At the beginning of the 21st century there are new expectations and challenges towards Technology Assessment (TA). Among these there is a new awareness on TA issues in education, in particular at universities. While TA was mainly an activity at extra-universitarian research institutions for a long time now there are new developments and initiative towards integrating TA issues in university courses. We will first give an insight into the international development. Secondly we will focus on the “TA and education” landscape in Germany and Portugal in more detail, followed by a description of new and emerging forms of cooperation between Portugal and Germany in this field which might serve as a model or an example for further cooperation between other partners.

At the beginning of the 21st century there are new expectations and challenges towards Technology Assessment (TA). Among these there is a new awareness on TA issues in education, in particular at universities. While TA was mainly an activity at extra-universitarian research institutions for a long time now there are new developments and initiative towards integrating TA issues in university courses. We will first give an insight into the international development. Secondly we will focus on the “TA and education” landscape in Germany and Portugal in more detail, followed by a description of new and emerging forms of cooperation between Portugal and Germany in this field which might serve as a model or an example for further cooperation between other partners.

The present invention relates to a system and process for detection and/or qualitative and quantitative identification of the biological material, such as specific sequences of nucleic acids or proteins as antibodies, present in biological samples. The system is composed by one or more light sources (1) combined with one or more integrated optical photo sensors, or not, and various electronic components (4), necessary for obtaining/ processing of the signal emitted by the metal nanoprobes functionalized with a solution of biological composite, as well as also a micro-controller and a microprocessor, fixed or portable. This photosensor structure is able to detect and to quantify the colour variations produced by metal nanoprobes, being this preferentially gold, functionalized by oligonucleotides complementary to specific DNA/RNA sequences, proteins, as for instance antibodies and/or antigens related with certain disease, or other sample or solution of biological composite, that are to be investigated. The detection and quantification process is based on the response of a photosensor, singular or integrated, based on thin film technology of amorphous, nanocrystalline or microcrystalline silicon and their alloys, as well as the new active ceramic semiconductors, amorphous and not amorphous.

Nitric Oxide Reductase (NOR) is an integral membrane protein performing the reduction of NO to N(2)O. NOR is composed of two subunits: the large one (NorB) is a bundle of 12 transmembrane helices (TMH). It contains a b type heme and a binuclear iron site, which is believed to be the catalytic site, comprising a heme b and a non-hemic iron. The small subunit (NorC) harbors a cytochrome c and is attached to the membrane through a unique TMH. With the aim to perform structural and functional studies of NOR, we have immunized dromedaries with NOR and produced several antibody fragments of the heavy chain (VHHs, also known as nanobodies). These fragments have been used to develop a faster NOR purification procedure, to proceed to crystallization assays and to analyze the electron transfer of electron donors. BIAcore experiments have revealed that up to three VHHs can bind concomitantly to NOR with affinities in the nanomolar range. This is the first example of the use of VHHs with an integral membrane protein. Our results indicate that VHHs are able to recognize with high affinity distinct epitopes on this class of proteins, and can be used as versatile and valuable tool for purification, functional study and crystallization of integral membrane proteins.

O presente invento relaciona-se com um método para controlo de reac{\c c}ões enzimáticas de síntese de ácidos nucleicos, recorrendo a nucleótidos funcionalizados com derivados de 4-metilcumarinas (1) protectores e fotolábeis. Quando ligado aos nucleótidos (2), o grupo cumarinico (3) impede que estes sejam utilizados como substrato por parte das enzimas, impossibilitando a ocorrência de reac{\c c}ão. Através de irradia{\c c}ão com radia{\c c}ão electromagnética, o grupo cumarinico é libertado, ficando o nucleótido disponível para a reac{\c c}ão. Desta forma, as reac{\c c}ões enzimáticas de síntese de ácidos nucleicos podem ser controladas através da luz.

Neste trabalho pretende-se estudar a variabilidade da frequência cardíaca através da implementação de vários métodos de análise de electrocardiogramas. Na primeira parte descrevem-se os métodos de análise espectral, bem como a representação tempo-frequência. A decomposição de sinais para detectar e isolar os batimentos cardíacos do electrocardiograma é implementada. Descrevem-se algoritmos de isolamento para detecção dos eventos interessantes e pontos fiduciais que os constituem. Na segunda parte estima-se a variação da frequência cardíaca e estabelece-se a sua relação com as variações temporais e espectrais das características encontradas pelos métodos implementados na primeira parte do trabalho.

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Aldehyde oxidoreductase from Desulfovibrio gigas (DgAOR) is a member of the xanthine oxidase (XO) family of mononuclear Mo-enzymes that catalyzes the oxidation of aldehydes to carboxylic acids. The molybdenum site in the enzymes of the XO family shows a distorted square pyramidal geometry in which two ligands, a hydroxyl/water molecule (the catalytic labile site) and a sulfido ligand, have been shown to be essential for catalysis. We report here steady-state kinetic studies of DgAOR with the inhibitors cyanide, ethylene glycol, glycerol, and arsenite, together with crystallographic and EPR studies of the enzyme after reaction with the two alcohols. In contrast to what has been observed in other members of the XO family, cyanide, ethylene glycol, and glycerol are reversible inhibitors of DgAOR. Kinetic data with both cyanide and samples prepared from single crystals confirm that DgAOR does not need a sulfido ligand for catalysis and confirm the absence of this ligand in the coordination sphere of the molybdenum atom in the active enzyme. Addition of ethylene glycol and glycerol to dithionite-reduced DgAOR yields rhombic Mo(V) EPR signals, suggesting that the nearly square pyramidal coordination of the active enzyme is distorted upon alcohol inhibition. This is in agreement with the X-ray structure of the ethylene glycol and glycerol-inhibited enzyme, where the catalytically labile OH/OH(2) ligand is lost and both alcohols coordinate the Mo site in a eta(2) fashion. The two adducts present a direct interaction between the molybdenum and one of the carbon atoms of the alcohol moiety, which constitutes the first structural evidence for such a bond in a biological system.

The clothing sector in several countries is still seen, in many aspects as a traditional sector with some average characteristics, nevertheless is a very important sector in terms of labour market. Globalization and de-localization are having a strong impact in the organisation of work and in occupational careers. Very few companies are able to keep a position in the market without changes in organisation of work and workers, founding different ways to face this reality according to size, capital and position. We could find two main paths: one where companies outsource production to another territory, close and/or dismissal the workers; other path, where companies up skilled their capacities. This paper will present some results from the European project WORKS – Work organisation and restructuring in the knowledge society (6th Framework Programme), focusing the Portuguese case studies in several clothing companies in a comparative analysis with some other European countries.

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The orange-coloured protein (ORP) from Desulfovibrio gigas is a 12 kDa protein that contains a novel mixed-metal sulfide cluster of the type [S(2)MoS(2)CuS(2)MoS(2)]. Diffracting crystals of the apo form of ORP have been obtained. Data have been collected for the apo form of ORP to 2.25 A resolution in-house and to beyond 2.0 A resolution at ESRF, Grenoble. The crystals belonged to a trigonal space group, with unit-cell parameters a = 43, b = 43, c = 106 A.

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A conductometric biosensor for nitrite detection was developed using cytochrome c nitrite reductase (ccNiR) extracted from Desulfovibrio desulfuricans ATCC 27774 cells immobilized on a planar interdigitated electrode by cross-linking with saturated glutaraldehyde (GA) vapour in the presence of bovine serum albumin, methyl viologen (MV), Nafion, and glycerol. The configuration parameters for this biosensor, including the enzyme concentration, ccNiR/BSA ratio, MV concentration, and Nafion concentration, were optimized. Various experimental parameters, such as sodium dithionite added, working buffer solution, and temperature, were investigated with regard to their effect on the conductance response of the biosensor to nitrite. Under the optimum conditions at room temperature (about 25 degrees C), the conductometric biosensor showed a fast response to nitrite (about 10s) with a linear range of 0.2-120 microM, a sensitivity of 0.194 microS/microM [NO(2)(-)], and a detection limit of 0.05 microM. The biosensor also showed satisfactory reproducibility (relative standard deviation of 6%, n=5). The apparent Michaelis-Menten constant (K(M,app)) was 338 microM. When stored in potassium phosphate buffer (100mM, pH 7.6) at 4 degrees C, the biosensor showed good stability over 1 month. No obvious interference from other ionic species familiar in natural waters was detected. The application experiments show that the biosensor is suitable for use in real water samples.

The characterization of a novel Mo-Fe protein (MorP) associated with a system that responds to Mo in Desulfovibrio alaskensis is reported. Biochemical characterization shows that MorP is a periplasmic homomultimer of high molecular weight (260 +/- 13 kDa) consisting of 16-18 monomers of 15321.1 +/- 0.5 Da. The UV/visible absorption spectrum of the as-isolated protein shows absorption peaks around 280, 320, and 570 nm with extinction coefficients of 18700, 12800, and 5000 M(-1) cm(-1), respectively. Metal content, EXAFS data and DFT calculations support the presence of a Mo-2S-[2Fe-2S]-2S-Mo cluster never reported before. Analysis of the available genomes from Desulfovibrio species shows that the MorP encoding gene is located downstream of a sensor and a regulator gene. This type of gene arrangement, called two component system, is used by the cell to regulate diverse physiological processes in response to changes in environmental conditions. Increase of both gene expression and protein production was observed when cells were cultured in the presence of 45 microM molybdenum. Involvement of this system in Mo tolerance of sulfate reducing bacteria is proposed.

With the emergence of a global division of labour, the internationalisation of markets and cultures, the growing power of supranational organisations and the spread of new information technologies to every field of life, it starts to appear a different kind of society, different from the industrial society, and called by many as ‘the knowledge-based economy’, emphasizing the importance of information and knowledge in many areas of work and organisation of societies. Despite the common trends of evolution, these transformations do not necessarily produce a convergence of national and regional social and economic structures, but a diversity of realities emerging from the relations between economic and political context on one hand and the companies and their strategies on the other. In this sense, which future can we expect to the knowledge economy? How can we measure it and why is it important? This paper will present some results from the European project WORKS – Work organisation and restructuring in the knowledge society (6th Framework Programme), focusing the future visions and possible future trends in different countries, sectors and industries, given empirical evidences of the case studies applied in several European countries, underling the importance of foresight exercises to design policies, prevent uncontrolled risks and anticipate alternatives, leading to different ‘knowledge economies’ and not to the ‘knowledge economy’.

With the emergence of a global division of labour, the internationalisation of markets and cultures, the growing power of supranational organisations and the spread of new information technologies to every field of life, it starts to appear a different kind of society, different from the industrial society, and called by many as ‘the knowledge-based economy’, emphasizing the importance of information and knowledge in many areas of work and organisation of societies. Despite the common trends of evolution, these transformations do not necessarily produce a convergence of national and regional social and economic structures, but a diversity of realities emerging from the relations between economic and political context on one hand and the companies and their strategies on the other. In this sense, which future can we expect to the knowledge economy? How can we measure it and why is it important? This paper will present some results from the European project WORKS – Work organisation and restructuring in the knowledge society (6th Framework Programme), focusing the future visions and possible future trends in different countries, sectors and industries, given empirical evidences of the case studies applied in several European countries, underling the importance of foresight exercises to design policies, prevent uncontrolled risks and anticipate alternatives, leading to different ‘knowledge economies’ and not to the ‘knowledge economy’.

O desenvolvimento de um pacote de software para lidar facilmente com electrocardiogramas de alta resolução tornou-se importante para pesquisa na área de electrocardiografia. O desenvolvimento de novas técnicas para detecção de potenciais tardios e outros problemas associados a arritmias cardíacas têm sido objecto de estudo ao longo dos anos. No entanto, ainda existe a lacuna de um pacote de software que facilmente permita implementar algumas destas inovadoras técnicas de uma forma integrada, possibilitando avaliar técnicas clássicas como o protocolo de Simson para a detecção de sinais não estacionários (potenciais tardios). Algumas destas inovadoras técnicas envolvem a detecção tempo-frequência usando escalogramas ou a análise espectral usando metodologias wavelet-packet, sendo implementadas no software desenvolvido com flexibilidade e versatilidade suficientes para que futuramente sirva de plataforma de pesquisa para o refinamento destas mesmas técnicas no que toca ao processamento de sinais de electrocardiogramas de alta resolução. O software aqui desenvolvido foi também desenhado de forma a suportar dois tipos de ficheiros diferentes provenientes de outros tantos sistemas de aquisição. Os sistemas suportados são o ActiveTwo da Biosemi e o USBamp da g.tec.

The following contribution considers whether global restructuring creates new forms of the division of labor. On the basis of empirical data from a comparative project in 14 European countries, the author supports the hypothesis that in addition to the ongoing process of the internationalization of work, there are ‘hidden’ effects at the local level. From the perspective of three occupational clusters, dynamics can be observed which have differing impacts on the occupational groups. Thus, there is a simultaneous process of restructuring and redefining skills, labor processes and the working organization which forms the daily reality of working men and women.

Nanotechnology is a multidisciplinary field that brings together diverse fields of research and development such as engineering, biology, physics and chemistry. Formal definitions of nanotechnology refer to man-made devices, components and structures in the 1-100 nm range in at least one dimension. Advances in nanoscience are having a significant impact on many scientific fields, boosting the development of a variety of important technologies. Nanotechnology offers an unprecedented opportunity to interact with cancer cells in real time at the molecular and cellular scale. Because of their small size, nanoscale devices can readily interact with biomolecules on both the surface of cells and inside of cells. The concerted development of nanoscale devices, structures and components have provided essential breakthroughs in monitoring and fighting cancer at the earliest stages of the cancer process. Nanotechnology offers a wealth of tools that may provide researchers with new and innovative ways to diagnose and treat cancer - new imaging agents; systems for real-time assessments of therapeutic and surgical efficacy; multifunctional, targeted devices capable of bypassing biological barriers to deliver multiple therapeutic agents directly to cancer cells and tissues that play a critical role in cancer growth and metastasis; agents that can monitor predictive molecular changes allowing for preventive action against precancerous cells becoming malignant; minimizing costs for multiplex analysis. Nanotechnology, if properly integrated with conventional cancer research, may provide extraordinary prospects towards better diagnosis and effective therapy.

FRET can be used as a strategy to assign different simultaneous events in the same sample but {"}cross-talk{"} problems are a limitation. Here we present a contribution for the better understanding of the {"}cross-talk{"} in FRET experiments that include several pairs in the same sample. Using oligonucleotide probes labeled with fluorescent dyes which can be selectively excited at a specific wavelength, and using target oligonucleotides tagged with a fluorescent dye with specific characteristics that allow only it to emit light upon selective excitation of a specific probe by energy transfer (FRET), we aim to identify the exact probe-target hybridized pair. When using three donors to probe the presence of complementary targets, only 20% of possible donor/acceptor combinations give straightforward signals readily identifiable with the sample composition, while in the remaining cases severe cross-excitation prevents the direct identification of the sample composition. To correctly resolve the samples identity, we developed a theoretical model that enables the unequivocal attribution of a sample composition to a given set of fluorescence signals, in the presence of three donors.

Nitric Oxide Reductase (NOR) is an integral membrane protein performing the reduction of NO to N(2)O. NOR is composed of two subunits: the large one (NorB) is a bundle of 12 transmembrane helices (TMH). It contains a b type heme and a binuclear iron site, which is believed to be the catalytic site, comprising a heme b and a non-hemic iron. The small subunit (NorC) harbors a cytochrome c and is attached to the membrane through a unique TMH. With the aim to perform structural and functional studies of NOR, we have immunized dromedaries with NOR and produced several antibody fragments of the heavy chain (VHHs, also known as nanobodies (TM)). These fragments have been used to develop a faster NOR purification procedure, to proceed to crystallization assays and to analyze the electron transfer of electron donors. BIAcore experiments have revealed that up to three VHHs can bind concomitantly to NOR with affinities in the nanomolar range. This is the first example of the use of VHHs with an integral membrane protein. Our results indicate that VHHs are able to recognize with high affinity distinct epitopes on this class of proteins, and can be used as versatile and valuable tool for purification, functional study and crystallization of integral membrane proteins.

High blood pressure or hypertension is a condition affecting many individuals and represents a controllable risk factor for cardiovascular diseases such as coronary heart disease and stroke. A non-pharmacological approach to manage these includes the application of food components with antihypertensive activity. Milk protein-derived peptides have been exploited as natural hypotensive agents, namely the peptides {Val-Pro-Pro} {(VPP)} and {Ile-Pro-Pro} {(IPP)}, already commercialized in functional foods as a potential alternative to synthetic drugs. These bioactive peptides inhibit in vitro and in vivo the Angiotensin I-converting enzyme {(ACE)}, a protein with an important role in blood pressure regulation. In this work, we attempted to elucidate the possible mode of interaction between the peptides and {ACE}, including mechanisms of binding to the cofactor Zn2+, and further contrast this with the known mode of inhibition exerted by synthetic drugs {(Captopril}, Enalaprilat and Lisinopril). The bioactive peptide {Ala-Leu-Pro-Met-His-Ile-Arg} {(ALPMHIR)}, also known to inhibit the enzyme {ACE} but with a lower efficiency than {VPP} and {IPP}, was utilized in the docking studies for comparison. It was observed that the best docking poses obtained for {VPP} and {IPP} were located at the {ACE} catalytic site with very high resemblance to the drugs mode of interaction, including the coordination with Zn2+. As for {ALPMHIR}, the best docking poses were located in the narrow {ACE} channel outside the catalytic site, representing higher affinity energies and fewer resemblances with the interaction established by drugs.