n/a

Respiratory nitric oxide reductase (NOR) was purified from membrane extract of Pseudomonas (Ps.) nautica cells to homogeneity as judged by polyacrylamide gel electrophoresis. The purified protein is a heterodimer with subunits of molecular masses of 54 and 18 kDa. The gene encoding both subunits was cloned and sequenced. The amino acid sequence shows strong homology with enzymes of the cNOR class. Iron/heme determinations show that one heme c is present in the small subunit (NORC) and that approximately two heme b and one non-heme iron are associated with the large subunit (NORB), in agreement with the available data for enzymes of the cNOR class. Mossbauer characterization of the as-purified, ascorbate-reduced, and dithionite-reduced enzyme confirms the presence of three heme groups (the catalytic heme b(3) and the electron transfer heme b and heme c) and one redox-active non-heme Fe (Fe-B). Consistent with results obtained for other cNORs, heme c and heme b in Ps. nautica cNOR were found to be low-spin while FeB was found to be high-spin. Unexpectedly, as opposed to the presumed high-spin state for heme b(3), the Mossbauer data demonstrate unambiguously that heme b(3) is, in fact, low-spin in both ferric and ferrous states, suggesting that heme b(3) is six-coordinated regardless of its oxidation state. EPR spectroscopic measurements of the as-purified enzyme show resonances at the g similar to 6 and g similar to 2-3 regions very similar to those reported previously for other cNORs. The signals at g = 3.60, 2.99, 2.26, and 1.43 are attributed to the two charge-transfer low-spin ferric heme c and heme b. Previously, resonances at the g similar to 6 region were assigned to a small quantity of uncoupled high-spin Fe-III heme b(3). This assignment is now questionable because heme b(3) is low-spin. On the basis of our spectroscopic data, we argue that the g = 6.34 signal is likely arising from a spin spin coupled binuclear center comprising the low-spin Fe-III heme b(3) and the high-spin Fe-B(III). Activity assays performed under various reducing conditions indicate that heme b(3) has to be reduced for the enzyme to be active. But, from an energetic point of view, the formation of a ferrous heme-NO as an initial reaction intermediate for NO reduction is disfavored because heme [FeNO](7) is a stable product. We suspect that the presence of a sixth ligand in the Fe-II-heme b(3) may weaken its affinity for NO and thus promotes, in the first catalytic step, binding of NO at the Fe-B(II) site. The function of heme b(3) would then be to orient the Fe-B-bound NO molecules for the formation of the N-N bond and to provide reducing equivalents for NO reduction.

n/a

The universal semi-empirical equation has been commonly used to quantitatively describe the energy dependence of the secondary electron yield (SEY). It is even used as a first reliability test for experimental data. The equation is based on the assumption that the stopping power is constant along the electron trajectory. In this article, we derive a novel analytical expression based on a more advanced model which considers linear stopping power dependence on penetration depth. Although coinciding with the universal equation at low energies, the novel function has lower intensity in the higher energy range. The models were compared with experimental SEY data of different metals, taken from literature, as well as freshly cleaved highly oriented pyrolytic graphite, measured in the frame of this work. It is confirmed that the novel expression better describes the experimental data.

We present a new approach for real-time monitoring of PCR amplification of a specific sequence from the human c-MYC proto-oncogene using a Ta 2O 5 electrolyte-insulator-semiconductor (EIS) sensor. The response of the fabricated EIS sensor to cycle DNA amplification was evaluated and compared to standard SYBR-green fluorescence incorporation, showing it was possible to detect DNA concentration variations with 30mV/??M sensitivity. The sensor's response was then optimized to follow in real-time the PCR amplification of c-MYC sequence from a genomic DNA sample attaining an amplification profile comparable to that of a standard real-time PCR. Owing to the small size, ease of fabrication and low-cost, the developed Ta 2O 5 sensor may be incorporated onto a microfluidic device and then used for real-time PCR. Our approach may circumvent the practical and economical obstacles posed by current platforms that require an external fluorescence detector difficult to miniaturize and incorporate into a lab-on-chip system. ?? 2011 Elsevier B.V.

A completely new strategy for cell culture focusing on the design of three-dimensional (3D) smart surfaces by supercritical fluid technology has been developed. This approach might overcome the limitations on cell expansion and proliferation of currently existing techniques. An alternative technology, based on supercritical carbon dioxide, was used to polymerize poly(N- isopropylacrylamide) (PNIPAAm) and to foam poly(d,l-lactic acid) (P D,L LA), creating a thermosensitive 3D structure which has proven to have potential as a substrate for cell growth and expansion. We demonstrated that the thermosensitive matrices promoted cell detachment, thus P D,L LA scaffolds have the potential to be used as substrates for cell growth and expansion avoiding enzymatic and mechanical methods of cell harvesting. The harvested cells were replated to evaluate their viability, which was not compromised. A major advantage of this technology is the fact that the prepared materials can be recovered and reused. Therefore, the same substrate can be recycled and reused for different batches. An indirect impact of the technology developed is related to the field of biotechnology, as this novel technology for cell expansion can be applied to any adherent cell cultures. © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

n/a

The application of diffraction gratings to solar cells is a promising approach to superseding the light trapping limits of conventional Lambertian structures. In this paper a mathematical formalism is derived for calculating the absorption that can be expected in a solar cell equipped with a diffraction grating, which can be applied to any lattice geometry and grating profile. Furthermore, the formalism is used to calculate the upper limit of total absorption that can theoretically be achieved using a diffraction grating. The derived formalism and limits are valid when the solar cell thickness is greater than the coherence length of the illuminating solar spectrum. Comparison is made to the upper limit achievable using an angularly selective Rugate filter, which is also calculated. Both limits are found to be considerably higher than the Lambertian limit within the range of sunlight concentration factors practically employed in photovoltaic systems (1–1000×). The upper limit of absorption using the diffraction grating is shown to be equal to the thermodynamic limit for all absorbances and concentration factors. The limit for the Rugate filter is generally lower, but tends to the thermodynamic limit for lower cell absorbances. Copyright © 2011 John Wiley {&} Sons, Ltd.

We prove necessary and sufficient conditions for the applicability of the finite section method to an arbitrary operator in the Banach algebra generated by the operators of multiplication by piecewise continuous functions and the convolution operators with symbols in the algebra generated by piecewise continuous and slowly oscillating Fourier multipliers on \(L^p(\mathbb{R})\), \(1 < p < \infty\).

n/a

The post-graduation in the field of Technology Assessment (TA) is recent and that are several and different ways to be organised. Most experiences are related with the Masters diplom level (2nd cycle of graduation in high education). Just one in PhD level is explicit in the field of TA, and some other PhD courses include also TA topics in their programme structure. In this chapter we will analyse the problems related with the design of a post-graduation (MA, MSc or PhD) programme in the field of TA using as reference some international experiences. Hereby, the main conclusion seems to address labour market needs in the specialised knowledge of TA, of technology management or technology innovation. In this sense TA should be included as “minor” into post-graduation courses which may range from engineering disciplines to social sciences. As a graduation programme it can fill an expertise gap between technicians,engineers, scientists and the strategic decision makers or policy makers.

n/a

n/a

n/a

n/a

n/a

No abstract is available for this item.

n/a

n/a

n/a

n/a

n/a

Pseudoazurins are small type 1 copper proteins that are involved in the flow of electrons between various electron donors and acceptors in the bacterial periplasm, mostly under denitrifying conditions. The previously determined structure of Paracoccus pantotrophus pseudoazurin in the oxidized form was improved to a nominal resolution of 1.4 angstrom, with R and R-free values of 0.188 and 0.206, respectively. This high-resolution structure makes it possible to analyze the interactions between the monomers and the solvent structure in detail. Analysis of the high-resolution structure revealed the structural regions that are responsible for monomer-monomer recognition during dimer formation and for protein-protein interaction and that are important for partner recognition. The pseudoazurin structure was compared with other structures of various type 1 copper proteins and these were grouped into families according to similarities in their secondary structure; this may be useful in the annotation of copper proteins in newly sequenced genomes and in the identification of novel copper proteins.

n/a

Research activities at European level on the concept of new working environments offers considerable attention to the challenges of the increased competencies of people working together with automated technologies. Since the decade of 1980 the development of approaches for the humanization of work organization, and for the development of participative organizational options induced to new proposals related to the development of complex and integrated automated systems. From such parallel conceptual development emerged the concept of “anthropocentric robotic systems” and quickly it covered also other fields of automation. More recently, the debate also covers issues related to working perception of people dealing with autonomous systems (e.g. Autonomous robotics) in tasks related to production planning, to programming and to process control. In fact, today one can understand the wider use of the anthropocentrism concept of production architectures, when understanding the new quality of these systems. In this chapter the author analyses the evolution of these issues related to governance of ICT applied to manufacturing and industrial services in research programmes strengthening very much the ‘classical’ concept of anthropocentric-based systems. It is emerging a new value of the intuitive capacities and human knowledge in the optimization and flexibilization of the manufacturing processes. While this would be a pre-condition to understand the human-robot communication needs, there is also a need to take into consideration the qualitative variables in the definition and design of robotic systems, jobs and production systems.