Welche Faktoren bedingen Implementationserfolg, auch und gerade über die rechtliche Umsetzung hinaus? Diese Frage wird von der EU-Implementationsforschung bislang nicht zufriedenstellend beantwortet. Fähigkeiten und Kapazitäten sowie Motivationen scheinen aber eine Rolle zu spielen. Die Implementation von EU-Informationsinstrumenten zur Energieeffizienz in Deutschland stellt für die Untersuchung dieser Frage einen idealen Fall dar: Ein gleichzeitig organisatorisch hochkomplexes Feld mit erwartbar geringen Implementationsfähigkeiten trifft auf eine politisch eher nicht-konfrontative Umsetzungsaufgabe mit vermutet guter Motivation. Die Ergebnisse aus den Fallstudien zu Gebäudeenergieausweisen, Energielabels auf Produkten und Energieaudits für große Unternehmen zeigten klar den hohen Erklärungswert des Faktors "Motivation", wohingegen "Fähigkeiten" kein eigenständiger Faktor sind. Mangelnde Fähigkeiten wurden von den Implementierern sehr geschickt adressiert und beseitigt, wenn das notwendig oder wünschenswert war. Dabei wurden alle Fälle von Wirtschaftsinteressen dominiert, aber auch von Europäisierungsmodi im Sinne von hierarchischem Druck, der als mindestens gleichwertiger Faktor neben dem Faktor Motivation steht.

Eukaryotic genes are mostly composed of a series of exons intercalated by sequences with no coding potential called introns. These sequences are generally removed from primary transcripts to form mature RNA molecules in a post-transcriptional process called splicing. An efficient splicing of primary transcripts is an essential step in gene expression and its misregulation is related to numerous human diseases. Thus, to better understand the dynamics of this process and the perturbations that might be caused by aberrant transcript processing, it is important to quantify splicing efficiency. In this thesis, I introduce SPLICE-q, a fast and user-friendly Python tool for genome-wide SPLICing Efficiency quantification. It supports studies focusing on the implications of splicing efficiency in transcript processing dynamics. SPLICE-q uses aligned reads from RNA-Seq to quantify splicing efficiency for each intron individually and allows the user to select different levels of restrictiveness concerning the introns’ overlap with other genomic elements, such as exons from other genes. I demonstrate SPLICE-q’s application using three use cases including two different species and methodologies. These analyses illustrate that SPLICE-q can detect a progressive increase of splicing efficiency throughout a time course of nascent RNA-Seq and it might be useful when it comes to understanding cancer progression beyond mere gene expression levels. Furthermore, I provide an in-depth study of time course nascent BrU-Seq data to address questions concerning differences in the speed of splicing and the underlying biological features that might be associated with it. SPLICE-q and its documentation are publicly available at: https://github.com/vrmelo/SPLICE-q.

With continually growing fields of applications for ENM and immense technological and economic potential, it needs to be kept in mind that nanotechnology also entails potential risks due to their high surface to volume ratio, some nanomaterials are chemically more reactive. What happens to the nanomaterials after being taken up by humans? How can one explain, and properly predict for example the unexpectedly fast clearance of BaSO4 from the lung, despite its insolubility in water? To determine the fate and the hazard potential of a nanomaterial within the human organism or the environment, it is mandatory to understand how an ENM interacts with physiological media. Essential for the understanding of the behavior is the determination of dissolution, transformation and dosimetry of ENM in relevant media. This work addressed the extrinsic properties of ENM for grouping and read across by assessing the dissolution and transformation in an abiotic flow-through dissolution setup, as well as investigation the limitations of current in vitro dosimetry setups.

Jährlich erreichen das Konsiliarlabor für Pockenviren des Robert Koch-Instituts zahlreiche Proben von Patienten mit pockentypischen und pockenähnlichen Hautveränderungen zur Diagnostik von Ortho-, Para- und Molluscipockenviren. Nicht immer können die eingesandten Proben eindeutig diagnostiziert werden und der Ursprung der pockentypischen Hautläsionen bleibt unklar. Für die Diagnostik von Viren gibt es verschiedene molekularbiologische Methoden. Während mit der Polymerase-Kettenreaktion (PCR) spezifisch auf einzelne Erreger gescreent werden kann, wird mit der Hochdurchsatz-Sequenzierung (HTS) ein unspezifischer Blick in die gesamte DNA und RNA der untersuchten Probe und damit alle darin enthaltenen Erreger ermöglicht. Seit 2014 hat der MinION in vielen Laboren Einzug gehalten, ein handtellergroßer Hochdurchsatz-Sequenzierer, welcher die Analyse von DNA-Molekülen in Echtzeit ermöglicht. Diese Technologie hat völlig neue Möglichkeiten in der Erregerdiagnostik eröffnet, jedoch sind die Sensitivität der Methode und die Qualität der generierten Sequenzen noch immer gering. In dieser Studie wurden 49 Patientenproben mittels Illumina HTS sowie mit einer neu entwickelten Methode (VAmpSeq) auf die verursachenden Erreger der pockenähnlichen Hautläsionen untersucht. Dabei handelt es sich bei der VAmpSeq (Virus Amplification based Sequencing)-Methode um eine Multiplex-PCR mit anschließender MinION-Sequenzierung. Dies kombiniert die Vorteile der PCR (Sensitivität, Spezifität) und der HTS (ungerichtetes, simultanes Screening) und ermöglicht eine sensitive und simultane Diagnostik und Differentialdiagnostik von pockenähnlichen Hautläsionen. Ergänzend wurde die VAmpSeq-Methode weiterentwickelt (PAmpliSeq – Pox Amplification based Sequencing), um neben der Detektion von Viren auch eine Vollgenomsequenzierung von Orthopockenviren zu ermöglichen. Für die Analyse in Echtzeit während der MinION-Sequenzierung wurde eine Oberflächenanwendung entwickelt (VAmpSeeker und PAmpliSeeker), um die Interpretation der generierten Daten auch ohne bioinformatische Kenntnisse zu ermöglichen. Mittels Illumina HTS und der VAmpSeq-Methode konnten in den Patientenproben Herpes simplex-Virus Typ 1, Molluscum-contagiosum-Virus Typ 1 und Typ 2 sowie Orf-Virus diagnostiziert werden. Zudem konnte mit der Illumina HTS ein neuartiges (Para)pockenvirus, mit der VAmpSeq-Methode in weiteren Proben das Epstein-Barr-Virus und Parvovirus B19 identifiziert werden. Mit der VAmpSeq-Methode konnte damit in dieser Arbeit ein neues diagnostisches Verfahren entwickelt werden, welches es ermöglicht, auf mehrere Erreger in kurzer Zeit zu screenen und welches bereits für zahlreiche Anwendungen im Laboralltag weiterentwickelt wurde.

Recent development in seismological instrumentation has made possible the deployment of temporary and permanent seismic networks with large numbers of seismometer stations. These networks register an extensive amount of seismic data that is then available to the scientific community for carrying out different types of seismological studies. Producing reliable earthquake catalogs, based on the implementation of efficient automatic event location procedures, which can handle such extensive amount of seismic data, is one of the critical steps prior to perform most studies. Precise and maximally complete event catalogs become essential in regions on the Earth where tectonic plate subduction takes place, such as the Chilean convergent margin. In subduction zones, seismicity analysis can help to shed light on the different tectonic processes involved in the generation of great tsunamigenic megathrust earthquakes, which are usually devastating for the inhabitants of these hazardous regions. Particularly, insights on the conditions characterizing the frictional behavior of a subduction interplate fault, can be investigated in detail from seismicity. This ultimately leads to a better assessment of the earthquake hazard potential in a subduction zone. In my thesis, I apply a multistage automatic earthquake detection and location workflow to generate a high-resolution earthquake catalog for the northern Chile region. This automatic workflow is applied to seismic data recorded by the Integrated Plate Boundary Observatory Chile (IPOC) permanent seismic network, as well as by a number of complementary temporary deployments, adding up to >100 seismic stations. The resulting event catalog contains ~19,000 foreshocks, aftershocks and background seismicity occurring in the time interval between one month preceding and nine months following the 1 April 2014 M8.1 Iquique earthquake. I analyse the seismicity features, in combination with modelled coseismic slip and modelled static stress changes, as well as geodetically-derived afterslip and interseismic locking models, in order to investigate the small-scale frictional heterogeneities on the plate interface and interpret the seismotectonic behavior of the overlying continental forearc in northern Chile. Results from this analysis point towards a primarily along-dip segmentation of the frictional behavior along the plate interface. In the downdip direction, an aseismic velocity-strengthening segment of the interface and a frictionally transitional region underlay, respectively, an aseismic frontal prism and a transitional zone in the outer continental wedge. Deeper on the plate interface, the seismogenic segment coincides with the highest coseismic slip underlying the inner continental wedge. The velocity-weakening segment connects downdip to a frictionally heterogeneous region where aftershock seismicity (interpreted as conditionally stable) and highest afterslip patches (velocity-strengthening) anticorrelate. A sparsity of events, which may be a generic feature along the Chilean subduction, separates this heterogenous segment from the deepest interplate events, where presumably frictionally transitional behaviour predominates. With the aim of improving the efficiency and simplifying the implementation of the earthquake catalog generation workflow, I additionally present DeepPhasePick, an automatic method entirely based on systematically optimized deep neural networks that carries out the first stages involved in this workflow: detection and picking of P and S phases originating from local earthquakes. DeepPhasePick makes use of a convolutional neural network architecture to perform the phase detection on three-component seismograms. It then uses two recurrent neural networks to conduct the phase picking on the vertical component for P phases and on the two-horizontal components for S phases. The phase time onsets and their corresponding onset uncertainties returned by the phase picking stage can feed a phase associator algorithm in the next step of the earthquake location workflow. DeepPhasePick architectures are optimized and trained using >30,000 manually-picked seismic records extracted from two sets of event waveforms occurring ~7 and ~17 years before the 2014 M8.1 Iquique earthquake, in a region of northern Chile that partially overlaps with the area covered by the automatically-derived ~19,000 earthquake catalog described above. The algorithm is then tested on different test sets: a set of manually-picked records independent from the training set, >1,000,000 automatically-picked records taken from the ~19,000 earthquake catalog, and a set of >200,000 automatically-picked records taken from another recently published earthquake catalog for the northern Chile region. Results from these tests show that DeepPhasePick is able to detect seismic phases with high accuracy, as well as to predict phase time onsets with a precision comparable to the more conventional phase picking methods, be they manual or automatic. Moreover, I demonstrate that DeepPhasePick’s detection and picking abilities perform effectively not only on the largely lower-seismic noise data recorded in northern Chile, but also generalize to higher-seismic noise data recorded from a different tectonic regime in an urban region of Albania.

Prostate cancer is one of the most widespread types of cancer in men and at the forth position of common cancer types in 2018 according to the World Health Organization report of 2020. The prostate-specific membrane antigen (PSMA) is highly expressed on prostate cancer cells and it is the important target for imaging and therapy of prostate cancer. In spite of the considerable progress in the improvement of existing tracers and the development of novel tracers for positron emission tomography (PET), the single-photon emission computed tomography (SPECT) with its workhorse 99mTc remains the dominating technique in diagnostic nuclear medicine. The fundament for labelling reactions with 99mTc is an exact knowledge of the coordination chemistry of the radioactive transition metal. The related structural studies are commonly done with the longlived isotope 99Tc, a weak β- emitter. This is a survey of the coordination capabilities of (η5-cyclopentadienyl)tris(dimethyl phosphito-P)cobaltate(III), {LOMe}−, the so-called Kläui ligand, to technetium with consideration of all common oxidation states of this transition metal and moreover, to further important metalions for medical approaches. The coordination chemistry of different thiourea ligands with rhenium and technetium are explored as well. The background of this work is the quest for stable metal cores for ongoing nuclear medical labeling experiments.

In epithelia, large amounts of water pass by transcellular and paracellular pathways, driven by the osmotic gradient built up by solutes’ movement. The transcellular pathway has been molecularly characterized by the discovery of aquaporin membrane channels. Unlike this, the existence of a paracellular pathway for water through the tight junctions (TJ) was discussed controversially for many years until two molecular components of paracellular water transport, claudin-2, and claudin-15, were identified. A main protein of the tricellular TJ (tTJ), tricellulin, was shown to be downregulated in ulcerative colitis leading to increased permeability to macromolecules. In addition to tricellulin, the family of angulin proteins is known to be able to recruit tricellulin to the tTJ. Recently, Gong and colleagues found that angulin-2 knockout increases water transport on isolated mouse kidney tubules, and its overexpression reduces the water transport in MDCK II cells. Whether or not tricellulin regulates water transport or angulin-1 also mediates a direct effect on water permeability independent of tricellulin, or act indirectly via tricellulin regulation, is unknown yet. To answer our research question, two epithelial cell lines featuring properties of the tight and intermediate-tight epithelium, MDCK C7 and HT-29/B6, respectively, were stably transfected with shRNA targeting tricellulin or sgRNA along with CRISPR/Cas9 targeting angulin-1, proteins of the tTJ essential for the barrier against passage of solutes up to 10 kDa. Interestingly, tricellulin knockdown and angulin-1 knockout reduced the transepithelial resistance (TER) in both cell lines and increased 4-kDa FITC-dextran permeability, especially in HT-29/B6 cell line. In addition, the expression and location of different TJ proteins in control cells and in knockdown or knockout clones were investigated. In MDCK C7 cells, tricellulin knockdown and angulin-1 knockout downregulated the expression of some tight junction proteins, while on HT-29/B6 cells, the opposite effect was observed; some TJ proteins were upregulated. Finally, water flux was induced by osmotic gradients using mannitol, 4-kDa, and 40-kDa dextran or albumin and measured in both cell lines. In MDCK C7 cells, tricellulin knockdown and angulin-1 knockout increased the water flux compared to that of vector controls, indicating a direct role of tricellulin in regulating water permeability in a tight epithelial cell line; nevertheless, in HT-29/B6 cells, water flux was unchanged between the control and the tricellulin knockdown or angulin-1 knockout clones. We conclude that tricellulin and angulin-1, the latter acting indirectly via tricellulin displacement, increase water permeability at reduced expression only in MDCK C7 cells, i.e., in the tight epithelium.

Die kumulative Dissertation setzt sich mit der Frage auseinander, warum selbst strukturell benachteiligte Personen in Organisationen diskriminierende Praktiken häufig reproduzieren. Außerdem erkundet sie geeignete Werkzeuge, um als Forschende subtile Formen der Reproduktion sozialer Ungleichheit in und durch Organisationen zu erkennen. Es wird diskutiert, warum Forschende dabei nicht nur kritisch selbstverständliche (Diversitäts-)Kategorien hinterfragen, sondern auch selbstreflektiert und politisch vorgehen sollten. Außerdem wird aufgezeigt, wie verschiedene Machtperspektiven helfen können, soziale Ungleichheit in ihrer Komplexität zu verstehen. Denn soziale Ungleichheit ist mehr als ungleiche Ressourcenverteilung und daraus resultierende mangelnde, gesellschaftliche Teilhabe. Schließlich wird am Beispiel von Altersungleichheiten verdeutlicht, dass benachteiligte Beschäftigte aus Angst vor Einkommens- oder Jobverlust sowie aufgrund von Männlichkeitsvorstellungen stereotype Altersbilder und benachteiligende Praktiken reproduzieren. Materielle Verhältnisse und Geschlechtsidentitäten prägen folglich Widerstandsformen und -möglichkeiten.

Protein structure and function are intimately connected. To deduce the mechanisms underlying specific functions, it is therefore of high interest to investigate structural changes during a reaction. Recently, the development of serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) has attracted a great deal of attention by enabling time-resolved (TR) experiments at atomic spatial and femtosecond temporal resolution, thereby allowing unprecedented insight into protein dynamics.

The high intensity of the XFEL pulse destroys any sample that has been exposed to the focused beam. A new protein crystal thus needs to be supplied for each pulse. This is typically achieved using a continuously flowing jet. For light-triggered reactions, an optical pulse starts the reaction in crystals of photosensitive proteins and the X-ray pulse then interrogates the system after a given time interval. For such experiments there are two main issues: First, appropriate conditions have to be found for triggering the reaction of interest. Second, the measurement of weak signals is severely limited by the low data collection rate (≤ 120 Hz) at first-generation XFELs. Moreover high sample consumption is an issue at these X-ray sources.

The goals of this thesis were therefore twofold: In the first part, techniques were developed to enable studying the ultrafast isomerization following photon absorption by bacteriorhodopsin in a TR-SFX experiment. Extending these results, light-matter interactions changing the incident excitation intensity were quantified based on experiments and calculations. This allowed establishing guidelines how to generally determine appropriate excitation conditions in SFX employing light triggering. These findings are fundamental to avoid multiphoton artefacts arising from excessive excitation and are thus essential for studying biological reactions which take place almost exclusively in the single photon regime.

In the second part of this thesis, opportunities and challenges of SFX experiments at next-generation XFELs were explored. These new machines generate X-ray pulses at MHz peak repetition rate and promise significantly higher throughput and more efficient sample usage. However, the short spacing between pulses introduces new challenges: it needs to be ensured that fresh sample is supplied sufficiently fast for each X-ray pulse. Moreover, it has been shown that the XFEL pulse launches shock waves in the sample carrying jet. These may damage sample probed by subsequent pulses. Here, first experiments at MHz peak repetition rate were conducted to investigate both issues. It was demonstrated that data collection of undamaged sample is indeed possible at 1.1 MHz repetition rate. At shorter pulse intervals (corresponding to 4.5 and 9.2 MHz), shock wave induced damage may lead to a significant loss in diffraction resolution of the crystal and even to structural changes in the protein.

Together, the results of this thesis delineate the limitations of (TR-) SFX due to XFEL induced shock damage and pave the way towards exploiting the promising capabilities of MHz XFELs, in particular for studying biologically relevant light-triggered reactions in proteins.

The ultimate purpose of this project was to elucidate the function and underlying mechanism of neurofibromin I during postnatal skeletal muscle development. Neurofibromin I is a tumor suppressor protein that works as a GTPase activator (GAP) through negative regulate the canonical Ras-MAPK-ERK signaling. Mutation of the Nf1 gene leads to a reduction of neurofibromin I, which causes an autochrosomal dominated genetic disorder named neurofibromatosis type I (NF1). Clinical features have been reported in NF1 patients, including pathological changes of the musculoskeletal system, particularly reduction of muscle mass and muscle strength. In this project, it is the first time that a muscle-specific Nf1 knock out mouse model was used for functional analysis of Nf1 during postnatal muscle development. Nf1Myf5 mice showed a gradual loss of muscle weight, which recapitulates the patient phenotype. Nf1 deletion in muscle progenitors leads to muscle stem cell pool depletion. Nf1/ERK/NO/Delta-Notch signaling is involved in the regulation of this phenotype. The results suggest that loss of Nf1 in muscle progenitors results in the hyper activation of Ras/ERK signaling, and ERK signaling via Nitric Oxide leads to a stronger stimulation of the Delta-Notch pathway. During early postnatal muscle development, activation of the Notch pathway drives progenitors out of cell cycle and go gradually to quiescent. For Nf1Myf5 muscle progenitors stronger Delta-Notch pathway contribute to the earlier transition of this process and satellite cell pool depletion. The quiescent signature of Nf1 deleted progenitors was identified from transcription of quiescent genes, reduction of glycolysis activity and also epigenetic change. Nf1 knockout mice showed a metabolism defect of glucose glycolysis and sever energy deficiency. As compensatory, energy sensor AMPK stimulated fatty acid oxidative phorsphorlyation which might through Pparg/Pgc1a signaling, accompanied by suppressed protein synthesis rate and increased the degradation process. In addition, a fiber type transition from fast to intermediate slow fibers was also detected. As Nf1 functions specifically before myoblasts differentiation. Thus the metabolic myopathy caused by Nf1 deletion should be generated from muscle progenitors. One possible explanation might be Nf1 deletion caused the epigenetic modification status change, in a certain way the change can be remembered by the nucleus of satellite cells even after they differentiated into mature muscle fibers. The reduction of glycolysis genes expression in both progenitors and mature muscle fibers is a good indicator. However, a further intensive study still needs to be performed to find the potential target mechanism.

This thesis mainly focused on the regulation of stem cells behavior by chemical and physical signals of carbon nanomaterial modified fibrous scaffolds. The cell adhesion on scaffolds is indirect via a surface layer of adsorbed proteins, thus the scaffold surface treatments could regulate the protein adsorption amount, cells adherence efficiency. The general property of scaffolds could further affect nutrient/waste exchange, protein synthesis, intracellular matrix construction and cell differentiation eventually, which is very important in stem cells-based tissue engineering, especially in organ and tissue damage. In this work, iPSCs and MSCs were used to investigate the effect on differentiation potential towards neuron and osteogenesis respectively via the chemical and physical cues of carbon nanomaterials-based scaffolds. The first project prepared a multivalent polyanion-dispersed CNTs modified fibrous scaffolds to integrate the chemical and physical in stem cell regulation research. The CNTs are dispersed and functionalized by biocompatible and multivalent hyperbranched polyglycerol sulfate (hPGS) noncovalently. After air plasma treatment of electrospun fibrous polycaprolactone (PCL) scaffolds, the HPGS modified CNT, namely CNT-HPGS were coated on the PCL fiber surface to combine the chemical and physical cues. Results suggests that CNT-HPGS modified fibrous scaffold is suitable for stem cells adherence and growth, because the combination of CNT and multivalent HPGS on scaffolds surface could offer anchoring points for proteins, growth factors and cytokines, which is very important for stem cells behavior. Meanwhile, the modified scaffolds promote the neural differentiation efficiency due to the special physical property of carbon nanotubes. Moreover, the aligned fibrous scaffolds could orient the elongation direction of grown neurites. Thus, the promoted protein adhesion property of CNT-HPGS contribute to the stem cells growth microenvironment and provide a novel method to construct functional scaffolds for stem therapy research. Since brutal plasma treatment could lead to polymer degradation and the longtime effects on surface may not be permanent, surface grafting of biocompatible and multivalent HPGS could be a suitable choice. In the second project, the HPGS was covalently conjugated onto the graphene oxide (GO) nanosheet by using nitrene through a 2+1 cycloaddition reaction. The physical property of graphene oxide and chemical property of HPGS were combined to mediate the stem cells growth and differentiation. Then the GO-HPGS nanosheets functionalized nanofibrous scaffolds were applied to mediate the proliferation, lineage specification, and differentiation of iPSC. Results suggest that coated scaffolds could promote differentiation and maturity of iPSC towards neural differentiation. This study addressed the stability of the dispersion and promote the stem cell lineage specification maturity, which integrate the chemical and physical cues to facilitate the targeted differentiation of iPSC. In the third project, we constructed a novel nanocarbon-structured fibrous scaffold for stem cell research, and the physical cues of carbon nanomaterials were MOF-derived nanocarbons. The porous carbon nanostructure could the promote the adhesion of proteins and growth factors, moreover, the caging property of the carbon nanostructure achieve the gradual release of chemical cues Zn2+ ions, which provide novel pathway for activation of signal pathways and guiding MSCs towards osteogenic differentiation process. This study designed stem cell scaffolds could help achieve multifunctional property, for example, simultaneously enhanced osteogenic and anti-infective capabilities. In conclusion, this work combined the physical cues of carbon nanomaterials and other chemical cues to investigate the stem cell behavior, including the IPS cells towards neural differentiation and MSCs towards osteogenic differentiation process. The fabrication of scaffolds presented a novel avenue for the future development of carbon nanomaterials in tissue regeneration, bionic, biomedical, and bioelectronics.

Fibrodysplasia Ossificans Progressiva (FOP) is a rare disease characterized by episodic bone formation outside of the skeleton (ectopic bone) in soft tissues in a process called heterotopic ossification (HO). Independent of the location and initial triggers, ectopic bone in FOP forms via a complex-multi-stage process, which mimics the developmental process of endochondral bone formation. Bone is a highly vascularized organ and its formation requires a tightly controlled interplay of osteogenesis and angiogenesis. Blocking or loss of pro-angiogenic factors (e.g. VEGFA) impairs bone formation by disturbed angiogenesis and decreased ossification. FOP patient biopsies revealed that HO lesions are highly angiogenic with aberrations in vascular markers and morphology suggesting the involvement of pathological angiogenesis and Endothelial to Mesenchymal Transition (EndMT). However, it remains elusive, which molecular mechanisms orchestrate the vasculature in ectopic bone formation and which signals are responsible for the aberrant vascular phenotype in FOP lesions. FOP is caused by mutations in the BMP type I receptor ALK2 leading to hypersensitive signaling and aberrant SMAD1/5 activation by ActivinA. In FOP mice, blocking of ActivinA prevents HO indicating a central role of this ligand in the disease. Whether signaling responses of Activin or BMP ligands cause the vascular phenotype in human FOP biopsies is unknown. The endothelium represents a fundamental component of the vasculature by forming the initial tubular structure. In the present study a patient derived endothelial cell (EC) model was generated by using induced Pluripotent Stem Cells (iPSCs) in a combinatorial cryopreservation and differentiation method. FOP iECs recapitulated the pathomechanism of aberrant ActivinA/SMAD1/5 signaling and additionally showed aberrant BMP/SMAD2 responses in an ALK2 dependent manner. ActivinA transduced a BMP like response only in FOP iECs and a comprehensive transcriptome analysis identified a FOP-specific genetic profile interlinking ActivinA with BMP/NOTCH pathways, blood vessel formation and EndMT. Collectively, the results propose a model in which ActivinA primes FOP ECs in favor of pathological angiogenesis and EndMT in presence of angiogenic and inflammatory factors as 2nd triggers. ActivinA induced only partial EndMT and alterations in angiogenesis were only observed in presence of VEGFA suggesting a second hit model for pathological endothelial function in FOP lesions. Thus, I propose that the ActivinA induced FOP specific transcriptome (1st trigger) pre-patterns the FOP endothelium and thereby challenges endothelial identity and plasticity, which leads to pathological angiogenesis and EndMT in presence of 2nd triggers. Drug testing in the FOP endothelial disease model demonstrated that the kinase inhibitor Saracatinib rescued aberrant signaling responses and the ActivinA-induced transcriptome in FOP iECs to WT levels, suggesting a preventive effect on pathological vascularization in pre-osseous FOP lesions.

Vaccines are considered as one of the most significant achievements of modern medicine. The first vaccine that protected animals against cancer is a vaccine designed for Marek’s disease virus. The most frequently used vaccine is the live-attenuated CVI988/Rispens (CVI) strain, which efficiently protects chickens against MD and prevents tumorigenesis. Interestingly, CVI expresses at least two isoforms of meq, the major oncogene of MDV. Meq is a basic leucine zipper (b-ZIP) protein consistently expressed in all MDV tumor and latently infected cells. We demonstrated that the longer isoform of meq strongly enhanced virus-induced pathogenesis and tumorigenesis, indicating that other mutations in the CVI genome contribute to virus attenuation. On the contrary, the shorter isoform completely abrogated pathogenesis, demonstrating that changes in the meq gene can indeed play a key role in virus attenuation. Although vaccinated chickens are protected against developing MD symptoms, the development of vaccines has raised questions about the potential consequences of vaccine-driven evolution of viruses. MDV is continually evolving towards higher virulence despite generations of vaccination. Circulating field strains have acquired numerous genomic mutations in the last 60 years. However, the evolutionary adaptations responsible for the vaccine breaks remained elusive. Distinct mutations in the meq oncogene arose every time that new vaccines were introduced that ultimately provided an evolutionary advantage. We tested recombinant viruses harbouring meq isoforms from different field strains in vivo. Here, we demonstrate that a few distinct mutations in the virus-encoded oncogene meq are responsible for the increase in virulence and oncogenicity. The viruses expressing the lower virulent meq isoforms showed reduced pathogenicity while in contrast the higher virulent meq isoforms dramatically increased pathogenesis in unvaccinated hosts. Only viruses harbouring the highest virulent meq isoform were able to break the vaccine barrier and cause tumors in vaccinated hosts - likely by overcoming innate cellular responses. Concomitantly, the polymorphisms in meq enhanced virus shedding into the environment putting naïve animals at greater risk.

In der deutschen Sprache sind die Begriffe „Opfer“ und „Suendenbock“ doppeldeutig, was in anthropologischen Theorien ueber Ursachen von Gewalt und Krieg dazu gefuehrt hat, dass Loesungen im paedagogischen und politischen Kontext unauffindbar waren, um Gewalt und Krieg zu beenden. Im Gegenteil: Gewalt und Krieg im 21. Jahrhundert sind deswegen weiterhin die „Geissel“ für die Menschheit (Charta der Vereinten Nationen, Praeambel), weil sowohl Suendenbocktheorien (zum Beispiel von Rene Girard und Hyam Maccoby) als auch Ethiken der Gewaltfreiheit (zum Beispiel von Leo Tolstoi und Mahatma Gandhi) ignoriert werden. Diese wissenschaftliche Untersuchung dient primaer dazu, für zukuenftige paedagogische Forschung kategoriale Klaerung zu ermoeglichen, und sekundaer dazu, die Rezeption der Theorien von Girard und Maccoby zu ergaenzen.

The interaction of biomolecules with UV light gives rise to a wide variety of fundamental processes in nature. For example, it is well known that biological molecules feature various relaxation pathways that efficiently dissipate the gained excess energy to their environment before any harmful, i.e., irreversible photoreactions can take place. The study of such processes may therefore extend the knowledge about why the biological matter is built exactly the way it is found on Earth today. In the present thesis, novel quantum-classical methods for the simulation of light-induced excited state dynamics in complex (bio)molecular systems have been developed and applied to selected examples. In addition, the methodology has been employed to simulate time-resolved spectroscopic observables.

In particular, the field-induced surface-hopping (FISH) approach has been extended to include terms beyond the electric field--electric dipole coupling as well as the interaction with circularly polarized light (CPL). The developed methodology has been applied to the smallest chiral amino acid, alanine. It has been found that asymmetric fragmentation and recombination in excited electronic states can be induced by irradiation with CPL. This leads to an enrichment of one of the stereogenic isomers up to 1.7%, where the sign depends on the helicity of the irradiating light.

The quantum-classical methodology has been further developed to study the photodynamics in multi-chromophoric molecular aggregates. In the newly developed multi-chromophoric FISH (McFISH) method, quantum-classical simulations in the frame of the extended Frenkel exciton model have been combined with QM/MM techniques in order to simulate the photodynamics of a double-stranded DNA decamer. In accordance with the experimental observations, a multi-exponential excited state decay involving the formation of long-lived delocalized as well as ultrafast decaying localized states resembling those of the bare nucleobases has been found.

Furthermore, a generally applicable method for the simulation of pump-probe time-resolved infrared (TRIR) spectra based on molecular dynamics simulations in the frame of the Wigner phase space representation of quantum mechanics has been developed. The obtained spectra are based on ab-initio potential energy surfaces and therefore include all anharmonicities. The developed methodology has been employed to investigate the migration dynamics of a single water molecule around the --CONH-- peptide linkage in two peptide analogs serving as models for biological proteins. The simulated spectra are in excellent agreement with the experimental observations, and the simulations have allowed for an atomistic interpretation of the measured time scales, migration pathways, and their fingerprints in the experimental ps-TRIR spectra.

This cumulative dissertation contains three self-contained research papers related to the topic of Education Economics in Brazil, with each chapter consisting of a single paper. While chapter 1 addresses the variation in intergenerational education mobility across Brazilian states, the chapters 2 and 3 deal with the impact evaluation of the teacher bonus of the state of São Paulo implemented in 2008. Chapter 1, entitled „The geography of mobility” applies the most recently published mobility data for Brazil to present a comprehensive and up-to-date analysis of mobility chances in Brazilian society. The study aims at expanding the literature on the “Great Gatsby curve” by incorporating a national investigation of the relationship between income inequality and intergenerational mobility. In addition, the chapter investigates a specific mechanism behind the “Great Gatsby curve,” namely whether income inequality does affect the (final) education outcomes of children from socially vulnerable families. Using cross-sectional data from nationally representative household surveys and econometric models, this paper provides two main empirical findings to the literature. First, the “Great Gatsby curve” also holds true within a single country; or, in other words, the Brazilian states with the highest levels of income disparity present also the lowest values for intergenerational education mobility. And, second, a possible reason for this association is the negative impact of inequality on education: each additional point in the 75/10 ratio of income inequality decreases the chances of achieving secondary education by 5.4 percent for children from low-educated parents. In chapter 2, entitled “Does a productivity bonus pay off?” I explore the panel nature of the GERES database and apply value-added models for the investigation of the bonus scheme’s impact in order to control the estimation strategy for the students’ previous knowledge and ability. Using a difference-in-difference approach in which students from state schools are the treatment and their peers from municipal schools the comparison group, the article concludes that the implementation of performance-based bonuses for teachers in the state schools of São Paulo has led to no statistically significant impact on the performance of students in Math and Portuguese. In the first year of the bonus program, the test scores in treatment schools were slightly lower than those in comparison schools. But none of this variation was statistically significant. The results from alternative specifications, placebo tests, and robustness checks also support these core findings. The final chapter, entitled “Addressing changes in professional behavior by teacher bonus” applies an explanatory sequential design by combining data from a questionnaire survey and qualitative interviews. The main motivation behind this analysis was addressing the lack of empirical evidence in the literature showing the mechanisms through which the teacher bonus program can lead to an increase in the academic achievements of students. Then, the paper focuses on the individual perceptions and viewpoints of teachers to investigate a possible causal link between the implementation of the merit-pay program in the education system and the improvement of professional practices and behaviors in schools. According to the opinions of the teachers themselves, the bonus scheme created no additional incentives for working in the state schools of São Paulo or for the improvement of teaching activities; it had furthermore a negative side effect, namely an increase in dishonest behavior in the schools. The only positive impact of the bonus program felt by the teachers was the reduction of work absenteeism.

Diese Arbeit untersucht den deutschen kolonialen Diskurs um 1700 anhand eines dreidimensionalen Diskursmodells. Im Zentrum stehen dabei die Texte von Otto Friedrich von der Gröben, die unter anderem die Gründung der brandenburg-preußischen Stützpunktkolonie Groß-Friedrichsburg im heutigen Ghana thematisieren. Sie waren bisher noch nicht Gegenstand einer umfassenden literaturwissenschaftlichen Untersuchung. Neben den drei umfangreichen Werken Gröbens, der „Orientalischen Reise-Beschreibung“, deren Anhang, der „Guineischen Reise-Beschreibung“, und dem Versepos „Bergone“, konnten auch einige seiner Briefe und Gedichte in die Betrachtung einbezogen werden, die zuvor von der Forschung über Gröben wenig Beachtung gefunden haben. Dazu zählt ein Epicedium, das ihm erstmals zugeschrieben werden konnte. Zusätzlich stützt sich die Diskursanalyse auf weitere zeitgenössische Texte, die Teil des deutschen kolonialen Diskurses um 1700 sind, sowie auf ausgewählte Prätexte und Rezeptionszeugnisse. Gröbens Biographie wurde ausführlich nachgezeichnet, wodurch autobiographische Angaben in seinen Texten mit konkreten Daten versehen und teilweise korrigiert werden konnten. Erkenntnisse über die produktive Dimension des Diskurses konnten durch die Beschäftigung mit dem Paratext und den materiellen Aspekten der einzelnen Fassungen der Reiseberichte und des Versepos gewonnen werden. So zählen die Widmungen ebenso wie die Ausstattung der einzelnen Exemplare zu denjenigen Praktiken des Diskurses, mit denen sich Gröben in der sozialen Dimension des Diskurses als adliger Kavalier nach dem Vorbild von Baldassare Castigliones „Il Libro del Cortegiano“ entwirft. Diesem Selbstbild dienen auch die Verwendung von Marginalien in der „Orientalischen Reise-Beschreibung“, das Motto der „Guineischen Reise-Beschreibung“ und weitere intertextuelle Bezüge. Anhand dieser Ergebnisse und weiterer Überlegungen zur produktiven Dimension des Diskurses konnte erwiesen werden, daß es sich bei Gröbens Texten um komplexe literarische Arbeiten handelt, die somit neu bewertet werden müssen. Ein weiterer Fokus liegt auf der Produktion von Wissenssystemen über Westafrika durch die Texte Gröbens und anderer europäischer Reisender, wie dem Schiffschirurgen Johann Peter Oettinger. Dazu wurden die dargestellten Interaktionen der brandenburgischen Akteure mit den anderen europäischen Akteuren, die im Kolonialhandel an der Westküste Afrikas aktiv waren, sowie mit den westafrikanischen Akteuren untersucht. Es konnte festgestellt werden, daß die Brandenburger an der westafrikanischen Küste soziale Praktiken vorfanden, die die anderen europäischen Akteure vor dem Eintreffen der Brandenburger dort mit ihren westafrikanischen Handelspartnern ausgehandelt hatten und an die sich die Brandenburger anpassen mußten. Dennoch gelang es Gröben auf der sozialen Ebene des Diskurses, die Brandenburger als gleichberechtigt mit den europäischen Akteuren und als überlegen gegenüber den westafrikanischen Akteuren zu inszenieren. Es konnten die textuellen Strategien aufgezeigt werden, mit denen Gröbens Texte dieses Wissen über die kolonialen Akteure im Westafrikahandel produzieren.

Human population is growing steadily (Goujon, 2019) and with this growth landscapes have been altered trough anthropogenic activities (Chase & Chase, 2016). An increasing amount of natural and semi-natural habitat is being transformed to provide residential space and associated infrastructure. Habitat transformation and loss of habitat connectivity exposes wildlife to new challenging conditions and novel environmental pressures including noise, chemical and light pollution (Grimm et al., 2008). Not all species can cope with these extensive and rapid changes. Urbanisation is identified as one of the main reasons for biodiversity loss (Altherr, 2007; Concepción et al., 2015; Luck & Smallbone, 2010; McKinney, 2006). Vertebrate loss is typically considered to be worst in urbanised areas because of intense and long-term disturbances that permanently alter habitats and depreciate food webs (Lombardi et al., 2017; McKinney, 2008). Nevertheless, there are always animal species that have adjusted to city life, so-called urban dwellers. An outstanding example of a successful urban dweller is the red fox (Vulpes vulpes). Foxes appear to be increasingly moving into human settlements throughout their range. Examples include Oslo, Norway (Christensen, 1985), Arhus, Denmark (Nielsen, 1990), Toronto, Canada (Adkins & Stott, 1998), Zurich, Switzerland (Gloor, 2002), or (in my focal area around) Berlin, Germany (Börner et al., 2009). Detailed knowledge of animal communities, food and competition relationships among the species, as well as species movement patterns and health status allows us to better understand the dynamics and predict the resilience of an ecosystem (Leibold et al., 2004). We need to know which characteristics allow species to persist in urban areas to prevent human-wildlife conflicts and promote biodiversity in cities. Identifying the biological traits favouring synurbisation is decisive to inform current management as well as to generate predictions for the future. In order to understand why the red fox is so successful in our anthropogenic world, we have to study different aspects of its ecology in, both, rural and urban settings. Therefore, this thesis investigates the diet, parasite spectrum and resting behaviour of red foxes along an urbanisation gradient in Berlin and Brandenburg (Germany). The diversity of these topics required the application of different analytical methods. For the investigation of the dietary strategies of rural and urban foxes, food niches were discovered using stable isotope analysis and compared with potentially available food items using Bayesian isotope mixing models (chapter 1). To study the diversity of helminths in the intestinal tract of foxes along a rural-urban gradient and to uncover environmental drivers of helminth communities I applied rarefaction curves, joint species distribution modelling (jSDM) and non-metric multidimensional scaling (NMDS) to helminth presence-absence data obtained by DNA metabarcoding. Finally, I compared the resting behaviour of foxes from Brandenburg and city foxes from Berlin using high resolution GPS and acceleration data. The assignment of behaviours based on the acceleration data served to determine temporal patterns of resting behaviour. Recurse analysis and the application of clustering algorithms allowed the identification of resting sites and their use. Our investigation on the red fox diet (Chapter 1) shows that dietary range of urban red foxes is smaller compared with that of rural conspecifics. Furthermore, higher δ13C values and lower δ15N values of urban foxes suggest relatively high input of anthropogenic food sources of urban foxes. Low within-individual variation compared to the between-individual variation lead to the conclusion that generalist fox populations consist of individual food specialists in urban and rural populations. Main results of the parasitological investigation (Chapter 2) show that the helminth diversity in the city Berlin is lower compared to surrounding rural Brandenburg and male red foxes tend to have higher helminth diversity than females. Diet features can drive helminth communities in red fox populations. Additionally, with increasing human population density, helminths transmitted via diet are less prevalent than pet-related helminths. Finally, I investigated habitat-dependent differences in resting patterns of red foxes from Berlin and Brandenburg (Chapter 3) and results revealed that urban foxes tend to rest more, with their resting behaviour concentrated during the day compared to rural red foxes. This increase in daily rest behaviour is reflected in an increased number of rest events. Moreover, the long-term resting events of city foxes last longer than those of foxes from Brandenburg. Even if rural foxes spend less time resting, rural red foxes of Brandenburg tend to have more resting sites compared to Berlin foxes. Overall, dietary specialisation and the use of anthropogenic food resources, in particular, have an overarching impact on the ecology of urban foxes. If proper food supply has such an extensive influence on the ecology, behaviour and lifestyle of red foxes, management strategies should focus on this topic. Reduced food availability would probably increase the competitive pressure within the fox population, reduce population density and thus also the contact rate between humans, domestic animals and foxes. Human-wildlife conflicts in the city could thus be reduced and the general acceptance of wild animals in the city increased. This could ultimately lead to the sharing of urban areas by humans and wildlife.

Die vorliegende Arbeit zielt darauf ab, die starke Hinwendung zum Monolog, die sich im europäischen Theater und Theaterschreiben seit den 1960er Jahren konstatieren lässt, zu thematisieren, theaterhistorisch einzuordnen und ästhetisch zu verorten. Den Gegenstand der Arbeit stellen nicht-dramatische bzw. postdramatische Theatertexte und -aufführungen dar, die sich zeitlich von den 1960er Jahren bis zur Gegenwart erstrecken und in denen die monologische Form als Organisationsprinzip zugrunde gelegt wird. In der vorliegenden Dissertation werden Theatertexte von Thomas Bernhard, Philippe Minyana, Samuel Beckett, Dea Loher, Dimitris Dimitriadis, Sarah Kane, Rainald Goetz, Forced Entertainment, Falk Richter, Elfriede Jelinek und René Pollesch und Inszenierungen von Andreas Kriegenburg, Thomas Ostermeier, Falk Richter, Jossi Wieler, Rimini Protokoll und René Pollesch besprochen und analysiert. Dabei dient der von Bachtins dialogischer Theorie übernommene aber neu konzeptualisierte Begriff der Monologizität als theoretische Kategorie, um die unterschiedlichen monologischen Theater- und Theatertextformen auf eine gemeinsame konzeptuelle Basis hin zu untersuchen und ihre wichtigsten gemeinsamen Form- und Strukturelemente herausarbeiten.

Seit dem Tode Mao Zedongs im Jahr 1976 kämpft die Kommunistische Partei Chinas um einen innerparteilichen und gesellschaftlichen Konsens zum Umgang mit ihren eigenen „historischen Fehlern". Immer wieder werden offizielle Narrative sowohl durch zivilgesellschaftliche Akteure als auch aus dem Inneren der Partei heraus in Frage gestellt bzw. kritisiert. Anhand der Entwicklung unterschiedlicher offizieller und inoffizieller Narrative zur Kampagne gegen Rechtsabweichler (1957–1958) lässt sich erkennen, dass der Kampf um das kollektive Gedächtnis, insbesondere im China unter Xi Jinping, eine langanhaltende zentrale politische Herausforderung für die nachhaltige Stabilisierung der Herrschaftslegitimation der KPCh darstellt.