In this work it is demonstrated that the characterization of c-type haem containing proteins by electrochemical techniques needs to be cautiously performed when using pyrolytic graphite electrodes. An altered form of the cytochromes, which has a redox potential 300 mV lower than that of the native state and displays peroxidatic activity, can be induced by interaction with the pyrolytic graphite electrode. Proper control experiments need to be performed, as altered conformations of the enzymes containing c-type haems can show activity towards the enzyme substrate. The work was focused on the study of the activation mechanism and catalytic activity of cytochrome c peroxidase from Paracoccus pantotrophus. The results could only be interpreted with the assignment of the observed non-turnover and catalytic signals to a non-native conformation state of the electron-transferring haem. The same phenomenon was detected for Met-His monohaem cytochromes (mitochondrial cytochrome c and Desulfovibrio vulgaris cytochrome c-553), as well as for the bis-His multihaem cytochrome c(3) from Desulfovibrio gigas, showing that this effect is independent of the axial coordination of the c-type haem protein. Thus, the interpretation of electrochemical signals of c-type (multi)haem proteins at pyrolytic graphite electrodes must be carefully performed, to avoid misassignment of the signals and incorrect interpretation of catalytic intermediates.

We are facing an era, where pressures on health costs are extremely high, and the reforms in health system are almost constant. But over time, one factor remains unchanged – Technology continues being the sustenance of health care. Manufacturers, clinicians, patients, diagnostic and therapeutic technicians, hospital managers, government leaders, among others, either in public or private sector, are increasingly demanding in the sustained seek for information that support its decisions. Those decisions are about different types of issues: if, or how the technology can be developed, whether a technology should or should not enter the market, whether to acquire and use certain technology, and so forth. Such demand is well implied in the growth and development of Health Technology Assessment (HTA). This specialised field is commonly understood according to the International Network of Agencies for Health Technology Assessment (INAHTA, 2003) as an multidisciplinary analysis and decisional field, which studies the implications of clinical, social, ethical and economic development, dissemination and use of health technologies, without neglecting its political analysis (Goodman, 2004). The political decisions made based on HTA reports should be based on scientific evidence, linking efforts between the technical, economic and political dimensions, resourcing to a participatory vision, so that we can translate the best possible decision (Novaes 2006). On the other hand, the success of these decisions depends critically on the skills of the researcher to convey wisdom and confidence in applying rules of argumentation (Grunwald, 2007). In this paper we analyse the technical and methodological aspects of HTA, seen as a tool for evaluating health procedures and techniques. And we analyse the needs for skills and qualifications development of the actors involved in this process.

This review focuses on the novel CuZ center of nitrous oxide reductase, an important enzyme owing to the environmental significance of the reaction it catalyzes, reduction of nitrous oxide, and the unusual nature of its catalytic center, named CuZ. The structure of the CuZ center, the unique tetranuclear copper center found in this enzyme, opened a novel area of research in metallobiochemistry. In the last decade, there has been progress in defining the structure of the CuZ center, characterizing the mechanism of nitrous oxide reduction, and identifying intermediates of this reaction. In addition, the determination of the structure of the CuZ center allowed a structural interpretation of the spectroscopic data, which was supported by theoretical calculations. The current knowledge of the structure, function, and spectroscopic characterization of the CuZ center is described here. We would like to stress that although many questions have been answered, the CuZ center remains a scientific challenge, with many hypotheses still being formed.

Identifying redox partners and the interaction surfaces is crucial for fully understanding electron flow in a respiratory chain. In this study, we focused on the interaction of nitrous oxide reductase (N(2)OR), which catalyzes the final step in bacterial denitrification, with its physiological electron donor, either a c-type cytochrome or a type 1 copper protein. The comparison between the interaction of N(2)OR from three different microorganisms, Pseudomonas nautica, Paracoccus denitrificans, and Achromobacter cycloclastes, with their physiological electron donors was performed through the analysis of the primary sequence alignment, electrostatic surface, and molecular docking simulations, using the bimolecular complex generation with global evaluation and ranking algorithm. The docking results were analyzed taking into account the experimental data, since the interaction is suggested to have either a hydrophobic nature, in the case of P. nautica N(2)OR, or an electrostatic nature, in the case of P. denitrificans N(2)OR and A. cycloclastes N(2)OR. A set of well-conserved residues on the N(2)OR surface were identified as being part of the electron transfer pathway from the redox partner to N(2)OR (Ala495, Asp519, Val524, His566 and Leu568 numbered according to the P. nautica N(2)OR sequence). Moreover, we built a model for Wolinella succinogenes N(2)OR, an enzyme that has an additional c-type-heme-containing domain. The structures of the N(2)OR domain and the c-type-heme-containing domain were modeled and the full-length structure was obtained by molecular docking simulation of these two domains. The orientation of the c-type-heme-containing domain relative to the N(2)OR domain is similar to that found in the other electron transfer complexes.

The deactivation reaction of the [CoCp(1,4-sigma-C(4)-[Ph](4))PPh(3)] catalyst for the cyclotrimerization of acetylenes has been kinetico-mechanistically studied under different temperature, pressure, and solvent conditions. The results indicate a dramatic change in mechanism from conventional to ionic liquid solvents due to the polarity of the medium.

The activation mechanism of Pseudomonas stutzeri cytochrome c peroxidase (CCP) was probed through the mediated electrochemical catalysis by its physiological electron donor, P. stutzeri cytochrome c-551. A comparative study was carried out, by performing assays with the enzyme in the resting oxidized state as well as in the mixed-valence activated form, using cyclic voltammetry and a pyrolytic graphite membrane electrode. In the presence of both the enzyme and hydrogen peroxide, the peak-like signal of cytochrome c-551 is converted into a sigmoidal wave form characteristic of an E(r)C'(i) catalytic mechanism. An intermolecular electron transfer rate constant of (4 +/- 1) x 10(5) M(-1) s(-1) was estimated for both forms of the enzyme, as well as a similar Michaelis-Menten constant. These results show that neither the intermolecular electron transfer nor the catalytic activity is kinetically controlled by the activation mechanism of CCP in the case of the P. stutzeri enzyme. Direct enzyme catalysis using protein film voltammetry was unsuccessful for the analysis of the activation mechanism, since P. stutzeri CCP undergoes an undesirable interaction with the pyrolytic graphite surface. This interaction, previously reported for the Paracoccus pantotrophus CCP, induces the formation of a non-native conformation state of the electron-transferring haem, which has a redox potential 200 mV lower than that of the native state and maintains peroxidatic activity.

The effect of different mild post-annealing treatments in air, at 270 degrees C, for 4-6 min, on the optical, electrical, structural and chemical properties of copper sulphide (CuxS) thin films deposited at room temperature are investigated. CuxS films, 70nm thick, are deposited on glass substrates by vacuum thermal evaporation from a Cu2S:S (50:50 wt.%) sulphur rich powder mixture. The as-deposited highly conductive crystalline CuS (covellite) films show high carrier concentration (similar to 10(22) cm(-3)), low electrical resistivity (similar to 10(-4) Omega cm) and inconclusive p-type conduction. After the mild post-annealing, these films display increasing values of resistivity (similar to 10(-3) to similar to 10(-2) Omega cm) with annealing time and exhibit conclusive p-type conduction. An increase of copper content in CuxS phases towards the semiconductive Cu2S (chalcocite) compound with annealing time is reported, due to re-evaporation of sulphur from the films. However, the latter stoichiometry was not obtained, which indicates the presence of vacancies in the Cu lattice. In the most resistive films a Cu2O phase is also observed, diminishing the amount of available copper to combine with sulphur, and therefore the highest values of optical transmittance are reached (65%). The appearance on the surface of amorphous sulphates with annealing time increase is also detected as a consequence of sulphur oxidation and replacement of sulphur with oxygen. All annealed films are copper deficient in regards to the stoichiometric Cu2S and exhibit stable p-type conductivity. (C) 2011 Elsevier B.V. All rights reserved.

The effect of different mild post-annealing treatments in air, at 270 degrees C, for 4-6 min, on the optical, electrical, structural and chemical properties of copper sulphide (CuxS) thin films deposited at room temperature are investigated. CuxS films, 70nm thick, are deposited on glass substrates by vacuum thermal evaporation from a Cu2S:S (50:50 wt.%) sulphur rich powder mixture. The as-deposited highly conductive crystalline CuS (covellite) films show high carrier concentration (similar to 10(22) cm(-3)), low electrical resistivity (similar to 10(-4) Omega cm) and inconclusive p-type conduction. After the mild post-annealing, these films display increasing values of resistivity (similar to 10(-3) to similar to 10(-2) Omega cm) with annealing time and exhibit conclusive p-type conduction. An increase of copper content in CuxS phases towards the semiconductive Cu2S (chalcocite) compound with annealing time is reported, due to re-evaporation of sulphur from the films. However, the latter stoichiometry was not obtained, which indicates the presence of vacancies in the Cu lattice. In the most resistive films a Cu2O phase is also observed, diminishing the amount of available copper to combine with sulphur, and therefore the highest values of optical transmittance are reached (65%). The appearance on the surface of amorphous sulphates with annealing time increase is also detected as a consequence of sulphur oxidation and replacement of sulphur with oxygen. All annealed films are copper deficient in regards to the stoichiometric Cu2S and exhibit stable p-type conductivity. (C) 2011 Elsevier B.V. All rights reserved.

n/a

Domain Specic Languages (DSLs) are claimed to contribute
to increment productivity, while reducing the required maintenance and
programming expertise. In this context, the usability of these languages
becomes a major issue: if the language is not easy to learn and use, it is
unlikely to be successfully adopted.

It is important to foster high quality DSLs during its engineering process.
We argue that a systematic approach based on User Interface Experi-
mental validation techniques should be used to assess the impact of the
introduction of DSLs in the productivity of DSL user. Productivity can
be fostered by assessing important usability attributes early in the lan-
guage construction . This work's contribution, besides highlighting the
problem of the absence of systematic approaches for experimental vali-
dation of DSLs in general, is to identify existing evaluation approaches
that can be adapted from the eld of User Interfaces.

KAOS is one of the most well-known goal-oriented requirements engineering approaches. Nevertheless, building large KAOS models sometimes results in incomplete and/or complex requirements models that are difficult to understand and maintain. These shortcomings often lead to an increase in costs of product development and evolution. Therefore, for large-scale systems, the ability to manage the complexity and completeness of KAOS models is essential. In this paper, we propose a metrics suite for supporting the quantitative assessment of KAOS models complexity and completeness, in order to support their early identification. We apply the metrics to an example taken from a health club system specification.

Scenario modeling can be realized through different perspectives. In UML, scenarios are often modeled with activity models, in an early stage of development. Later, sequence diagrams are used to detail object interactions. The migration from activity diagrams to sequence diagrams is a repetitive and error-prone task. Model-Driven Development (MDD) can help streamlining this process, through transformation rules. Since the information in the activity model is insufficient to generate the corresponding complete sequence model, manual refinements are required. Our goal is to compare the relative effort of building the sequence diagrams manually with that of building them semi-automatically. Our results show a decrease in the number of operations required to build and refine the sequence model of approximately 64% when using MDD, when compared to the manual approach.