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This work focuses on a fundamental problem in modern numerical rela- tivity: Extracting gravitational waves in a coordinate and gauge independent way to nourish a unique and physically meaningful expression. We adopt a new procedure to extract the physically relevant quantities from the numerically evolved space-time. We introduce a general canonical form for the Weyl scalars in terms of fundamental space-time invariants, and demonstrate how this ap- proach supersedes the explicit definition of a particular null tetrad. As a second objective, we further characterize a particular sub-class of tetrads in the Newman-Penrose formalism: the transverse frames. We establish a new connection between the two major frames for wave extraction: namely the Gram-Schmidt frame, and the quasi-Kinnersley frame. Finally, we study how the expressions for the Weyl scalars depend on the tetrad we choose, in a space-time containing distorted black holes. We apply our newly developed method and demonstrate the advantage of our approach, compared with methods commonly used in numerical relativity.
The study investigates the water resources and aquifer dynamics of the igneous fractured aquifer-system of the Troodos Mountains in Cyprus, using a coupled, finite differences water balance and groundwater modelling approach. The numerical water balance modelling forms the quantitative framework by assessing groundwater recharge and evapotranspiration, which form input parameters for the groundwater flow models. High recharge areas are identified within the heavily fractured Gabbro and Sheeted Dyke formations in the upper Troodos Mountains, while the impervious Pillow Lava promontories - with low precipitation and high evapotranspiration - show unfavourable recharge conditions. Within the water balance studies, evapotranspiration is split into actual evapotranspiration and the so called secondary evapotranspiration, representing the water demand for open waters, moist and irrigated areas. By separating the evapotranspiration of open waters and moist areas from the one of irrigated areas, groundwater abstraction needs are quantified, allowing the simulation of single well abstraction rates in the groundwater flow models. Two sets of balanced groundwater models simulate the aquifer dynamics in the presented study: First, the basic groundwater percolation system is investigated using two-dimensional vertical flow models along geological cross-sections, depicting the entire Troodos Mountains up to a depth of several thousands of metres. The deeply percolating groundwater system starts in the high recharge areas of the upper Troodos, shows quasi stratiform flow in the Gabbro and Sheeted Dyke formations, and rises to the surface in the vicinity of the impervious Pillow Lava promontories. The residence times mostly yield less than 25 years, the ones of the deepest fluxes several hundreds of years. Moreover, inter basin flow and indirect recharge of the Circum Troodos Sedimentary Succession are identified. In a second step, the upper and most productive part of the fractured igneous aquifer-system is investigated in a regional, horizontal groundwater model, including management scenarios and inter catchment flow studies. In a natural scenario without groundwater abstractions, the recovery potential of the aquifer is tested. Predicted future water demand is simulated in an increased abstraction scenario. The results show a high sensitivity to well abstraction rate changes in the Pillow Lava and Basal Group promontories. The changes in groundwater heads range from a few tens of metres up to more than one hundred metres. The sensitivity in the more productive parts of the aquifer-system is lower. Inter-catchment flow studies indicate that - besides the dominant effluent conditions in the Troodos Mountains - single reaches show influent conditions and are sub-flown by groundwater. These fluxes influence the local water balance and generate inter catchment flow. The balanced groundwater models form thus a comprehensive modelling system, supplying future detail models with information concerning boundary conditions and inter-catchment flow, and allowing the simulation of impacts of landuse or climate change scenarios on the dynamics and water resources of the Troodos aquifer-system.
Two-particle excitations, such as spin and charge excitations, play a key role in high-Tc cuprate superconductors (HTSC). Due to the antiferromagnetism of the parent compound the magnetic excitations are supposed to be directly related to the mechanism of superconductivity. In particular, the so-called resonance mode is a promising candidate for the pairing glue, a bosonic excitation mediating the electronic pairing. In addition, its interactions with itinerant electrons may be responsible for some of the observed properties of HTSC. Hence, getting to the bottom of the resonance mode is crucial for a deeper understanding of the cuprate materials . To analyze the corresponding two-particle correlation functions we develop in the present thesis a new, non-perturbative and parameter-free technique for T=0 which is based on the Variational Cluster Approach (VCA, an embedded cluster method for one-particle Green's functions). Guided by the spirit of the VCA we extract an effective electron-hole vertex from an isolated cluster and use a fully renormalized bubble susceptibility chi0 including the VCA one-particle propagators.Within our new approach, the magnetic excitations of HTSC are shown to be reproduced for the Hubbard model within the relevant strong-coupling regime. Exceptionally, the famous resonance mode occurring in the underdoped regime within the superconductivity-induced gap of spin-flip electron-hole excitations is obtained. Its intensity and hourglass dispersion are in good overall agreement with experiments. Furthermore, characteristic features such as the position in energy of the resonance mode and the difference of the imaginary part of the susceptibility in the superconducting and the normal states are in accord with Inelastic Neutron Scattering (INS) experiments. For the first time, a strongly-correlated parameter-free calculation revealed these salient magnetic properties supporting the S=1 magnetic exciton scenario for the resonance mode. Besides the INS data on magnetic properties further important new insights were gained recently via ARPES (Angle-Resolved Photoemission-Spectroscopy) and Raman experiments which disclosed a quite different doping dependence of the antinodal compared to the near-nodal gap. This thesis provides an approach to the Raman response similar to the magnetic case for inspecting this gap dichotomy. In agreement with experiments and one-particle data obtained in the VCA, we recover the antinodal gap decreasing and the near-nodal gap increasing as a function of doping. Hence, our results prove the Hubbard model to account for these salient gap features. In summary, we develop a two-particle cluster approach which is appropriate for the strongly-correlated regime and contains no free parameter. Our results obtained with this new approach combined with the phase diagram and the one-particle excitations obtained in the VCA strongly constitute a Hubbard model description of HTSC cuprate materials.
In this thesis, we present novel approaches for formation driving of nonholonomic robots and optimal trajectory planning to reach a target region. The methods consider a static known map of the environment as well as unknown and dynamic obstacles detected by sensors of the formation. The algorithms are based on leader following techniques, where the formation of car-like robots is maintained in a shape determined by curvilinear coordinates. Beyond this, the general methods of formation driving are specialized and extended for an application of airport snow shoveling. Detailed descriptions of the algorithms complemented by relevant stability and convergence studies will be provided in the following chapters. Furthermore, discussions of the applicability will be verified by various simulations in existing robotic environments and also by a hardware experiment.
Mycobacterium tuberculosis is the causative agent of tuberculosis and responsible for more than eight million new infections and about two million deaths each year. Novel chemotherapeutics are urgently needed to treat the emerging threat of multi drug resistant and extensively drug resistant strains. Cell wall biosynthesis is a widely used target for chemotherapeutic intervention in bacterial infections. In mycobacteria, the cell wall is comprised of mycolic acids, very long chain fatty acids that provide protection and allow the bacteria to persist in the human macrophage. The type II fatty acid biosynthesis pathway in Mycobacterium tuberculosis synthesizes fatty acids with a length of up to 56 carbon atoms that are the precursors of the critical mycobacterial cell wall components mycolic acids. KasA, the mycobacterial ß-ketoacyl synthase and InhA, the mycobacterial enoyl reductase, are essential enzymes in the fatty acid biosynthesis pathway and validated drug targets. In this work, KasA was expressed in Mycobacterium smegmatis, purified and co-crystallized in complex with the natural thiolactone antibiotic thiolactomycin (TLM). High-resolution crystal structures of KasA and the C171Q KasA variant, which mimics the acyl enzyme intermediate of the enzyme, were solved in absence and presence of bound TLM. The crystal structures reveal how the inhibitor is coordinated by the enzyme and thus specifically pinpoint towards possible modifications to increase the affinity of the compound and develop potent new drugs against tuberculosis. Comparisons between the TLM bound crystal structures explain the preferential binding of TLM to the acylated form of KasA. Furthermore, long polyethylene glycol molecules are bound to KasA that mimic a fatty acid substrate of approximately 40 carbon atoms length. These structures thus provide the first insights into the molecular mechanism of substrate recognition and reveal how a wax-like substance can be accommodated in a cytosolic environment. InhA was purified and co-crystallized in complex with the slow, tight binding inhibitor 2-(o-tolyloxy)-5-hexylphenol (PT70). Two crystal structures of the ternary InhA-NAD+-PT70 were solved and reveal how the inhibitor is bound to the substrate binding pocket. Both structures display an ordered substrate binding loop and corroborate the hypothesis that slow onset inhibition is coupled to loop ordering. Upon loop ordering, the active site entrance is more restricted and the inhibitor is kept inside more tightly. These studies provide additional information on the mechanistic imperatives for slow onset inhibition of enoyl ACP reductases.
Genome sequence analysis A combination of genome analysis application has been established here during this project. This offers an efficient platform to interactively compare similar genome regions and reveal loci differences. The genes and operons can be rapidly analyzed and local collinear blocks (LCBs) categorized according to their function. The features of interests are parsed, recognized, and clustered into reports. Phylogenetic relationships can be readily examined such as the evolution of critical factors or a certain highly-conserved region. The resulting platform-independent software packages (GENOVA and inGeno), have been proven to be efficient and easy to handle in a number of projects. The capabilities of the software allowed the investigation of virulence factors, e.g., rsbU, strains’ biological design, and in particular pathogenicity feature storage and management. We have successfully investigated the genomes of Staphylococcus aureus strains (COL, N315, 8325, RN1HG, Newman), Listeria spp. (welshimeri, innocua and monocytogenes), E.coli strains (O157:H7 and MG1655) and Vaccinia strains (WR, Copenhagen, Lister, LIVP, GLV-1h68 and parental strains). Metabolic network analysis Our YANAsquare package offers a workbench to rapidly establish the metabolic network of such as Staphylococcous aureus bacteria in genome-scale size as well as metabolic networks of interest such as the murine phagosome lipid signalling network. YANAsquare recruits reactions from online databases using an integrated KEGG browser. This reduces the efforts in building large metabolic networks. The involved calculation routines (METATOOL-derived wrapper or native Java implementation) readily obtain all possible flux modes (EM/EP) for metabolite fluxes within the network. Advanced layout algorithms visualize the topological structure of the network. In addition, the generated structure can be dynamically modified in the graphic interface. The generated network as well as the manipulated layout can be validated and stored (XML file: scheme of SBML level-2). This format can be further parsed and analyzed by other systems biology software, such as CellDesigner. Moreover, the integrated robustness-evaluation routine is able to examine the synthesis rates affected by each single mutation throughout the whole network. We have successfully applied the method to simulate single and multiple gene knockouts, and the affected fluxes are comprehensively revealed. Recently we applied the method to proteomic data and extra-cellular metabolite data of Staphylococci, the physiological changes regarding the flux distribution are studied. Calculations at different time points, including different conditions such as hypoxia or stress, show a good fit to experimental data. Moreover, using the proteomic data (enzyme amounts) calculated from 2D-Gel-EP experiments our study provides a way to compare the fluxome and the enzyme expression. Oncolytic vaccinia virus (VACV) We investigated the genetic differences between the de novo sequence of the recombinant oncolytic GLV-1h68 and other related VACVs, including function predictions for all found genome differences. Our phylogenetic analysis indicates that GLV-1h68 is closest to Lister strains but has lost several ORFs present in its parental LIVP strain, including genes encoding CrmE and a viral Golgi anti-apoptotic protein, v-GAAP. Functions of viral genes were either strain-specific, tissue-specific or host-specific comparing viral genes in the Lister, WR and COP strains. This helps to rationally design more optimized oncolytic virus strains to benefit cancer therapy in human patients. Identified differences from the comparison in open reading frames (ORFs) include genes for host-range selection, virulence and immune modulation proteins, e.g. ankyrin-like proteins, serine proteinase inhibitor SPI-2/CrmA, tumor necrosis factor (TNF) receptor homolog CrmC, semaphorin-like and interleukin-1 receptor homolog proteins. The contribution of foreign gene expression cassettes in the therapeutic and oncolytic virus GLV-1h68 was studied, including the F14.5L, J2R and A56R loci. The contribution of F14.5L inactivation to the reduced virulence is demonstrated by comparing the virulence data of GLV-1h68 with its F14.5L-null and revertant viruses. The comparison suggests that insertion of a foreign gene expression cassette in a nonessential locus in the viral genome is a practical way to attenuate VACVs, especially if the nonessential locus itself contains a virulence gene. This reduces the virulence of the virus without compromising too much the replication competency of the virus, the key to its oncolytic activity. The reduced pathogenicity of GLV-1h68 was confirmed by our experimental collaboration partners in male mice bearing C6 rat glioma and in immunocompetent mice bearing B16-F10 murine melanoma. In conclusion, bioinformatics and experimental data show that GLV-1h68 is a promising engineered VACV variant for anticancer therapy with tumor-specific replication, reduced pathogenicity and benign tissue tropism.
This thesis analyzes the relationship between market concentration and efficiency of the market outcome in a differentiated good context from different points of view. The first chapter introduces the objectives of competition policy and antitrust authorities and outlines the importance of market concentration. Chapter 2 analyzes the relationship between social surplus and market heterogeneity in a differentiated Cournot oligopoly. Market heterogeneity is due to differently efficient firms, each of them producing one variety of a differentiated good. All firms exhibit constant but different marginal costs without fixed costs. Consumers preferences are given by standard quadratic utility originated by Dixit (1979). Since preferences are quasi-linear social surplus is the measure for Pareto-optimality. The main finding is that consumer suprlus as well as producer surplus increase with the variance of marginal costs. The third chapter analyzes the relationship between the cost structure and market concentration measured by the Herfindahl-Hirschman Index. Market concentration increases with the variance of marginal costs as well as the mean of marginal costs. Chapter four analyzes welfare implications of present antitrust enforcement policy on basis of the same theoretical model. European as well as the US Merger Guidelines presume a negative impact of market concentration on the competitiveness of the market and, therefore, on the efficiency of the market outcome. The results of the previous chapters indicate that this assumption is false. The main finding is that post-merger joint profit of the insider increase with the size of the merger. Moreover, there is a negative relationship between the size of the merger and efficiency of the market outcome. Present antitrust enforcement policy increases the disparity of output levels and enforces the removal of the least efficient firm of the market. The welfare gains can be traced back on these two effects. Therefore, neither a minimum of market concentration nor a maximum of product diversity is necessarily welfare enhancing even in absence of fixed costs.
Like many other social insect societies, honeybees collectively share the resources they gather by feeding each other. These feeding contacts, known as trophallaxis, are regarded as the fundamental basis for social behavior in honeybees and other social insects for assuring the survival of the individual and the welfare of the group. In honeybees, where most of the trophallactic contacts are formed in the total darkness of the hive, the antennae play a decisive role in initiation and maintenance of the feeding contact, because they are sensitive to gustatory stimuli. The sequences of behaviors performed by the receiver bees at the beginning of a feeding contact includes the contact of one antenna with the mouthparts of a donor bee where the regurgitated food is located. The antennal motor action is characterized by behavioral asymmetry, which is novel among communicative motor actions in invertebrates. This preference of right over left antenna is without exception even after removal of the antennal flagellum. This case of laterality in basic social interaction might have its reason in the gustatory asymmetry in the antennae, because the right antenna turns out to be significantly more sensitive to stimulation with sugar water of various concentrations than the left one. Trophallactic contacts which guarantee a constant access to food for every individual in the hive are vitally important to the honeybee society, because honeybees are heterothermic insects which actively regulate their thoracic temperature. Even though the individual can regulate its body temperature, its heating performance is strictly limited by the amount of sugar ingested. The reason for this is that honeybees use mostly the glucose in their hemolymph as the energy substrate for muscular activity, and the heat producing flight muscles are among the metabolically most active tissues known. The fuel for their activity is honey; processed nectar with a sugar content of ~80% stored in the honeycomb. The results show that the sugar content of the ingested food correlates positively with the thoracic temperature of the honeybees even if they are caged and show no actual heating-related behavior as in brood warming or heating in the centre of the winter cluster. Honeybees actively regulate their brood temperature by heating to keep the temperature between 33 °C to 36 °C if ambient temperatures are lower. Heating rapidly depletes the worker’s internal energy; therefore the heating performance is limited by the honey that is ingested before the heating process. This study focused on the behavior and the thoracic temperature of the participants in trophallactic food exchanges on the brood comb. The brood area is the centre of heating activity in the hive, and therefore the region of highest energy demand. The results show that the recipients in a trophallactic food exchange have a higher thoracic temperature during feeding contacts than donors, and after the feeding contact the former engage in brood heating more often. The donor bees have lower thoracic temperature and shuttle constantly between honey stores and the brood comb, where they transfer the stored honey to heating bees. In addition, the results show a heat-triggered mechanism that enables donor and recipient to accomplish trophallactic contacts without delay in the total darkness of the hive in the brood area as the most energy consuming part of the hive. Providing heat-emitting workers with small doses of high performance fuel contributes to an economic distribution of resources consistent with the physiological conditions of the bees and the ecological requirements of the hive, resulting in a highly economical resource management system which might be one of the factors favouring the evolution of perennial bee colonies in temperate regions. The conclusion of these findings suggests a resource management strategy that has evolved from submissive placation behavior as it is seen in honeybees, bumblebees and other hymenopterans. The heat-triggered feedback mechanism behind the resource management of the honeybee´s thermoregulatory behavior reveals a new aspect of the division of labor and a new aspect of communication, and sheds new light on sociality in honeybees.
The spectroscopic properties of molecular aggregates have been investigated by means of quantum dynamical calculations. Thereby both linear and nonlinear spectroscopic techniques have been taken into account. For the simulation of absorption and CD-spectra, coupling effects were regarded as well as the relative orientation of the monomer units in order to determine the parameters by reproducing measured spectra. For a more detailled description, results from quantum chemical calculations have also been included. Furthermore, investigations on nonlinear spectroscopy of molecular dimers have been performed.
Platelet activation induces cytoskeletal rearrangements involving a change from discoid to spheric shape, secretion, and eventually adhesion and spreading on immobilized ligands. Small GTPases of the Rho family, such as Rac1 and Cdc42, are known to be involved in these processes by facilitating the formation of lamellipodia and filopodia, respectively. This thesis focuses on the role Rac1 and Cdc42 for platelet function and formation from their precursor cells, the megakaryocytes (MKs), using conditional knock-out mice. In the first part of the work, the involvement of Rac1 in the activation of the enzyme phospholipase (PL) C2 in the signaling pathway of the major platelet collagen receptor glycoprotein (GP) VI was investigated. It was found that Rac1 is essential for PLC2 activation independently of tyrosine phosphorylation of the enzyme, resulting in a specific platelet activation defect downstream of GPVI, whereas signaling of other activating receptors remains unaffected. Since Rac1-deficient mice were protected from arterial thrombosis in two different in vivo models, the GTPase might serve as a potential target for the development of new drugs for the treatment and prophylaxis of cardio- and cerebrovascular diseases. The second part of the thesis deals with the first characterization of MK- and platelet-specific Cdc42 knock-out mice. Cdc42-deficient mice displayed mild thrombo-cytopenia and platelet production from mutant MKs was markedly reduced. Unexpectedly, Cdc42-deficient platelets showed increased granule content and release upon activation, leading to accelerated thrombus formation in vitro and in vivo. Furthermore, Cdc42 was not generally required for filopodia formation upon platelet activation. Thus, these results indicate that Cdc42, unlike Rac1, is involved in multiple signaling pathways essential for proper platelet formation and function. Finally, the outcome of combined deletion of Rac1 and Cdc42 was studied. In contrast to single deficiency of either GTPase, platelet production from double-deficient MKs was virtually abrogated, resulting in dramatic macrothrombocytopenia in the animals. Formed platelets were largely non-functional leading to a severe hemostatic defect and defective thrombus formation in double-deficient mice in vivo. These results demonstrate for the first time a functional redundancy of Rac1 and Cdc42 in the hematopoietic system.
Regulation of effector T cells is an important mechanism to control organ-specific inflammation. Thereby regulatory T cells (Treg cells) are essential for maintaining peripheral immune tolerance and for establishing parenchyma immune homeostasis. A novel population of natural human Treg characterized by the constitutive expression of the immune-tolerogenic human HLA-G molecule has been identified. In the first part of the study, we elucidated the mechanism(s) by which CD4+ HLA-Gpos Treg modulates their cellular targets namely autologous HLA-G negative responder T cells (HLAGneg Tresp). Using a suppression system free of antigen-presenting cells (APC), we demonstrate a T-T cell interaction resulting in suppression of HLA-Gneg Tresp. We could also show that this suppression was independent of cell-cell contact. Importantly, stimulus of T cell receptor (TCR) on HLA-Gpos Treg facilitated their suppressive capacity. We also observed that removal of HLA-Gpos Treg from the established co-cultures could restore the ability of HLA-Gneg Tresp to proliferate upon TCR re-stimulation, indicating that the suppression was reversible. Further, HLA-Gpos Treg–mediated suppression was critically depending on the secretion of IL-10 but not TGF-β. Taken together, this part of the work provides an in-depth characterization of the mechanisms of how HLA-Gpos Treg suppresses T responder cells in direct T-T interactions. Understanding the suppressive mechanism used by HLA-Gpos Treg may help to develop therapeutic strategies to modulate regulatory arms of T-cell suppression. In the second part of this study, the potential role of HLA-Gpos Treg in the pathophysiological process of Multiple Sclerosis (MS), a prototypic autoimmune inflammatory central nervous system (CNS), has been investigated. We found that HLA-Gpos Treg are enriched in the cerebrospinal fluid (CSF) from MS patients, but not in non-inflammatory controls. CSFderived HLA-Gpos Treg showed predominance of central memory (CD45RA-CD27+) phenotype, exhibited markers of activation (ICOS), and had significantly higher expression of the inflammatory chemokine receptor CCR5. Importantly, these cells demonstrated as potent suppressors to autologous CD4+ T-cell proliferation. Using an in vitro model of human blood brain barrier, we showed that HLA-Gpos Treg have a strong propensity to migrate, which could be facilitated by MIP1α and RANTES (ligands of CCR5) but not MIP3β (a ligand of CCR7). The HLA-Gpos Treg migration triggered by chemokines was also associated with a gain of suppressive capacity upon cellular transmigration. In contrast to CD4+CD25+ naturally occurring FoxP3-expressing Treg, HLA-Gpos Treg from patients with MS did not exhibit impaired function, suggesting that HLA-Gpos Treg are selectively recruited to the sites of CNS inflammation in an effort to combat destructive inflammation during MS. Our results contribute to the understanding of the role and function of HLA-Gpos Treg and provide an important example of “beneficial” T-cell inflammation in CNS autoimmunity- interesting both from a patho/-physiological and a therapeutically point of view.
Macromolecular complexes, also termed molecular machines, facilitate a large spectrum of biological reactions and tasks crucial to the survival of cells. These complexes are composed of either protein only, or proteins bound to nucleic acids (DNA or RNA). Prominent examples for each class are the proteosome, the nucleosome and the ribosome. How such units are assembled within the context of a living cell is a central question in molecular biology. Earlier studies had indicated that even very large complexes such as ribosomes could be reconstituted from purified constituents in vitro. The structural information required for the formation of macromolecular complexes, hence, lies within the subunits itself and, thus, allow for self- assembly. However, increasing evidence suggests that in vivo many macromolecular complexes do not form spontaneously but require assisting factors (“assembly chaperones”) for their maturation. In this thesis the assembly of RNA-protein (RNP) complexes has been studied by a combination of biochemical and structural approaches. A resourceful model system to study this process is the biogenesis pathway of the uridine-rich small nuclear ribonucleoproteins (U snRNPs) of the spliceosome. This molecular machine catalyzes pre-mRNA splicing, i.e. the removal of non-coding introns and the joining of coding exons to functional mRNA. The composition and architecture of U snRNPs is well defined, also, the nucleo-cytoplasmic transport events enabling the formation of these particles in vivo have been analyzed in some detail. Furthermore, recent studies suggest that the formation of U snRNPs in vivo is mediated by an elaborate assembly machinery consisting of protein arginine methyltransferase (PRMT5)- and survival motor neuron (SMN)-complexes. The elucidation of the reaction mechanism of cellular U snRNP assembly would serve as a paradigm for our understanding of how RNA-protein complexes are formed in the cellular environment. The following key findings were obtained as part of this study: 1) Efforts were made to establish a full inventory of the subunits of the SMN-complex. This was achieved by the biochemical definition and characterization of an atypical component of this complex, the unrip protein. This protein is associated with the SMN-complex exclusively in the cytoplasm and influences its subcellular localization. 2) With a full inventory of the components in hand, the architecture of the SMN-complex was defined on the basis of an interaction map of all subunits. This study elucidated that the proteins SMN, Gemin7 and Gemin8 form a backbone, onto which the remaining subunits adhere in a modular manner. 3) The two studies mentioned above formed the basis to elucidate the reaction mechanism of cellular U snRNP assembly. Initially, an early phase in the SMN-assisted formation of U snRNPs was analyzed. Two subunits of the U7 snRNP (LSm10 and 11) were found to interact with the PRMT5-complex, without being methylated. This report suggests that the stimulatory role of the PRMT5-complex is independent of its methylation activity. 4) Key reaction intermediates in U snRNP assembly were found and characterized by a combination of biochemistry and structural studies. Initially, a precursor to U snRNPs with a sedimentation coefficient of 6S is formed by the pICln subunit of the PRMT5-complex and Sm proteins. This intermediate was shown to constitute a kinetic trap in the U snRNP assembly reaction. Progression towards the assembled U snRNP depends on the activity of the SMN-complex, which acts as a catalyst. The formation of U snRNPs is shown to be structurally similar to the way clamps are deposited onto DNA to tether poorly processive polymerases. 5) The human SMN-complex is composed of several subunits. However, it is unknown whether all subunits of this entity are essential for U snRNP assembly. A combination of bioinformatics and biochemistry was applied to tackle this question. By mining databases containing whole-genome assemblies, the SMN-Gemin2 heterodimer is recognized as the most ancestral form of the SMN-complex. Biochemical purification of the Drosophila melanogaster SMN-complex reveals that this complex is composed of the same two subunits. Furthermore, evidence is provided that the SMN-Gemin2 heterodimer is necessary and sufficient to promote faithful U snRNP assembly. Future studies will adress further details in the reaction mechanism of cellular U snRNP assembly. The results obtained in this thesis suggest that the SMN and Gemin2 subunits are sufficient to promote U snRNP formation. What then is the function of the remaining subunits of the SMN-complex? The reconstitution schemes established in this thesis will be instrumental to address this question. Furthermore, additional mechanistic insights into the U snRNP assembly reaction will require the elucidation of structures of the assembly machinery trapped at various states. The prerequisite for these structural studies, the capability to generate homogenous complexes in sufficient amounts, has been accomplished in this thesis.
The olfactory system of leafcutting ants: neuroanatomy and the correlation to social organization
(2009)
In leaf-cutting ants (genera Atta and Acromyrmex), the worker caste exhibits a pronounced size-polymorphism, and division of labor is largely dependent on worker size (alloethism). Behavioral studies have shown a rich diversity of olfactory-guided behaviors, and the olfactory system seems to be highly developed and very sensitive. To allow fine-tuned behavioral responses to different tasks, adaptations within the olfactory system of different sized workers are expected. In a recent study, two different phenotypes of the antennal lobe of Atta vollenweideri workers were found: MG- and RG-phenotype (with and without a macroglomerulus, MG). The existence of the macroglomerulus is correlated to the body size of workers, with small workers showing the RG-phenotype and large workers showing the MG-phenotype. In the MG, the information about the releaser component of the trail-pheromone is processed. In the first part of my PhD-project, I focus on quantifying behavioral differences between different sized workers in Atta vollenweideri. The study analyzes the trail following behavior; which can be generally performed by all workers. An artificial trail consisting of the releaser component of the trail-pheromone in decreasing concentration was used to test the trail-following performance of individual workers. The trail-following performance of the polymorphic workers is depended of the existence of the MG in the antennal lobe. Workers possessing the MG-phenotype were significantly better in following a decreasing trail then workers showing the RG-phenotype. In the second part I address the question if there are more structural differences, besides the MG, in the olfactory system of different sized workers. Therefore I analyze whether the glomerular numbers are related to worker size. The antennal lobes of small workers contain ~390 glomeruli (low-number; LN-phenotype), and in large workers I found a substantially higher number of ~440 glomeruli (high-number; HN-phenotype). All LN-phenotype workers and some of the small HN-phenotype workers do not possess an MG (LN-RG-phenotype and HN-RG-phenotype) at all, whereas the remaining majority of HN-phenotype workers do possess an MG (HN-MG-phenotype). Mass-stainings of antennal olfactory receptor neurons revealed that the sensory tracts divide the antennal lobe into six clusters of glomeruli (T1-T6). In the T4-cluster ~50 glomeruli are missing in the LN-phenotype workers. Selective staining of single sensilla and their associated receptor neurons showed that T4-glomeruli are innervated by receptor neurons from the main type of olfactory sensilla, the Sensilla trichodea curvata which are also projecting to glomeruli in all other clusters. The other type of olfactory sensilla, the Sensilla basiconica, exclusively innervates T6-glomeruli. Quantitative analyses revealed a correlation between the number of Sensilla basiconica and the volume of T6 glomeruli in different sized workers. The results of both behavioral and neuroanatomical studies in Atta vollenweideri suggest that developmental plasticity of antennal-lobe phenotypes promotes differences in olfactory-guided behavior which may underlie task specialization within ant colonies. The last part of my project focuses on the evolutionary origin of the macroglomerulus and the number of glomeruli in the antennal lobe. I compared the number, volumes and position of the glomeruli of the antennal lobe of 25 different species from all three major Attini groups (lower, higher and leaf-cutting Attini). The antennal lobes of all investigated Attini comprise a high number of glomeruli (257-630). The highest number was found in Apterostigma cf. mayri. This species is at a basal position within the Attini phylogeny, and a high number of glomeruli might have been advantageous in the evolution of the advanced olfactory systems of this Taxa. The macroglomerulus can be found in all investigated leaf-cutting Attini, but in none of the lower and higher Attini species. It is found only in large workers, and is located close to the entrance of the antennal nerve in all investigated species. The results indicate that the presence of a macroglomerulus in large workers of leaf-cutting Attini is a derived overexpression of a trait in the polymorphic leaf-cutting species. It presumably represents an olfactory adaptation to elaborate foraging and mass recruitment systems, and adds to the complexity of division of labor and social organization known for this group.
This study presents new petrological results obtained from high-grade metamorphic rocks of the Beit Bridge, Mahalapye and Phikwe Complexes, which constitute the Central Zone of the Limpopo Belt in southern Africa. These results provide detailed information about the prograde and retrograde pressure-temperature (P-T) evolution of the three investigated complexes and, in concert with geochronological data, form the basis for the development of a coherent geodynamic model for the evolution of the Limpopo’s Central Zone. The P-T paths were inferred by the thorough investigation of silica-saturated and silica- undersaturated metapelitic and metabasic rocks, comprising six sillimanite-garnet-cordierite gneisses, four (garnet)-biotite-plagioclase gneisses, two garnet-orthopyroxene-biotite-Kfeldspar-plagioclase gneisses, one garnet- cordierite-orthoamphibole fels, one garnet-biotite amphibolite, and one garnet-clinopyroxene amphibolite. P-T points and P-T evolutions were derived by the application of conventional geothermobarometers, and quantitative phase diagrams in the systems Na2O - CaO - K2O - FeO - MgO - Al2O3 - SiO2 - H2O - TiO2 - O (NCKFMASHTiO), and MnO - TiO2 - Na2O - CaO - K2O - FeO - MgO - Al2O3 - SiO2 - H2O (MnTiNCKFMASH) - using the computer software THERMOCALC and THERIAK-DOMINO. The petrological information, in particular those obtained by comparison between observed and thermodynamically calculated mineral assemblages, zonations and modes, in combination with new and existing geochronological data provide evidence that rocks from the three investigated complexes underwent slightly different P-T evolutions at different times. The samples from the Bulai Pluton area (Beit Bridge Complex) provide evidence for a Neoarchean high-grade metamorphic event at ~2.64 Ga (M2), with peak P-T conditions of ~850°C at 8-9 kbar, and a decompression-cooling path to ~750°C at 5-6 kbar. This metamorphic evolution perhaps took place in a magmatic arc setting. In contrast, samples from the Mahalapye and Phikwe Complex document a Palaeoproterozoic event at ~2.03-2.05 Ga (M3), and were subject to different styles of prograde metamorphism. Metamorphic rocks from the Mahalapye Complex experienced a high-temperature low-pressure (HT-LP) metamorphic overprint, accompanied by the emplacement of voluminous granite bodies between 2.06 and 2.02 Ga, and provide evidence for a slightly prograde decompression from ~650°C/7 kbar to ~800°C/5.5 kbar. In contrast, the metamorphic rocks from the Phikwe Complex provide evidence for a simultaneous pressure and temperature increase from ~600°C/6 kbar to ~750°C/8 kbar, in the absence of significant Palaeoproterozoic magmatism. The HT-LP metamorphic evolution of the Mahalapye Complex is interpreted to be initiated by the underplating of hot mafic melts, either formed in response to SE-subduction during the Kheis-Magondi orogeny, and/or by contemporaneous mantle plume activities related to the formation of the Bushveld Complex. In contrast, the prograde pressure and temperature increase reflected by the rocks from the Phikwe Complex rather reflects successive crustal stacking at ~2.03 Ga. This stacking, which is also reported from many other units throughout the Limpopo Belt, is interpreted to result from the final convergence between the Kaapvaal and Zimbabwe Cratons, perhaps caused by SE-directed compression in response to the Kheis-Magondi orogeny between ~2.06 and 1.90 Ga.
Plants must respond to multiple stimuli in a natural environment. Therefore they need the ability to rapidly reorganise and specifically build up appropriate metabolites to adapt to their environment. Abiotic cues, such as ambient solar radiation, influence the next trophic level directly, but also an altered plant composition triggered by these environmental cues can have an effect on the behaviour of herbivores. The aim of this study was to test effects of the important ultraviolet (UV) radiation on plants and on plant-insect interactions using multi-level investigations. The focus was on the conduction of controlled experiments with broccoli plants in highly engineered greenhouses covered with innovative materials, which only differed in their UV-B transmission. For the first time in this controlled environment the plant-mediated UV-B effects on phloem-feeding aphids were studied. Broccoli plants (Brassica oleracea L. convar. botrytis, Brassicaceae) were under filter tents either exposed to (inclusion, +UV) or not exposed to (exclusion, -UV) UV-A / UV-B radiation. In greenhouses covered with new, innovative materials transmitting high (80%), medium (23%) or low (4%) levels of ambient solar UV-B radiation, in particular the influence of UV-B radiation on broccoli was examined. Plants respond highly specific to environmental stimuli such as UV-B radiation and herbivory. UV-B radiation has a strong impact on the plants’ architecture and flavonoid contents, which can in turn influence plant-insect interactions. Phloem-feeding aphids can be negatively affected by UV-B mediated plant changes. However, a direct effect of UV radiation on the behaviour of herbivores is also evident. Mainly the number, composition and quality of herbivorous species as well as an exceeding of a certain infestation threshold determine the mode of plant changes. In conclusion, UV-B radiation has the potential to harden plants against herbivores and simultaneously increases the concentrations of valuable secondary metabolites for human nutrition in important crop species such as broccoli.
The bidirectional influence of parenchymal cells and cells of the immune system, especially of antigen-presenting and CD8\(^+\) T cells, in situations of putative auto- immune pathogenicity and degeneration was the main topic of this thesis. In the first part, the influence of human muscle cells on antigen-presenting cells was investigated. In inflammatory myopathies prominent infiltrates of immune cells containing T cells and antigen-presenting cells like macrophages and dendritic cells are present. The hypothesis was that human myoblasts have an inhibiting influence on these antigen-presenting cells under homeostatic conditions. A dysfunction or impairment under inflammatory circumstances might contribute to the development of myopathic conditions. The surface analysis of dendritic cells cocultured with myoblasts showed that immature dendritic cells could be driven into a reversible semi- mature state with significantly elevated levels of CD80. These dendritic cells were additionally characterized by their inhibiting function on T-cell proliferation. It was also shown that the lysates of healthy myoblasts could strongly enhance the phagocytic ability of macrophages, which could help with muscle regeneration and which might be disturbed in myositis patients. The second part of this thesis was about the clonal specificity of CD8\(^+\) T cells in a mouse model with genetically induced over-expression of PLP in oligodendrocytes. Here, we could show that the cytotoxic T lymphocytes, which had previously been shown to be pathogenic, were clonally expanded in the CNS of the transgenic mice. The amino acid sequences of the corresponding receptor chains were not identical, yet showed some similarities, which could mean that these clones recognize similar antigens (or epitopes of the same antigen). The knockout of PD-1 in this setting allowed for an analysis of the importance of tissue immune regulation. It became evident that the absence of PD-1 induced a larger number of clonal expansions in the CNS, hinting towards a reduced threshold for clonal disturbance and activation in these T cells. The expansions were, however, not pathogenic by themselves. Only in the presence of tissue damage and an antigenic stimulus (in our case the overexpression of PLP), the PD-1 limitation exacerbated the immune pathogenicity. Therefore, only in the presence of a “tissue damage signal”, the dyshomeostasis of T cells lacking PD-1 achieved high pathogenetic relevance. Finally, we investigated the pathogenetic role of CD8 T cells in Rasmussen encephalitis, a rare and chronic neurological disease mainly affecting children. The analysis of the T-cell receptor repertoire in Rasmussen encephalitis patients in the peripheral CD4\(^+\) and CD8\(^+\) T-cell compartments as well as the brain revealed the involvement of T cells in the pathogenicity of this disease. Many clonal expansions in the brain matched CD8\(^+\) T-cell expansions in the periphery on the sequence level. These putatively pathogenic clones could be visualized by immunohistochemistry in the brain and were found in close proximity to astrocytes and neurons. Additionally, the expanded clones could be found in the periphery of patients for at least one year.
The work of the previous chapters describes the role of Nipah virus (NiV) V and W proteins regarding their role in interferon antagonism and regulation of viral replication. Previous publications have shown that NiV encodes IFN antagonist activity in its V, W and C protein (Park et al., 2003b; Rodriguez et al., 2002). In order to study the effect of both NiV proteins in the context of a virus infection, recombinant Newcastle disease viruses (rNDVs) expressing NiV V or NiV W were constructed. As a control virus served rNDV expressing NDV V proteins, which behaved like wildtype NDV. Growth kinetic experiments demonstrated that rNDVs expressing NiV V or W grew to higher titers than rNDV expressing NDV V in human A549 cells. This result suggested that both NiV V and W were able to render the avian virus, which normally does not replicate well in human cells, into a better growing virus. This hypothesis was supported by the fact that all rNDVs grew similarly in avian DF1 or Vero cells. When rNDV-infected A549 cells were specifically stained for NiV V or W protein it was observed that V is localized in the cytoplasm whereas W could be predominantly found in the nucleus. This observation was in agreement with previous studies reporting a nucleus export signal (NES) for NiV V and a nuclear localization signal (NLS) for NiV W (Rodriguez et al., 2004; Shaw et al., 2005). The specific localization of each NiV protein has also been shown to contribute to different functions in terms of IFN antagonism (Shaw et al., 2005). Here, NiV V and W proteins caused a severe attenuation of the immune response in rNDV-infected human A549 and dendritic cells. The transcription of type I interferons and ISGs was significantly downregulated in the presence of NiV V and W proteins. As a consequence of the transcriptional block, there was also an inhibition at the level of translation (as seen for A549 cells) and the secretion of IFNs and cytokines/chemokines (as seen for DCs). In contrast, NDV V protein induced a host immune response. Both NiV V and W also displayed a strong inhibitory effect on the function DCs. DCs represent a very important cell class because they link the innate immune response to the adaptive immune response (Banchereau & Steinman, 1998). By downregulating the production and secretion of important cytokines/chemokines that are important for the activation of B and T lymphocytes, NiV V and W were able to disrupt that link. Interestingly, NiV W seemed to be a stronger inhibitor than NiV V in both A549 cells and DCs. Overall, it was demonstrated that NiV V and W were able to prevent the induction of the innate and adaptive host immune response cascade by inhibiting the transcription of immune genes in DCs and A549 cells. The second part of this work addressed the question whether NiV V and W proteins have a regulatory role in viral replication. This has been previously reported for Nipah virus itself (Sleeman et al., 2008) and other viruses (Atreya et al., 1998; Horikami et al., 1996; Witko et al., 2006). In order to study the ability of the V and W proteins of NiV to regulate viral transcription and/or replication, an existing NiV minireplicon assay was used (Halpin et al., 2004). Here, it was shown that NiV V and W (but not C) proteins significantly downregulated NiV minireplicon activity. The common N terminal region was shown to harbor the inhibitory activity. Co-immunoprecipitation experiments showed that both NiV V and W (but not C) were able to interact with NiV N, one component of the NiV polymerase. This result was supported by immunofluorescence experiments that revealed co-localization of NiV N with V and W. The binding of NiV V or W to NiV N occurred via their N terminus and more specifically amino acids 1-50. This suggested that V and W might inhibit viral replication by interacting with the viral polymerase resulting in a loss of function. Exact mechanisms still have to be elucidated.
In this work, a behavioural analysis of different mutants of the fruit fly Drosophila melanogaster has been carried out. Primarily, the gap climbing behaviour (Pick & Strauss, 2005) has been assayed as it lends itself for the investigation of decision making processes and the neuronal basis of adaptive behaviour. Furthermore it shows how basic motor actions can be combined into a complex motor behaviour. Thanks to the neurogenetic methods, Drosophila melanogaster has become an ideal study object for neurobiological questions. Two different modules of climbing control have been examined in detail. For the decision making, the mutant climbing sisyphus was analysed. While wild-type flies adapt the initiation of climbing behaviour to the width of the gap and the probability for a successful transition. climbing sisyphus flies initiate climbing behaviour even at clearly insurmountable gap widths. The climbing success itself is not improved in comparison to the wild-type siblings. The mutant climbing sisyphus is a rare example of a hyperactive mutant besides many mutants that show a reduced activity. Basic capabilities in vision have been tested in an optomotor and a distance-estimation paradigm. Since they are not affected, a defect in decision making is most probably the cause of this behavioural aberration. A second module of climbing control is keeping up orientation towards the opposite side of the gap during the execution of climbing behaviour. Mutants with a structural defect in the protocerebral bridge show abnormal climbing behaviour. During the climbing attempt, the longitudinal body axis does not necessarily point into the direction of the opposite side. Instead, many climbing events are initiated at the side edge of the walking block into the void and have no chance to ever succeed. The analysed mutants are not blind. In one of the mutants, tay bridge1 (tay1) a partial rescue attempt used to map the function in the brain succeeded such that the state of the bridge was restored. That way, a visual targeting mechanism has been activated, allowing the flies to target the opposite side. When the visibility of the opposing side was reduced, the rescued flies went back to a tay1 level of directional scatter. The results are in accord with the idea that the bridge is a central constituent of the visual targeting mechanism. The tay1 mutant was also analysed in other behavioural paradigms. A reduction in walking speed and walking activity in this mutant could be rescued by the expression of UAS-tay under the control of the 007Y-GAL4 driver line, which concomitantly restores the structure of the protocerebral bridge. The separation of bridge functions from functions of other parts of the brain of tay1 was accomplished by rescuing the reduced optomotor compensation in tay1 by the mb247-GAL4>UAS-tay driver. While still having a tay1-like protocerebral bridge, mb247-GAL4 rescue flies are able to compensate at wild-type levels. An intact compensation is not depended on the tay expression in the mushroom bodies, as mushroom body ablated flies with a tay1 background and expression of UAS-tay under the control of mb247-GAL4 show wild-type behaviour as well. The most likely substrate for the function are currently unidentified neurons in the fan-shaped body, that can be stained with 007Y-GAL4 and mb247-GAL4 as well.
The taphonomic and paleoecologic aspects of the Upper Hauterivian to Lower Barremian Agua de la Mula Member of the Agrio Formation (Neuquén Basin, Argentina) were studied in the frame of the sequence stratigraphic paradigm. The Agua de la Mula Member, a ca. 600 m thick succession of highly cyclic marine sediments was surveyed at two localities. Detailed bed-by-bed sedimentologic, stratigraphic, ichnologic, taphonomic and paleoecologic data collection allowed a precise paleoenvironmental, stratigraphic, taphonomic and synecologic interpretation, in a controlled sequence stratigraphic framework. The main architectural stratigraphic component is the Starvation-Dilution Sequence, interpreted as a the effect of a sixth-order, Milankovitch precession-driven cycle. Dilution hemisequences are siliciclastic-dominated and show evidence of depth changes. Starvation hemisequences show a diverse variation of mixed carbonate-siliciclastic facies that is linked to sequence stratigraphy. Ammonite-based biostratigraphy was revised and new knowledge proposed. The stratigraphic framework was improved by combining biostratigraphy, sequence stratigraphy and event stratigraphy. Nine main sequences were described, linked to other stratigraphic markers and correlated with other sequence stratigraphic charts. Several orders of cyclicity were inferred. Third- and fourth-order sequences are the major sequences, not subordinated to higher hierarchies (lower order). Precession, obliquity, and short and long eccentricity cycles of the Milankovitch band are proposed. Among the different sequence stratigraphic models the transgression-regression model fits the majority of the sequences described in this work. The depositional-sequence model could be applied only to the first third-order sequence, in which the true sequence boundary is identifiable. Starvation-dilution sequences, however, are composed by to components that are not completely explained by those models. Starvation hemisequences developed in intermediate to deep settings record the transgressive phase as well as the earLy regressive one without visible stratigraphic boundaries. 112 samples with 22,572 individuals were grouped into fifteen fossil associations and one assemblage that reflect the interaction of different factors: age, position in major, medium and starvation dilution sequences and, linked to sequence stratigraphy, depth, oxygen availability, rate of terrigenous input, water agitation, and substrate conditions. Temporary possible reduction in oxygen content is inferred based on all sources of available evidence. Organic buildups are briefly described and their development interpreted in terms of the sequence stratigraphic framework. Vertical patterns of replacement of fossil associations are described and related to sequence stratigraphy. Five types of skeletal concentrations represent the diversity of coquinas decribed in this study. Type 1, 2, 4 and 5 correspond to starvation hemisequences deposited in progressively shallower settings, from basin to inner ramp. Type 3 is embedded into dilution hemisequences and inferred to be linked to shell bed type I of Kidwell (1985). Types 1 and 2 correspond to transgression, maximum flooding and early regression without distinction. Type 4A as well as Type 5 are interpreted as onlap shell beds (Kidwell 1991a) or early TST shell beds (Fürsich and Pandey 2003). Type 4B corresponds to the MFZ shell bed (Fürsich and Pandey 2003) or mid-cycle shell bed (Abbott 1997), while Type 4C to the downlap shell bed (Kidwell 1991a). Time-averaging of shell beds was assessed with precision as the time involved in the deposition of the starvation hemisequences could be inferred. All shell beds comprise within-habitat assemblages forming within a few thousand years, with little environmental condensation. The fossilization of the marine calcareous shells is modelled as a series of steps called windows: environmental, destructional, burial and diagenetic. The “diagenetic window” is the most relevant. Connected to this it is proposed that carbonate dissolution is the primary control on the development of shell beds, as has been proposed before (Fürsich 1982; Fürsich and Pandey 2003). The interpretative power resulting from combining several lines of evidence, e.g., facies analysis, sequence stratigraphy, biostratigraphy, trace fossil analysis, paleoecology and taphonomy, and unravelling their multiple relationships, are the most relevant conclusions of this study.
The observation of neutrino masses and lepton mixing has highlighted the incompleteness of the Standard Model of particle physics. In conjunction with this discovery, new questions arise: why are the neutrino masses so small, which form has their mass hierarchy, why is the mixing in the quark and lepton sectors so different or what is the structure of the Higgs sector. In order to address these issues and to predict future experimental results, different approaches are considered. One particularly interesting possibility, are Grand Unified Theories such as SU(5) or SO(10). GUTs are vertical symmetries since they unify the SM particles into multiplets and usually predict new particles which can naturally explain the smallness of the neutrino masses via the seesaw mechanism. On the other hand, also horizontal symmetries, i.e., flavor symmetries, acting on the generation space of the SM particles, are promising. They can serve as an explanation for the quark and lepton mass hierarchies as well as for the different mixings in the quark and lepton sectors. In addition, flavor symmetries are significantly involved in the Higgs sector and predict certain forms of mass matrices. This high predictivity makes GUTs and flavor symmetries interesting for both, theorists and experimentalists. These extensions of the SM can be also combined with theories such as supersymmetry or extra dimensions. In addition, they usually have implications on the observed matter-antimatter asymmetry of the universe or can provide a dark matter candidate. In general, they also predict the lepton flavor violating rare decays mu -> e gamma, tau -> mu gamma and tau -> e gamma which are strongly bounded by experiments but might be observed in the future. In this thesis, we combine all of these approaches, i.e., GUTs, the seesaw mechanism and flavor symmetries. Moreover, our request is to develop and perform a systematic model building approach with flavor symmetries and to search for phenomenological implications. This provides a new perspective in model building since it allows us to screen models by its predictions on the theoretical and phenomenological side, i.e., we can apply further model constraints to single out a desired model. The results of our approach are, e.g., diverse lepton flavor and GUT models, a systematic scan of lepton flavor violation, new mass matrices, a new understanding of lepton mixing angles, a general extension of the idea of quark-lepton complementarity theta_12=pi/4-epsilon/sqrt{2} and for the first time the QLC relation in an SU(5) GUT.
Semiconductor Quantum Dots (QDs) have been attracting immense interest over the last decade from both basic and application-orientated research because of their envisioned use as fundamental building blocks in non-classical device architectures. Their presumable ease of integration into existing semiconductor technology has bought them the reputation of being cost-efficiently scalable and renders them a place among the top candidates in a wide range of proposed quantum logic and quantum information processing schemes. These include the highly acclaimed use of QD as triggered sources of single pairs of entangled photons, which is a key ingredient of most of the intensivly investigated optical quantum cryptography operations. A big obstacle towards these goals are the pronounced asymmetries that are intrinsically present in all currently availabe semiconductor QD systems. They are a natural by-product that stems from the employed self-assembled growth methods and manifest in various forms such as shape-asymmetry, inhomogeneous strain distribution within the QD and concomittant piezo-elecric fields. These asymmetries in return give rise to distinct anisotropies in the optical properties of QDs, which in fact render their optical response classic. For device oriented research these anisotropies are therefore typically considered unwanted and actively researched to be controlled. They are, however, interesting from a fundamental point of view, as anisotropic QDs basically provide a testbed system for fundamental atom-like quantum physics with non-centrosymmetric potentials. As shall be shown in the current work, this gives rise to novel and interesting physics in its own right. Employing photoluminescence spectroscopy (PL) we investigate the optical anisotropy of the radiative recombination of excitons confined to CdSe/ZnSe QDs. This is done by angle-dependent polarization-resolved PL. We demonstrate experimentally that the electron-hole exchange interaction in asymmetric QDs gives rise to an effective conversion of the optical polarization from linear to circular and vice versa. The experiment is succesfully modeled in the frame of an exciton pseudospin-formalism that is based on the exchange induced finestructure splitting of the radiative excitonic states and unambiguously proves that the observed polarization conversion is the continuous-wave equivalent to quantum beats between the exchange split states in the time domain. These results indicate that QDs may offer extended functionality beyond non-classical light sources in highly integrated all-optical device schemes, such as polarization converters or modulators. In a further extension we apply the exciton pseudospin-formalism to optical alignment studies and demonstrate how these can be used to directly measure the otherwise hidden symmetry distribution over an ensemble of QDs. This kind of measurement may be used on future optical studies in order to link optical data more directly to structural investigations, as it yields valuable information on capped QDs that cannot be looked at directly by topological methods. In the last part of this work we study the influence of an in-plane magnetic field on the optical anisotropy. We find that the optical axis of the linear polarization component of the photoluminescence signal either rotates in the opposite direction to that of the magnetic field or remains fixed to a given crystalline direction. A qualitative theoretical analysis based on the exciton pseudospin Hamiltonian unambiguously demonstrates that these effects are induced by isotropic and anisotropic contributions to the heavy-hole Zeeman term, respectively. The latter is shown to be compensated by a built-in uniaxial anisotropy in a magnetic field B=0.4 T, resulting in an optical response that would be expected for highly symmetric QDs. For a comprehensive quantitative analysis the full heavy-hole exciton k.p-Hamiltonian is numerically calculated and the resulting optical polarization is modeled. The model is able to quantitatively describe all experimental results using a single set of parameters. From this model it is explicitly seen that a optical response characteristic for high symmetry QDs may be obtained from an ensemble of asymmetric QDs without a crossing of the zero-field bright exciton states, which was required for application of QDs in non-classical light sources. It is clearly demonstrated that any scheme using in-plane magnetic fields to symmetrize the optical response has to take into account at least four optically active states instead of the two observed in the absence of magnetic fields. These findings may explain some of the major disagreement on recent entanglement studies in asymmetric QDs, as models that do not take the above result into account cannot be a priori expected to provide reliable results on excitonic Bell states.
Since its popularization due to Randall and Sundrum (RS) one decade ago, and in connection with the $AdS/CFT$ correspondence in particular, 5D warped background spacetime has been one of the most fruitful new ideas in physics beyond the standard model (SM), leading to new insights into symmetry breaking and the properties of strongly interacting theories inaccessible to direct perturbative calculations, while at the same time relating gravity to phenomenological model building. This has, among others, led to a renewed interest in models of electroweak symmetry breaking without physical scalar fields in the guise of so-called 'warped higgsless' models, which could provide an alternative to the famed Higgs mechanism of electroweak symmetry breaking which is part of the Standard Model of particle physics. However, little emphasis was put on reconciling these models with the strong evidence from astrophysical observations that one or several new, as yet unknown, stable particle species exist which form the cold dark matter content of the universe. The nature of dark matter and electroweak symmetry breaking are among the most prominent puzzles subject to experimental scrutiny at the Tevatron, direct search experiments, and in the near future at the LHC, which compels us the believe that both issues should be addressed together in any alternative scenario beyond the Standard Model. In this thesis we have investigated phenomenological implications which arise for cosmology and collider physics when the electroweak symmetry breaking sector of warped higgsless models is extended to include warped supersymmetry with conserved $R$ parity. The goal was to find the simplest supersymmetric extension of these models which still has a realistic light spectrum including a viable dark matter candidate. To accomplish this, we have used the same mechanism which is already at work for symmetry breaking in the electroweak sector to break supersymmetry as well, namely symmetry breaking by boundary conditions. While supersymmetry in five dimensions contains four supercharges and is therefore directly related to 4D $\mathcal{N}=2$ supersymmetry, half of them are broken by the background leaving us with ordinary $\mathcal{N}=1$ theory in the massless sector after Kaluza-Klein expansion. We thus use boundary conditions to model the effects of a breaking mechanism for the remaining two supercharges. The simplest viable scenario to investigate is a supersymmetric bulk and IR brane without supersymmetry on the UV brane. Even though parts of the light spectrum are effectively projected out by this mechanism, we retain the rich phenomenology of complete $\mathcal{N}=2$ supermultiplets in the Kaluza-Klein sector. While the light supersymmetric spectrum consists of electroweak gauginos which get their $\mathcal{O}(100\mbox{ GeV})$ masses from IR brane electroweak symmetry breaking, the light gluinos and squarks are projected out on the UV brane. The neutralinos, as mass eigenstates of the neutral bino-wino sector, are automatically the lightest gauginos, making them LSP dark matter candidates with a relic density that can be brought to agreement with WMAP measurements without extensive tuning of parameters. For chargino masses close to the experimental lower bounds at around $m_{\chi^+}\approx 100\dots 110$ GeV, the dark matter relic density points to LSP masses of around $m_\chi\approx 90$ GeV. At the LHC, the standard particle content of our model shares most of the key features of known warped higgsless models. We have performed Monte Carlo simulations of warped higgsless LSP and NLSP production at a benchmark point using \nameomega/\namewhizard, concentrating on $\ptmiss$ in association with third generation quarks. After background reduction cuts on the quark momenta and angles, we get hadronic cross sections of $\sigma>100\mbox{ fb}$ at $14\mbox{ TeV}$ with characteristic $\ptmiss$ distributions for $\chi\chi t\overline{t}$ final states, while the final states with $b\overline{b}$ pairs have much lower event rates and shapes which are hard to discern in experiments. Our results suggest that the discovery of warped higgsless LSP dark matter at the LHC via missing energy is within reach for the first few $\mbox{ fb}^{-1}$ at $14$ TeV if $b$ and in particular $t$ identification is reliable.
Studies on platelet cytoskeletal dynamics and receptor regulation in genetically modified mice
(2009)
Platelets are produced by bone marrow megakaryocytes in a process involving actin dynamics. Actin-depolymerizing factor (ADF) and cofilin are actin-binding proteins that act as key regulators in actin turnover by promoting filament severing and depolymerization. The overall significance of ADF/cofilin function and actin turnover in platelet formation is presently unclear. In the first part of this thesis, platelet formation and function were studied in mice constitutively lacking ADF and/or mice with a conditional deficiency (Cre/loxP) in n-cofilin. To delete cofilin exclusively in megakaryocytes and platelets, cofilinfl/fl mice were crossed with PF4 (platelet factor 4)-Cre mice. While a single-deficiency in ADF or n-cofilin resulted in no or only a minor platelet formation defect, respectively, a double-deficiency in ADF and n-cofilin led to an almost complete loss of platelets. Bone marrow megakaryocytes of ADF/n-cofilin-deficient mice showed defective platelet zone formation. Interestingly, in vitro and ex vivo megakaryocyte differentiation revealed reduced proplatelet formation and absence of platelet-forming swellings. These data establish that ADF and n-cofilin have redundant but essential roles in the terminal step of platelet formation in vitro and in vivo. In the second part of the thesis, mechanisms underlying cellular regulation of the major platelet collagen receptor, glycoprotein VI (GPVI), were studied. GPVI mediates platelet activation on exposed subendothelial collagens at sites of vascular injury, and thereby contributes to normal hemostasis but also to occlusion of diseased vessels in the setting of myocardial infarction or stroke. Thus, GPVI is an attractive target for anti-thrombotic therapy, particularly because previous studies have shown that anti-GPVI antibodies induce irreversible down-regulation of the receptor in circulating platelets by internalization and ectodomain shedding. Metalloproteinases of the ADAM (a disintegrin and metalloproteinase domain) family are suspected to mediate this ectodomain shedding, but in vivo evidence for this is lacking. To study the mechanism of GPVI regulation in vivo, two mouse lines, Gp6 knock-out and Adam10fl/fl, PF4-Cre mice, were generated and in addition low TACE (TNFalpha converting enzyme) mice were analyzed. It was shown that GPVI can be cleaved in vitro by ADAM10 or TACE depending on the shedding-inducing signaling pathway. Moreover, GPVI was down-regulated in vivo upon antibody injection in ADAM10-deficient and low TACE mice suggesting that either both or an additional metalloproteinase is involved in GPVI regulation in vivo.
Land plants must control the transpiration water stream and balance it with carbon dioxide uptake for optimal photosynthesis. A highly specialized type of plant cell called guard cells have evolutionary appeared which are suited for this complicated purpose. Guard cells are located by pairs on aerated plant surface and form stomata – structural units, which represent highly regulated “watergate” (Roelfsema and Hedrich, 2005). Guard cells sense many environmental and internal plant-derived stimuli and by changing degree of their swelling tightly regulate diffusion of water vapor and other gases. Cell processes taking place in stomata during their movements had been a subject of intensive investigation for more than three decades (Schroeder et al., 2001; Assmann and Shimazaki, 1999). With use of electrophysiological technique the basic processes underlying stomatal movements were described (Thiel et al., 1992; Dietrich et. al., 2001; Roelfsema and Hedrich, 2005). Another set of questions arised between plant biologists is how the signals affecting stomatal aperture are transduced in guard cells starting from perception by receptor structures and ending on the osmodynamic motor components. Introduction of fluorescent microspectroscopy technique allowed to characterize some Ca2+ and H+-based signaling events, taking place in the cytoplasm during stomata function. Most of the processes, taking place in stomata were characterized in guard cell preparations, such as strips of isolated leaf epidermis or guard cell protoplasts, - cells with enzymaticaly digested cell walls. Some experimental observations although point that reactions of guard cells located in their natural environment, leaves of intact plants can differ from those could be registered in preparations. These deviations might be explained by the modulation of guard cell function by apoplastic factors originating from surrounding tissues like mesophyll or leaf epidermis (Roelfsema and Hedrich, 2002). On the other hand registration of physiological responses in prepared tissues may also contain possible artifacts, related to the preparation procedures. The aim of the experimental work presented here was to investigate the cell signaling events, taking place in guard cells upon plant stress hormone abscisic acid (ABA) and some other stimuli action. Abscisic acid is a compound that synthesized in plant roots upon drought and closes stomata in the leaf to prevent the plant organism from excessive water loss. Previous studies on guard cell of isolated epidermis and guard cell protoplasts showed, that ABA induces stomatal closure via activation of plasma membrane anion channels (Grabov et al., 1997; Pei et al, 1997). Anion channels are known to be activated by elevated 2 concentrations of cytoplasmic Ca2+ [Ca2+]cyt (Schroeder and Hagiwara, 1989; Hedrich et al., 1990). Application of Ca2+-sensitive fluorescent probes revealed [Ca2+]cyt increases in guard cells upon ABA action (McAinsh et al., 1990). This observation led to suggestion that [Ca2+]cyt directly participate in the transduction of ABA signal in guard cells. Although no direct evidences for co-occurrence of [Ca2+]cyt rises and following activation of anion channels upon ABA action was not presented until yet. Results of experimental work performed on intact Vicia faba, Commelina communis and Nicotiana plumbagnifolia plants showed that guard cells of intact plant leaves respond with transient activation of plasma membrane anion channels upon perception of ABA. Kinetics of the response is highly reproducible and seemed to be conserved between species. Although despite clear generation of anion current transients, no [Ca2+]cyt increases could be recorded with using fluorescent probe Fura-2 microinjected into the cytoplasm. Together with results of later study on intact Nicotiana tabacum guard cells, reported obligatory [Ca2+]cyt increases which were desynchronized with anion current transients (Marten et al., 2007b) this, may indicate that [Ca2+]cyt increases are not necessary component of ABA signal transduction pathway. Together with absence of the effect of cytoplasm-delivered Ca2+- mobilizing agents IP3, IP6 and NAADP on anion currents these data may suppose that role of [Ca2+]cyt in ABA signaling must be reassessed. Further interest represented characterization of [Ca2+]cyt signaling and homeostasis in intact guard cells comparing with those in prepared cells. Experiments revealed strong deviations in [Ca2+]cyt behavior between different measuring systems. While guard cells of intact plants were able to strictly maintain [Ca2+]cyt level upon experimental shifting of [Ca2+]cyt level in either direction of elevation or decrease, cells of isolated epidermis showed complete absence of such ability. Guard cell protoplasts showed even weaker [Ca2+]cyt regulation ability and were capable of low physiological [Ca2+]cyt levels maintaining only at depolarized membrane potentials. Apart to these differences, prepared guard cells showed also for-time less activation of anion currents by experimentally imposed [Ca2+]cyt increases. These data strongly suggest that registered in guard cell preparations [Ca2+]cyt signals may contain significant part of artifacts and must be carefully used for the building of models of guard cells signaling. Further experimental investigations are strongly required for understanding guard cell functioning, especially with relation of vacuoles participation. The experimental work was done by the author in the period from october 2001 until november 2004 under supervision of Professor Dr. Rainer Hedrich in laboratory of molecular plant physiology and biophysics at Julius-Maximillians University of Würzburg, Würz3 burg, Federal Republic of Germany. Scientific coordinator of the Ph. D. project is Dr. Max Robert Gustaaf Roelfsema, University of Würzburg. Most of experimental results, presented here (chapter III) are also published elsewhere (Roelfsema et al., 2004; Langer et al., 2004; Levchenko et al., 2005, 2008). Chapter I intend to shortly introduce the reader into the field of guard cell research and point out the current level of understanding regarding this branch of plant research. Special attention is given to description of guard cell ion channels, their function and regulation, including the mechanisms of Ca2+-, H+- and phosphorylation-based signaling. This section is preceded by a short history of guard cell research and explains the actuality of presented work. In chapter II experimental techniques, methods and data processing approaches, used in the presented work are described. Technique used for electrophysiological registrations on intact plant leaves were used before and described in more details by Roelfsema et al. (2001). Fluorescent microspectroscopy technique was for the first time applied to intact plant leaves in this work and described in more details including calibration of Fura-2 based measurements. Chapter III presents the major results of the experimental work. In chapter IV the experimental results are discussed and put into context with current knowledge of guard cell function knowledge. Finally, remarks on perspectives of guard cell signaling research are drawn.
The Upper Bajocian-Bathonian Kashafrud Formation is a thick package of siliciclastic sediments that crops out in NE Iran from the southeast, near the Afghanistan border, to north- northwestern areas around the city of Mashhad. The thickness ranges from less than 300 m in a deltaic succession (Kuh-e-Radar) to more than 2500 m in the Maiamay area, but the normal thickness in Ghal-e-Sangi, Kol-e-Malekabad, and Fraizi areas is about 1200-1300 m. It is the fill of an elongated basin, which extended for more than 200 km in NW-SE direction and a width of at least 50 km along the southern margin of the Koppeh Dagh. Prior to this study, little information existed about the sedimentary environments and other characters, especially the geometry of the basin. Exact biostratigraphic data from the top of the Kashafrud Formation were rare. Based on the macrofauna from the lower part of the overlying Chamanbid Formation the upper boundary of the Kashafrud Formation had been attributed to the Late Bathonian and/or Early Callovian, but now the upper limit of the Kashafrud Formation is defined as Late Bathonian in age, based on ammonite biostratigraphy. Except for chapter one, which deals with the introduction and related sub-titles, in the following chapters, step by step, field observations and data were surveyed according to the questions to solve. In order to reconstruct the facies architecture and the geometry of the basin, a number of sections have been logged in detail (see chapter 3, “The sections”). The exact biostratigraphic setting is discussed in chapter 4 (“Biostratigraphy”). Sedimentary environments range from non-marine alluvial fans and braided rivers in the basal part of the succession to deltas, storm-dominated shelf, slope and deep-marine basin. The latter comprises the largest part of the basin fill, consisting of monotonous mudstones, siltstones and proximal to distal turbidities. The only continuous carbonate unit (~30 m) locally formed at Tappenader. Other localities in which thin fossil-bearing carbonate strata occur are Torbat-e-Jam (benthic fauna) and, to a lesser extent, Ghal-e-Sangi. These rare shallow-water carbonates, which also contain corals, represent only short intervals (see chapter 5,” Facies association and sedimentary environments”). Relative changes in sea level were reconstructed on the basis of deepening- and shallowing-upward trends. Sequence boundaries and parasequences have been distinguished and analyzed in chapter 6 (“Sequence stratigraphy”). In most areas, the basin rapidly evolved from a shallow marine, transgressive succession to a deep-marine, basinal succession. The only area where shallow conditions persisted from the Late Bajocian to the Late Bathonian, and even into the Early Callovian is the Kuh-e-Radar area which corresponds to a fan-delta setting. A trace fossil analysis has been carried out to obtain additional evidence on the bathymetry of the basin (see chapter 7, “Ichnology”). Altogether 29 ichnospecies belonging to 15 ichnogenera have been identified, as well as 10 ichnogenera, which were determined only at genus level. They can be grouped in the well-known “Seilacherian ichnofacies”. Very high subsidence rates and strong lateral thickness variations suggest that the Kashafrud Formation is a rift related basin that formed as the eastern extension of the South Caspian Basin. The basin evolution is reviewed, the eastern and western continuations of the basin were checked in the field and also in the literature (see chapter 8, “Basin evolution”). In all, the present study provided new insights into the development of the Kashafrud Formation, e.g. more biostratigraphic data from the base and the top of the succession, a relatively complete picture of the trace fossil associations, a better recognition and reconstruction of the sedimentary environments in different parts of the basin. Finally this research project will be a good basis for further investigations, especially towards the west, as parts of the Kashafrud Formation are source rocks of a hydrocarbon reservoir in NE Iran.
Several lines of evidence implicate a dysregulation of tryptophan hydroxylase (TPH)-dependent serotonin (5-HT) synthesis in emotions and stress and point to their potential relevance to the etiology and pathogenesis of various neuropsychiatric disorders. However, the differential expression pattern of the two isoforms TPH1 and TPH2 which encode two forms of the rate-limiting enzyme of 5-HT synthesis is controversial. Here, a comprehensive spatio-temporal analysis clarifies TPH1 and TPH2 expression during pre- and postnatal development of the mouse brain and in adult human brain as well as in peripheral organs including the pineal gland. Four different methods (real time PCR, in situ hybridization, immunohistochemistry and Western blot analysis) were performed to systematically control for tissue-, species- and isoform-specific expression on both the pre- and posttranslational level. TPH2 expression was consistently detected in the raphe nuclei, as well as in fibres in the deep pineal gland and in the gastrointestinal tract. Although TPH1 expression was found in these peripheral tissues, no significant TPH1 expression was detected in the brain, neither during murine development, nor in mouse and human adult brain. Also under conditions like stress and clearing the tissue from blood cells, no changes in expression levels were detectable. Furthermore, the reuptake of 5-HT into the presynaptic neuron by the serotonin transporter (SERT) is the major mechanism terminating the neurotransmitter signal. Thus, mice with a deletion in the Sert gene (Sert KO mice) provide an adequate model for human affective disorders to study lifelong modified 5-HT homeostasis in interaction with stressful life events. To further explore the role of TPH isoforms, Tph1 and Tph2 expression was studied in the raphe nuclei of Sert deficient mice under normal conditions as well as following exposure to acute immobilization stress. Interestingly, no statistically significant changes in expression were detected. Moreover, in comparison to Tph2, no relevant Tph1 expression was detected in the brain independent from genotype, gender and treatment confirming expression in data from native animals. Raphe neurons of a brain-specific Tph2 conditional knockout (cKO) model were completely devoid of Tph2-positive neurons and consequently 5-HT in the brain, with no compensatory activation of Tph1 expression. In addition, a time-specific Tph2 inducible (i) KO mouse provides a brain-specific knockdown model during adult life, resulting in a highly reduced number of Tph2-positive cells and 5-HT in the brain. Intriguingly, expression studies detected no obvious alteration in expression of 5-HT system-associated genes in these brain-specific Tph2 knockout and knockdown models. The findings on the one hand confirm the specificity of Tph2 in brain 5-HT synthesis across the lifespan and on the other hand indicate that neither developmental nor adult Tph2-dependent 5-HT synthesis is required for normal formation of the serotonergic system, although Tph1 does not compensate for the lack of 5-HT in the brain of Tph2 KO models. A further aim of this thesis was to investigate the expression of the neuropeptide oxytocin, which is primarily produced in the hypothalamus and released for instance in response to stimulation of 5-HT and selective serotonin reuptake inhibitors (SSRIs). Oxytocin acts as a neuromodulator within the central nervous system (CNS) and is critically involved in mediating pain modulation, anxiolytic-like effects and decrease of stress response, thereby reducing the risk for emotional disorders. In this study, the expression levels of oxytocin in different brain regions of interest (cortex, hippocampus, amygdala, hypothalamus and raphe nuclei) from female and male wildtype (WT) and Sert KO mice with or without exposure to acute immobilization stress were investigated. Results showed significantly higher expression levels of oxytocin in brain regions which are involved in the regulation of emotional stimuli (amygdala and hippocampus) of stressed male WT mice, whereas male Sert KO as well as female WT and Sert KO mice lack these stress-induced changes. These findings are in accordance with the hypothesis of oxytocin being necessary for protection against stress, depressive mood and anxiety but suggest gender-dependent differences. The lack of altered oxytocin expression in Sert KO mice also indicates a modulation of the oxytocin response by the serotonergic system and provides novel research perspectives with respect to altered response of Sert KO mice to stress and anxiety inducing stimuli.
In this thesis, soft x-ray absorption spectroscopy (XAS) and resonant inelastic x-ray scattering (RIXS) studies of the electronic structure of selected organic molecules and liquids were carried out. The first part focuses on the used experimental techniques and the development of the instrumentation necessary for these studies, namely a soft x-ray spectrometer, and a temperature-controlled flow-through liquid cell. The former was optimized by a special analytical ray tracing method developed exclusively for this purpose. Due to its high transmission, the spectrometer facilitates a novel experimental approach of recording comprehensive 'RIXS maps', which are 2-dimensional plots of x-ray scattering intensities as a function of both, excitation and emission photon energy. The liquid cell extends these possibilities to the study of liquids, especially the interaction of molecules in liquids and their chemical reactions under well-controlled conditions. Organic molecules have attracted considerable attention in the last decade. The intense research activities related to these materials have two main motivations: on the one hand, organic molecules have a technological application as building blocks of organic semiconductors, while, on the other hand, organic molecules are the functional elements in biological systems. In order to cost-effectively produce optimized organic electronic devices, a fundamental knowledge of the electronic properties of the organic molecules interface is necessary. Therefore, many studies of the electronic structure of potential candidates for organic electronics exist. Two of these candidates, namely C60 and well-ordered multilayers PTCDA on a Ag(111) surface are investigated in this thesis. For the study of C60 molecules, a comprehensive 'RIXS map' was recorded and analyzed. The RIXS map taken in only 25 minutes allows a quantitative analysis of energy losses, yielding for example the HOMO-LUMO distance. It also identifies a core-excitonic state and facilitates a quantitative comparison of its binding energy with that of valence excitons in C60. Furthermore, decay channel-selective partial fluorescence yield XAS spectra can be extracted from the RIXS map, yielding information on the population of the core-excitonic state as a function of excitation energy. As a second model system of organic molecules relevant for organic electronics, PTCDA was chosen. The complex electronic structure of the occupied states of a highly ordered, flat-lying PTCDA multilayer on a Ag(111) surface was investigated by symmetry-resolved resonant x-ray emission spectroscopy. The rapidly occurring beam damage effects were characterized on the basis of irradiation-time dependent series of C and O x-ray emission spectra. Upon varying the excitation energy and emission geometry, atom- and symmetry-specific carbon K emission spectra with negligible beam damage effects were obtained that allow to distinguish between electronic states with sigma and pi symmetry. A density functional theory calculation of the PTCDA molecule reproduces the energy positions of the most prominent emission features remarkably well. In addition, the energy positions of the sigma and pi emissions agree well with the calculated energies of the respective orbitals. In order to shed light on the second aspect of organic molecules, namely their role in biological systems, first a detailed investigation of the electronic structure and proton dynamics of liquid water as the medium of most chemical and biochemical reactions was carried out. Therefore, a comprehensive oxygen K RIXS map of liquid water was recorded and analyzed in great detail. A temperature-dependent comparison with XAS and RIXS data of D2O, NaOH, and NaOD leads to the conclusion, that ultra-fast dissociation takes place in liquid water on the timescale of the oxygen 1s core hole lifetime, resulting in a characteristic spectral contribution in the RIXS spectra. The dissociation is promoted by intact hydrogen bonds with neighboring molecules. In consequence, the rate of dissociation directly depends on the initial hydrogen bond configuration. In the next step towards biologically relevant systems, the nitrogen K edges of the amino acids glycine and histidine were investigated in powderous form as well as in their native environment, namely in aqueous solution. X-ray absorption and emission spectra of the aqueous solutions were analyzed at pH-values of 6 and for glycine also at pH 12 and compared to the spectra of powders. A pH-value of 12 causes deprotonation of the amino group, leading to significant changes in the nitrogen spectra as compared to pH 6. The results from these four examples demonstrate that a wealth of novel information can be obtained by using the new experimental tools developed in this thesis, namely a highly sensitive x-ray spectrometer and a flow-through liquid cell.
Aim of Investigation: The neurological manifestations of Fabry’s disease, a rare, X-linked, multisystem disorder caused by alpha-galactosidase A deficiency and globotriosylceramide (Gb3) accumulation, include both peripheral and central nervous system symptoms. Here we evaluated a prospectively recruited cohort of patients with Fabry’s disease for pain, small nerve fiber function, and skin innervation. Methods: 66 patients (31 male and 35 female) were enrolled,31 patients were on ERT. All patients underwent quantitative sensory testing (QST), electrophysiological examination, and extra- and transcranial Doppler sonography. For pain and mood assessment standardized questionnaires were used. Skin biopsies were performed at the left distal leg in 38 patients for intraepidermal nerve fiber density (IENFD) assessment. Results: Age at examination did not differ significantly between women (40.2+/-16.2 years) and men (38.9+/-13.8; n.s.). 29/31 male and 19/35 female patients complained of acroparesthesias or neuropathic pain. QST abnormalities indicative of small fiber impairment were found in 26/31 male and 28/35 female patients. Electrophysiological examination of large fibers and autonomic fibers revealed pathological findings in 11/31 male and 3/35 female patients. All patients had normal Doppler sonography results. Indicators for depression were present in 14/31 male and 10/35 female patients. 20/31 male and 18/35 female patients had a skin biopsy, the IENFD was significantly reduced in male (2.0+/-2.8 fibers/mm) compared with female patients (6.7 +/- 4.4 fibers/mm). In 10 patients free from neurological symptoms, QST and IENFD abnormalities were still detected. Follow up examination after one year in 12 patients under ERT (2.1+/-1.7 years) showed improvement in some symptoms and in QST and neurophysiology in six patients with normal renal function. 20/35 female patients older than 40 y had concomitant diseases, while none of the 18 younger female patients did. The corresponding radio in male patients was 5/19 (>=40y) and 2/13 (<40y) respectively. Conclusions: Neuropathic pain and sensory deficits of the distal extremities are common in patients with Fabry’s disease. QST and IENFD analysis are important for early diagnosis of nerve involvement in Fabry’s disease. Small fiber function may improve under ERT in patients without severe renal impairment.
Abstract: Inbreeding depression, asymmetries in costs or benefits of dispersal, and the mating system have been identified as potential factors underlying the evolution of sex-biased dispersal. We use individual-based simulations to explore how the mating system and demographic stochasticity influence the evolution of sex-specific dispersal in a metapopulation with females competing over breeding sites, and males over mating opportunities. Comparison of simulation results for random mating with those for a harem system (locally, a single male sires all offspring) reveal that even extreme variance in local male reproductive success (extreme male competition) does not induce male-biased dispersal. The latter evolves if the between-parch variance in reproductive success is larger for males than females. This can emerge due to demographic stochasticity if the habitat patches are small. More generally, members of a group of individuals experiencing higher spatio-temporal variance in fitness expectations may evolve to disperse with greater probability than others.
Background: Male killing endosymbionts manipulate their arthropod host reproduction by only allowing female embryos to develop into infected females and killing all male offspring. Because the resulting change in sex ratio is expected to affect the evolution of sex-specific dispersal, we investigated under which environmental conditions strong sex-biased dispersal would emerge, and how this would affect host and endosymbiont metapopulation persistence. Results: We simulated host-endosymbiont metapopulation dynamics in an individual-based model, in which dispersal rates are allowed to evolve independently for the two sexes. Prominent male-biased dispersal emerges under conditions of low environmental stochasticity and high dispersal mortality. By applying a reshuffling algorithm, we show that kin-competition is a major driver of this evolutionary pattern because of the high within-population relatedness of males compared to those of females. Moreover, the evolution of sex-specific dispersal rescues metapopulations from extinction by (i) reducing endosymbiont fixation rates and (ii) by enhancing the extinction of endosymbionts within metapopulations that are characterized by low environmental stochasticity. Conclusion: Male killing endosymbionts induce the evolution of sex-specific dispersal, with prominent male-biased dispersal under conditions of low environmental stochasticity and high dispersal mortality. This male-biased dispersal emerges from stronger kin-competition in males compared to females and induces an evolutionary rescue mechanism.
From the introduction: It is not always easy to define what a word means. We can choose between a variety of possibilities, from simply pointing at the correct object as we say its name to lengthy definitions in encyclopaedias, which can sometimes fill multiple pages. Although the former approach is pretty straightforward and is also very important for first language acquisition, it is obviously not a practical solution for defining the semantics of the whole lexicon. The latter approach is more widely accepted in this context, but it turns out that defining dictionary and encyclopaedia entries is not an easy task. In order to simplify the challenge of defining the meaning of words, it is of great advantage to organize the lexicon in a way that the structure in which the words are integrated gives us information about the meaning of the words by showing their relation to other words. These semantic relations are the focal point of this paper. In the first chapter, different ways to describe meaning will be discussed. It will become obvious why semantic relations are a very good instrument to organizing the lexicon. The second chapter deals with WordNet, an electronic lexical database which follows precisely this approach. We will examine the semantic relations which are used in WordNet and we will study the distinct characteristics of each of them. Furthermore, we will see which contribution is made by which relation to the organization of the lexicon. Finally, we will look at the downside of the fact that WordNet is a manually engineered network by examining the shortcomings of WordNet. In the third chapter, an alternative approach to linguistics is introduced. We will discuss the principles of corpus linguistics and, using the example of the British National Corpus, we will consider possibilities to extract semantic relations from language corpora which could help to overcome the deficiencies of the knowledge based approach. In the fourth chapter, I will describe a project the goal of which is to extend WordNet by findings from cognitive linguistics. Therefore, I will discuss the development process of a piece of software that has been programmed in the course of this thesis. Furthermore, the results from a small‐scale study using this software will be analysed and evaluated in order to check for the success of the project.
Leonia cymosa (Violaceae) is a small tree from the under story of the Amazonian rain forest. I investigated the seed dispersal ecology of L. cymosa in plots of old growth terra firme forest located within the Cuyabeno Faunistic Reserve in north-eastern Ecuador. This species offered good conditions to examine the variation of traits of individual trees and the way they are linked with fruit removal from each tree. With this study I aimed to address the question whether frugivores exert selection pressures on fruits and the fruiting regime of fleshy fruited plants. The mean height of a fruiting L. cymosa was 6.6 m (range: 2 - 12.6 m). The median tree density was 11.8 trees per hectare. Trees grew in clusters consisting of different numbers of trees of different heights. L. cymosa flowered two times a year, in late February to March and in October. The respective fruiting seasons occurred in August/September and between March and May. The fruit pulp of L. cymosa contained the sugars fructose, glucose, and sucrose, the total soluble sugar being the first important nutritional compound of the fruit pulp. The second important compound was proteins. No lipids were found in the fruit pulp. The variation of nutritional quality of the fruits was high within trees. Nonetheless, significant differences were found among trees in all nutrient constituents studied. The maximum of ripe fruits produced per season by a single tree was 427. Median productivity of the trees was 45 ripe fruits throughout the fruiting season in 1999 (n=57) and 36 ripe fruits in 2000 (n=92). The maximum standing crop of fruits in a tree was 324 fruits (counted in 2000). Black mantle tamarins, Saguinus nigricollis (Callitrichidae), and squirrel monkeys, Saimiri sciureus (Cebidae), and possibly an unknown nocturnal frugivore consumed the fruits of L. cymosa at my study site. Green-rumped acouchis (Myoprocta pratti, Dasyproctidae) consumed fallen fruits and seeds underneath the trees. Black mantle tamarins and squirrel monkeys differed widely in their effectiveness as seed dispersers. Black mantle tamarins swallowed the seeds together with the fruit pulp and defecated intact seeds far away from the mother tree. Squirrel monkeys opened the fruits to suck and gnaw on the fruit pulp, and then dropped seeds to the forest floor below the tree crowns. Each of my study plots fell into the core home range of one group each of S. nigricollis and S. sciureus. Thus, the frugivore assemblage is small and disperser availability is limited for the individual tree of L. cymosa. In a sample of 6 trees of comparable and high fruit crop size, the total of ripe fruits removed from a tree throughout the whole fruiting season by the reliable seed disperser S. nigricollis was neither significantly correlated with the content of any of the nutrients measured in the fruit pulp (fructose, glucose, sucrose, total protein; pulp does not contain lipids), nor with total metabolisable energy, seed to pulp weight ratio, or water content of the fruit pulp. Feeding preferences for single sugars determined by other laboratory studies were not confirmed by this field study. The reliable seed disperser S. nigricollis does not seem to exert selective pressure on the nutrient content of the fruits of L. cymosa. Seasonal fruit crop size was the main predictor of all aspects of fruit removal by the effective disperser of L. cymosa, Saguinus nigricollis, as well as by the non-disperser, Saimiri sciureus. Trees with larger seasonal fruit crop size had a higher probability to have fruits removed by the disperser than those with small seasonal fruit crop sizes. They also had a higher number of fruits removed by the seed disperser. However, the proportion of fruits removed by the disperser decreased with increasing seasonal fruit crop size. In contrast, probability of fruit removal, the number of fruits removed, and the proportion of fruits removed by the non-disperser increased with increasing seasonal fruit crop sizes. The observed differences between disperser and non-disperser are due to differences in feeding capacity, group size and foraging behavior. Tamarins were less likely to harvest Leonia trees that were not or less completely covered by surrounding vegetation. This probably reflects a behavior to avoid predation by forest raptors. At high con-specific fruit abundance in the neighborhood, the proportion of fruits removed by tamarins was reduced. This suggests competition of trees for the disperser. My study revealed selection of the disperser on seasonal fruit crop size of L. cymosa. My results are consistent with the “fruit crop size hypothesis”. FCSH appears to constitute a valid framework also in the monkey-dispersed L. cymosa. My findings also show that factors beyond the tree’s control influenced fruit removal from Leonia trees. Disperser-mediated selection may be constrained (yet not impeded) by neighborhood conditions.
Around 10.000 – 150.000 endogenous DNA damage-induced lesions occur in a human body per day and cell. Accumulation of unrepaired lesions can lead to aneuploidy and the loss of genomic integrity which in turn contributes to tumor formation. Therefore, an efficient DNA damage response has to be initiated, in the end leading to cell cycle inhibition and induction of repair. Since it is known that a recently characterized human multiprotein complex named LINC (or human dREAM) together with B-MYB is involved in the regulation of G2/M gene expression (Plk1, cyclin B1, cdc2 etc.), its function in the DNA damage response was analyzed in this study. In growing cells B-MYB is associated to the LIN core complex which consists of 5 different proteins named LIN-9, LIN-54, LIN-52, LIN-37 and RbAp48. After induction of DNA damage B-MYB leaves the complex and binding of E2F4 and p130 to LINC is induced. Importantly, the upstream pathway leading to LINC rearrangement is dependent on the activation of p53 and p21. Interestingly, p53 -/- cells solely have the potential to block in the G2 phase of the cell cycle, thereby making them vulnerable for errors during G2 arrest induction or maintenance. Here I demonstrate that LINC rearrangement is absent in p53 -/- cells and that B-MYB/LINC binding to target gene promoters is increased. This in turn leads to an increased G2/M gene expression after DNA damage induction and triggers premature cell cycle re-entry (checkpoint adaptation). Significantly, B-MYB expression is increased in p53 mutated primary breast cancer tumors and correlates with poor prognosis and reoccurrence probably due to its function in checkpoint adaptation. This study gives evidence that inhibition of B-MYB gene expression or B-MYB function in p53 mutant tumors could be a good choice for adjuvant therapy.
Phytohormones are known for their pivotal roles in promoting normal growth and development of the plants and contributing to the mechanism of defense. Although an over simplification, however, they may be categorized as stress specific and growth promoting. SA and JA/Ethylene are implicated in stress responses while auxins, cytokinins and gibberellins are involved in developmental processes. Phytohormones from the above perspective got much attention in the last few decades; however their reciprocal role is currently in focus. It is because of the reason that plant pathogens cause overall hormonal imbalance at host pathogen interface and alter host physiology for the sake of pathogenecity. Despite their importance in growth and development, cytokinins are among the most neglected phytohormones that are usually noticed as consequence rather than a cause of pathogen infection. Results presented in this thesis are based on the hypothesis that elevated levels of CKs embody plants with resistance against hemibiotrophic pathogens. To explore a connection between the spread of P. syringae and its tobacco host, CKs over producing transgenic plants were investigated whereby bacterial IPT gene was expressed under the control of pathogen inducible, tetracycline inducible and developmentally inducible promoters. To further validate the out-come of transgenic plants, various types of cytokinins were exogenously fed to detached tobacco leaves. Mentioned transgenics and exogenous CKs feeding approaches unanimously resulted in, “more cytokinins less disease symptoms” and vice versa. This state of cytokinins mediated resistance was further substantiated with various cellular, signaling, biochemical and microbial approaches wherein levels of SA and JA remained unaffected. Conversely, PR1 gene expression was strongly up-regulated in enhanced cytokinins accumulating samples. Moreover, less accumulation of ROS was observed in IPT expressing sites of the plants as compared to their corresponding controls. Additionally, we neither noticed any direct effect of cytokinins on the growth of P. syringae pv. tabaci nor found presence of anti-microbial peptides in cytokinins enriched extracts. Interestingly, enhanced accumulation of phtyoalexins in elevated CKs status of the plant proved to be a possible gesture in jeopardizing the spread of pathogen. Contrarily, no reduction was observed in the spread of fungal necrotrophic pathogen Sclerotinia sclerotiorum when leaves of elevated CKs were inoculated. Besides host-pathogen interaction in perspective of elevated cytokinins, impact of modulated sugar status of the plant on the spread of pathogen was also investigated. For this purpose, previously generated modulated invertase enzyme tobacco transgenic plants were analyzed. We showed that repression and de-repression of CIN1 gene under the control of tetracycline inducible-promoter did not affect the growth of P. syrinage pv. tabaci in Tet::CIN1 transgenic plants. Moreover, invertase inhibitor tobacco lines expressing NtCIF gene under the control of the same promoter failed to exhibit differential pathogenic responses in induced and non induced status of the plant. Similar was the case of tomato transgenic plants expressing NtCIF gene under the control of invertase gene Lin6 promoter in Lin6:: NtCIF plants for P.syringae pv. tomato DC 3000. Interestingly, when challenged Lin6:: NtCIF tomato plants with Botrytis cinerea, severe disease symptoms were observed on transgenic leaves as compared to control plants. To dissect a potential link between cytokinins and sugar metabolism with its effect on the growth of pathogen, invertase transgenic plants with elevated CKs were probed. When expressed exogenous IPT gene under the control of pathogen inducible promoter (4xJERE::IPT) in transgenic background of Tet::CIN1, we observed localized differences in symptom development for P.syringae pv. tabaci. Similarly, when exogenously fed with kinetin, detached leaves of Tet::CIN1 exhibited retarded growth of P.syringae pv. tabaci as compared to the tetracycline induced leaves. These results led to the conclusion that extracellular invertase may not play an essential role in cytokinins mediated disease resistance against hemibiotrophic pathogens.
Abstract: 1. The response of dispersal towards evolution largely depends on its heritability for which upper limits are determined by the trait's repeatability. 2. In the Linyphiid spider E. atra, we were able to separate long- and short-distance dispersal behaviours (respectively ballooning and rappelling) under laboratory conditions. By performing repeated behavioural trials for females, we show that average dispersal trait values decrease with increasing testing days. By comparing mated and unmated individuals during two periods (before and after mating for the mated group, and the same two periods for the unmated group), we show that mating has no effect on the mean displayed dispersal behaviour or its within-individual variation. Repeatabilities were high and consistent for ballooning motivation, but not for rappelling. 3. Ballooning motivation can be regarded as highly individual-specific behaviour, while general pre-dispersal and rappelling behaviours showed more individual variation. Such difference in repeatability between long-and short-distance dispersal suggests that short-and long-distance dispersal events are triggered by different ecological and evolutionary mechanisms.
The bacterial pathogen Legionella pneumophila replicates intracellularly in protozoa, but can also cause severe pneumonia, called Legionnaires' disease. The bacteria invade and proliferate in the alveolar macrophages of the human lung. L. pneumophila bacteria exhibit a biphasic life cycle: replicative bacteria are avirulent; in contrast, transmissive bacteria express virulence traits and flagella. Primarily aim of this thesis was to evaluate the impact of the regulatory proteins FleQ, FleR, and RpoN in flagellar gene regulation. Phenotypic analysis, Western blot and electron microscopy of regulatory mutants in the genes coding for FleQ, RpoN and FleR demonstrated that flagellin expression is strongly repressed and that these mutants are non-flagellated in transmissive phase. Transcriptomic studies of these putative flagellar gene expression regulators demonstrated that fleQ controls the expression of numerous flagellar biosynthetic genes. Together with RpoN, FleQ controls transcription of 14 out of 31 flagellar class II genes, coding for the basal body, hook, and regulatory proteins. Unexpectedly, 7 out of 15 late flagellar genes class III and IV) are expressed dependent on FleQ but independent of RpoN. Thus, in contrast to the commonly accepted view that enhancer binding proteins as FleQ always interact with RpoN to initiate transcription, our results strongly indicate that FleQ of L. pneumophila regulates gene expression RpoN-dependent as well as RpoN-independent. Moreover, transcriptome analysis of a fleR mutant strain elucidated that FleR does not regulate the flagellar class III genes as previously suggested. Instead FleR regulates together with RpoN numerous protein biosynthesis and metabolic genes. Based on these experimental results our modified model for the transcriptional regulation of flagellar genes in L. pneumophila is that flagellar class II genes are controlled by FleQ and RpoN, while flagellar class III and IV genes are controlled in a fleQ-dependent but rpoN-independent manner. Although all L. pneumophila strains share the same complex life style, various pathotypes have evolved. This is reflected by the genomes, which contain e.g. genomic islands. The genomic island Trb-1 of L. pneumophila Corby, carries all genes necessary for a type-IV conjugation system, an integrase gene and a putative oriT site. The second aim of this thesis was to investigate the implication of this genomic island in conjugative DNA transfer. Using conjugation assays we showed that the oriT site located on Trb-1 is functional and contributes to conjugation between different L. pneumophila strains. As this is the first oriT site of L. pneumophila known to be functional our results provide evidence that conjugation is a major mechanism for the evolution of new pathotypes in L. pneumophila.
Photosynthesis is the most fundamental process of life on earth. The biological production of oxygen in plant photosynthesis occurs in photosystem II (PSII). Here two water molecules are coupled in a four-electron oxidation to one O2 molecule, catalyzed by a tetranuclear manganese complex, known as the oxygen-evolving complex (OEC). In this thesis, density-functional theory (DFT) methods were validated and subsequently employed to study structures, spin-density distributions and EPR parameters of mono-, di-, and tetranuclear complexes with regard to the OEC. The goal was to draw conclusions on the molecular and electronic structure of the OEC.
Simulations (MASim) and non-rational behaviour. This non-rational behaviour is here based on the Prospect Theory [KT79] (PT), which is compared to the rational behaviour in the Expected Utility Theory [vNM07] (EUT). This model was used to design a modified Q-Learning [Wat89, WD92] algorithm. The PT based Q-Learning was then integrated into a proposed agent architecture. Because much attention is given to a limited interpretation of Simon's definition of bounded-rationality, this interpretation is broadened here. Both theories, rationality and the non-rationality, are compared and the discordance in their results discussed. The main contribution of this work is to show that an alternative is available to the EUT that is more suitable for human decision-makers modelling. The evidences show that rationality is not appropriated for modelling persons. Therefore, instead of fine-tuning the existent model the use of another one is proposed and evaluated. To tackle this, the route choice problem was adopted to perform the experiments. To evaluate the proposed model three traffic scenarios are simulated and their results analysed.
The genus Borrelia belongs to the spirochete phylum, an ancient evolutionary branch of the domain bacteria that is only afar related to Gram-negative bacteria. Borreliae can be subdivided into the agents of the two borrelian-caused human diseases, Lyme disease and relapsing fever. Both disease patterns are closely related to the peculiar biology of Borrelia species and exhibit a wide spectrum of diverse clinical manifestations. Due to the small 0.91 Mb chromosome, borreliae have a lack of biosynthetic capacity. Thus, all Borrelia species are highly dependent on nutrients provided by their hosts. The transport of nutrients and other molecules across the outer membrane is enabled by pore-forming proteins, so-called porins. Porins are water-filled channels and can be subdivided into two different classes, general diffusion pores and substrate-specific porins. In terms of the Lyme disease agent Borrelia burgdorferi, three putative porins were characterized in previous studies: P13, Oms28 and P66. In contrast to Lyme disease species, the porin knowledge of relapsing fever Borrelia is low, which means that not any porin has actually been described for representatives of these agents. Thus, the general aim of this thesis was to provide insight into the porin content of both, Lyme disease and relapsing fever spirochetes. This aim could be achieved by isolating and identifying porins from Borrelia outer membranes and by biophysically characterizing them in artificial lipid membranes. In one chapter of this study, the first identification and characterization of a relapsing fever porin is presented. The pore-forming protein was isolated from outer membranes of Borrelia duttonii, Borrelia hermsii and Borrelia recurrentis and designated Oms38, for “outer membrane-spanning protein of 38 kDa”. Biophysical characterization of Oms38 was achieved by using the black lipid bilayer method and demonstrated that Oms38 forms small, water-filled channels with a single-channel conductance of 80 pS in 1 M KCl. The Oms38 channel did not exhibit voltage-dependent closure and is slightly selective for anions with a permeability ratio of cations over anions of 0.41 in KCl. Subsequently, a protein homologous to Oms38 was identified in the Lyme disease agents Borrelia burgdorferi, Borrelia garinii and Borrelia afzelii. The pore-forming protein of these species exhibits high sequence homology to Oms38 and similar biophysical properties, i.e. it forms pores of 50 pS in 1 M KCl. Interestingly, titration experiments revealed that this pore could be partly blocked by dicarboxylic anions, which means that this protein does not form a general diffusion pore but a channel with a binding-site specific for those compounds. Consequently, this porin was termed DipA, for “dicarboxylate-specific porin A”. In another set of experiments, it was shown that the porin P66 is present in both Lyme disease and relapsing fever species. Therefor, the outer membranes of the Lyme disease species Borrelia burgdorferi, Borrelia afzelii, Borrelia garinii and the relapsing fever species Borrelia duttonii, Borrelia recurrentis and Borrelia hermsii were closer investigated. Except of the P66 homologue of Borrelia hermsii P66 of all species was highly active in artificial lipid membranes, forming pores with huge single-channel conductances between 9 and 11 nS in 1 M KCl. Moreover, the channel diameter and the constitution of Borrelia burgdorferi P66 were investigated in detail. Therefor, the P66 single-channel conductance in the presence of different nonelectrolytes with known hydrodynamic radii was analyzed in black lipid bilayers. The effective diameter of the P66 channel lumen was determined to be ~1.9 nm. Furthermore, as derived from multi-channel experiments the P66-induced membrane conductance could be blocked by certain nonelectrolytes, such as PEG 400, PEG 600 and maltohexaose. Additional blocking experiments on the single-channel level revealed seven subconducting states and indicated a heptameric constitution of the P66 channel. This indication could be confirmed by Blue native PAGE analysis which demonstrated that P66 units form a complex with a corresponding mass of approximately 440 kDa. Taking together, this thesis describes detailed biochemical and biophysical investigations of both Lyme disease and relapsing fever Borrelia porins and represents an important step forward in understanding the outer membrane pathways for nutrient uptake of these strictly host-dependent, pathogenic spirochetes. Furthermore, it provides some knowledge of the outer-membrane protein composition of Borrelia spirochetes. A profound knowledge of surface-exposed proteins, such as porins, is one precondition for the production of a successful vaccine and the drug design against the two borrelian-caused diseases.
Charcot-Marie-Tooth disease (CMT) is a cohort of human hereditary disorders of the peripheral nervous system (PNS) which exhibit symptoms like sensory dysfunction, muscle weakness and gait disturbances. Different mutations are described as causation for this neuropathy, such as a duplication of chromosome 17 comprising the gene for the peripheral myelin protein-22 (PMP22). Based on different animal models former studies identified immune cells, i.e. macrophages and T-lymphocytes, as crucial mediators of pathology in these neuropathies. In this study, PMP22-overexpressing mice (PMP22tg, C61), serving as a model for a specific type of CMT – CMT1A – were crossbred with immune-deficient mutant mice to examine the impact of the immune system on nerve pathology. Crossbreeding of PMP22tg mice with recombination activating gene-1 (RAG-1) deficient mice, lacking mature T- and B-lymphocytes, caused no striking alterations of pathogenesis in peripheral nerves of mutant mice. In contrast, crossbreeding of PMP22tg myelin mutants with mice deficient in the chemokine monocyte chemoattractant protein-1 (MCP-1, CCL2) caused an amelioration of the demyelinating phenotype of peripheral nerves when MCP-1 was either reduced or completely absent. Furthermore, functional investigations, i.e. neurographic recordings and examinations of the grip strength of the extremities, revealed an amelioration in PMP22tg/MCP-1-/- mice in regard to a symptomatic improvement in the compound action muscle potential (CMAP) and stronger grip strength of the hindlimbs. Interestingly, peripheral nerves of PMP22tg mice showed an irregular distribution of potassium channels in presence of MCP-1, whereas the absence of MCP-1 in the myelin mutants rescued the ion channel distribution and resulted in a more wild type-like phenotype. Having shown the impact of MCP-1 as an important mediator of nerve pathology in PMP22/MCP-1 double mutants, the regulation of this chemokine became an important target for potential treatment strategies. We found that the signaling cascade MEK1/2/ERK1/2 was more strongly activated in peripheral nerves of PMP22tg mice compared to nerves of wild type mice. This activation corresponded to an increase in MCP-1 mRNA expression in peripheral nerves at the same age. Furthermore, a MEK1/2-inhibitor was used in vivo to confirm the regulation of MCP-1 by the MEK1/2/ERK1/2 pathway. After a treatment period of three weeks, a clear reduction of ERK1/2-phosphorylation as well as a reduction of MCP-1 mRNA expression was observed, accompanied by a decline in macrophage number in peripheral nerves of PMP22tg mice. These observations suggest that the expression of MCP-1 is crucial for the neuropathological progression in a mouse model for CMT1A. Therefore, this chemokine could provide a basis for a putative treatment strategy of inherited neuropathies.
In future telecommunication systems, we observe an increasing diversity of access networks. The separation of transport services and applications or services leads to multi-network services, i.e., a future service has to work transparently to the underlying network infrastructure. Multi-network services with edge-based intelligence, like P2P file sharing or the Skype VoIP service, impose new traffic control paradigms on the future Internet. Such services adapt the amount of consumed bandwidth to reach different goals. A selfish behavior tries to keep the QoE of a single user above a certain level. Skype, for instance, repeats voice samples depending on the perceived end-to-end loss. From the viewpoint of a single user, the replication of voice data overcomes the degradation caused by packet loss and enables to maintain a certain QoE. The cost for this achievement is a higher amount of consumed bandwidth. However, if the packet loss is caused by congestion in the network, this additionally required bandwidth even worsens the network situation. Altruistic behavior, on the other side, would reduce the bandwidth consumption in such a way that the pressure on the network is released and thus the overall network performance is improved. In this monograph, we analyzed the impact of the overlay, P2P, and QoE paradigms in future Internet applications and the interactions from the observing user behavior. The shift of intelligence toward the edge is accompanied by a change in the emerging user behavior and traffic profile, as well as a change from multi-service networks to multi-networks services. In addition, edge-based intelligence may lead to a higher dynamics in the network topology, since the applications are often controlled by an overlay network, which can rapidly change in size and structure as new nodes can leave or join the overlay network in an entirely distributed manner. As a result, we found that the performance evaluation of such services provides new challenges, since novel key performance factors have to be first identified, like pollution of P2P systems, and appropriate models of the emerging user behavior are required, e.g. taking into account user impatience. As common denominator of the presented studies in this work, we focus on a user-centric view when evaluating the performance of future Internet applications. For a subscriber of a certain application or service, the perceived quality expressed as QoE will be the major criterion of the user's satisfaction with the network and service providers. We selected three different case studies and characterized the application's performance from the end user's point of view. Those are (1) cooperation in mobile P2P file sharing networks, (2) modeling of online TV recording services, and (3) QoE of edge-based VoIP applications. The user-centric approach facilitates the development of new mechanisms to overcome problems arising from the changing user behavior. An example is the proposed CycPriM cooperation strategy, which copes with selfish user behavior in mobile P2P file sharing system. An adequate mechanism has also been shown to be efficient in a heterogeneous B3G network with mobile users conducting vertical handovers between different wireless access technologies. The consideration of the user behavior and the user perceived quality guides to an appropriate modeling of future Internet applications. In the case of the online TV recording service, this enables the comparison between different technical realizations of the system, e.g. using server clusters or P2P technology, to properly dimension the installed network elements and to assess the costs for service providers. Technologies like P2P help to overcome phenomena like flash crowds and improve scalability compared to server clusters, which may get overloaded in such situations. Nevertheless, P2P technology invokes additional challenges and different user behavior to that seen in traditional client/server systems. Beside the willingness to share files and the churn of users, peers may be malicious and offer fake contents to disturb the data dissemination. Finally, the understanding and the quantification of QoE with respect to QoS degradations permits designing sophisticated edge-based applications. To this end, we identified and formulated the IQX hypothesis as an exponential interdependency between QoE and QoS parameters, which we validated for different examples. The appropriate modeling of the emerging user behavior taking into account the user's perceived quality and its interactions with the overlay and P2P paradigm will finally help to design future Internet applications.
Background
Suicide and non-fatal suicidal behaviour are significant public health issues in Europe requiring effective preventive interventions. However, the evidence for effective preventive strategies is scarce. The protocol of a European research project to develop an optimized evidence based program for suicide prevention is presented.
Method
The groundwork for this research has been established by a regional community based intervention for suicide prevention that focuses on improving awareness and care for depression performed within the European Alliance Against Depression (EAAD). The EAAD intervention consists of (1) training sessions and practice support for primary care physicians,(2) public relations activities and mass media campaigns, (3) training sessions for community facilitators who serve as gatekeepers for depressed and suicidal persons in the community and treatment and (4) outreach and support for high risk and self-help groups (e.g. helplines). The intervention has been shown to be effective in reducing suicidal behaviour in an earlier study, the Nuremberg Alliance Against Depression. In the context of the current research project described in this paper (OSPI-Europe) the EAAD model is enhanced by other evidence based interventions and implemented simultaneously and in standardised way in four regions in Ireland, Portugal, Hungary and Germany.
The enhanced intervention will be evaluated using a prospective controlled design with the primary outcomes being composite suicidal acts (fatal and non-fatal), and with intermediate outcomes being the effect of training programs, changes in public attitudes, guideline-consistent media reporting. In addition an analysis of the economic costs and consequences will be undertaken, while a process evaluation will monitor implementation of the interventions within the different regions with varying organisational and healthcare contexts.
Discussion
This multi-centre research seeks to overcome major challenges of field research in suicide prevention. It pools data from four European regions, considerably increasing the study sample, which will be close to one million. In addition, the study will gather important information concerning the potential to transfer this multilevel program to other health care systems. The results of this research will provide a basis for developing an evidence-based, efficient concept for suicide prevention for EU-member states.
The main purpose of volcano-seismology concerns the qualitative and quantitative description of one or more unknown seismic source(s) located at some unknown depth beneath a volcano. Even if many different volcanoes show similar seismic signal characteristics, up to now it was not possible to find a standard seismic source model for volcanoes, as the double-couple in earthquake seismology. Volcanoes with a continuous activity, like Stromboli (Italy), represent for the volcano seismologist a perfect natural laboratory to address this question. This thesis treats the study of explosion-quakes and volcanic tremor recorded on Stromboli in a broadband frequency range, and discusses the location and the possible mechanisms of the seismic source(s). Seismic and infrasonic recordings of explosion-quake from Stromboli showed that the high-frequency phase propagates with a velocity of approximately 330 m/s. The seismic source can be explained as an explosion at the top of the magma column generated by rising gas bubbles. The seismic P-wave and the air-wave are both generated in the same point at the same time. The different path lengths and velocities for the seismic wave and the air-wave result in a difference in arrival times dt, that could be used to deduce the magma level and sound speed in the eruption column inside the conduit. Stations installed near the active crater reveal that infrasonic and seismic recordings of the short-period tremor (> 1 Hz) share the same spectral content and show similar energy fluctuations. Therefore, the short-period volcanic tremor at Stromboli originates from the continuous out-bursting of small gas bubbles in the upper part of the magmatic column. The spectrum of the long-period tremor recorded at Stromboli consists of three main peaks with periods at 4.8 s, 6 s and 10 s, and amplitudes varying with the regional meteorological situation. Hence, they are not generated by a close volcanic source but rather by ocean microseisms (OMS). The passage of a local cyclone seems to be the seismic source for spectral energy at 4.8 s and 10 s, which represent the Double Frequency and the Primary Frequency of the OMS, respectively. Concerning the 6 s peak, a cyclone near the British Isles could act as a seismic source. Seismic data from the first broadband array deployed on Stromboli showed surprisingly simple waveforms, indicating an initially contracting source mechanism. The analysis of particle motion and the application of seismic array techniques allowed the location of a seismic source in the shallow part of the volcano. Eruption parameters and seismic source characteristics of the April 5, 2003 Stromboli eruption have been estimated using different inversion approaches. The paroxysm was triggered by a shallow slow thrust-faulting dislocation event with a moment magnitude of Mw = 3.0 and possibly associated with a crack that formed previously by dike extrusion. At least one blow-out phase during the paroxysmal explosion could be identified from seismic signals with an equivalent moment magnitude of Mw = 3.7. It can be represented by a vertical linear vector dipole and two weaker horizontal linear dipoles in opposite direction, plus a vertical force. Seismic measurements performed during controlled and reproducible blow-out experiments with a gas volume entrapped in basaltic melt revealed the following: Monochromatic seismic signals suggest a blow-out in a more ductile regime, whereas broader frequency content indicates rupture in a more brittle environment. The longer the crucible, the weaker the seismic signals. An increase in pressure results in a stronger fragmentation, but not in a higher ejection velocity of the plug neither in a higher seismic amplitude. Even if the very long period observations like the tilt signal could not be simulated in the laboratory, the blow-out experiments simulate very well the short-period seismic signals recorded at Stromboli volcano.
In the generalized Nash equilibrium problem not only the cost function of a player depends on the rival players' decisions, but also his constraints. This thesis presents different iterative methods for the numerical computation of a generalized Nash equilibrium, some of them globally, others locally superlinearly convergent. These methods are based on either reformulations of the generalized Nash equilibrium problem as an optimization problem, or on a fixed point formulation. The key tool for these reformulations is the Nikaido-Isoda function. Numerical results for various problem from the literature are given.
Contents List of Publications 1 Introduction 2 Basic concepts and instrumentation 2.1 Mathematical description of femtosecond laser pulses 2.2 Optical quantities and measurements 2.2.1 Intensity 2.2.2 Absorbance and Beer-Lambert law 2.3 Laser system 2.4 General software framework for scientific data acquisition and simulation 2.4.1 Core components 2.4.2 Program for executing a single measurement sequence 2.4.3 Scan program 2.4.4 Evolutionary algorithm optimization program 2.4.5 Applications of the software framework 2.5 Summary 3 Generation of ultrabroadband femtosecond pulses in the visible 3.1 Nonlinear optics 3.1.1 Nonlinear polarization and frequency conversion 3.1.2 Phase matching 3.2 Optical parametric amplification 3.3 Noncollinear optical parametric amplifier 3.4 Considerations and experimental design of NOPA 3.4.1 Options for broadening the NOPA bandwidth 3.4.2 Experimental setup 3.5 NOPA pulse characterization 3.5.1 Second harmonic generation frequency-resolved optical gating 3.5.2 Transient grating frequency-resolved optical gating 3.6 Compression and shaping methods for NOPA pulses 3.6.1 Grating compressor 3.6.2 Prism compressor 3.6.3 Chirped mirrors 3.6.4 Detuned zero dispersion compressor 3.6.5 Deformable mirror pulse shaper 3.6.6 Liquid crystal pulse shaper 3.7 Liquid crystal pulse shaper 3.7.1 Femtosecond pulse shapers 3.7.2 Experimental design and parameters 3.7.3 Optical setup of the LC pulse shaper 3.7.4 Calibrations of the pulse shaper 3.8 Adaptive pulse compression 3.8.1 Closed loop pulse compression 3.8.2 Open loop pulse compression 3.9 Conclusions 4 Coherent optical two-dimensional spectroscopy 4.1 Introduction 4.2 Theory of third order nonlinear optical spectroscopies 4.2.1 Response function, electric fields, and signal field 4.2.2 Signal detection with spectral interferometry 4.2.3 Evaluation of two-dimensional spectra and phasing 4.2.4 Selection and classification of terms in induced nonlinear polarization 4.2.5 Oscillatory character of measured signal 4.3 Previous experimental implementations 4.4 Inherently phase-stable setup using conventional optics only 4.4.1 Manipulation of pulse pairs as a basis for stability 4.4.2 Experimental setup 4.4.3 Measurement procedure 4.4.4 Data evaluation 4.5 First experimental results 4.5.1 Demonstration of phase stability 4.5.2 2D spectrum of Nile Blue at room temperature 4.6 Summary and outlook 5 Product accumulation for ultrasensitive femtochemistry 5.1 The problem of sensitivity in femtochemistry 5.2 Accumulation for increased sensitivity 5.2.1 Comparison of conventional and accumulative sensitivity 5.2.2 Schematics and illustrative example 5.3 Experimental setup 5.4 Calibration and modeling of accumulation 5.5 Experiments on indocyanine green 5.5.1 Calibration of the setup 5.5.2 Chirped pulse excitation 5.5.3 Adaptive pulse shaping 5.6 Conclusions 6 Ultrafast photoconversion of the green fluorescent protein 6.1 Green fluorescent protein 6.2 Experimental setup for photoconversion of GFP 6.3 Calibration of the setup for GFP 6.3.1 Model for concentration dynamics of involved GFP species 6.3.2 Estimate of sensitivity 6.4 Excitation power study 6.5 Time-resolved two-color experiment 6.6 Time-delayed unshaped 400 nm – shaped 800 nm pulse excitation 6.6.1 Inducing photoconversion with chirped pulses 6.6.2 Photoconversion using third order phase pulses 6.7 Conclusions 7 Applications of the accumulative method to chiral systems 7.1 Introduction 7.2 Chiral asymmetric photochemistry 7.2.1 Continuous-wave circularly polarized light 7.2.2 Controlled asymmetric photochemistry using femtosecond laser pulses 7.3 Sensitive and fast polarimeter 7.3.1 Polarimeter setup 7.3.2 Detected signal I(t) 7.3.3 Angular amplification 7.3.4 Performance of the polarimeter 7.4 Molecular systems and mechanisms for enantioselective quantum control 7.4.1 Binaphthalene derivatives 7.4.2 Photochemical helicene formation 7.4.3 Spiropyran/merocyanine chiroptical molecular switches 7.5 Summary 8 Summary Zusammenfassung Bibliography Acknowledgements
Recent progresses and developments in molecular biology provide a wealth of new but insufficiently characterised data. This fund comprises amongst others biological data of genomic DNA, protein sequences, 3-dimensional protein structures as well as profiles of gene expression. In the present work, this information is used to develop new methods for the characterisation and classification of organisms and whole groups of organisms as well as to enhance the automated gain and transfer of information. The first two presented approaches (chapters 4 und 5) focus on the medically and scientifically important enterobacteria. Its impact in medicine and molecular biology is founded in versatile mechanisms of infection, their fundamental function as a commensal inhabitant of the intestinal tract and their use as model organisms as they are easy to cultivate. Despite many studies on single pathogroups with clinical distinguishable pathologies, the genotypic factors that contribute to their diversity are still partially unknown. The comprehensive genome comparison described in Chapter 4 was conducted with numerous enterobacterial strains, which cover nearly the whole range of clinically relevant diversity. The genome comparison constitutes the basis of a characterisation of the enterobacterial gene pool, of a reconstruction of evolutionary processes and of comprehensive analysis of specific protein families in enterobacterial subgroups. Correspondence analysis, which is applied for the first time in this context, yields qualitative statements to bacterial subgroups and the respective, exclusively present protein families. Specific protein families were identified for the three major subgroups of enterobacteria namely the genera Yersinia and Salmonella as well as to the group of Shigella and E. coli by applying statistical tests. In conclusion, the genome comparison-based methods provide new starting points to infer specific genotypic traits of bacterial groups from the transfer of functional annotation. Due to the high medical importance of enterobacterial isolates their classification according to pathogenicity has been in focus of many studies. The microarray technology offers a fast, reproducible and standardisable means of bacterial typing and has been proved in bacterial diagnostics, risk assessment and surveillance. The design of the diagnostic microarray of enterobacteria described in chapter 5 is based on the availability of numerous enterobacterial genome sequences. A novel probe selection strategy based on the highly efficient algorithm of string search, which considers both coding and non-coding regions of genomic DNA, enhances pathogroup detection. This principle reduces the risk of incorrect typing due to restrictions to virulence-associated capture probes. Additional capture probes extend the spectrum of applications of the microarray to simultaneous diagnostic or surveillance of antimicrobial resistance. Comprehensive test hybridisations largely confirm the reliability of the selected capture probes and its ability to robustly classify enterobacterial strains according to pathogenicity. Moreover, the tests constitute the basis of the training of a regression model for the classification of pathogroups and hybridised amounts of DNA. The regression model features a continuous learning capacity leading to an enhancement of the prediction accuracy in the process of its application. A fraction of the capture probes represents intergenic DNA and hence confirms the relevance of the underlying strategy. Interestingly, a large part of the capture probes represents poorly annotated genes suggesting the existence of yet unconsidered factors with importance to the formation of respective virulence phenotypes. Another major field of microarray applications is gene expression analysis. The size of gene expression databases rapidly increased in recent years. Although they provide a wealth of expression data, it remains challenging to integrate results from different studies. In chapter 6 the methodology of an unsupervised meta-analysis of genome-wide A. thaliana gene expression data sets is presented, which yields novel insights in function and regulation of genes. The application of kernel-based principal component analysis in combination with hierarchical clustering identified three major groups of contrasts each sharing overlapping expression profiles. Genes associated with two groups are known to play important roles in Indol-3 acetic acid (IAA) mediated plant growth and development as well as in pathogen defence. Yet uncharacterised serine-threonine kinases could be assigned to novel functions in pathogen defence by meta-analysis. In general, hidden interrelation between genes regulated under different conditions could be unravelled by the described approach. HMMs are applied to the functional characterisation of proteins or the detection of genes in genome sequences. Although HMMs are technically mature and widely applied in computational biology, I demonstrate the methodical optimisation with respect to the modelling accuracy on biological data with various distributions of sequence lengths. The subunits of these models, the states, are associated with a certain holding time being the link to length distributions of represented sequences. An adaptation of simple HMM topologies to bell-shaped length distributions described in chapter 7 was achieved by serial chain-linking of single states, while residing in the class of conventional HMMs. The impact of an optimisation of HMM topologies was underlined by performance evaluations with differently adjusted HMM topologies. In summary, a general methodology was introduced to improve the modelling behaviour of HMMs by topological optimisation with maximum likelihood and a fast and easily implementable moment estimator. Chapter 8 describes the application of HMMs to the prediction of interaction sites in protein domains. As previously demonstrated, these sites are not trivial to predict because of varying degree in conservation of their location and type within the domain family. The prediction of interaction sites in protein domains is achieved by a newly defined HMM topology, which incorporates both sequence and structure information. Posterior decoding is applied to the prediction of interaction sites providing additional information of the probability of an interaction for all sequence positions. The implementation of interaction profile HMMs (ipHMMs) is based on the well established profile HMMs and inherits its known efficiency and sensitivity. The large-scale prediction of interaction sites by ipHMMs explained protein dysfunctions caused by mutations that are associated to inheritable diseases like different types of cancer or muscular dystrophy. As already demonstrated by profile HMMs, the ipHMMs are suitable for large-scale applications. Overall, the HMM-based method enhances the prediction quality of interaction sites and improves the understanding of the molecular background of inheritable diseases. With respect to current and future requirements I provide large-scale solutions for the characterisation of biological data in this work. All described methods feature a highly portable character, which allows for the transfer to related topics or organisms, respectively. Special emphasis was put on the knowledge transfer facilitated by a steadily increasing wealth of biological information. The applied and developed statistical methods largely provide learning capacities and hence benefit from the gain of knowledge resulting in increased prediction accuracies and reliability.
Humans and other animals share choice preference for smaller-but-sooner over later-but-larger rewards, indicating that the subjective value of a reward is discounted as a function of time. This phenomenon referred to as delay discounting (DD), represents one facet of impulsivity which is inherently connected with reward processing and, within a certain range, adaptive. Maladaptive levels, however, can lead to suboptimal decision-making and represent important characteristics of psychopathologies such as attention-deficit/hyperactivity disorder (ADHD). In line with a proposed influence of dysregulated dopamine (DA) levels on impulsivity, neural structures involved in DD (the ventral-striatum [VS]; orbitofrontal cortex [OFC]) are highly innervated by dopaminergic neurons. However, studies explicitly testing the triadic interplay of dopaminergic neurotransmission, impulsivity and brain activation during intertemporal choice are missing. Therefore, the first study of the thesis examined the effect of different DA-bioavailability levels, indicated by a genetic polymorphism (Val158Met) in the gene of the catechol-O-methyltransferase, on the association of delay discounting and OFC activation. OFC response to monetary rewards that varied by delay-to-delivery was recorded with functional near-infrared spectroscopy (fNIRS) in a sample of 49 healthy human subjects. The results suggest a DA-related enhancement in OFC function from low (low DA level) to partial (intermediate DA level) and full (high DA level) reward delay sensitivity. Furthermore, DA-bioavailability was shown to moderate the association of neural reward delay sensitivity and impulsivity: OFC reward delay sensitivity was strongly correlated with impulsivity at intermediate DA-levels, but not at low or high DA-levels where impulsivity was related to delay-independent OFC amplitudes. It is concluded that DA-level should be considered as a crucial factor whenever impulsivity-related brain activation, in particular to reward delay, is examined in healthy subjects. Dysfunctional reward processing, accompanied by a limited ability to tolerate reward delays (delay aversion), has been proposed as an important feature in ADHD putatively caused by striatal hypo-dopaminergia. Therefore, the aim of the second study of this thesis was to examine subcortical processing of reward delays and to test for neural indicators of a negative emotional response to delay periods. Using functional magnetic resonance imaging (fMRI), brain activation in adult patients with ADHD (n=14) and healthy control subjects (n=12) was recorded during the processing of immediate and delayed rewards. Compared with healthy control subjects, hyporesponsiveness of the VS reward system was evident in patients with ADHD for both immediate and delayed rewards. In contrast, delayed rewards evoked hyperactivation in the dorsal caudate nucleus and the amygdala of ADHD patients, corroborating the central predictions of the delay aversion hypothesis. In combination both studies support the conception of a close link between delay discounting, brain activation and dopaminergic neurotransmission. The results implicate that studies on neural correlates of DD have to account for the DA-bioavailability level and for a negative emotional response to reward delays.
The main focus of this thesis was the synthesis and analysis of multifunctional oligopeptides. The study of their non-covalent interactions with various counterparts revealed interesting new results, leading to both methodological and application related progress. The first project of this thesis concentrated on the in-depth analysis of the peptide receptor CBS-Lys-Lys-Phe-NH2 to acquire a better understanding of its binding mode upon complexation with a substrate. In this context it was possible to develop—in cooperation with the group of Prof. Sebastian Schlücker—a direct and label free spectroscopic detection of immobilized compounds which are often found in combinatorial libraries. This new screening method utilizes the advantages of the surface enhanced Raman spectroscopy and allowed for the first time a surface mapping of a single polystyrene bead for the identification of peptides in femtomolar concentrations. Hence, this method allows a very fast and sensitive detection of resin bound compounds. The development of this promising new approach set the starting point for future experiments to enable on-bead library screenings and to investigate the complex formation of immobilized compounds. After the comprehensive analysis of the basic structural features of small peptide receptors in the first part of this thesis, the second big block focused on its in vitro evaluation using biological relevant targets. Therefore, several different modifications of the initial peptide structures were synthesized. These modifications provided a molecular toolkit for the tailor made synthesis of structures individually designed for the respective target. The first tests addressed the interaction with Alzheimer’s related amyloid fibrils. During these experiments, the successful SPPS syntheses of tri- and tetravalent systems were achieved. The comparison of the multivalent form with the corresponding monovalent version was then under special investigations. These concentrated mainly on the interaction with various bacteria strains, as well as with different parasites. To localize the compounds within the organisms, the synthesis of fluorescence labelled versions was achieved. In addition, several compounds were tested by the Institute for Molecular Infection Biology of the University of Würzburg for their antibacterial activity. This thorough evaluation of the biological activity generated precious information about the influence of small structural changes in the peptide receptors. Especially the distinct influence of the multivalency effect and the acquired synthetic skills led to the development of an advanced non-covalent recognition event, as described in the final project of this thesis. The last part of this thesis discussed the development of a novel inhibitor for the serine protease beta-tryptase based on a tailor-made surface recognition event. It was possible to study and analyze the complex interaction with the unique structure of tryptase, that features a tetrameric frame and four catalytic cleavage sites buried deep inside of the hollow structure. However, the point of attack were not the four binding pockets, as mostly described in the literature, but rather the acidic areas around the cleavage sites and at the two circular openings. These should attract peptides with basic residues, which then can block the accessibility to the active sites. A combinatorial library of 216 tetravalent peptide compounds was synthesized to find the best structural composition for the non-covalent inhibition of beta-tryptase. For the screening of the library a new on-bead assay was applied. With this method a simultaneous readout of the total inhibition of all library members was possible, thus allowing a fast and direct investigation of the still resin bound inhibitors. Several additional experiments in solution unveiled the kinetics of the inhibition process. In conclusion, both mono- and multivalent inhibitors interact in a non-destructive and reversible way with the tryptase.