Increased levels of unconjugated bilirubin, the end product of heme catabolism, impair crucial aspects of nerve cell function. In previous studies, we demonstrated that bilirubin toxicity may be due to cell death by apoptosis. To characterize the sequence of events leading to neurotoxicity, we exposed developing rat brain astrocytes and neurons to unconjugated bilirubin and investigated whether changes in membrane dynamic properties can mediate apoptosis. Bilirubin induced a rapid, dose-dependent increase in apoptosis, which was nevertheless preceded by impaired mitochondrial metabolism. Using spin labels and electron paramagnetic resonance spectroscopy analysis of whole cell and isolated mitochondrial membranes exposed to bilirubin, we detected major membrane perturbation. By physically interacting with cell membranes, bilirubin induced an almost immediate increase in lipid polarity sensed at a superficial level. The enhanced membrane permeability coincided with an increase in lipid fluidity and protein mobility and was associated with significant oxidative injury to membrane lipids. In conclusion, apoptosis of nerve cells induced by bilirubin is mediated by its primary effect at physically perturbing the cell membrane. Bilirubin directly interacts with membranes influencing lipid polarity and fluidity, protein order, and redox status. These data suggest that nerve cell membranes are primary targets of bilirubin toxicity.

n/a

Resonance Raman (RR) spectroscopy is used to examine porphyrin substrate, product, and inhibitor interactions with the active site of murine ferrochelatase (EC 4.99.1.1), the terminal enzyme in the biosynthesis of heme. The enzyme catalyzes in vivo Fe2+ chelation into protoporphyrin IX to give heme. The RR spectra of native ferrochelatase show that the protein, as isolated, contains varying amounts of endogenously bound high- or low-spin ferric heme, always at much less than 1 equiv. RR data on the binding of free-base protoporphyrin IX and its metalated complexes (Fe(III), Fe(II), and Ni(II)) to active wild-type protein were obtained at varying ratios of porphyrin to protein. The binding of ferric heme, a known inhibitor of the enzyme, leads to the formation of a low-spin six-coordinate adduct. Ferrous heme, the enzyme's natural product, binds in the ferrous high-spin five-coordinate state. Ni(II) protoporphyrin, a metalloporphyrin that has a low tendency toward axial ligation, becomes distorted when bound to ferrochelatase. Similarly for free-base protoporphyrin, the natural substrate of ferrochelatase, the RR spectra of porphyrin-protein complexes reveal a saddling distortion of the porphyrin. These results corroborate and extend our previous findings that porphyrin distortion, a crucial step of the catalytic mechanism, occurs even in the absence of bound metal substrate. Moreover, RR data reveal the presence of an amino acid residue in the active site of ferrochelatase which is capable of specific axial ligation to metals.

Crystallographic studies of the hydrogenases (Hases) from Desulfovibrio gigas (Dg) and Desulfovibrio vulgaris Miyazaki (DvM) have revealed heterodinuclear nickel-iron active centers in both enzymes. The structures, which represent the as-isolated (unready) Ni-A (S = (1)/(2)) enzyme state, disclose a nonprotein ligand (labeled as X) bridging the two metals. The bridging atom was suggested to be an oxygenic (O(2)(-) or OH(-)) species in Dg Hase and an inorganic sulfide in DvM Hase. To determine the nature and chemical characteristics of the Ni-X-Fe bridging ligand in Dg Hase, we have performed 35 GHz CW (17)O ENDOR measurements on the Ni-A form of the enzyme, exchanged into H(2)(17)O, on the active Ni-C (S = (1)/(2)) form prepared by H(2)-reduction of Ni-A in H(2)(17)O, and also on Ni-A formed by reoxidation of Ni-C in H(2)(17)O. In the native state of the protein (Ni-A), the bridging ligand does not exchange with the H(2)(17)O solvent. However, after a reduction/reoxidation cycle (Ni-A --> Ni-C --> Ni-A), an (17)O label is introduced at the active site, as seen by ENDOR. Detailed analysis of a 2-D field-frequency plot of ENDOR spectra taken across the EPR envelope of Ni-A((17)O) shows that the incorporated (17)O has a roughly axial hyperfine tensor, A((17)O) approximately [5, 7, 20] MHz, discloses its orientation relative to the g tensor, and also yields an estimate of the quadrupole tensor. The substantial isotropic component (a(iso)((17)O) approximately 11 MHz) of the hyperfine interaction indicates that a solvent-derived (17)O is indeed a ligand to Ni and thus that the bridging ligand X in the Ni-A state of Dg Hase is indeed an oxygenic (O(2)(-) or OH(-)) species; comparison with earlier EPR results by others indicates that the same holds for Ni-B. The small (57)Fe hyperfine coupling seen previously for Ni-A (A((57)Fe) approximately 0.9 MHz) is now shown to persist in Ni-C, A((57)Fe) approximately 0.8 MHz. However, the (17)O signal is lost upon reductive activation to the Ni-C state; reoxidation to Ni-A leads to the reappearance of the signal. Consideration of the electronic structure of the EPR-active states of the dinuclear center leads us to suggest that the oxygenic bridge in Ni-A(B) is lost in Ni-C and is re-formed from solvent upon reoxidation to Ni-A. This implies that the reductive activation to Ni-C opens Ni/Fe coordination sites which may play a central role in the enzyme's activity.

Spectroscopy coupled with density functional calculations has been used to define the spin state, oxidation states, spin distribution, and ground state wave function of the mu4-sulfide bridged tetranuclear CuZ cluster of nitrous oxide reductase. Initial insight into the electronic contribution to N2O reduction is developed, which involves a sigma superexchange pathway through the bridging sulfide.

n/a

Recent studies continue to indicate the existence of a narrow relationship between flexible work organizations and the economic growth, in particular, Sweden and Germany. The measure of this relationship is many times the result of the value added per worker. Therefore, the causes of economic growth must be perceived from the interior of the company (work organization, technology, infrastructures, product design). On the other hand, the capacity of innovation can be perceived by the market through new products and services, and still significant changes with the introduction of new equipment and design of an efficient work organization. In this article it is analyzed the evolution of the productivity and employment levels in Portugal and other European countries, over all, during the decade of 90. An analysis for sector is made still. One verifies often that economic growth can be without employment growth. However, the growth can be obtained when if it reaches bigger added value and the efficiency can be verified when there are diminished costs for a same period of time, although that the labor productivity is also the pointer of the quality of life in a economy, therefore it represents the value produced by the work. Moreover, the total factors productivity is the measure of the technological and organizational progress (don’t includes only the technological investments). It is tried to get and to analyze the available statistics on these dimension in Portugal, concluding that Portugal presents an enterprise structure predominantly based on small and very small companies, a great number of which is market by a weak potential of adaptability, innovation and sustainability. It becomes urgent to take the non-material factors that integrate the productivity dimension, as factors of intervention for a benchmarking that allows a process of sustainable growth.

This article aims to study hydrogen production/consumption in Desulfovibrio (D.) desulfuricans strain New Jersey, a sulfate reducer isolated from a medium undergoing active biocorrosion and to compare its hydrogen metabolism with two other Desulfovibrio species, D. gigas and D. vulgaris Hildenborough. Hydrogen production was followed during the growth of these three bacterial species under different growth conditions: no limitation of sulfate and lactate, sulfate limitation, lactate limitation, pyruvate/sulfate medium and in the presence of molybdate. Hydrogen production/consumption by D. desulfuricans shows a behavior similar to that of D. gigas but a different one from that of D. vulgaris, which produces higher quantities of hydrogen on lactate/sulfate medium. The three species are able to increase the hydrogen production when the sulfate became limiting. Moreover, in a pyruvate/sulfate medium hydrogen production was lower than on lactate/sulfate medium. Hydrogen production by D. desulfuricans in presence of molybdate is extremely high. Hydrogenases are key enzymes on production/consumption of hydrogen in sulfate reducing organisms. The specific activity, number and cellular localization of hydrogenases vary within the three Desulfovibrio species used in this work, which could explain the differences observed on hydrogen utilization.

The Portuguese National Action Plan for Employment (NAP) is the main instrument for the labour market policy. It was adopted in 1998. The NAP transposes to the Portuguese reality the contents of such guidelines, with the adequate adjustments required by the national specificities, establishing objectives, quantified targets and action deadlines, as well as defining new programmes and measures. Therefore, the objectives of the labour market policy were focusing on the promotion of an adequate transition of the youngsters in active life, on the promotion of social and professional insertion and fight against long term unemployment and exclusion, on the improvement of basic and professional qualification of the working population in a perspective of lifelong training, namely as a way to prevent unemployment phenomena, and on the preventive management and follow-up of sectoral restructuring processes.

Recent studies continue to indicate the existence of a narrow relationship between flexible work organizations and the economic growth, in particular, Sweden and Germany. The measure of this relationship is many times the result of the value added per worker. Therefore, the causes of economic growth must be perceived from the interior of the company (work organization, technology, infrastructures, product design). On the other hand, the capacity of innovation can be perceived by the market through new products and services, and still significant changes with the introduction of new equipment and design of an efficient work organization. In this article it is analyzed the evolution of the productivity and employment levels in Portugal and other European countries, over all, during the decade of 90. An analysis for sector is made still. One verifies often that economic growth can be without employment growth. However, the growth can be obtained when if it reaches bigger added value and the efficiency can be verified when there are diminished costs for a same period of time, although that the labor productivity is also the pointer of the quality of life in a economy, therefore it represents the value produced by the work. Moreover, the total factors productivity is the measure of the technological and organizational progress (don’t includes only the technological investments). It is tried to get and to analyze the available statistics on these dimension in Portugal, concluding that Portugal presents an enterprise structure predominantly based on small and very small companies, a great number of which is market by a weak potential of adaptability, innovation and sustainability. It becomes urgent to take the non-material factors that integrate the productivity dimension, as factors of intervention for a benchmarking that allows a process of sustainable growth.

The Portuguese National Action Plan for Employment (NAP) is the main instrument for the labour market policy. It was adopted in 1998. The NAP transposes to the Portuguese reality the contents of such guidelines, with the adequate adjustments required by the national specificities, establishing objectives, quantified targets and action deadlines, as well as defining new programmes and measures. Therefore, the objectives of the labour market policy were focusing on the promotion of an adequate transition of the youngsters in active life, on the promotion of social and professional insertion and fight against long term unemployment and exclusion, on the improvement of basic and professional qualification of the working population in a perspective of lifelong training, namely as a way to prevent unemployment phenomena, and on the preventive management and follow-up of sectoral restructuring processes.

A propose for this article is to present information on the emergence of the knowledge field of sociology of work and labor studies in Portugal. The period of 1974-1984 produced an interesting bibliography in the fields of social stratification, labor conflicts and social history. However, and since the mid-1980s, significant changes have emerged. There took place an increased theoretical diversity and also there was a major preoccupation with the development of a more professional teaching of the discipline. At this time, the first graduated specialists in Sociology appeared in the labor market since 1980, working in municipalities, industrial firms, public health institutions, schools, or even in research and development (R&D) units. The Portuguese association in this field (APSIOT) organized many scientific meetings, debates with unionists, managers and politicians, beside the regular publication of review “Organiza{\c c}ões e Trabalho” (Organization and Work) since 1989. It started the diffusion of a professional culture of sociologists who associate science and their occupation concerning themselves not only with scientific quality, but also with technical competence and social responsibility. At the same time, one could assist to the participation of sociologists in the social and organizational molding of new technologies in order to promote alternative production systems.

A propose for this article is to present information on the emergence of the knowledge field of sociology of work and labor studies in Portugal. The period of 1974-1984 produced an interesting bibliography in the fields of social stratification, labor conflicts and social history. However, and since the mid-1980s, significant changes have emerged. There took place an increased theoretical diversity and also there was a major preoccupation with the development of a more professional teaching of the discipline. At this time, the first graduated specialists in Sociology appeared in the labor market since 1980, working in municipalities, industrial firms, public health institutions, schools, or even in research and development (R&D) units. The Portuguese association in this field (APSIOT) organized many scientific meetings, debates with unionists, managers and politicians, beside the regular publication of review “Organizações e Trabalho” (Organization and Work) since 1989. It started the diffusion of a professional culture of sociologists who associate science and their occupation concerning themselves not only with scientific quality, but also with technical competence and social responsibility. At the same time, one could assist to the participation of sociologists in the social and organizational molding of new technologies in order to promote alternative production systems.

n/a

Molecular photoreactors consisting of polyamine chains (receptors) bearing terminal naphthalene units (antennae) are described. The receptors are used to bind the substrate hexacyanocobaltate(III) and the antennae to transfer energy to the complex and thus promote a photoaquation reaction.

Intramolecular electron (eT) and energy transfer (ET) have shown to occur in a covalently linked donor-acceptor (CLDA) system consisting of a naphthalene donor covalently linked through a polyamine chain connector to an anthracene acceptor; the connector has been chosen in order to switch ON or OFF the energy flux as a function of its protonation state as well as by co-ordination to Zn2+. The largest energy transfer efficiency (eta = 0.61) occurs for the fully protonated form (pH < 2), while at pH > 9 (eT) from the lone pairs of the nitrogens to the excited fluorophore takes place, leading to complete quenching of the emission. On the other hand at neutral and basic pH values. coordination of Zn2+ prevents the eT quenching allowing the ET process to occur. (C) 2002 Elsevier Science B.V. All rights reserved.

The thermoacidophilic archaeon Acidianus ambivalens contains a monomeric 47 kDa type-II NADH dehydrogenase (NDH), which contains a covalently bound flavin. In this work, by a combination of several methods, namely 31P-nuclear magnetic resonance and fluorescence spectroscopies, it is proven that this enzyme contains covalent FMN, a novelty among this family of enzymes, which were so far thought to mainly have the flavin dinucleotide form. Discrimination between several possible covalent flavin linkages was achieved by spectral and fluorescence experiments, which identified an 8α-N(1)-histidylflavin-type of linkage. Analysis of the gene-deduced amino acid sequence of type-II NDH showed no transmembranar helices and allowed the definition of putative dinucleotide and quinone binding motifs. Further, it is suggested that membrane anchoring can be achieved via amphipatic helices.

n/a

The unambiguous assignment of the nuclear magnetic resonance (NMR) signals of the α-substituents of the haems in the tetrahaem cytochrome isolated from Shewanella frigidimarina NCIMB400, was made using a combination of homonuclear and heteronuclear experiments. The paramagnetic 13C shifts of the nuclei directly bound to the porphyrin of each haem group were analysed in the framework of a model for the haem electronic structure. The analysis yields g-tensors for each haem, which allowed the assignment of some electron paramagnetic resonance (EPR) signals to specific haems, and the orientation of the magnetic axes relative to each haem to be established. The orientation of the axial ligands of the haems was determined semi-empirically from the NMR data, and the structural results were compared with those of the homologous tetrahaem cytochrome from Shewanella oneidensis MR-1 showing significant similarities between the two proteins.