Untersuchungen zu biologischen Schadstoffeffekten bei Nordseefischen: Langzeitdaten zum Auftreten von Lebertumoren bei der Kliesche (Limanda limanda)

Data are presented on the spatial distribution and long-term temporal trends in the occurrence of neoplastic liver lesions of North Sea dab (Limanda limanda) in the period 1988 to 2001, obtained in the framework of the regular fish disease monitoring programme carried out by the German Federal Research Centre for Fishery. Highest prevalences consistently recorded in the first part of the period occurred in the German Bight, at the Dogger Bank and at sampling sites off Humber and Wash. In contrast, stations in the northern North Sea (e.g. in the Firth of Forth area) were always characterised by low prevalences. Particularly during the first years of observation, a clear and general decrease in prevalence could be observed, that was most pronounced in the hot spot areas German Bight and Dogger Bank compared to the Firth of Forth. Current data reveal that, due to the decrease, spatial differences between sampling sites are now much less obvious than at the beginning of the studies. Limited chemical data available suggest that the temporal decrease in prevalence of liver tumours and their precursor stages generally correspond with the decrease in organic contaminants in dab livers as well as in water and sediments. However, the spatial variation in prevalence recorded can not entirely be explained by contaminant levels. For instance, the prevalence was continuously low in the Firth of Forth area wheras the contaminant levels in dab were comparatively high. The opposite feature was observed at the Dogger Bank. Future studies will therefore address the role of other host- and site-specific factors potentially involved in the aetiology of the disease.

Phosphorylation of the carboxy-terminal domain of histone H1: effects on secondary structure and DNA condensation

Linker histone H1 plays an important role in chromatin folding. Phosphorylation by cyclin-dependent kinases is the main post-translational modification of histone H1. We studied the effects of phosphorylation on the secondary structure of the DNA-bound H1 carboxy-terminal domain (CTD), which contains most of the phosphorylation sites of the molecule. The effects of phosphorylation on the secondary structure of the DNA-bound CTD were site-specific and depended on the number of phosphate groups. Full phosphorylation significantly increased the proportion of -structure and decreased that of -helix. Partial phosphorylation increased the amount of undefined structure and decreased that of -helix without a significant increase in -structure. Phosphorylation had a moderate effect on the affinity of the CTD for the DNA, which was proportional to the number of phosphate groups. Partial phosphorylation drastically reduced the aggregation of DNA fragments by the CTD, but full phosphorylation restored to a large extent the aggregation capacity of the unphosphorylated domain. These results support the involvement of H1 hyperphosphorylation in metaphase chromatin condensation and of H1 partial phosphorylation in interphase chromatin relaxation. More generally, our results suggest that the effects of phosphorylation are mediated by specific structural changes and are not simply a consequence of the net charge.

Nutzungskonflikte zwischen Windparks und Fischerei in der Nordsee – was die marine Raumordnung noch nicht berücksichtigt

The current approval procedure for wind farm proposals in the German EEZ only considers site specific conflict analysis between the wind farm and fisheries. Due to the relatively small spatial coverage of single sites potential opportunity losses to the fisheries are always considered as low or negligible. Cumulative effects on fisheries that will occur once all proposed wind farms are in place are not yet considered adequately. However, those cumulative effects will be quite substantial because, in particular, opportunities to catch such valuable species as flatfish will be considerably reduced.

Directed evolution of terpene synthases for non-natural substrates

Terpenes represent about half of known natural products, with terpene synthases catalyzing reactions to increase the complexity of substrates and generate cyclizations of the linear diphosphate substrates, therefore forming rings and stereocenters. With their diverse functionality, terpene synthases may be highly evolvable, with the ability to accept a wide range of non-natural compounds and with high product selectivity. Our hypothesis is that directed evolution of terpene synthases can be used to increase selectivity of the synthase on a specific substrate. In the first part of the work presented herein, three natural terpene synthases, Cop2, BcBOT2, and SSCG_02150, were tested for activity against the natural substrate and a non-natural substrate, called Surrogate 1, and the relative activities on both the natural and non-natural substrates were compared. In the second part of this work, a terpene synthase variant of BcBOT2 that has been evolved for thermostability, was used for directed evolution for increased activity and selectivity on the non-natural substrate referred to as Surrogate 2. Mutations for this evolution were introduced using random mutagenesis, with error prone polymerase chain reactions, and using site-specific saturation mutagenesis, in which an NNK library is designed with a specific active site amino acid targeted for mutation. The mutant enzymes were then screened and selected for enhancement of the desired functionality. Two neutral mutants, 19B7 W367F and 19B7 W118Q, were found to maintain activity on Surrogate 2, as measured by the screen.

Synthesis and spectroscopy of ruthenium-modified nucleic acids

Redox-active probes are designed and prepared for use in DNA-mediated electron transfer studies. These probes consist of ruthenium(II) complexes bound to nucleosides that possess metal-binding ligands. Low- and high-potential oxidants are synthesized from these modified nucleosides and display reversible one-electron electrochemical behavior. The ruthenium-modified nucleosides exhibit distinct charge-transfer transitions in the visible region that resemble those of appropriate model complexes. Resonance Raman and time-resolved emission spectroscopy are used to characterize the nature of these transitions.

The site-specific incorporation of these redox-active probes into oligonucleotides is explored using post-synthetic modification and solid-phase synthetic methods. The preparation of the metal-binding nucleosides, their incorporation into oligonucleotides, and characterization of the resulting oligonucleotides is described. Because the insertion of these probes into modified oligonucleotides using post-synthetic modification is unsuccessful, solid-phase synthetic methods are explored. These efforts lead to the first report of 3'-metallated oligonucleotides prepared completely by automated solid-phase synthesis. Preliminary efforts to prepare a bis-metallated oligonucleotide by automated synthesis are described.

The electrochemical, absorption, and emissive features of the ruthenium-modified oligonucleotides are unchanged from those of the precursor metallonucleoside. The absence of any change in these properties upon incorporation into oligonucleotides and subsequent hybridization suggests that the incorporated ruthenium(II) complex is a valuable probe for DNA-mediated electron transfer studies.

Discounting the value of natural resources in costbenefit analysis: a case study for policy making

4 p.

The application of metallointercalators in recognition of and charge transport in nucleic acids

Metal complexes that utilize the 9,10-phenanthrene quinone diimine (phi) moiety bind to DNA through the major groove. These metallointercalators can recognize DNA sites and perform reactions on DNA as a substrate. The site-specific metallointercalator Λ-1-Rh(MGP)_2phi^(5+) competitively disrupts the major groove binding of a transcription factor, yAP-1, from an oligonucleotide that contains a common binding site. The demonstration that metal complexes can prevent transcription factor binding to DNA site-specifically is an important step in using metallointercalators as therapeutics.

The distinctive photochemistry of metallointercalators can also be applied to promote long range charge transport in DNA. Experiments using duplexes with regions 4 to 10 nucleotides long containing strictly adenine and thymine sequences of varying order showed that radical migration is more dependent on the sequence of bases, and less dependent on the distance between the guanine doublets. This result suggests that mechanistic proposals of long range charge transport must involve all the bases.

RNA/DNA hybrids show charge migration to guanines from a remote site, thus demonstrating that nucleic acid stacking other than B-form can serve as a radical bridge. Double crossover DNA assemblies also provide a medium for charge transport at distances up to 100 Å from the site of radical introduction by a tethered metal complex. This radical migration was found to be robust to mismatches, and limited to individual, electronically distinct base stacks. In single DNA crossover assemblies, which have considerably greater flexibility, charge migration proceeds to both base stacks due to conformational isomers not present in the rigid and tightly annealed double crossovers.

Finally, a rapid, efficient, gel-based technique was developed to investigate thymine dimer repair. Two oligonucleotides, one radioactively labeled, are photoligated via the bases of a thymine-thymine interface; reversal of this ligation is easily visualized by gel electrophoresis. This assay was used to show that the repair of thymine dimers from a distance through DNA charge transport can be accomplished with different photooxidants.

Thus, nucleic acids that support long range charge transport have been shown to include A-track DNA, RNA/DNA hybrids, and single and double crossovers, and a method for thymine dimer repair detection using charge transport was developed. These observations underscore and extend the remarkable finding that DNA can serve a medium for charge transport via the heteroaromatic base stack.

A Detailed Analysis of Transcriptional Regulators Affecting the Saccharomyces cerevisiae Heat-shock Gene SSA1

The yeast Saccharomyces cerevisiae contains a family of hsp70 related genes. One member of this family, SSA1, encodes a 70kD heat-shock protein which in addition to its heat inducible expression has a significant basal level of expression. The first 500 bp upstream of the SSA1 start point of transcription was examined by DNAse I protection analysis. The results reveal the presence of at least 14 factor binding sites throughout the upstream promoter region. The function of these binding sites has been examined using a series of 5' promoter deletions fused to the recorder gene lacZ in a centromere-containing yeast shuttle vector. The following sites have been identified in the promoter and their activity in yeast determined individually with a centromere-based recorder plasmid containing a truncated CYC1 /lacZ fusion: a heat-shock element or HSE which is sufficient to convey heat-shock response on the recorder plasmid; a homology to the SV40 'core' sequence which can repress the GCN4 recognition element (GCRE) and the yAP1 recognition element (ARE), and has been designated a upstream repression element or URE; a 'G'-rich region named G-box which can also convey heatshock response on the recorder plasmid; and a purine-pyrimidine alternating sequence name GT-box which is an activator of transcription. A series of fusion constructs were made to identify a putative silencer-like element upstream of SSA1. This element is position dependent and has been localized to a region containing both an ABF1 binding site and a RAP1 binding site. Five site-specific DNA-binding factors are identified and their purification is presented: the heat-shock transcription factor or HSTF, which recognizes the HSE; the G-box binding factor or GBF; the URE recognition factor or URF; the GT-box binding factor; and the GC-box binding factor or yeast Sp1.

An introduction to RIVPACS

RIVPACS (River InVertebrate Prediction And Classification System) is a software package developed by the Institute of Freshwater Ecology (IFE). The primary application is to assess the biological quality of rivers within the UK. RIVPACS offers site-specific predictions of the macroinvertebrate fauna to be expected in the absence of major environmental stress. The expected fauna is derived by RIVPACS using a small suite of environmental characteristics. The biological evaluation is then obtained by comparing the fauna observed at the site with the expected fauna. RIVPACS also includes a site classification based on the macroinvertebrate fauna of the component reference sites. New sites, judged by their fauna to be of high biological quality, may be allocated to classification groups within the fixed RIVPACS classification. This has potential for evaluating sites for conservation. In this chapter, the origins and history of the RIVPACS approach are described, including major scientific and operational developments over the life of the project. RIVPACS III is described in detail and predictions at different taxonomic levels are demonstrated. The value of the reference dataset for river management and conservation is examined, and the chapter concludes with a brief consideration of some future challenges.

Del boceto al objeto a través del BIT en la obra "La materia del tiempo", de Richard Serra

423 p.

Curadoria, pedagogia e colaboração social

Na virada para o século XXI artistas e curadores em bienais e outras exposições internacionais estão adotando formas colaborativas e educacionais de fazer arte e curadoria envolvendo artistas e não artistas, comunidades, escolas e outras instâncias sociais. Essas iniciativas criam encontros interdisciplinares onde imbricações entre arte e não-arte são colocadas em questão, considerando contextos sociopolíticos mundiais, a globalização e outros temas pós-colonialistas. A dissertação examina as aproximações e atravessamentos de culturas e identidades, questões de centro e periferia, local e global gerando entre-lugares e o interesse por propostas de site-oriented, geradoras de situações de convivência, compartilhamento de experiências coletivas e prática educativa. A pesquisa reflete sobre as intenções dos artistas, curadores, a participação do público e o diálogo entre diferentes discursos críticos e campos do conhecimento em projetos com essas características. O estudo de caso é o projeto Cadernos de Viagem, projeto de residência artística e exposição, realizado na 8 Bienal do Mercosul, em Porto Alegre, Brasil, 2011. Diversos teóricos como Homi Bhabha, Jacques Rancière, Nicolas Bourriaud, Claire Bishop, Grant Kester, Miwon Kwon, Moacir dos Anjos e Félix Guattari, são abordados, entre outros

Ongoing rehabilitation of coastal communities in Aceh province : a new project

This article presents an overview of the project on Rehabilitation of Fisheries and Aquaculture in Tsunami-affected Coastal Communities in Aceh Province. Building on the research results from the recently completed projects detailed in the previous articles, this project shall synthesize information on coastal fishing communities and resources in order to develop site-specific management options to support rehabilitation of fisheries and aquaculture.

Ativação do estresse oxidativo e da resposta antioxidante por ExoU de Pseudomonas aeruginosa

ExoU, uma citotoxina produzida pelo patógeno oportunista Pseudomonas aeruginosa e translocada para o citosol de células hospedeiras via sistema de secreção do tipo III, é associada à gravidade de infecções agudas. No presente trabalho, o efeito de ExoU na ativação do estresse oxidativo e da resposta antioxidante foi avaliado em culturas de células epiteliais respiratórias humanas infectadas com a cepa PA103 de P. aeruginosa (produtora de ExoU), com a mutante deletada no gene exoU, PA103∆exoU, ou com a mutante complementada com exoU sem atividade tipo fosfolipase A2, PA103∆UT/S142A. Análises das dosagens de hidroperóxidos lipídicos e isoprostanos, considerados marcadores de estresse oxidativo, revelaram que ExoU promoveu um aumento em suas concentrações. Foi observado, também, que ExoU estimulou a produção de espécies reativas de oxigênio, óxido nítrico e peroxinitrito nas células infectadas, assim como a expressão de iNOS e eNOS, mas não de nNOS. Além disso, ExoU foi responsável pelo aumento da atividade de SOD1 e pela redução dos níveis de GSH, mas não afetou a atividade da catalase ou de NQO1. No modelo in vivo, a dosagem de malondialdeído, um subproduto da lipoperoxidação de membranas, evidenciou uma maior produção deste composto no pulmão de camundongos infectados pela cepa produtora de ExoU, em comparação ao pulmão de camundongos infectados pela cepa mutante. Em conjunto, estes resultados mostram que ExoU ativa a produção de espécies reativas de oxigênio e nitrogênio, levando à peroxidação lipídica e modulando o sistema de defesa antioxidante

Quase imperceptível: arte participativa e ficção

Esta dissertação levantou um conjunto de questões relacionadas à reflexão do meu processo artístico, tendo como ponto de partida as imagens poéticas da casa, do dormir, do desabrigo, da invisibilidade e da visibilidade que culminaram em trabalhos acerca dos moradores de rua, da acessibilidade e do uso da cidade. Para analisar as experiências artísticas participativas realizadas em âmbito urbano, foi apresentado um estudo da produção de artistas como Flávio de Carvalho, Lygia Clark e Hélio Oiticica, assim como os conceitos de site specific e site oriented (KWON 1998). Acerca dos trabalhos que envolvem relações e propostas de convivência, foi acessado o livro Estética Relacional (BOURRIAUD 2009) e o texto Antagonismos e Estética Relacional (BISHOP 2004). O ato de ouvir e as histórias narradas durante as propostas participativas aproximaram o estudo da ideia de autoficção (DOUBROVSKY 1977) e da escrita de si (FOULCAULT 2002). A relação com o desenho foi observada como produção de sentido, assim como foi problematizado o papel do artista a partir da observação do papel do mediador, revelando possibilidades outras de definição com base nos conceitos de participação total (OITICICA 1967), e artista-etc. (BASBAUM 2013).

A stochastic model of temporal variations in monthly temperature, precipitation, snowfall, and resulting snowpack

We report a Monte Carlo representation of the long-term inter-annual variability of monthly snowfall on a detailed (1 km) grid of points throughout the southwest. An extension of the local climate model of the southwestern United States (Stamm and Craig 1992) provides spatially based estimates of mean and variance of monthly temperature and precipitation. The mean is the expected value from a canonical regression using independent variables that represent controls on climate in this area, including orography. Variance is computed as the standard error of the prediction and provides site-specific measures of (1) natural sources of variation and (2) errors due to limitations of the data and poor distribution of climate stations. Simulation of monthly temperature and precipitation over a sequence of years is achieved by drawing from a bivariate normal distribution. The conditional expectation of precipitation. given temperature in each month, is the basis of a numerical integration of the normal probability distribution of log precipitation below a threshold temperature (3°C) to determine snowfall as a percent of total precipitation. Snowfall predictions are tested at stations for which long-term records are available. At Donner Memorial State Park (elevation 1811 meters) a 34-year simulation - matching the length of instrumental record - is within 15 percent of observed for mean annual snowfall. We also compute resulting snowpack using a variation of the model of Martinec et al. (1983). This allows additional tests by examining spatial patterns of predicted snowfall and snowpack and their hydrologic implications.