@phdthesis{Perscheid2013, author = {Perscheid, Michael}, title = {Test-driven fault navigation for debugging reproducible failures}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-68155}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {The correction of software failures tends to be very cost-intensive because their debugging is an often time-consuming development activity. During this activity, developers largely attempt to understand what causes failures: Starting with a test case that reproduces the observable failure they have to follow failure causes on the infection chain back to the root cause (defect). This idealized procedure requires deep knowledge of the system and its behavior because failures and defects can be far apart from each other. Unfortunately, common debugging tools are inadequate for systematically investigating such infection chains in detail. Thus, developers have to rely primarily on their intuition and the localization of failure causes is not time-efficient. To prevent debugging by disorganized trial and error, experienced developers apply the scientific method and its systematic hypothesis-testing. However, even when using the scientific method, the search for failure causes can still be a laborious task. First, lacking expertise about the system makes it hard to understand incorrect behavior and to create reasonable hypotheses. Second, contemporary debugging approaches provide no or only partial support for the scientific method. In this dissertation, we present test-driven fault navigation as a debugging guide for localizing reproducible failures with the scientific method. Based on the analysis of passing and failing test cases, we reveal anomalies and integrate them into a breadth-first search that leads developers to defects. This systematic search consists of four specific navigation techniques that together support the creation, evaluation, and refinement of failure cause hypotheses for the scientific method. First, structure navigation localizes suspicious system parts and restricts the initial search space. Second, team navigation recommends experienced developers for helping with failures. Third, behavior navigation allows developers to follow emphasized infection chains back to root causes. Fourth, state navigation identifies corrupted state and reveals parts of the infection chain automatically. We implement test-driven fault navigation in our Path Tools framework for the Squeak/Smalltalk development environment and limit its computation cost with the help of our incremental dynamic analysis. This lightweight dynamic analysis ensures an immediate debugging experience with our tools by splitting the run-time overhead over multiple test runs depending on developers' needs. Hence, our test-driven fault navigation in combination with our incremental dynamic analysis answers important questions in a short time: where to start debugging, who understands failure causes best, what happened before failures, and which state properties are infected.}, language = {en} } @phdthesis{Duensing2013, author = {Duensing, Nina}, title = {Transport processes in the arbuscular mycorrhizal symbiosis}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-68210}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {The nutrient exchange between plant and fungus is the key element of the arbuscular mycorrhizal (AM) symbiosis. The fungus improves the plant's uptake of mineral nutrients, mainly phosphate, and water, while the plant provides the fungus with photosynthetically assimilated carbohydrates. Still, the knowledge about the mechanisms of the nutrient exchange between the symbiotic partners is very limited. Therefore, transport processes of both, the plant and the fungal partner, are investigated in this study. In order to enhance the understanding of the molecular basis underlying this tight interaction between the roots of Medicago truncatula and the AM fungus Rhizophagus irregularis, genes involved in transport processes of both symbiotic partners are analysed here. The AM-specific regulation and cell-specific expression of potential transporter genes of M. truncatula that were found to be specifically regulated in arbuscule-containing cells and in non-arbusculated cells of mycorrhizal roots was confirmed. A model for the carbon allocation in mycorrhizal roots is suggested, in which carbohydrates are mobilized in non-arbusculated cells and symplastically provided to the arbuscule-containing cells. New insights into the mechanisms of the carbohydrate allocation were gained by the analysis of hexose/H+ symporter MtHxt1 which is regulated in distinct cells of mycorrhizal roots. Metabolite profiling of leaves and roots of a knock-out mutant, hxt1, showed that it indeed does have an impact on the carbohydrate balance in the course of the symbiosis throughout the whole plant, and on the interaction with the fungal partner. The primary metabolite profile of M. truncatula was shown to be altered significantly in response to mycorrhizal colonization. Additionally, molecular mechanisms determining the progress of the interaction in the fungal partner of the AM symbiosis were investigated. The R. irregularis transcriptome in planta and in extraradical tissues gave new insight into genes that are differentially expressed in these two fungal tissues. Over 3200 fungal transcripts with a significantly altered expression level in laser capture microdissection-collected arbuscules compared to extraradical tissues were identified. Among them, six previously unknown specifically regulated potential transporter genes were found. These are likely to play a role in the nutrient exchange between plant and fungus. While the substrates of three potential MFS transporters are as yet unknown, two potential sugar transporters are might play a role in the carbohydrate flow towards the fungal partner. In summary, this study provides new insights into transport processes between plant and fungus in the course of the AM symbiosis, analysing M. truncatula on the transcript and metabolite level, and provides a dataset of the R. irregularis transcriptome in planta, providing a high amount of new information for future works.}, language = {en} } @phdthesis{Dannehl2013, author = {Dannehl, Claudia}, title = {Fragments of the human antimicrobial LL-37 and their interaction with model membranes}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-68144}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {A detailed description of the characteristics of antimicrobial peptides (AMPs) is highly demanded, since the resistance against traditional antibiotics is an emerging problem in medicine. They are part of the innate immune system in every organism, and they are very efficient in the protection against bacteria, viruses, fungi and even cancer cells. Their advantage is that their target is the cell membrane, in contrast to antibiotics which disturb the metabolism of the respective cell type. This allows AMPs to be more active and faster. The lack of an efficient therapy for some cancer types and the evolvement of resistance against existing antitumor agents make AMPs promising in cancer therapy besides being an alternative to traditional antibiotics. The aim of this work was the physical-chemical characterization of two fragments of LL-37, a human antimicrobial peptide from the cathelicidin family. The fragments LL-32 and LL-20 exhibited contrary behavior in biological experiments concerning their activity against bacterial cells, human cells and human cancer cells. LL-32 had even a higher activity than LL-37, while LL-20 had almost no effect. The interaction of the two fragments with model membranes was systematically studied in this work to understand their mode of action. Planar lipid films were mainly applied as model systems in combination with IR-spectroscopy and X-ray scattering methods. Circular Dichroism spectroscopy in bulk systems completed the results. In the first approach, the structure of the peptides was determined in aqueous solution and compared to the structure of the peptides at the air/water interface. In bulk, both peptides are in an unstructured conformation. Adsorbed and confined to at the air-water interface, the peptides differ drastically in their surface activity as well as in the secondary structure. While LL-32 transforms into an α-helix lying flat at the water surface, LL-20 stays partly unstructured. This is in good agreement with the high antimicrobial activity of LL-32. In the second approach, experiments with lipid monolayers as biomimetic models for the cell membrane were performed. It could be shown that the peptides fluidize condensed monolayers of negatively charged DPPG which can be related to the thinning of a bacterial cell membrane. An interaction of the peptides with zwitterionic PCs, as models for mammalian cells, was not clearly observed, even though LL-32 is haemolytic. In the third approach, the lipid monolayers were more adapted to the composition of human erythrocyte membranes by incorporating sphingomyelin (SM) into the PC monolayers. Physical-chemical properties of the lipid films were determined and the influence of the peptides on them was studied. It could be shown that the interaction of the more active LL-32 is strongly increased for heterogeneous lipid films containing both gel and fluid phases, while the interaction of LL-20 with the monolayers was unaffected. The results indicate an interaction of LL-32 with the membrane in a detergent-like way. Additionally, the modelling of the peptide interaction with cancer cells was performed by incorporating some negatively charged lipids into the PC/SM monolayers, but the increased charge had no effect on the interaction of LL-32. It was concluded, that the high anti-cancer activity of the peptide originates from the changed fluidity of cell membrane rather than from the increased surface charge. Furthermore, similarities to the physical-chemical properties of melittin, an AMP from the bee venom, were demonstrated.  }, language = {en} } @phdthesis{Rothe2013, author = {Rothe, Monique}, title = {Response of intestinal Escherichia coli to dietary factors in the mouse intestine}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-66387}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {Diet is a major force influencing the intestinal microbiota. This is obvious from drastic changes in microbiota composition after a dietary alteration. Due to the complexity of the commensal microbiota and the high inter-individual variability, little is known about the bacterial response at the cellular level. The objective of this work was to identify mechanisms that enable gut bacteria to adapt to dietary factors. For this purpose, germ-free mice monoassociated with the commensal Escherichia coli K-12 strain MG1655 were fed three different diets over three weeks: a diet rich in starch, a diet rich in non-digestible lactose and a diet rich in casein. Two dimensional gel electrophoresis and electrospray tandem mass spectrometry were applied to identify differentially expressed proteins of E. coli recovered from small intestine and caecum of mice fed the lactose or casein diets in comparison with those of mice fed the starch diet. Selected differentially expressed bacterial proteins were characterised in vitro for their possible roles in bacterial adaptation to the various diets. Proteins belonging to the oxidative stress regulon oxyR such as alkyl hydroperoxide reductase subunit F (AhpF), DNA protection during starvation protein (Dps) and ferric uptake regulatory protein (Fur), which are required for E. coli's oxidative stress response, were upregulated in E. coli of mice fed the lactose-rich diet. Reporter gene analysis revealed that not only oxidative stress but also carbohydrate-induced osmotic stress led to the OxyR-dependent expression of ahpCF and dps. Moreover, the growth of E. coli mutants lacking the ahpCF or oxyR genes was impaired in the presence of non-digestible sucrose. This indicates that some OxyR-dependent proteins are crucial for the adaptation of E. coli to osmotic stress conditions. In addition, the function of two so far poorly characterised E. coli proteins was analysed: 2 deoxy-D gluconate 3 dehydrogenase (KduD) was upregulated in intestinal E. coli of mice fed the lactose-rich diet and this enzyme and 5 keto 4 deoxyuronate isomerase (KduI) were downregulated on the casein-rich diet. Reporter gene analysis identified galacturonate and glucuronate as inducers of the kduD and kduI gene expression. Moreover, KduI was shown to facilitate the breakdown of these hexuronates, which are normally degraded by uronate isomerase (UxaC), altronate oxidoreductase (UxaB), altronate dehydratase (UxaA), mannonate oxidoreductase (UxuB) and mannonate dehydratase (UxuA), whose expression was repressed by osmotic stress. The growth of kduID-deficient E. coli on galacturonate or glucuronate was impaired in the presence of osmotic stress, suggesting KduI and KduD to compensate for the function of the regular hexuronate degrading enzymes under such conditions. This indicates a novel function of KduI and KduD in E. coli's hexuronate metabolism. Promotion of the intracellular formation of hexuronates by lactose connects these in vitro observations with the induction of KduD on the lactose-rich diet. Taken together, this study demonstrates the crucial influence of osmotic stress on the gene expression of E. coli enzymes involved in stress response and metabolic processes. Therefore, the adaptation to diet-induced osmotic stress is a possible key factor for bacterial colonisation of the intestinal environment.}, language = {en} } @phdthesis{Patz2013, author = {Patz, Ronny}, title = {Information flows in the context of EU policy-making : affiliation networks and the post-2012 reform of the EU's Common Fisheries Policy}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-70732}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {Information flows in EU policy-making are heavily dependent on personal networks, both within the Brussels sphere but also reaching outside the narrow limits of the Belgian capital. These networks develop for example in the course of formal and informal meetings or at the sidelines of such meetings. A plethora of committees at European, transnational and regional level provides the basis for the establishment of pan-European networks. By studying affiliation to those committees, basic network structures can be uncovered. These affiliation network structures can then be used to predict EU information flows, assuming that certain positions within the network are advantageous for tapping into streams of information while others are too remote and peripheral to provide access to information early enough. This study has tested those assumptions for the case of the reform of the Common Fisheries Policy for the time after 2012. Through the analysis of an affiliation network based on participation in 10 different fisheries policy committees over two years (2009 and 2010), network data for an EU-wide network of about 1300 fisheries interest group representatives and more than 200 events was collected. The structure of this network showed a number of interesting patterns, such as - not surprisingly - a rather central role of Brussels-based committees but also close relations of very specific interests to the Brussels-cluster and stronger relations between geographically closer maritime regions. The analysis of information flows then focused on access to draft EU Commission documents containing the upcoming proposal for a new basic regulation of the Common Fisheries Policy. It was first documented that it would have been impossible to officially obtain this document and that personal networks were thus the most likely sources for fisheries policy actors to obtain access to these "leaks" in early 2011. A survey of a sample of 65 actors from the initial network supported these findings: Only a very small group had accessed the draft directly from the Commission. Most respondents who obtained access to the draft had received it from other actors, highlighting the networked flow of informal information in EU politics. Furthermore, the testing of the hypotheses connecting network positions and the level of informedness indicated that presence in or connections to the Brussels sphere had both advantages for overall access to the draft document and with regard to timing. Methodologically, challenges of both the network analysis and the analysis of information flows but also their relevance for the study of EU politics have been documented. In summary, this study has laid the foundation for a different way to study EU policy-making by connecting topical and methodological elements - such as affiliation network analysis and EU committee governance - which so far have not been considered together, thereby contributing in various ways to political science and EU studies.}, language = {en} } @phdthesis{Theves2013, author = {Theves, Matthias}, title = {Bacterial motility and growth in open and confined environments}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-70313}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {In the presence of a solid-liquid or liquid-air interface, bacteria can choose between a planktonic and a sessile lifestyle. Depending on environmental conditions, cells swimming in close proximity to the interface can irreversibly attach to the surface and grow into three-dimensional aggregates where the majority of cells is sessile and embedded in an extracellular polymer matrix (biofilm). We used microfluidic tools and time lapse microscopy to perform experiments with the polarly flagellated soil bacterium Pseudomonas putida (P. putida), a bacterial species that is able to form biofilms. We analyzed individual trajectories of swimming cells, both in the bulk fluid and in close proximity to a glass-liquid interface. Additionally, surface related growth during the early phase of biofilm formation was investigated. In the bulk fluid, P.putida shows a typical bacterial swimming pattern of alternating periods of persistent displacement along a line (runs) and fast reorientation events (turns) and cells swim with an average speed around 24 micrometer per second. We found that the distribution of turning angles is bimodal with a dominating peak around 180 degrees. In approximately six out of ten turning events, the cell reverses its swimming direction. In addition, our analysis revealed that upon a reversal, the cell systematically changes its swimming speed by a factor of two on average. Based on the experimentally observed values of mean runtime and rotational diffusion, we presented a model to describe the spreading of a population of cells by a run-reverse random walker with alternating speeds. We successfully recover the mean square displacement and, by an extended version of the model, also the negative dip in the directional autocorrelation function as observed in the experiments. The analytical solution of the model demonstrates that alternating speeds enhance a cells ability to explore its environment as compared to a bacterium moving at a constant intermediate speed. As compared to the bulk fluid, for cells swimming near a solid boundary we observed an increase in swimming speed at distances below d= 5 micrometer and an increase in average angular velocity at distances below d= 4 micrometer. While the average speed was maximal with an increase around 15\% at a distance of d= 3 micrometer, the angular velocity was highest in closest proximity to the boundary at d=1 micrometer with an increase around 90\% as compared to the bulk fluid. To investigate the swimming behavior in a confinement between two solid boundaries, we developed an experimental setup to acquire three-dimensional trajectories using a piezo driven objective mount coupled to a high speed camera. Results on speed and angular velocity were consistent with motility statistics in the presence of a single boundary. Additionally, an analysis of the probability density revealed that a majority of cells accumulated near the upper and lower boundaries of the microchannel. The increase in angular velocity is consistent with previous studies, where bacteria near a solid boundary were shown to swim on circular trajectories, an effect which can be attributed to a wall induced torque. The increase in speed at a distance of several times the size of the cell body, however, cannot be explained by existing theories which either consider the drag increase on cell body and flagellum near a boundary (resistive force theory) or model the swimming microorganism by a multipole expansion to account for the flow field interaction between cell and boundary. An accumulation of swimming bacteria near solid boundaries has been observed in similar experiments. Our results confirm that collisions with the surface play an important role and hydrodynamic interactions alone cannot explain the steady-state accumulation of cells near the channel walls. Furthermore, we monitored the number growth of cells in the microchannel under medium rich conditions. We observed that, after a lag time, initially isolated cells at the surface started to grow by division into colonies of increasing size, while coexisting with a comparable smaller number of swimming cells. After 5:50 hours, we observed a sudden jump in the number of swimming cells, which was accompanied by a breakup of bigger clusters on the surface. After approximately 30 minutes where planktonic cells dominated in the microchannel, individual swimming cells reattached to the surface. We interpret this process as an emigration and recolonization event. A number of complementary experiments were performed to investigate the influence of collective effects or a depletion of the growth medium on the transition. Similar to earlier observations on another bacterium from the same family we found that the release of cells to the swimming phase is most likely the result of an individual adaption process, where syntheses of proteins for flagellar motility are upregulated after a number of division cycles at the surface.}, language = {en} } @phdthesis{Berg2013, author = {Berg, Gregor}, title = {Virtual prototypes for the model-based elicitation and validation of collaborative scenarios}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-69729}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {Requirements engineers have to elicit, document, and validate how stakeholders act and interact to achieve their common goals in collaborative scenarios. Only after gathering all information concerning who interacts with whom to do what and why, can a software system be designed and realized which supports the stakeholders to do their work. To capture and structure requirements of different (groups of) stakeholders, scenario-based approaches have been widely used and investigated. Still, the elicitation and validation of requirements covering collaborative scenarios remains complicated, since the required information is highly intertwined, fragmented, and distributed over several stakeholders. Hence, it can only be elicited and validated collaboratively. In times of globally distributed companies, scheduling and conducting workshops with groups of stakeholders is usually not feasible due to budget and time constraints. Talking to individual stakeholders, on the other hand, is feasible but leads to fragmented and incomplete stakeholder scenarios. Going back and forth between different individual stakeholders to resolve this fragmentation and explore uncovered alternatives is an error-prone, time-consuming, and expensive task for the requirements engineers. While formal modeling methods can be employed to automatically check and ensure consistency of stakeholder scenarios, such methods introduce additional overhead since their formal notations have to be explained in each interaction between stakeholders and requirements engineers. Tangible prototypes as they are used in other disciplines such as design, on the other hand, allow designers to feasibly validate and iterate concepts and requirements with stakeholders. This thesis proposes a model-based approach for prototyping formal behavioral specifications of stakeholders who are involved in collaborative scenarios. By simulating and animating such specifications in a remote domain-specific visualization, stakeholders can experience and validate the scenarios captured so far, i.e., how other stakeholders act and react. This interactive scenario simulation is referred to as a model-based virtual prototype. Moreover, through observing how stakeholders interact with a virtual prototype of their collaborative scenarios, formal behavioral specifications can be automatically derived which complete the otherwise fragmented scenarios. This, in turn, enables requirements engineers to elicit and validate collaborative scenarios in individual stakeholder sessions - decoupled, since stakeholders can participate remotely and are not forced to be available for a joint session at the same time. This thesis discusses and evaluates the feasibility, understandability, and modifiability of model-based virtual prototypes. Similarly to how physical prototypes are perceived, the presented approach brings behavioral models closer to being tangible for stakeholders and, moreover, combines the advantages of joint stakeholder sessions and decoupled sessions.}, language = {en} } @phdthesis{Santan2013, author = {Santan, Harshal Diliprao}, title = {Synthesis and characterization of thermosensitive hydrogels derived from polysaccharides}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-69793}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {In this work, thermosensitive hydrogels having tunable thermo-mechanical properties were synthesized. Generally the thermal transition of thermosensitive hydrogels is based on either a lower critical solution temperature (LCST) or critical micelle concentration/ temperature (CMC/ CMT). The temperature dependent transition from sol to gel with large volume change may be seen in the former type of thermosensitive hydrogels and is negligible in CMC/ CMT dependent systems. The change in volume leads to exclusion of water molecules, resulting in shrinking and stiffening of system above the transition temperature. The volume change can be undesired when cells are to be incorporated in the system. The gelation in the latter case is mainly driven by micelle formation above the transition temperature and further colloidal packing of micelles around the gelation temperature. As the gelation mainly depends on concentration of polymer, such a system could undergo fast dissolution upon addition of solvent. Here, it was envisioned to realize a thermosensitive gel based on two components, one responsible for a change in mechanical properties by formation of reversible netpoints upon heating without volume change, and second component conferring degradability on demand. As first component, an ABA triblockcopolymer (here: Poly(ethylene glycol)-b-poly(propylene glycol)-b-poly(ethylene glycol) (PEPE) with thermosensitive properties, whose sol-gel transition on the molecular level is based on micellization and colloidal jamming of the formed micelles was chosen, while for the additional macromolecular component crosslinking the formed micelles biopolymers were employed. The synthesis of the hydrogels was performed in two ways, either by physical mixing of compounds showing electrostatic interactions, or by covalent coupling of the components. Biopolymers (here: the polysaccharides hyaluronic acid, chondroitin sulphate, or pectin, as well as the protein gelatin) were employed as additional macromolecular crosslinker to simultaneously incorporate an enzyme responsiveness into the systems. In order to have strong ionic/electrostatic interactions between PEPE and polysaccharides, PEPE was aminated to yield predominantly mono- or di-substituted PEPEs. The systems based on aminated PEPE physically mixed with HA showed an enhancement in the mechanical properties such as, elastic modulus (G′) and viscous modulus (G′′) and a decrease of the gelation temperature (Tgel) compared to the PEPE at same concentration. Furthermore, by varying the amount of aminated PEPE in the composition, the Tgel of the system could be tailored to 27-36 °C. The physical mixtures of HA with di-amino PEPE (HA·di-PEPE) showed higher elastic moduli G′ and stability towards dissolution compared to the physical mixtures of HA with mono-amino PEPE (HA·mono-PEPE). This indicates a strong influence of electrostatic interaction between -COOH groups of HA and -NH2 groups of PEPE. The physical properties of HA with di-amino PEPE (HA·di-PEPE) compare beneficially with the physical properties of the human vitreous body, the systems are highly transparent, and have a comparable refractive index and viscosity. Therefore,this material was tested for a potential biological application and was shown to be non-cytotoxic in eluate and direct contact tests. The materials will in the future be investigated in further studies as vitreous body substitutes. In addition, enzymatic degradation of these hydrogels was performed using hyaluronidase to specifically degrade the HA. During the degradation of these hydrogels, increase in the Tgel was observed along with decrease in the mechanical properties. The aminated PEPE were further utilised in the covalent coupling to Pectin and chondroitin sulphate by using EDC as a coupling agent. Here, it was possible to adjust the Tgel (28-33 °C) by varying the grafting density of PEPE to the biopolymer. The grafting of PEPE to Pectin enhanced the thermal stability of the hydrogel. The Pec-g-PEPE hydrogels were degradable by enzymes with slight increase in Tgel and decrease in G′ during the degradation time. The covalent coupling of aminated PEPE to HA was performed by DMTMM as a coupling agent. This method of coupling was observed to be more efficient compared to EDC mediated coupling. Moreover, the purification of the final product was performed by ultrafiltration technique, which efficiently removed the unreacted PEPE from the final product, which was not sufficiently achieved by dialysis. Interestingly, the final products of these reaction were in a gel state and showed enhancement in the mechanical properties at very low concentrations (2.5 wt\%) near body temperature. In these hydrogels the resulting increase in mechanical properties was due to the combined effect of micelle packing (physical interactions) by PEPE and covalent netpoints between PEPE and HA. PEPE alone or the physical mixtures of the same components were not able to show thermosensitive behavior at concentrations below 16 wt\%. These thermosensitive hydrogels also showed on demand solubilisation by enzymatic degradation. The concept of thermosensitivity was introduced to 3D architectured porous hydrogels, by covalently grafting the PEPE to gelatin and crosslinking with LDI as a crosslinker. Here, the grafted PEPE resulted in a decrease in the helix formation in gelatin chains and after fixing the gelatin chains by crosslinking, the system showed an enhancement in the mechanical properties upon heating (34-42 °C) which was reversible upon cooling. A possible explanation of the reversible changes in mechanical properties is the strong physical interactions between micelles formed by PEPE being covalently linked to gelatin. Above the transition temperature, the local properties were evaluated by AFM indentation of pore walls in which an increase in elastic modulus (E) at higher temperature (37 °C) was observed. The water uptake of these thermosensitive architectured porous hydrogels was also influenced by PEPE and temperature (25 °C and 37 °C), showing lower water up take at higher temperature and vice versa. In addition, due to the lower water uptake at high temperature, the rate of hydrolytic degradation of these systems was found to be decreased when compared to pure gelatin architectured porous hydrogels. Such temperature sensitive architectured porous hydrogels could be important for e.g. stem cell culturing, cell differentiation and guided cell migration, etc. Altogether, it was possible to demonstrate that the crosslinking of micelles by a macromolecular crosslinker increased the shear moduli, viscosity, and stability towards dissolution of CMC-based gels. This effect could be likewise be realized by covalent or non-covalent mechanisms such as, micelle interactions, physical interactions of gelatin chains and physical interactions between gelatin chains and micelles. Moreover, the covalent grafting of PEPE will create additional net-points which also influence the mechanical properties of thermosensitive architectured porous hydrogels. Overall, the physical and chemical interactions and reversible physical interactions in such thermosensitive architectured porous hydrogels gave a control over the mechanical properties of such complex system. The hydrogels showing change of mechanical properties without a sol-gel transition or volume change are especially interesting for further study with cell proliferation and differentiation.}, language = {en} } @phdthesis{Ritz2013, author = {Ritz, Julia}, title = {Discourse-givenness of noun phrases : theoretical and computational models}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-70818}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {This thesis gives formal definitions of discourse-givenness, coreference and reference, and reports on experiments with computational models of discourse-givenness of noun phrases for English and German. Definitions are based on Bach's (1987) work on reference, Kibble and van Deemter's (2000) work on coreference, and Kamp and Reyle's Discourse Representation Theory (1993). For the experiments, the following corpora with coreference annotation were used: MUC-7, OntoNotes and ARRAU for Englisch, and TueBa-D/Z for German. As for classification algorithms, they cover J48 decision trees, the rule based learner Ripper, and linear support vector machines. New features are suggested, representing the noun phrase's specificity as well as its context, which lead to a significant improvement of classification quality.}, language = {en} } @phdthesis{Dethloff2013, author = {Dethloff, Frederik}, title = {In vivo 13C stable isotope tracing of single leaf development in the cold}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-70486}, school = {Universit{\"a}t Potsdam}, year = {2013}, abstract = {Measuring the metabolite profile of plants can be a strong phenotyping tool, but the changes of metabolite pool sizes are often difficult to interpret, not least because metabolite pool sizes may stay constant while carbon flows are altered and vice versa. Hence, measuring the carbon allocation of metabolites enables a better understanding of the metabolic phenotype. The main challenge of such measurements is the in vivo integration of a stable or radioactive label into a plant without perturbation of the system. To follow the carbon flow of a precursor metabolite, a method is developed in this work that is based on metabolite profiling of primary metabolites measured with a mass spectrometer preceded by a gas chromatograph (Wagner et al. 2003; Erban et al. 2007; Dethloff et al. submitted). This method generates stable isotope profiling data, besides conventional metabolite profiling data. In order to allow the feeding of a 13C sucrose solution into the plant, a petiole and a hypocotyl feeding assay are developed. To enable the processing of large numbers of single leaf samples, their preparation and extraction are simplified and optimised. The metabolite profiles of primary metabolites are measured, and a simple relative calculation is done to gain information on carbon allocation from 13C sucrose. This method is tested examining single leaves of one rosette in different developmental stages, both metabolically and regarding carbon allocation from 13C sucrose. It is revealed that some metabolite pool sizes and 13C pools are tightly associated to relative leaf growth, i.e. to the developmental stage of the leaf. Fumaric acid turns out to be the most interesting candidate for further studies because pool size and 13C pool diverge considerably. In addition, the analyses are also performed on plants grown in the cold, and the initial results show a different metabolite pool size pattern across single leaves of one Arabidopsis rosette, compared to the plants grown under normal temperatures. Lastly, in situ expression of REIL genes in the cold is examined using promotor-GUS plants. Initial results suggest that single leaf metabolite profiles of reil2 differ from those of the WT.}, language = {en} }