This is the second interim report of the research project "Information Society, Work and the Generation of New Forms of Social Exclusion" (SOWING). It is based on a firm survey conducted in the eight regions participating in the research project — Flanders (Belgium), Lazio (Italy), Niederösterreich (Austria), Portugal, the Republic of Ireland, the Stuttgart area (Germany), the Tampere region (Finland) and the West London area (U.K.). The aim of this report is to present a broad overview of the collected data. In general, only simple statistical methods have been applied. The report focuses on a regional comparison; however, the data have also been analysed by firm size, measured by quantity of staff, and industrial sector. It should be seen as a first step in the data analysis; it may also give some hints for a more strategic analysis of the survey data.

This is the second interim report of the research project "Information Society, Work and the Generation of New Forms of Social Exclusion" (SOWING). It is based on a firm survey conducted in the eight regions participating in the research project — Flanders (Belgium), Lazio (Italy), Niederösterreich (Austria), Portugal, the Republic of Ireland, the Stuttgart area (Germany), the Tampere region (Finland) and the West London area (U.K.). The aim of this report is to present a broad overview of the collected data. In general, only simple statistical methods have been applied. The report focuses on a regional comparison; however, the data have also been analysed by firm size, measured by quantity of staff, and industrial sector. It should be seen as a first step in the data analysis; it may also give some hints for a more strategic analysis of the survey data.

Resonance Raman (RR) spectroscopy, molecular mechanics (MM) calculations, and normal-coordinate structural decomposition (NSD) have been used to investigate the conformational differences in the hemes in ferricytochromes c3. NSD analyses of heme structures obtained from X-ray crystallography and MM calculations of heme-peptide fragments of the cytochromes c3 indicate that the nonplanarity of the hemes is largely controlled by a fingerprint peptide segment consisting of two heme-linked cysteines, the amino acids between the cysteines, and the proximal histidine ligand. Additional interactions between the heme and the distal histidine ligand and between the heme propionates and the protein also influence the heme conformation, but to a lesser extent than the fingerprint peptide segment. In addition, factors that influence the folding pattern of the fingerprint peptide segment may have an effect on the heme conformation. Large heme structural differences between the baculatum cytochromes c3 and the other proteins are uncovered by the NSD procedure [Jentzen, W., Ma, J.-G., and Shelnutt, J. A. (1998) Biophys. J. 74, 753-763]. These heme differences are mainly associated with the deletion of two residues in the covalently linked segment of hemes 4 for the baculatum proteins. Furthermore, some of these structural differences are reflected in the RR spectra. For example, the frequencies of the structure-sensitive lines (nu4, nu3, and nu2) in the high-frequency region of the RR spectra are lower for the Desulfomicrobium baculatum cytochromes c3 (Norway 4 and 9974) than for the Desulfovibrio (D.) gigas, D. vulgaris, and D. desulfuricans strains, consistent with a more ruffled heme. Spectral decompositions of the nu3 and nu10 lines allow the assignment of the sublines to individual hemes and show that ruffling, not saddling, is the dominant factor influencing the frequencies of the structure-sensitive Raman lines. The distinctive spectra of the baculatum strains investigated are a consequence of hemes 2 and 4 being more ruffled than is typical of the other proteins.

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As a Norwegian sociologist pointed out recently at the Encontro Internacional de Vilamoura on Fishing, “the fisheries management is the management of people, not fish" This statement may surprise many specialists, but it puts once again a series of questions and problems in their true place: society, social relationships, individuals. It is necessary to adopt a new attitude, a new type of intervention, a new vision, which may mean “community management”, a system of co-management, new models of business organisation and consumer behaviour. Towards this end, sociology can and should contribute with its analytical instruments, with its set of scientific reflections and controversies, to the enrichment of the knowledge about a complex reality in profound change, such as that of the socio-economic fisheries system.

As a Norwegian sociologist pointed out recently at the Encontro Internacional de Vilamoura on Fishing, “the fisheries management is the management of people, not fish" This statement may surprise many specialists, but it puts once again a series of questions and problems in their true place: society, social relationships, individuals. It is necessary to adopt a new attitude, a new type of intervention, a new vision, which may mean “community management”, a system of co-management, new models of business organisation and consumer behaviour. Towards this end, sociology can and should contribute with its analytical instruments, with its set of scientific reflections and controversies, to the enrichment of the knowledge about a complex reality in profound change, such as that of the socio-economic fisheries system.

The equilibria in the systems VO2+ + L (L = Gly-L-Asp, L-Asp-Gly, N-acetyl-L-aspartic acid or succinic acid) have been studied at 25 degrees C and 0.2 mol dm(3) K(CI) medium by a combination of potentiometric and spectroscopic methods (ESR, circular dichroism and visible absorption). Formation constants were calculated from pH-metric data with total oxovanadium(Iv) concentrations of(0.6-4) x 10(-3) mol dm(-3) and ligand-to-metal (L:M) ratios of 2-8 (AspGly) or 4-15: 1 (other systems). The position of the Asp residue in the peptide chain affects the co-ordination mode of the ligands: while in the GlyAsp system bis complexes start to form at pH less than 2, for AspGly only 1 : 1 complexes form, with relatively high CD signal. The co-ordination behaviour of N-acetyl-L-aspartic and succinic acids is similar. The results of potentiometric and spectroscopic methods are self consistent. Isomeric structures are discussed for each stoichiometry proposed and the results compared with those for L-aspartic acid and dipeptides with non-coordinating side chains.

Adenosine triphosphate sulfurylase catalyzes the formation of adenosine 5'-phosphosulfate from adenosine triphosphate and sulfate. The enzyme plays a crucial role in sulfate activation, the key step for sulfate utilization, and has been purified from crude extracts of Desulfovibrio desulfuricans ATCC 27774 and Desulfovibrio gigas. Both proteins are homotrimers [141 kDa (3 x 47) for D. desulfuricans and 147 kDa (3 x 49) for D. gigas] and have been identified, for the first time, as metalloproteins containing cobalt and zinc. EXAFS reveals that either cobalt or zinc binds endogenously at presumably equivalent metal binding sites and is tetrahedrally coordinated to one nitrogen and three sulfur atoms. Furthermore, the electronic absorption spectra display charge-transfer bands at 335 and 370 nm consistent with sulfur coordination to cobalt, and as expected for a distorted tetrahedral cobalt geometry, d-d bands are observed at 625, 666, and 715 nm. This geometry is supported by the observation of high-spin Co2+ EPR signals at g approximately 6.5.

Mossbauer and EPR spectroscopies were used to characterize the Fe clusters in an Fe-S protein isolated from Desulfovibrio desulfuricans (ATCC 27774). This protein was previously thought to contain hexanuclear Fe clusters, but a recent X-ray crystallographic measurement on a similar protein isolated from Desulfovibrio vulgaris showed that the protein contains two tetranuclear clusters, a cubane-type [4Fe-4S] cluster and a mixed-ligand cluster of novel structure [Lindley et al. (1997) Abstract, Chemistry of Metals in Biological Systems, European Research Conference, Tomar, Portugal]. Three protein samples poised at different redox potentials (as-purified, 40 and 320 mV) were investigated. In all three samples, the [4Fe-4S] cluster was found to be present in the diamagnetic 2+ oxidation state and exhibited typical Mossbauer spectra. The novel-structure cluster was found to be redox active. In the 320-mV and as-purified samples, the cluster is at a redox equilibrium between its fully oxidized and one-electron reduced states. In the 40-mV sample, the cluster is in a two-electron reduced state. Distinct spectral components associated with the four Fe sites of cluster 2 in the three oxidation states were identified. The spectroscopic parameters obtained for the Fe sites reflect different ligand environments, making it possible to assign the spectral components to individual Fe sites. In the fully oxidized state, all four iron ions are high-spin ferric and antiferromagnetically coupled to form a diamagnetic S = 0 state. In the one-electron and two-electron reduced states, the reducing electrons were found to localize, consecutively, onto two Fe sites that are rich in oxygen/nitrogen ligands. Based on the X-ray structure and the Mossbauer parameters, attempts could be made to identify the reduced Fe sites. For the two-electron reduced cluster, EPR and Mossbauer data indicate that the cluster is paramagnetic with a nonzero interger spin. For the one-electron reduced cluster, the data suggest a half-integer spin of 9/2. Characteristic fine and hyperfine parameters for all four Fe sites were obtained. Structural implications and the nature of the spin-coupling interactions are discussed.

In humans, the lymphomyeloid system has a fundamental role on iron metabolism promoting its recycling due to a continuous removal of effete red blood cells. Additionally, one of the most intriguing aspects of metalloporphyrins in biology is their effect on the immune system. However, the process of erythrocyte catabolism is still poorly understood and needs further research. In the present study, we attempt to investigate the nature and the possible physiologic role of Fe compounds released after erythrophagocytosis during the removal of red blood cells. Monocyte erythrophagocytosis in vitro experiments were done to characterize chemically the Fe compounds present inside the cells and in the culture supernatants. We tested the probable immunomodulatory functions of erythrophagocytosis products over lymphocyte cultures activated in vitro with T mitogens (alpha-CD3). Data obtained from atomic absorption spectroscopy confirmed the presence of Fe in the culture supernatants of monocyte cultures after erythrophagocytosis. Also, high-spin haem complexes derived from erythrocyte catabolism were detected by electron paramagnetic electronic resonance. Finally, in vitro activated lymphocyte proliferation experiments indicate the co-mitogenic properties of monocyte culture supernatants after red blood cells phagocytosis. Thus, the results of the present work provide evidence that culture monocyte supernatants after in vitro erythrophagocytosis contain Fe (III) high-spin haem complexes and show lymphocyte proliferation co-stimulatory properties.

We present a high-resolution bacterial contig map of 3.4 Mb of genomic DNA in human chromosome 21q11-q21, encompassing the region of elevated disomic homozygosity in Down Syndrome-associated abnormal myelopoiesis and leukemia, as well as the markers, which has shown a strong association with Alzheimer's Disease that has never been explained. The map contains 89 overlapping PACs, BACs, or cosmids in three contigs (850, 850, and 1500 kb) with two gaps (one of 140-210 kb and the second < 5 kb). To date, eight transcribed sequences derived by cDNA selection, exon trapping, and/or global EST sequencing have been positioned onto the map, and the only two genes so far mapped to this cytogenetic region, STCH and RIP140 have been precisely localized. This work converts a further 10% of chromosome 21q into a high-resolution bacterial contig map, which will be the physical basis for the long-range sequencing of this region. The map will also enable positional derivation of new transcribed sequences, as well as new polymorphic probes, that will help in elucidation of the role the genes in this region may play in abnormal myelopoiesis and leukemia associated with trisomy 21 and Alzheimer's Disease.

The autosomal dominant disorder Rieger syndrome (RIEG) shows genetic heterogeneity and has a phenotype characterized by malformations of the anterior segment of the eye, failure of the periumbilical skin to involute, and dental hypoplasia. The main locus for RIEG was mapped to the 4q25-q27 chromosomal segment using a series of cytogenetic abnormalities as well as by genetic linkage to DNA markers. Recently, a bicoid-related homeobox transcription factor gene called RIEG has been cloned, characterized, and proven to cause the 4q25 linked RIEG. Its mode of action in the pathogenesis of RIEG was not conclusively proven, since most etiological mutations detected. In the RIEG sequence caused amino acid substitutions or splice changes in the homeodomain. Through FISH analysis of a 460-kb sequence-ready map (PAC contig) around RIEG that we report in this paper, we demonstrate that the 4q25 linked RIEG disorder can arise from the haploid, whole-gene deletion of RIEG, but also from a translocation break 90 kb upstream from the gene. The data provide conclusive evidence that physical or functional haploinsufficiency of RIEG is the pathogenic mechanism for Rieger syndrome. The map also defines restriction fragments bearing sequences with a potential key regulatory role in the control of homeobox gene expression.

The implications of the dimeric state of cytochrome c550 for its binding to Paracoccus cytochrome c peroxidase and its delivery of the two electrons required to restore the active enzyme during catalysis have been investigated. The amino acid sequence of cytochrome c550 of Paracoccus denitrificans strain LMD 52.44 was determined and showed 21 differences from that of strain LMD 22.21. Based on the X-ray structure of the latter, a structure for the cytochrome c550 monomer from strain 52.44 is proposed and a dipole moment of 945 debye was calculated with an orientation close to the exposed haem edge. The behaviour of the cytochrome on molecular-exclusion chromatography is indicative of an ionic strength-dependent monomer (15 kDa)/dimer (30 kDa) equilibrium that can also be detected by 1H-NMR spectroscopy. The apparent mass of 50 kDa observed at very low ionic strength was consistent with the presence of a strongly asymmetric dimer. This was confirmed by cross-linking studies, which showed that a cross-linked species of mass 30 kDa on SDS behaved with an apparent mass of 50 kDa on molecular-exclusion chromatography. A programme which carried out and evaluated molecular docking of two monomers to give a dimer generated a most probable dimer in which the monomer dipoles lay almost antiparallel to each other. The resultant dipole moment of the dimer is therefore small. Although this finding calls into question the possibility of preorientation of a strongly asymmetrically charged cytochrome as it collides with a redox partner, the stoichiometry of complex formation with cytochrome c peroxidase as studied by 1H-NMR spectroscopy shows that it is the monomer that binds.

The di-haem cytochrome c peroxidase of Paracoccus denitrificans is a calcium binding dimer of 37.5 kDa subunits. It is responsible for reduction of H(2)O(2) to H(2)O with oxidation of cytochrome c(550) and is isolated in a fully oxidised state (inactive) in which one haem (centre I) is in a high-spin/low-spin equilibrium and high potential and the other (centre II) is low-spin and low potential. The enzyme undergoes direct electron transfer (without the need for mediators) with a 4,4'-dithiodipyridine-modified gold electrode and the response of both haem groups can be observed. By combination of the cyclic and pulse voltammetric data with the established spectroscopic information, it was demonstrated that entry of one electron to the high potential haem leads (in a mechanism involving strong haem-haem interactions) to a complex change of spin states and redox potentials of both haems in order to attain a "ready state" for binding, reduction and cleavage of the hydrogen peroxide. In the absence of endogenous calcium, haem communication can be completely disconnected and is recovered only when Ca(2+) is added, an essential step for the formation of the peroxidatic site. The intricate electrochemical behaviour of this enzyme was interpreted as a mechanism involving, both reduction and oxidation of the high potential haem, an interfacial electron transfer coupled to a homogenous chemical reaction (EC mechanism). We discuss two different models for the sequence of events leading to the appearance of the active pentacoordinated peroxidatic haem.

Desulforedoxin and the N-terminus of desulfoferrodoxin share a 36 amino acid domain containing a (Cys-S)(4) metal binding site. Recombinant forms of desulforedoxin, an N-terminal fragment of desulfoferrodoxin, and two desulforedoxin mutant proteins were reconstituted with Fe3+ Cd2+, and Zn2+ and relative metal ion affinities assessed by proton titrations. Protons compete with metal for protein ligands, a process that can be followed by monitoring the optical spectrum of the metal-protein complex as a function of pH. For all polypeptides, Fe3+ bound with the highest affinity, whereas the affinity of Zn2+ was greater than Cd2+ in desulforedoxin and the N-terminal fragment of desulfoferrodoxin, but this order was reversed in desulforedoxin mutant proteins. Metal binding in both mutants was significantly impaired. Furthermore, the Fe3+ complex of both mutants underwent a time-dependent bleaching process which coincided with increased reactivity of cysteine residues to Ellman's reagent and concomitant metal dissociation. It is hypothesized that this results from an autoredox reaction in which Fe3+ is reduced to Fe2+ with attendant oxidation of ligand thiols.

A novel sulphate-reducing bacterium (Ind 1) was isolated from a biofilm removed from a severely corroded carbon steel structure in a marine environment. Light microscopy observations revealed that cells were Gram-negative, rod shaped and very motile. Partial 16S rRNA gene sequencing and analysis of the fatty acid profile demonstrated a strong similarity between the new species and members from the Desulfovibrio genus. This was confirmed by the results obtained following purification and characterisation of the key proteins involved in the sulphate-reduction pathway. Several metal-containing proteins, such as two periplasmic proteins: hydrogenase and cytochrome c3, and two cytoplasmic proteins: ferredoxin and sulphite reductase, were isolated and purified. The latter proved to be of the desulfoviridin type which is typical of the Desulfovibrio genus. The study of the remaining proteins revealed a high degree of similarity with the homologous proteins isolated from Desulfovibrio gigas. However, the position of the strain within the phylogenetic tree clearly indicates that the bacterium is closely related to Desulfovibrio gabonensis, and these three strains form a separate cluster in the delta subdivision of the Proteobacteria. On the basis of the results obtained, it is suggested that Ind 1 belongs to a new species of the genus Desulfovibrio, and the name Desulfovibrio indonensis is proposed.

Desulforedoxin (Dx) is a simple homodimeric protein isolated from Desulfovibrio gigas (Dg) containing a distorted rubredoxin-like center with one iron coordinated by four cysteinyl residues (7.9 kDa with 36 amino acids per monomer). In order to probe the geometry and the H-bonding at the active site of Dx, the protein was reconstituted with 113Cd and the solution structure determined using 2D NMR methods. The structure of this derivative was initially compared with the NMR solution structure of the Zn form (Goodfellow BJ et al., 1996, J Biol Inorg Chem 1:341-353). Backbone amide protons for G4, D5, G13, L11 NH, and the Q14 NH side-chain protons, H-bonded in the X-ray structure, were readily exchanged with solvent. Chemical shift differences observed for amide protons near the metal center confirm the H-bonding pattern seen in the X-ray model (Archer M et al., 1995, J Mol Biol 251:690-702) and also suggest that H-bond lengths may vary between the Fe, Zn, and 113Cd forms. The H-bonding pattern was further probed using a heteronuclear spin echo difference (HSED) experiment; the results confirm the presence of NH-S H-bonds inferred from D2O exchange data and observed in the NMR family of structures. The presence of "H-bond mediated" coupling in Dx indicates that the NH-S H-bonds at the metal center have significant covalent character. The HSED experiment also identified an intermonomer "through space" coupling for one of the L26 methyl groups, indicating its proximity to the 113Cd center in the opposing monomer. This is the first example of an intermonomer "through space" coupling. Initial structure calculations produced subsets of NMR families with the S of C28 pointing away from or toward the L26 methyl: only the subset with the C28 sulfur pointing toward the L26 methyl could result in a "through space" coupling. The HSED result was therefore included in the structure calculations. Comparison of the Fe, Zn, and 113Cd forms of Dx suggests that the geometry of the metal center and the global fold of the protein does not vary to any great extent, although the H-bond network varies slightly when Cd is introduced. The similarity between the H-bonding pattern seen at the metal center in Dx, Rd (including H-bonded and through space-mediated coupling), and many zinc-finger proteins suggests that these H-bonds are structurally vital for stabilization of the metal centers in these proteins.

The new cylindrical molecule L containing two tetraazamacrocyclic rings linked by two azobenzene pillars displays photoelastic properties, Light absorption at 366 nm gives rise to trans --> cis isomerization of the azobenzene moieties producing two isomers containing one or two cis-azobenzenes, respectively, The three trans-trans (E-E), trans-cis (E-Z) and cis-cis (Z-Z) isomers have been identified and characterized by H-1 NMR spectroscopy, allowing the dependence of their formation percentages with irradiation time to be determined, The sequence of photochemical reactions E-E --> E-Z --> Z-Z allows almost complete conversion of the E-E into the Z-Z isomer at 366 nm and 298 K, Both thermal (k = 1.75 x 10(-5) s(-1) at 313 K) and photo-induced (at 436 and 313 nm) back-isomerization reactions have been studied, The protonation constants of the three isomers in equimolar solutions of water-DMSO indicate a decreasing basicity in the order E-E > E-Z > Z-Z, in agreement with increasing electrostatic repulsion between the positive charges caused by a reduction in the separation between the protonation sites occurring upon Z --> E isomerization.

The structural transformations and photochromic properties of the 7-hydroxyflavylium ion have been investigated by means of the pH jump technique and continuous and pulsed light excitation. The primary photoproduct of UV irradiation of the colorless trans-chalcone form, which is the predominant species at pH 4, is its colorless cis isomer, which rapidly disappears on a time scale of seconds through two competitive processes: i) back-reaction to yield the trans-chalcone form, and ii) formation of the colored flavylium ion and its conjugated quinoidal base. Over minutes or hours (depending on pH), the system reverts quantitatively to its original state. The rate constants and equilibrium constants of the various processes have been obtained and compared with those previously reported for the 4'-hydroxyflavylium and 4',7-dihydroxyflavylium ions. This comparison demonstrates the substituent effect on the rate and equilibrium constants; the effect on the rate constant of the cis-->trans thermal isomerization reaction is particularly strong. For the 7-hydroxyflavylium and 4',7-dihydroxyflavylium ions the pH of the solution plays the role of a tap for the color intensity generated by light excitation. This also means that this system can be viewed as a light-switchable pH indicator.

The acid-base properties as well as the photochemical reactivity of the co-ordination compound K-4[Co(CN)(5)(SO3)] in the presence of three polyammonium macrocyclic receptors were studied in aqueous solution. The pK(a) of the free complex (3.9) (sulfite deprotonation) changed to pK(a) <0.5 upon complexation with the receptors. The quantum yield for sulfite photoaquation of the free complex in the basic form (Phi = 0.85 +/- 0.09) decreased to 0.05 +/- 0.01, 0.12 +/- 0.03 and 0.45 +/- 0.09 in the presence of[24]aneN(8)H(8)(8+), [30]aneN(10)H(10)(10+) and [32]aneN(8)H(8)(8+), respectively. For the acidic form of the free complex (Phi = 0.40 +/- 0.05) the quantum yield was not affected by supercomplexation with [32]aneN(8)H(8)(8+). For the adducts formed from the other two macrocyclic receptors it was not possible to evaluate the quantum yields of the acidic forms, because protonation was not complete even at very high proton concentrations. The results were interpreted in terms of second-sphere interactions involving hydrogen bonding between the complex and the macrocycles. In the case of [32]aneN(8)H(8)(8+) the experimental results are compatible with a structure in which the cyanides are involved in hydrogen bonding but the sulfite ligand is not. In the two other supercomplexes the sulfite ligand seems to be involved in hydrogen bonding.

Reacting transition metal complexes in low oxidation states, containing one or two cyanide ligands, with methyltrioxorhenium(VII) leads to bridged mixed metal compounds in good yields. The Re(VII) core is then surrounded by five or six ligands, respectively. The strength of these CN bridges and thus the stability of the newly generated bimetallic compound strongly depends on the donor strength of the ligands surrounding of the Cr/Mo/W or Fe moiety. The stability of the mixed metal molecules is reflected in the temperature dependent behavior of their O-17-NMR spectra, in their IR (Re=O) stretching frequencies and force constants, as well as several other spectroscopic data. UV-vis absorption spectra show the appearance of charge transfer bands. In the case of the mixed Mo/Re complexes the Mo-95-NMR spectroscopy is also a helpful tool to examine the donor capability of the Mo moiety. The described compounds also show photosensitivity. (C) 1998 Elsevier Science S.A. All rights reserved.