasoning component. Implementation of such a component provides credence to this belief in that the context applications are able to derive the user context profiles. Furthermore, a context cache scheme is implemented to manage the cache on the client device in order to minimize processing requirements and other resources (bandwidth, CPU cycle, power). Java and MySQL platforms are used to implement the proposed architecture and to test scenarios derived from user’s daily activities. To meet the practical demands required of a testing environment without the impositions of a heavy cost for establishing such a comprehensive infrastructure, a software simulation using a free Yahoo search API is provided as a means to evaluate the effectiveness of the design approach in a most realistic way. The integration of Yahoo search engine into the context-aware architecture design proves how context aware application can meet user demands for tailored services and products in and around the user’s environment. The test results show that the overall design is highly effective, providing new features and enriching the mobile user’s experience through a broad scope of potential applications.

Otimização do sistema de supervisão e controle de uma planta de tratamento de resíduos utilizando plasma

A major and growing problems faced by modern society is the high production of waste and related effects they produce, such as environmental degradation and pollution of various ecosystems, with direct effects on quality of life. The thermal treatment technologies have been widely used in the treatment of these wastes and thermal plasma is gaining importance in processing blanketing. This work is focused on developing an optimized system of supervision and control applied to a processing plant and petrochemical waste effluents using thermal plasma. The system is basically composed of a inductive plasma torch reactors washing system / exhaust gases and RF power used to generate plasma. The process of supervision and control of the plant is of paramount importance in the development of the ultimate goal. For this reason, various subsidies were created in the search for greater efficiency in the process, generating events, graphics / distribution and storage of data for each subsystem of the plant, process execution, control and 3D visualization of each subsystem of the plant between others. A communication platform between the virtual 3D plant architecture and a real control structure (hardware) was created. The goal is to use the concepts of mixed reality and develop strategies for different types of controls that allow manipulating 3D plant without restrictions and schedules, optimize the actual process. Studies have shown that one of the best ways to implement the control of generation inductively coupled plasma techniques is to use intelligent control, both for their efficiency in the results is low for its implementation, without requiring a specific model. The control strategy using Fuzzy Logic (Fuzzy-PI) was developed and implemented, and the results showed satisfactory condition on response time and viability

Alinhamento estratégico da TI: o caso da UFRN

It is considered that the Strategic Alignment IT is the first step within the IT Governance process for any institution. Taking as initial point the recognition that the governance corporate has an overall view of the organizations, the IT Governance takes place as a sub-set responsible for the implementation of the organization strategies in what concerns the provision of the necessary tools for the achievement of the goals set in the Institutional Development Plan. In order to do so, COBIT specifies that such Governance shall be built on the following principles: Strategic Alignment, Value Delivery, Risk Management, Performance Measurement. This paper aims at the Strategic Alignment, considered by the authors as the foundation for the development of the entire IT Governance core. By deepening the technical knowledge of the management system development, UFRN has made a decisive step towards the technical empowerment needed to the “Value Delivery”, yet, by perusing the primarily set processes to the “Strategic Alignment”, gaps that limited the IT strategic view in the implementation of the organizational goals were found. In the qualitative study that used documentary research with content analysis and interviews with the strategic and tactical managers, the view on the role of SINFO – Superintendência de Informática was mapped. The documentary research was done on public documents present on the institutional site and on TCU – Tribunal de Contas da União – documents that map the IT Governance profiles on the federal public service as a whole. As a means to obtain the documentary research results equalization, questionnaires/interviews and iGovTI indexes, quantitative tools to the standardization of the results were used, always bearing in mind the usage of the same scale elements present in the TCU analysis. This being said, similarly to what the TCU study through the IGovTI index provides, this paper advocates a particular index to the study area – SA (Strategic Alignment), calculated from the representative variables of the COBIT 4.1 domains and having the representative variables of the Strategic Alignment primary process as components. As a result, an intermediate index among the values in two adjacent surveys done by TCU in the years of 2010 and 2012 was found, which reflects the attitude and view of managers towards the IT governance: still linked to Data Processing in which a department performs its tasks according to the demand of the various departments or sectors, although there is a commission that discusses the issues related to infrastructure acquisition and systems development. With an Operational view rather than Strategic/Managerial and low attachment to the tools consecrated by the market, several processes are not contemplated in the framework COBIT defined set; this is mainly due to the inexistence of a formal strategic plan for IT; hence, the partial congruency between the organization goals and the IT goals.

Growth, Characterization, and Properties of Hybrid Graphene-Carbon Nanotube Films and Related Carbon Nanostructures

Graphene, first isolated in 2004 and the subject of the 2010 Nobel Prize in physics, has generated a tremendous amount of research interest in recent years due to its incredible mechanical and electrical properties. However, difficulties in large-scale production and low as-prepared surface area have hindered commercial applications. In this dissertation, a new material is described incorporating the superior electrical properties of graphene edge planes into the high surface area framework of carbon nanotube forests using a scalable and reproducible technology.

The objectives of this research were to investigate the growth parameters and mechanisms of a graphene-carbon nanotube hybrid nanomaterial termed “graphenated carbon nanotubes” (g-CNTs), examine the applicability of g-CNT materials for applications in electrochemical capacitors (supercapacitors) and cold-cathode field emission sources, and determine materials characteristics responsible for the superior performance of g-CNTs in these applications. The growth kinetics of multi-walled carbon nanotubes (MWNTs), grown by plasma-enhanced chemical vapor deposition (PECVD), was studied in order to understand the fundamental mechanisms governing the PECVD reaction process. Activation energies and diffusivities were determined for key reaction steps and a growth model was developed in response to these findings. Differences in the reaction kinetics between CNTs grown on single-crystal silicon and polysilicon were studied to aid in the incorporation of CNTs into microelectromechanical systems (MEMS) devices. To understand processing-property relationships for g-CNT materials, a Design of Experiments (DOE) analysis was performed for the purpose of determining the importance of various input parameters on the growth of g-CNTs, finding that varying temperature alone allows the resultant material to transition from CNTs to g-CNTs and finally carbon nanosheets (CNSs): vertically oriented sheets of few-layered graphene. In addition, a phenomenological model was developed for g-CNTs. By studying variations of graphene-CNT hybrid nanomaterials by Raman spectroscopy, a linear trend was discovered between their mean crystallite size and electrochemical capacitance. Finally, a new method for the calculation of nanomaterial surface area, more accurate than the standard BET technique, was created based on atomic layer deposition (ALD) of titanium oxide (TiO2).

Development of a New Client-Server Architecture for Context Aware Mobile Computing

This dissertation studies the context-aware application with its proposed algorithms at client side. The required context-aware infrastructure is discussed in depth to illustrate that such an infrastructure collects the mobile user’s context information, registers service providers, derives mobile user’s current context, distributes user context among context-aware applications, and provides tailored services. The approach proposed tries to strike a balance between the context server and mobile devices. The context acquisition is centralized at the server to ensure the usability of context information among mobile devices, while context reasoning remains at the application level. Hence, a centralized context acquisition and distributed context reasoning are viewed as a better solution overall. The context-aware search application is designed and implemented at the server side. A new algorithm is proposed to take into consideration the user context profiles. By promoting feedback on the dynamics of the system, any prior user selection is now saved for further analysis such that it may contribute to help the results of a subsequent search. On the basis of these developments at the server side, various solutions are consequently provided at the client side. A proxy software-based component is set up for the purpose of data collection. This research endorses the belief that the proxy at the client side should contain the context reasoning component. Implementation of such a component provides credence to this belief in that the context applications are able to derive the user context profiles. Furthermore, a context cache scheme is implemented to manage the cache on the client device in order to minimize processing requirements and other resources (bandwidth, CPU cycle, power). Java and MySQL platforms are used to implement the proposed architecture and to test scenarios derived from user’s daily activities. To meet the practical demands required of a testing environment without the impositions of a heavy cost for establishing such a comprehensive infrastructure, a software simulation using a free Yahoo search API is provided as a means to evaluate the effectiveness of the design approach in a most realistic way. The integration of Yahoo search engine into the context-aware architecture design proves how context aware application can meet user demands for tailored services and products in and around the user’s environment. The test results show that the overall design is highly effective,providing new features and enriching the mobile user’s experience through a broad scope of potential applications.

TERENO (Terrestrial Environmental Observatories) wild bee monitoring in six agriculturally dominated landscapes of Saxony-Anhalt (Germany)

In 2001 we started as part of the EU FP5 project Greenveins monitoring of insect communities in the normal landscape of Saxony-Anhalt (Germany), which is dominated by agricultural use. We selected four landscape sites of 4x4 km and recorded insects using combined flight traps, combining the ideas of window and yellow pan traps (see Duelli et al., 1999). Traps consist of a yellow funnel (25 cm diameter) filled with water (preserving agent added) and two perspex windows mounted in a way that they are crossed in the center. Within each square km of a site one trap was placed at ecotones between semi-natural habitats and agricultural fields (16 traps per site). Traps were operated in late spring-early summer (three sampling rounds) and late summer (three sampling rounds). Follow-up sampling started in 2010 as long-term monitoring within the TERENO project (www.tereno.net), contributing to the LTER network (Long-Term Ecosystem Research) in Germany (www.lter-d.de) and internationally as well (www.lter-europe.net). Metadata about the sites and related activities and data sets can be found in the DEIMS Repository for Research Sites and Datasets (https://data.lter-europe.net/deims/). In 2010 another two landscapes were added and yearly sampled in the same way. Due to long processing time of trapped insects data of follow-up years will be available about 18 months after trapping.

Värdeflödesanalys och Data Envelopment Analysis : En fallstudie för att värdera effektiviteten hos ett logistikföretag

Companies face new challenges almost every day. In order to stay competitive, it is important that companies strive for continuous development and improvement. By describing companies through their processes it is possible to get a clear overview of the entire operation, which can contribute, to a well-established overall understanding of the company. This is a case study based on Stort AB which is a small logistics company specialized in international transportation and logistics solutions. The purpose of this study is to perform value stream mapping in order to create a more efficient production process and propose possible improvements in order to reduce processing time. After performing value stream mapping, data envelopment analysis is used to calculate how lean Stort AB is today and how lean the company can become by implementing the proposed improvements. The results show that the production process can improve efficiency by minimizing waste produced by a bad workplace layout and over-processing. The authors suggested solution is to introduce standardized processes and invest in technical instruments in order to automate the process to reduce process time. According to data envelopment analysis the business is 41 percent lean at present and may soon become 55 percent lean and finally reach an optimum 100 percent lean mode if the process is automated.

Envelhecer… Embelecer… Envilecer: comprometimento ou desvinculação com a vida? Atividade e sociabilidade dos idosos da Alta de Coimbra

Este estudo surgiu com o intuito de compreender se os mais velhos continuam, após a transição para a reforma, a participar na esfera da economia, formal e informal, e da sociabilidade, analisando com mais pormenor no que se refere a esta última faceta o sentimento de solidão. Partindo dos diferentes enquadramentos teóricos sobre o contributo socioeconómico dos idosos o objetivo foi compreender, no campo empírico, a posição dos sujeitos relativamente à transição para a reforma e de que forma os mesmos se percecionavam em áreas ligadas à produtividade e sociabilidade. A amostra foi intencional (N=30) e abrangeu sujeitos com idade igual ou superior a sessenta e cinco anos, com pelo menos um ano de reforma e residentes na Alta de Coimbra. A entrevista semiestruturada e a observação direta foram os instrumentos selecionados para aprofundar as vivências dos participantes. No que respeita ao tratamento dos dados recorremos à técnica da análise de conteúdo, com base em Bardin, que nos permitiu captar a singularidade de cada sujeito, sem descurar a totalidade das observações. No final concluiu-se que os sujeitos entrevistados vivenciam a transição para a reforma de maneiras distintas, reforçando assim a heterogeneidade deste grupo, e que são dotados de um capital social e económico que urge potencializar, quer no seio da sociedade civil, quer de forma específica no âmbito da Gestão de Recursos Humanos. / This study aims to understand if, after they retire, the elderly continue to get involved in areas like economy, formal and informal, and sociability, analyzing in detail the feeling of loneliness. Bearing in mind the different theories about the elderly’s social and economic contribution, the goal was to empirically understand the attitude of these individuals towards their transition to retirement, and to see how they perceive themselves and their role in productivity and sociability. The sample was intentional (N=30) and has included people aged sixty five and over sixty five, who have been retired for at least one year, and living in Alta de Coimbra. The semisstructured interview and direct observation were used to know the participants’ way of live. In what comes to data processing, we followed the content analysis technique, based on Bardin, which allows us to capture the singularity of each individual, without forgetting the whole set of observations. In the end we have concluded that the interviewed individuals live the transition into retirement in different ways, thus reinforcing the heterogeneity of this group and their social and economic capital, which needs to be put to use, both in civil society and more specifically in the sphere of the human management resources.

Tributos indiretos na cadeia produtiva do camarão

The state of Rio Grande do Norte counts with a relevant potential in the shrimp farming supply chain. In the larviculture step the state responds for more than half of the national production. In the farming step it is the second largest producer. In the industrial step, its industries have almost 40% of the shrimp processing capacity of the northeast of Brazil. However, this country has the highest tax rate comparing with the main shrimp producer countries. Considering the influence of taxes in the competition among companies, the main goal of this research is to analyze the impact of indirect taxes in the above steps of the supply chain. To achieve it, it will be used the data of the 2011 Census of the Shrimp Farming and it will be applied the Herfindahl-Hirschman Index to identify the market form of those steps. In order to contribute with the characterization of the supply chain, CEO´s of farms and industries will be interviewed. The price-elasticity of the shrimp larvae, the in natura shrimp and the processed shrimp will be analyzed in order to verify the possibility that each one of those three steps has to pass-through the onus of the end of benefit over the ICMS. The data analysis shows that the larviculture step functions as a duopoly and, facing the end of that benefit, it will be able to pass-through most its onus to the farming step. On the other hand, this step functions similar to a perfect competing market, which diminishes its capacity to pass-through that onus to the processing step. This step operates as oligopoly with a lower concentration than the larviculture step but, due to the fact that it faces an oligopsony, it will end up assuming most of that onus, which will cause a decrease in the amount of processed shrimp. It is concluded that the end of that benefit would impact negatively, in this state, the supply chain at all, but mainly the farming and the industrial steps

Materiais à base de Sr(Ti,Nb)O3 para sensores lambda

Os sensores lambda resistivos possuem as vantagens de simplicidade e menor custo relativamente à utilização generalizada de sensores potenciométricos de oxigénio. Nesse sentido, os titanatos de estrôncio têm sido alvo de diversos estudos. Para a produção de uma relação inequívoca entre a condutividade destes materiais e a pressão parcial de oxigénio é necessária a adição de um dopante dador que suprime a condução eletrónica do tipo-p na região de pressões parciais de oxigénio próximas de ar. Contudo, a adição de um dopante dador produz respostas lentas destes materiais quando densos a variações da pressão parcial de oxigénio. Além da preparação usual dos pós por reação do estado sólido, foram preparadas diversas composições por mecanossíntese. Tal relaciona-se com o fato exaustivamente reportado de as amostras destes materiais, especialmente quando dopados com dadores, apresentarem comportamentos dependentes das condições de processamento. Teve ainda o intuito de avaliar a viabilidade da sua preparação por este método, e consequentemente verificar se este método de preparação, que presumivelmente produzirá pós com composição mais homogénea e mais reativos, permite alterar/manipular a resposta obtida por amostras com eles produzidas. Foram preparados diversos filmes, tipologia muito usada na produção de sensores resistivos, e amostras porosas com diversas composições à base de titanato de estrôncio produzidos com variadas condições de processamento. Foram realizadas diversas caracterizações sobre estes espécimes numa tentativa de melhor compreender as propriedades destes materiais e a dependência destas com parâmetros microestruturais como o tamanho de grão e a porosidade. Foi verificado que os exemplares de titanato de estrôncio não dopado, quer em filmes quer em amostras porosas, apresentam um comportamento elétrico semelhante ao apresentado por amostras densas deste material. Apurou-se ainda, que as suas características apresentam uma variação ténue com a alteração das condições de processamento. Já espécimes de titanato de estrôncio dopados com dador revelam uma forte dependência das suas propriedades com as condições de processamento utilizadas, nomeadamente, a temperatura de sinterização e o tempo de permanência a essa temperatura. Para o fabrico de sensores resistivos de oxigénio poderá ser preferível o recurso a amostras porosas pelo facto de mais facilmente se manipularem as suas características microestruturais e devido à exclusão dos problemas associados à interação entre o substrato de alumina e o filme. As composições não dopadas são as indicadas para esta função se a gama de pressões de oxigénio a avaliar for relativamente pouco extensa sendo aconselhadas as composições dopadas com dador se for pretendida uma medição da pressão parcial de oxigénio em zonas mais extensas correspondentes à queima com deficiência ou excesso de oxigénio. Mesmo em amostras de elevada porosidade poderá ocorrer resposta transiente do material dopado com dador.

Biofuels and high value added products from the yeast Rhodosporidium toruloides NCYC 921: Strategies towards a true cost-effective and environmentally sustainable integrated multiproduct driven biorefinery [Poster]

Single-cell oils (SCO) have been considered a promising source of 3rd generation biofuels mainly in the final form of biodiesel. However, its high production costs have been a barrier towards the commercialization of this commodity. The fast growing yeast Rhodosporidium toruloides NCYC 921 has been widely reported as a potential SCO producing yeast. In addition to its well-known high lipid content (that can be converted into biodiesel), is rich in high value added products such as carotenoids with commercial interest. The process design and integration may contribute to reduce the overall cost of biofuels and carotenoid production and is a mandatory step towards their commercialization. The present work addresses the biomass disruption, extraction, fractionation and recovery of products with special emphasis on high added valued carotenoids (beta-carotene, torulene, torularhodin) and fatty acids directed to biodiesel. The chemical structure of torularhodin with a terminal carboxylic group imposes an additional extra challenge in what concern its separation from fatty acids. The proposed feedstock is fresh biomass pellet obtained directly by centrifugation from a 5L fed-batch fermentation culture broth. The use of a wet instead of lyophilised biomass feedstock is a way to decrease processing energy costs and reduce downstream processing time. These results will contribute for a detailed process design. Gathered data will be of crucial importance for a further study on Life-Cycle Assessment (LCA).

The role of microRNA 21 in the development of in-stent restenosis

Coronary heart disease is a major cause of morbidity and mortality worldwide. Percutaneous coronary intervention (PCI) has become the most widely used method of coronary artery revascularisation. The use of stents to hold open atherosclerosis induced arterial narrowing has significantly reduced elastic recoil and acute vessel occlusion following balloon angioplasty. However, bare metal stents have been associated with in-stent restenosis attributed to vascular smooth muscle cell (VSMC) hyperplasia and excessive neointimal formation. The resultant luminal renarrowing may manifest clinically with the return of symptoms such as chest pain or shortness of breath. The development of drug eluting stents has significantly reduced the incidence of in-stent restenosis (ISR). Unfortunately the antiproliferative medications used not only inhibit VSMC proliferation but also re-endothelialisation of the stented vessel. In addition, the drug impregnated polymer coating has been associated with a chronic inflammatory response within the vessel wall predisposing patients to stent thrombosis. Thus the identification of novel therapies which promote vessel healing without excessive proliferative or inflammatory response may improve long term outcome and reduce the need for repeated revascularisation. MicroRNAs (miRs) are short (18-25 nucleotide) non-coding RNAs acting to regulate gene expression. By binding to the 3’untranslated region of mRNA they act to fine tune gene expression either by mRNA degradation or translational repression. Originally identified in coordinating tissue development microRNAs have also been shown to play important roles coordinating the inflammatory response and in numerous cardiovascular diseases. MiR-21 has been identified in human atherosclerotic plaques, arteriosclerosis obliterans and abdominal aortic aneurysms. In addition, its up regulation has been documented in preclinical models of vascular injury. This study sought to identify the role of miR-21 in the development of ISR. Utilising a small animal model of stenting and in vitro techniques, we sought to investigate its influence upon VSMC and immune cell response following stenting. 19 The refinement of a murine stenting model within the Baker laboratory and the electrochemical dissolution of the metal stent from within harvested vascular tissues significantly improved the ability to perform detailed histological analysis. In addition, identification of miRNAs using in situ hybridisation was achieved for the first time within stented tissue. Neointimal formation and ISR was significantly reduced in mice in which miR-21 had been genetically deleted. In addition, neointimal composition was found to be altered in miR-21 KO mice with reductions in VSMC and elastin content demonstrated. Importantly, no difference in re-endothelialisation was observed. In vitro analysis demonstrated that VSMCs from miR-21 KO mice had both reduced proliferative and migratory capacity following platelet derived growth factor stimulation. Molecular analysis revealed that these differences may, at least in part, be due to de-repression of programmed cell death 4 (PDCD4). PDCD4 is a known miR-21 target within VSMCs implicated in the suppression of proliferation and promotion of apoptosis. Unfortunately, initial attempts at antimiR mediated knockdown of miR-21 in vivo, failed to produce a similar change in the suppression of ISR. Furthermore, a significant alteration in macrophage polarisation state within the neointima of miR-21 WT and KO mice was noted. Immunohistochemical staining revealed a preponderance of anti-inflammatory M2 macrophages in KO mice. Analysis of bone marrow derived macrophages from miR-21 KO mice demonstrated an increased level of the peroxisome proliferation activating receptor-γ (PPARγ) which facilitates M2 polarisation. Importantly, significant alterations in numerous pro-inflammatory cytokines, which also have mitogenic effects, were also found following genetic deletion of miR-21. In Summary, this is the first study to look at miRs in the development of ISR. MiR-21 plays an important role in the development of ISR by influencing the proliferative response of VSMCs and modulating the immune response following stent deployment. Further attempts to modulate miR-21 expression following PCI may reduce ISR and the need for repeat revascularisation while also reducing the risk of stent thrombosis.

Pseudoneglect and visual attention networks

Pseudoneglect represents the tendency for healthy individuals to show a slight but consistent bias in favour of stimuli appearing in the left visual field. The bias is often measured using variants of the line bisection task. An accurate model of the functional architecture of the visuospatial attention system must account for this widely observed phenomenon, as well as for modulation of the direction and magnitude of the bias within individuals by a variety of factors relating to the state of the participant and/or stimulus characteristics. To date, the neural correlates of pseudoneglect remain relatively unmapped. In the current thesis, I employed a combination of psychophysical measurements, electroencephalography (EEG) recording and transcranial direct current stimulation (tDCS) in an attempt to probe the neural generator(s) of pseudoneglect. In particular, I wished to utilise and investigate some of the factors known to modulate the bias (including age, time-on-task and the length of the to-be-bisected line) in order to identify neural processes and activity that are necessary and sufficient for the lateralized bias to arise. Across four experiments utilising a computerized version of a perceptual line bisection task, pseudoneglect was consistently observed at baseline in healthy young participants. However, decreased line length (experiments 1, 2 and 3), time-on-task (experiment 1) and healthy aging (experiment 3) were all found to modulate the bias. Specifically, all three modulations induced a rightward shift in subjective midpoint estimation. Additionally, the line length and time-on-task effects (experiment 1) and the line length and aging effects (experiment 3) were found to have additive relationships. In experiment 2, EEG measurements revealed the line length effect to be reflected in neural activity 100 – 200ms post-stimulus onset over source estimated posterior regions of the right hemisphere (RH: temporo-parietal junction (TPJ)). Long lines induced a hemispheric asymmetry in processing (in favour of the RH) during this period that was absent in short lines. In experiment 4, bi-parietal tDCS (Left Anodal/Right Cathodal) induced a polarity-specific rightward shift in bias, highlighting the crucial role played by parietal cortex in the genesis of pseudoneglect. The opposite polarity (Left Cathodal/Right Anodal) did not induce a change in bias. The combined results from the four experiments of the current thesis provide converging evidence as to the crucial role played by the RH in the genesis of pseudoneglect and in the processing of visual input more generally. The reduction in pseudoneglect with decreased line length, increased time-on-task and healthy aging may be explained by a reduction in RH function, and hence contribution to task processing, induced by each of these modulations. I discuss how behavioural and neuroimaging studies of pseudoneglect (and its various modulators) can provide empirical data upon which accurate formal models of visuospatial attention networks may be based and further tested.

Importance of size-selective particle sampling for assessing occupational exposures: results from three different occupational settings

Sampling the total air concentration of particulate matter (PM) only provides a basic estimate of exposure that normally not allows correlating with the observed health effects. Therefore is of extreme importance to know the particles size distribution and, in more detail, the exposure to fine particles (≤ 2.5 µm). This particles dimension corresponds to the respirable fraction. This particle fraction can result, besides local effects, in systemic effects due to particle deposition and clearance from the lungs and transport within the organism. This study intended to describe occupational exposure to PM2.5 in three different units located near Lisbon and related with occupational exposure to organic dust, namely: swine and poultry feed production and waste management.