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Modern biological analysis techniques supply scientists with various forms of data. One category of such data are the so called "expression data". These data indicate the quantities of biochemical compounds present in tissue samples. Recently, expression data can be generated at a high speed. This leads in turn to amounts of data no longer analysable by classical statistical techniques. Systems biology is the new field that focuses on the modelling of this information. At present, various methods are used for this purpose. One superordinate class of these methods is machine learning. Methods of this kind had, until recently, predominantly been used for classification and prediction tasks. This neglected a powerful secondary benefit: the ability to induce interpretable models. Obtaining such models from data has become a key issue within Systems biology. Numerous approaches have been proposed and intensively discussed. This thesis focuses on the examination and exploitation of one basic technique: decision trees. The concept of comparing sets of decision trees is developed. This method offers the possibility of identifying significant thresholds in continuous or discrete valued attributes through their corresponding set of decision trees. Finding significant thresholds in attributes is a means of identifying states in living organisms. Knowing about states is an invaluable clue to the understanding of dynamic processes in organisms. Applied to metabolite concentration data, the proposed method was able to identify states which were not found with conventional techniques for threshold extraction. A second approach exploits the structure of sets of decision trees for the discovery of combinatorial dependencies between attributes. Previous work on this issue has focused either on expensive computational methods or the interpretation of single decision trees a very limited exploitation of the data. This has led to incomplete or unstable results. That is why a new method is developed that uses sets of decision trees to overcome these limitations. Both the introduced methods are available as software tools. They can be applied consecutively or separately. That way they make up a package of analytical tools that usefully supplement existing methods. By means of these tools, the newly introduced methods were able to confirm existing knowledge and to suggest interesting and new relationships between metabolites.
The Reversible Addition Fragmentation Chain Transfer (RAFT) process using the new RAFT agent benzyldithiophenyl acetate is shown to be a powerful polymerization tool to synthesize novel well-defined amphiphilic diblock copolymers composed of the constant hydrophobic block poly(butyl acrylate) and of 6 different hydrophilic blocks with various polarities, namely a series of non-ionic, non-ionic comb-like, anionic and cationic hydrophilic blocks. The controlled character of the polymerizations was supported by the linear increase of the molar masses with conversion, monomodal molar mass distributions with low polydispersities and high degrees of end-group functionalization. The new macro-surfactants form micelles in water, whose size and geometry strongly depend on their composition, according to dynamic and static light scattering measurements. The micellization is shown to be thermodynamically favored, due to the high incompatibility of the blocks as indicated by thermal analysis of the block copolymers in bulk. The thermodynamic state in solution is found to be in the strong or super strong segregation limit. Nevertheless, due to the low glass transition temperature of the core-forming block, unimer exchange occurs between the micelles. Despite the dynamic character of the polymeric micellar systems, the aggregation behavior is strongly dependent on the history of the sample, i.e., on the preparation conditions. The aqueous micelles exhibit high stability upon temperature cycles, except for an irreversibly precipitating block copolymer containing a hydrophilic block exhibiting a lower critical solution temperature (LCST). Their exceptional stability upon dilution indicates very low critical micelle concentrations (CMC) (below 4∙10<sup>-4 g∙L<sup>-1). All non-ionic copolymers with sufficiently long solvophobic blocks aggregated into direct micelles in DMSO, too. Additionally, a new low-toxic highly hydrophilic sulfoxide block enables the formation of inverse micelles in organic solvents. The high potential of the new polymeric surfactants for many applications is demonstrated, in comparison to reference surfactants. The diblock copolymers are weakly surface-active, as indicated by the graduate decrease of the surface tension of their aqueous solutions with increasing concentration. No CMC could be detected. Their surface properties at the air/water interface confer anti-foaming properties. The macro-surfactants synthesized are surface-active at the interface between two liquid phases, too, since they are able to stabilize emulsions. The polymeric micelles are shown to exhibit a high ability to solubilize hydrophobic substances in water.
Quantified Boolean formulas (QBFs) play an important role in theoretical computer science. QBF extends propositional logic in such a way that many advanced forms of reasoning can be easily formulated and evaluated. In this dissertation we present our ZQSAT, which is an algorithm for evaluating quantified Boolean formulas. ZQSAT is based on ZBDD: Zero-Suppressed Binary Decision Diagram , which is a variant of BDD, and an adopted version of the DPLL algorithm. It has been implemented in C using the CUDD: Colorado University Decision Diagram package. The capability of ZBDDs in storing sets of subsets efficiently enabled us to store the clauses of a QBF very compactly and let us to embed the notion of memoization to the DPLL algorithm. These points led us to implement the search algorithm in such a way that we could store and reuse the results of all previously solved subformulas with a little overheads. ZQSAT can solve some sets of standard QBF benchmark problems (known to be hard for DPLL based algorithms) faster than the best existing solvers. In addition to prenex-CNF, ZQSAT accepts prenex-NNF formulas. We show and prove how this capability can be exponentially beneficial.
This work explores the equilibrium structure and thermodynamic phase behavior of complexes formed by charged polymer chains (polyelectrolytes) and oppositely charged spheres (macroions). Polyelectrolyte-macroion complexes form a common pattern in soft-matter physics, chemistry and biology, and enter in numerous technological applications as well. From a fundamental point of view, such complexes are interesting in that they combine the subtle interplay between electrostatic interactions and elastic as well as entropic effects due to conformational changes of the polymer chain, giving rise to a wide range of structural properties. This forms the central theme of theoretical studies presented in this thesis, which concentrate on a number of different problems involving strongly coupled complexes, i.e. complexes that are characterized by a large adsorption energy and small chain fluctuations. In the first part, a global analysis of the structural phase behavior of a single polyelectrolyte-macroion complex is presented based on a dimensionless representation, yielding results that cover a wide range of realistic system parameters. Emphasize is made on the interplay between the effects due to the polyelectrolytes chain length, salt concentration and the macroion charge as well as the mechanical chain persistence length. The results are summarized into generic phase diagrams characterizing the wrapping-dewrapping behavior of a polyelectrolyte chain on a macroion. A fully wrapped chain state is typically obtained at intermediate salt concentrations and chain lengths, where the amount of polyelectrolyte charge adsorbed on the macroion typically exceeds the bare macroion charge leading thus to a highly overcharged complex. Perhaps the most striking features occur when a single long polyelectrolyte chain is complexed with many oppositely charged spheres. In biology, such complexes form between DNA (which carries the cell's genetic information) and small oppositely charged histone proteins serving as an efficient mechanism for packing a huge amount of DNA into the micron-size cell nucleus in eucaryotic cells. The resultant complex fiber, known as the chromatin fiber, appears with a diameter of 30~nm under physiological conditions. Recent experiments indicate a zig-zag spatial arrangement for individual DNA-histone complexes (nucleosome core particles) along the chromatin fiber. A numerical method is introduced in this thesis based on a simple generic chain-sphere cell model that enables one to investigate the mechanism of fiber formation on a systematic level by incorporating electrostatic and elastic contributions. As will be shown, stable complex fibers exhibit an impressive variety of structures including zig-zag, solenoidal and beads-on-a-string patterns, depending on system parameters such as salt concentration, sphere charge as well as the chain contour length (per sphere). The present results predict fibers of compact zig-zag structure within the physiologically relevant regime with a diameter of about 30~nm, when DNA-histone parameters are adopted. In the next part, a numerical method is developed in order to investigate the role of thermal fluctuations on the structure and thermodynamic phase behavior of polyelectrolyte-macroion complexes. This is based on a saddle-point approximation, which allows to describe the experimentally observed reaction (or complexation) equilibrium in a dilute solution of polyelectrolytes and macroions on a systematic level. This equilibrium is determined by the entropy loss a single polyelectrolyte chain suffers as it binds to an oppositely charged macroion. This latter quantity can be calculated from the spectrum of polyelectrolyte fluctuations around a macroion, which is determined by means of a normal-mode analysis. Thereby, a stability phase diagram is obtained, which exhibits qualitative agreement with experimental findings. At elevated complex concentrations, one needs to account for the inter-complex interactions as well. It will be shown that at small separations, complexes undergo structural changes in such a way that positive patches from one complex match up with negative patches on the other. Furthermore, one of the polyelectrolyte chains may bridge between the two complexes. These mechanisms lead to a strong inter-complex attraction. As a result, the second virial coefficient associated with the inter-complex interaction becomes negative at intermediate salt concentrations in qualitative agreement with recent experiments on solutions of nucleosome core particles.
Consumer attitudes towards genetically modified foods in Europe : structure and changeability
(2004)
Genetically modified foods have been at the center of debate in European consumer policy in the last two decades. Although the quasi-moratorium has been lifted in May 2004 and the road to the market is in principle reopened, strategies for product introduction are lacking. The aim of the research is to assess potential barriers in the area of consumer acceptance and suggest ways in which they can be overcome. After a short history of the genetically modified foods debate in Europe, the existing literature is reviewed. Although previous research converges in its central results, issues that are more fundamental have remained unresolved. Based on classical approaches in attitude research and modern theories of social cognition, a general model of the structure, function and dynamics of whole systems of attitudes is developed. The predictions of the model are empirically tested based on an attitude survey (N = 2000) and two attitude change experiments (N = 1400 and N = 750). All three studies were conducted in parallel in four EU member states. The results show that consumer attitudes towards genetically modified foods are embedded into a structured system of general socio-political attitudes. The system operates as a schema through which consumers form global evaluations of the technology. Specific risk and benefit judgments are mere epiphenomena of this process. Risk-benefit trade-offs, as often presupposed in the literature, do not appear to enter the process. The attitudes have a value-expressive function; their purpose is not just a temporary reduction of complexity. These properties render the system utterly resistant to communicative interventions. At the same time, it exerts stong anchoring effects on the processing of new information. Communication of benefit arguments can trigger boomerang effects and backfire on the credibility of the communicator when the arguments contrast with preexisting attitudes held by the consumer. Only direct sensory experience with high-quality products can partially bypass the system and lead to the formation of alternative attitude structures. Therefore, the recommended market introduction strategy for genetically modified foods is the simultaneous and coordinated launch of many high-quality products. Point of sale promotions should be the central instrument. Information campaigns, on the other hand, are not likely to have an effect on the product and technology acceptance of European consumers.
Arabidopsis thaliana trichomes exhibit strong birefringence under polarized light, a characteristic of cell walls containing large amounts of highly ordered cellulose microfibrils. The tbr1 mutant of Arabidopsis lacks trichome birefringence and is deficient in secondary cell wall cellulose synthesis (Potikha and Delmer, 1995). The TBR gene was identified by recombinational mapping, candidate gene sequencing and molecular complementation using genomic cosmid clones, as well as a p35S:TBR genomic DNA construct, fully rescuing the mutant phenotype in both cases. The only mutant allele available (tbr-1) carries a substitution (G to E) in a conserved aminoacid domain of the protein. TBR gene structure was proved to have a longer size than the one found to be annotated at the time of identification in the data-base. A full cDNA clone containing the full transcript was available and also complementation experiments using different gene fragments (annotated and suggested) leaded to the result that TBR gene is indeed, longer. TBR encodes a novel plant-specific protein with predicted plasma membrane localization, therefore being consistent with idea that is required for-, or is a novel component of a functional cellulose synthase complex. TBR is part of an Arabidopsis gene/protein family, (TBL-trichome birefringence like) which, depending on homology, comprises up to 20 members, none of which has a biological or biochemical function attributed. T-DNA insertion lines in TBR gene and two close homologues have been screened by PCR, but no homozygous were found and no trichomes phenotype was identified. Promoter-GUS lines were produced for TBR, as well as for its two closest homologues (one being a segmentally duplicated gene on chromosome III), using 1.6-2 kb of promoter sequence upstream of the annotated start codons. The TBR promoter was the only one of the three that yielded trichome expression, this probably explaining the phenotype of the TBR mutant. Moreover, TBR is expressed in leaves, in growing lateral roots, and in vascular tissues of young Arabidopsis seedlings and plantlets. Later on, the expression appears in inflorescens, stems, flowers and green siliques. This expression pattern is largely overlapping with those of the two analyzed homologues and it corresponds with data of RT-PCR expression profiling performed for TBR and the two analyzed homologues in different tissues, at different developmental stages. Biochemical analysis of cell wall (leaves and trichomes), as GC and MALDI-TOF, were performed, but revealed no major differences between tbr1 and wild type plants. Scanning electron microscopy analysis and cell wall polysaccharides antibody labeling showed a clear difference in the trichomes cell wall structure between mutant plant and wild type.
The scope of this study is to investigate the environmental change in the German part of the Elbe river basin, whereby the focus is on two water related problems: having too little water and having water of poor quality. The Elbe region is representative of humid to semi-humid landscapes in central Europe, where water availability during the summer season is the limiting factor for plant growth and crop yields, especially in the loess areas, where the annual precipitation is lower than 500 mm. It is most likely that water quantity problems will accelerate in future, because both the observed and the projected climate trend show an increase in temperature and a decrease in annual precipitation, especially in the summer. Another problem is nutrient pollution of rivers and lakes. In the early 1990s, the Elbe was one of the most heavily polluted rivers in Europe. Even though nutrient emissions from point sources have notably decreased in the basin due to reduction of industrial sources and introduction of new and improved sewage treatment facilities, the diffuse sources of pollution are still not sufficiently controlled. The investigations have been done using the eco-hydrological model SWIM (Soil and Water Integrated Model), which has been embedded in a model framework of climate and agro-economic models. A global scenario of climate and agro-economic change has been regionalized to generate transient climate forcing data and land use boundary conditions for the model. The model was used to transform the climate and land use changes into altered evapotranspiration, groundwater recharge, crop yields and river discharge, and to investigate the development of water quality in the river basin. Particular emphasis was given to assessing the significance of the impacts on the hydrology, taking into account in the analysis the inherent uncertainty of the regional climate change as well as the uncertainty in the results of the model. The average trend of the regional climate change scenario indicates a decrease in mean annual precipitation up to 2055 of about 1.5 %, but with high uncertainty (covering the range from -15.3 % to +14.8 %), and a less uncertain increase in temperature of approximately 1.4 K. The relatively small change in precipitation in conjunction with the change in temperature leads to severe impacts on groundwater recharge and river flow. Increasing temperature induces longer vegetation periods, and the seasonality of the flow regime changes towards longer low flow spells in summer. As a results the water availability will decrease on average of the scenario simulations by approximately 15 %. The increase in temperatures will improve the growth conditions for temperature limited crops like maize. The uncertainty of the climate trend is particularly high in regions where the change is the highest. The simulation results for the Nuthe subbasin of the Elbe indicate that retention processes in groundwater, wetlands and riparian zones have a high potential to reduce the nitrate concentrations of rivers and lakes in the basin, because they are located at the interface between catchment area and surface water bodies, where they are controlling the diffuse nutrient inputs. The relatively high retention of nitrate in the Nuthe basin is due to the long residence time of water in the subsurface (about 40 years), with good conditions for denitrification, and due to nitrate retention and plant uptake in wetlands and riparian zones. The concluding result of the study is that the natural environment and communities in parts of Central Europe will have considerably lower water resources under scenario conditions. The water quality will improve, but due to the long residence time of water and nutrients in the subsurface, this improvement will be slower in areas where the conditions for nutrient turn-over in the subsurface are poor.
In the present work, we discuss two subjects related to the nonequilibrium dynamics of polymers or biological filaments adsorbed to two-dimensional substrates. The first part is dedicated to thermally activated dynamics of polymers on structured substrates in the presence or absence of a driving force. The structured substrate is represented by double-well or periodic potentials. We consider both homogeneous and point driving forces. Point-like driving forces can be realized in single molecule manipulation by atomic force microscopy tips. Uniform driving forces can be generated by hydrodynamic flow or by electric fields for charged polymers. In the second part, we consider collective filament motion in motility assays for motor proteins, where filaments glide over a motor-coated substrate. The model for the simulation of the filament dynamics contains interactive deformable filaments that move under the influence of forces from molecular motors and thermal noise. Motor tails are attached to the substrate and modeled as flexible polymers (entropic springs), motor heads perform a directed walk with a given force-velocity relation. We study the collective filament dynamics and pattern formation as a function of the motor and filament density, the force-velocity characteristics, the detachment rate of motor proteins and the filament interaction. In particular, the formation and statistics of filament patterns such as nematic ordering due to motor activity or clusters due to blocking effects are investigated. Our results are experimentally accessible and possible experimental realizations are discussed.
Nitrogen is often a limiting factor for plant growth due to its heterogenous distribution in the soil and to seasonal and diurnal changes in growth rates. In most soils, NH4+ and NO3 – are the predominant sources of inorganic nitrogen that are available for plant nutrition. In this context, plants have evolved mechanisms that enable them to optimize nitrogen acquisition, which include transporters specialized in the uptake of nitrogen and susceptible to a regulation that responds to nitrogen limiting or excess conditions. Although the average NH4+ concentrations of soils are generally 100 to 1000 times lower than those of NO3 – (Marschner, 1995), most plants preferentially take up NH4+ when both forms are present because unlike NO3– , NH4+ has not to be reduced prior to assimilation and thus requires less energy for assimilation (Bloom et al., 1992). Apart from high uptake rates in roots, high intracellular ammonium concentrations also result from quantitatively important internal breakdown of amino acids (Feng et al., 1998), and originates in high quantities during photorespiration (Mattson et al., 1997, Pearson et al., 1998). Thus, NH4+ is a key component of nitrogen metabolism for all plants and can accumulate to varying concentrations in all compartments of the cell, including the cytosol, the vacuole and in the apoplast (Wells and Miller, 2000; Nielsen and Schjoerring, 1998). Two related families of ammonium transporters (AMT1 and AMT2), containing six genes which encode transporter proteins that are specific for ammonium had been identified prior to this thesis and some genes had partially been characterised in Arabidopsis (Gazzarrini et al., 1999; Sohlenkamp et al. 2002; Kaiser et al., 2002). However, these studies were not sufficient to assign physiological functions to the individual transporters and AMT1.4 and AMT1.5 had not been studied prior to this thesis. Given this background, it was considered desirable to acquire a deeper knowledge of the physiological functions of the six Arabidopsis ammonium transporters. To this end, tissue specific expression profiles of the individual wildtype AtAMT genes were performed by quantitative real time PCR (qRT-PCR) and promoter-GUS expression. Modern approaches such as the use of T-DNA insertional mutants and RNAi hairpin constructs were employed to reduce the expression levels of AMT genes. Transcript levels were determined, and physiological, biochemical and developmental analysis such as growth tests on different media and 14C-MA and NH4+ uptake studies with the isolated insertional mutants and RNAi lines were performed to deepen the knowledge of the individual functions of the six AMTs in Arabidopsis. In addition, double mutants of the insertional mutants were created to investigate the extent in which homologous genes could compensate for lost transporter functions. The results described in this thesis show that the six AtAMT genes display a high degree of specifity in their tissue specific expression and are likely to play complementary roles in ammonium uptake into roots, in shoots, and in flowers. AtAMT1.1 is likely to be a ‘work horse’ for cellular ammonium transport and reassimilation. A major role is probably the recapture of photorespiratory NH3/NH4+ escaping from the cytosol. In roots, it is likely to transport NH4+ from the apoplast into cortical cells. AtAMT1.3 and AtAMT1.5 appear to be specialised in the acquisition of external NH4+ from the soil. Furthermore, AtAMT1.5 plays an additional role in the reassimilation of NH3/NH4+ released during the breakdown of storage proteins in the cotyledons of germinating seedlings. It was difficult to distinguish a specialisation between the transporters AtAMt1.2 and AtAMt1.1, however the root and flower specific expression patterns are different and indicate alternative functions of both. AtAMT1.4 has a very distinct expression which is restricted to the vascular bundels of leaves and to pollen only, where it is likely to be involved in the loading of NH4+ into the cells.The AtAMT2.1 expression pattern is confined to vascular bundels and meristematic active tissues in leaves where ammonium concentrations can reach very high levels. Additionally, the Vmax of AtAMT2 increases with increasing external pH, contrasting to AtAMT1.1. Thus, AtAMT2.1 it might be specialised in ammonium transport in ammonium rich environments, where the functions of other transporters are limited, enabling cells to take up NH4+ over a wide range of concentrations. The root hair expression ascribes an additional role in NH3/NH4+ acquisition where it possibly serves as a transporter that is able to acquire ammonium from basic soils where other transporters become less effective.RNAi lines showing a reduction in AtAMT gene mRNA levels and NH4+ transport kinetics, grew slower and flowering time was delayed. This indicates that NH4+ is a crucial and limiting factor for plant growth.
Agricultural policy in the transition states of Central Eastern Europe is a very complex issue – ranging from privatisation of farm land, the establishment of agricultural markets to detailed questions of veterinary care, plant health and animal nutrition. Its main elements are the introduction of market liberalization, farm restructuring, privatisation, the reform of the sector and the creation of supporting market institutions and services.1 In this process central state agriculture administration plays a decisive role. This paper is summing up the research of the author on Slovak agricultural administration between 2002 and 2004. This work was part of a DFG-funded research project on “Genesis, Organization and Efficiency of the central-state Ministerial Administration in Central and Eastern Europe”. The project was analysing the processes, results and efficiency of administrative structures at central-state level in Estonia, Poland and Slovakia with reference to public administration in the policy fields of agriculture and telecommunications. The paper is reflecting the situation in the sector and its administration at the beginning of 2004. At first, an overview of the role of the agricultural sector in Slovak economy in the past and presence is provided (section I). Against this background, the development of the agricultural policy in the different periods since 1989 will be analysed, mainly what privatisation, accession to the EU and subsidy policy are concerned (section II). A detailed study of the developments in agricultural administration forms the next part of the paper (section III), i.e. the changes taking place in the ministry of agriculture and in the other institutions responsible for the implementation of agricultural policy. The role of interest groups in agriculture is briefly analysed (section IV). In the conclusions two different scenarios on the further development of Slovak agricultural administration will be deployed.