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{Nitrite reductase from the sulfate-reducing bacterium Desulfovibrio desulfuricans ATCC 27774 is a multihaem (type c) membrane-bound enzyme that catalyzes the dissimilatory conversion of nitrite to ammonia. Crystals of the oxidized form of this enzyme were obtained using PEG and CaCl2 as precipitants in the presence of 3-(decylmethylammonium)propane-1-sulfonate and belong to the space group P2(1)2(1)2(1), With unit-cell parameters a = 78.94

The socio-economic subsystem encompassing fisheries may be defined as including not only the harvesting sector but also several related activities occurring both upstream (shipbuilding, gear manufacture) and downstream (processing, distribution and trade). But these closely interrelated economic activities can also be set within a much broader system which would include the ecological, institutional and political influences which frame economic behaviour. The value of this broader conceptualisation is that it treats fisheries not as an isolated and independent economic activity but as part of a more holistic and complex system. This broader perspective is of particular significance when attempting to examine the concept of regional dependence. The socio-economic subsystem for fisheries is dominated by small and medium sized enterprises (SMEs). And Peniche emerges as one of Portugal’s most important fishing ports whether measured in terms of the volume of landings or the total numbers of fishermen. It also has one of the highest levels of fisheries dependence of all coastal municipalities in Portugal with over 20% of its workforce currently engaged in fisheries related employment, faces a daunting and uncertain future. The social fabric of fisheries dependent communities also suffers serious damage; once again, the technocratic approach to management has no solutions to offer. It is essential, therefore, to turn away from the existing approach and to develop instead new forms of intervention; in short, to provide a new vision. This implies change not only to the policy process but also in the attitudes of the social actors and in the preoccupations of fisheries related research. An integrated approach is required based on participative action and the development of an integrated information network.

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The required results of stochastic calculus are introduced as well as sufficient conditions for a diffusin to be ergodic. Invariant densities are obtained for two families of diffusion. These families belong the Ornstein-Uhlenbeck and Cox-Ingersoll & Ross diffusions. The moments of transition density of the Cox-Ingersoll & Ross diffusion were obtained and it was shown that this density converges to the invariant density. Lastly a technique, based on sample partition, is given for parameter estimation for ergodic diffusion. A numerical application of that technique for the Ornstein-Uhlenbeck diffusion is given. Final remarks and comments are included.

The experimental analysis of nine prestressed flat slab models under punching is described and the results are compared with the recommendations of EC2 and MC90. The tests were performed on specimens at a scale 1/3, prestressed with unbonded high strength steel. In the first set, the specimens (AR2 to AR5 and AR7) were only subjected to in-plane compression, to evaluate the effect of the in-plane forces on the punching resistance. The second set of tests (AR8 to AR11) intended to study the effect of the vertical component of the tendon forces near the column in the punching resistance. This work aims to improve the understanding of the behaviour of prestressed flat slabs under punching load and the evaluation of the punching resistance.

The present work reports experimental research carried out on repair and strengthening methods of flat slabs for punching. The repair and strengthening methods studied are: strengthening using transversal prestress, repair by substitution of the damaged concrete; and strengthening using steel beams as a column head, connected to the column and to the slab with epoxy resin and mechanical expansion anchors. Four experimental test slabs (AR1 to AR4) were produced and tested: two with transversal prestress, one only repaired and one strengthened with a steel column head. The execution process, its efficiency and design, are discussed.

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In PE films aged under electric field the crystallisation of water (and melting of ice) has been studied by quadrupolar NMR, this technique allows one to determine the concentration of water as low as 10(-4). It is shown that the pore dimensions of the tracks forming the water trees of the order of 2.5 nm, are independent of the ageing time. The mobility of water in these water trees and in porous glass, of similar pore dimensions, are compared. (C) 2000 Elsevier Science Ltd. All rights reserved.

In PE films aged under electric field the crystallisation of water (and melting of ice) has been studied by quadrupolar NMR, this technique allows one to determine the concentration of water as low as 10(-4). It is shown that the pore dimensions of the tracks forming the water trees of the order of 2.5 nm, are independent of the ageing time. The mobility of water in these water trees and in porous glass, of similar pore dimensions, are compared. (C) 2000 Elsevier Science Ltd. All rights reserved.

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The aerobic purification of Pseudomonas nautica 617 nitrous oxide reductase yielded two forms of the enzyme exhibiting different chromatographic behaviors. The protein contains six copper atoms per monomer, arranged in two centers named CUA and Cut. Cut could be neither oxidized nor further reduced under our experimental conditions, and exhibits a 4-line EPR spectrum (g(x)= 2.015, A(x) = 1.5 mT, g(y) = 2.071, A(y) = 2 mT, g(z) = 2.138, A(z) = 7 mT) and a strong absorption at similar to 640 nm. Cu-A can be stabilized in a reduced EPR-silent state and in an oxidized state with a typical 7-line EPR spectrum (g(x) g(y) = 2.021, A(x) = A(y) = 0 T, g(z) =0.178, A(z) = 4 mT) and absorption bands at 480, 540, and similar to 800 nm. The difference between the two purified forms of nitrous oxide reductase is interpreted as a difference in the oxidation state of the CuA center. In form A, CUA is predominantly oxidized (S = 1/2, Cu1.5+-Cu1.5+), while in form B it is mostly in the one-electron reduced state (S = 0, Cu1+-Cu1+). In both forms, Cu-Z remains reduced (S = 1/2). Complete crystallographic data at 2.4 Angstrom indicate that Cu-A is a binuclear site (similar to the site found in cytochrome c oxidase) and Cu-Z is a novel tetracopper cluster [Brown, K., et ai. (2000) Nat. Struct. Biol. (in press)]. The complete amino acid sequence of the enzyme was determined and comparisons made with sequences of other nitrous oxide reductases, emphasizing the coordination of the centers. A 10.3 kDa peptide copurified with both forms of nitrous oxide reductase shows strong homology with proteins of the heat-shock GroES chaperonin family.

Mycobacterium tuberculosis KatG is a multifunctional heme enzyme responsible for activation of the antibiotic isoniazid. A KatG(S315T) point mutation is found in >50% of isoniazid-resistant clinical isolates. Since isoniazid activation is thought to involve an oxidation reaction, the redox potential of KatG was determined using cyclic voltammetry, square wave voltammetry, and spectroelectrochemical titrations. Isoniazid activation may proceed via a cytochrome P450-like mechanism. Therefore, the possibility that substrate binding by KatG leads to an increase in the heme redox potential and the possibility that KatG(S315T) confers isoniazid resistance by altering the redox potential were examined. Effects of the heme spin state on the reduction potentials of KatG and KatG(S315T) were also determined. Assessment of the Fe3+/Fe2+ couple gave a midpoint potential of ca. -50 mV for both KatG and KatG(S315T). In contrast to cytochrome P450s, addition of substrate had no significant effect on either the KatG or KatG(S315T) redox potential. Conversion of the heme to a low-spin configuration resulted in a -150 to -200 mV shift of the KatG and KatG(S315T) redox potentials. These results suggest that isoniazid resistance conferred by KatG(S315T) is not mediated through changes in the heme redox potential. The redox potentials of isoniazid were also determined using cyclic and square wave voltammetry, and the results provide evidence that the ferric KatG and KatG(S315T) midpoint potentials are too low to promote isoniazid oxidation without formation of a high-valent enzyme intermediate such as compounds I and IT or oxyferrous KatG.