With an estimated 19 million new cancer cases each year and almost 10 million deaths worldwide, there is a huge need for an optimised development of new therapeutic anticancer compounds as well as prediction of cancer patient response at bedside. Pharmacometric modelling and simulation allow to optimally leverage the rich longitudinal data from clinical studies and the different oncology variables and endpoints as tumour response, biomarker concentrations and survival information to better optimise clinical decision-making in oncology at its different stages: the development of new therapeutic compounds with optimised dosing selection and during therapeutic use to predict patient prognosis for improved treatment decisions at bedside.

Beginning with the development of new therapeutic compounds, identification of the appropriate/optimal dosing for the confirmatory phase III trials that maximises efficacy and minimises toxicity remains the most challenging component of clinical drug development. For this reason, a better understanding of the impact of different dose levels on efficacy and toxicity is needed to characterise the dose-response relationship and offer a more rational derivation of optimal doses/dosing. Compared to traditional pairwise comparisons between different study arms of dose-finding studies, statistical and model-based approaches have been shown to best leverage phase II dose-finding study data and characterise the dose-response relationship. These different approaches have been endorsed by the US Food and Drug Administration (FDA) which has recently initiated “Project Optimus” with the aim to optimise dose finding in oncology. Therefore, project I aimed to compare the performance of two model-based approaches within the oncology setting: the recently proposed combined Likelihood Ratio Test (cLRT) which leverages longitudinal phase II data and has shown high power detecting a dose-response relationship—but is computationally expensive, and the Multiple Comparison Procedure (MCP), the earlier and more established approach that has gained the FDA’s qualification as “fit-for-purpose” for the design and analysis of phase II studies. A simulation-based framework of a dose-finding phase II study under different study design considerations was established and applied to investigate cLRT and MCP performance. The results showed that, in general, cLRT was associated with higher power (=1−type II error) compared with MCP (89.8% vs 27.0%); however, its type I error (i.e. false positive) was not well controlled (mean: 13%) compared with MCP (<4%). Moreover, cLRT power was less sensitive to the different study design variables (e.g. number of patient with respect to number of dose levels) in contrast with MCP. Therefore, based on these results, before cLRT can be recommended to analyse dose-finding studies in oncology, further investigation of its robustness to different model complexities and study design variables as well as investigation of conditions that would better control type I error are needed to justify its high power at the expense of its computational demands.

Equally important to the development of new therapeutic compounds with optimised dose selection, is the accurate and early prediction of patient prognosis to monitor patient response and improve treatment decisions at bedside. Non-small cell lung cancer (NSCLC), the leading cause of cancer-related death, represents a disease of high burden and poor prognosis. Therefore, project II was conducted with the aim to identify early predictors of efficacy for NSCLC patients to spare them the unnecessary exposure to toxicities and contribute to better prediction of treatment outcomes. Clinical data from patients with advanced NSCLC receiving first-line combination therapy with paclitaxel and a platinum-based drug, were leveraged to characterise and quantify the relationships between anticancer drug exposure, tumour dynamics and C-reactive protein (CRP) concentrations—as a measure of the inflammatory level, using pharmacometric modelling. Model-derived variables were then investigated as potential predictors of the most important and commonly adopted efficacy endpoints progression-free survival (PFS) and overall survival (OS) by means of parametric time-to-event modelling, with a special focus on the potential of early longitudinal biomarker information as a potential early prognostic predictor. The results of our modelling framework in which longitudinal CRP concentrations were leveraged for the first time for prognostic investigations, identified the inflammatory level at treatment cycle 3 (CRPcycle3), i.e. day 42 from start of treatment, and the extent of absolute reduction in the inflammatory level between treatment cycles 3 and 2 to be the most significant predictors of PFS. Besides the CRP-related metrics, baseline tumour size and presence/absence of liver lesions were found to be predictors of OS. Nevertheless, CRPcycle3 was by far of the highest impact. The identification of CRP at treatment cycle 3 points to the potential and more informative value of longitudinal biomarker data compared to the commonly applied approach in which only baseline (pre-treatment) measurements are investigated and which do not reflect the patient situation and dynamic evolution of the disease. Measuring longitudinal CRP as a routine biomarker allows for the monitoring of inflammatory levels and, along with its reduction across treatment cycles, presents a promising prognostic marker for the timely identification of patients at risk of therapeutic failure, early progression and/or short survival to spare them unnecessary toxicities and provide alternative treatment decisions.

Overall, the two projects presented in this thesis, acknowledged and addressed, by leveraging pharmacometrics methodologies, two critical needs within the scope of oncology drug development and therapeutics that require early and more optimised clinical decisions. First, was the need to optimally identify the drug effect for an accelerated and optimised development of efficacious compounds for oncologic indications. Based on a scientific understanding of the dose-response relationship during early clinical drug development, a more informed and optimised dosing selection can be achieved. Within the investigated approaches, MCP, a robust but less powered approach currently exists. However, for better power, systematic investigations of cLRT are needed to achieve a more robust performance. Through the establishment of a proper understanding of the dose-response relationship, decisions regarding optimal dosing selection can be better informed that would maximise the drug’s efficacy, safety and tolerability and would consequently reflect on more successful phase III trials and lower attrition rates. Second, was the need to accurately and timely predict treatment outcomes and prognosis to monitor patient response, and improve treatment decisions at bedside. Our developed modelling framework, successfully identified the minimally invasive and cost-effective biomarker, CRPcycle3, as a significant predictor of PFS and OS. It also proposed that monitoring the inflammatory level was of prognostic value. The application of this modelling framework goes beyond NSCLC and is envisioned to suit other treatment modalities such as immunotherapies or targeted therapies for prediction of patient response and treatment outcomes in different settings. Finally, with use of proper methodologies as pharmacometric modelling and simulation, different data were successfully leveraged to optimise dosing selection and patient monitoring for a pharmacometric-based optimisation of early clinical decision-making in oncology.

Objective Despite the availability of effective antibiotics and pneumococcal vaccines, lung barrier failure remains a major complication of pneumonia, with high mortality and morbidity rates. Loss of function of the endothelial cystic fibrosis transmembrane conductance regulator (CFTR) has been associated with lung barrier failure in pneumonia. Additionally, pharmacological restoration of CFTR function with ivacaftor has shown barrier-protective properties. The regulatory cascade of CFTR includes lysine-deficient protein kinase 1 (WNK1), which has been suggested as a possible mediator of barrier breakdown. Yet, the mechanisms by which CFTR contributes to lung barrier integrity and its functional interaction with WNK1 remain incompletely understood. To address this, this study has three main objectives: (i) to test whether in vivo WNK1 activation could mimic the effect of ivacaftor-mediated CFTR potentiation; (ii) to investigate the role of WNK1 and its downstream target kinases, SPS/Ste20-related proline-alanine-rich kinase (SPAK) and oxidative stress-responsive kinase 1 (OSR1), in pneumonia-induced lung barrier disruption, and (iii) to determine whether the regulation of WNK1-SPAK/OSR1 is cell-type specific and differs between airway epithelial and endothelial cells. Methods A murine model (C57BL/6J wild-type) of severe pneumococcal pneumonia was used to study WNK1 activation and its effects on vascular leakage and inflammatory response. Furthermore, a heterozygous knockout of WNK1 (Wnk1+/-) and a homozygous knock-in of SPAK (SpakL502A/L502A) were used to investigate the functional role of the kinases in the murine infection model. To further characterize cell-type specific regulation of WNK1-SPAK/OSR1 signaling, human primary pulmonary microvascular endothelial cells (HPMEC) and primary human pulmonary alveolar epithelial cells (HPAEpiC) were used. Transepithelial/transendothelial electrical resistance (TEER) was used to measure the cellular integrity after pharmacological inhibition of WNK1 or SPAK. In addition, the effect of WNK1 inhibition (WNK-In-11) and nonspecific WNK1 activation (temozolomide) in uninfected or S. pneumoniae-infected conditions and their consequence on cell-specific regulation were investigated by proteomic and phosphoproteomic profiling. Results Pharmacological WNK1 activation prior to S. pneumoniae infection partly prevented severe barrier disruption, improved body temperature, and reduced bacterial burden. Genetic deficiency of WNK1 and SPAK in mice, however, did not affect lung permeability distinctly. Whereas, under in vitro conditions, inhibition of WNK1 decreased cellular resistance HPMEC but not in HPAEpiC. Notably, SPAK inhibition decreased resistance in both cell types. Although no obvious changes in the global proteome were observed, the phosphoproteomic profiling revealed that inhibitory WNK1 phosphorylation is increased in HPMEC but not in HPAEpiC after targeted WNK1 inhibition and S. pneumoniae infection. Conclusion The results suggest an integral role for WNK1-SPAK/OSR1 signaling in lung barrier function. Activation of WNK1 in vivo was shown to have beneficial effects on barrier stability. Furthermore, the signaling was shown to have distinct and cell-type-specific characteristics. In addition to WNK1 and SPAK/OSR1, the regulation of pulmonary permeability is likely to depend on other kinases, including other members of the WNK family.

In recent years, adoptive T cell therapy has brought a revolutionary change in the treatment for B-cell leukemias, lymphomas and multiple myeloma. Along with T cells modified with chimeric antigen receptors (CARs), there is ongoing development of T cells modified with T cell receptors (TCRs). TCR-T cells are not restricted to recognizing surface antigens but can also recognize targets derived from intracellular antigens and specifically mutated antigens, including neoantigens. This increases the number of target antigens for developing precision immunotherapies with high tumor specificity and a favorable safety profile. Challenges in the development include identifying suitable tumor epitopes, isolating high-affinity TCRs, and expanding patient TCR-T cells in sufficient quantities with the preferred phenotype. To tackle these questions, the first objective was to isolate neoepitope-specific TCRs from different repertoires in a side-by-side investigation. These repertoires included patients’ peripheral blood lymphocytes (PBLs) and tumor-infiltrating lymphocytes (TILs), PBLs of healthy donors, and a humanized mouse model. Results of the study show the advantage and feasibility of stimulating healthy donor repertoires and humanized mice TCR repertoires to generate mutation-specific TCRs with different specificities, especially when patient material is sparse. It was also shown that no specific TCR was isolated for neoepitope used in this study from the patients’ own repertoire. Furthermore, it is desirable to target recurrent mutations. A high-affinity TCR specific to the HLA-B*07:02-restricted epitope of the common lymphoma-specific driver mutation MyD88 L265P was successfully isolated from the healthy donor repertoire. However, screening 24 donors was necessary to identify one high-avidity TCR that could recognize tumor cells endogenously expressing L265P. This highlights the influence of the individual TCR repertoire of the T cell donor, along with inherent immunogenicity of the epitope. Lastly, the expansion of gene-modified T cells is a crucial part of the TCR-T cell manufacturing process. This is especially true for patients with insufficient numbers of T cells, or for T cells with insufficient expansion during the standard culture process. The feasibility of prolonging the T cell culture processes with minimal alteration to the standard process was demonstrated in the last presented study. These findings need to be confirmed on a GMP-compliant manufacturing platform but hold the promise to increase accessibility of adoptive T cell therapy (ATT) for certain patients.

Autoimmune diseases affect approximately 5-8% of the population worldwide and are characterized by defective or misdirected immune responses. Studies have shown that most autoimmune diseases result from a complex interplay of several factors, including genetic susceptibility, environmental stimuli and defects in immune regulation. However, in some cases, autoimmune diseases can also be caused by mutations in a single gene. The underlying mechanisms of those rare autoimmune diseases are often poorly understood, leading to inadequate management of disease symptoms and to a reduced life expectancy and quality of life for patients. Research on rare diseases provides the unique opportunity to study the function of the affected genes in humans, which not only contributes to a better understanding of the human immune system, but is also relevant for identifying new therapeutic targets for autoimmune diseases. In this thesis, we investigated mechanisms of immune regulation and function by studying three rare diseases. First, we studied the function of the alpha kinase ALPK1 in immune regulation by analyzing the immune cell composition and function of a patient with ROSAH syndrome, which is caused by a mutation in ALPK1. We found that the gain-of-function mutation in ALPK1 leads to an increased expression of TNFα and IL-6 by myeloid cells which triggers the auto-inflammatory symptoms observed in ROSAH patients. Consequently, blockade of TNFα resulted in a reduction of auto-inflammatory symptoms in our ROSAH patient. Furthermore, we investigated the role of mutations in the IL-36 receptor antagonist (IL36RN) in the pathogenesis of inflammatory bowel disease to better understand the function of IL-36 signaling in intestinal homeostasis and inflammation. We demonstrated that pathogenic IL36RN mutations are present in patients with Crohn’s disease (CD) and that anti-IL-36R therapy resulted in a reduced intestinal inflammation in one IL36RN-mutated patient. Thus, our data suggest that defects in IL-36 signaling might be involved in the pathogenesis of CD in a subgroup of patients and that blocking IL-36 signaling might represent a personalized treatment for these patients. Finally, we studied the role of the leptin-melanocortin signaling pathway in immune regulation by performing an in-depth immune cell profiling of patients with monogenic obesity. We observed a decreased frequency of pro-inflammatory myeloid cells in patients with mutations in the leptin receptor (LEPR), an increased frequency of effector T cells in patients with mutations in proopiomelanocortin (POMC) and an increased frequency of B cells in patients with mutation in the melanocortin 4 receptor (MC4R). These data indicate that LEPR, POMC and MC4R are not only important for appetite regulation but also have specific functions in the immune system. In summary, the data presented here highlight the importance of rare diseases in investigating the mechanisms of immune regulation and function and to identify new therapeutic targets for the treatment of autoimmune diseases. Based on our research performed here, we also developed a pipeline for characterizing rare autoimmune diseases, thus enabling a more precise diagnosis and identification of targeted treatment options for patients.

Diese Arbeit hat als Ziel die Bedeutung der 31sten Schrift (Plot.5.8) des Neuplatonikers Plotin (205– 275) für das Verständnis der sogenannten „Theologie des Aristoteles“ (ThA) aufzuzeigen. Der Titel von Plotins in die ThA eingearbeiteten Schrift lautet: „Wie könnte jemand die Schönheit des Nous und die von dessen Kosmos schauen?“ Die ältesten erhaltenen Fassungen der ThA sind in einer arabischen Kurzfassung (Kf) aus dem 15ten Jh., einer vollständig erhaltenen lateinischen Langfassung (Lf) von 1519 und in judäo-arabischen Fragmenten der Lf aus dem 14ten Jh. überliefert. Auf eine Schlüsselfunktion der 31sten Schrift deutet ihre Aufteilung auf zwei Einzel-Traktate und auf den Schlussteil der ThA in der lateinischen Lf hin. Diese gilt auch für die Kf der ThA, wie eine Wiederherstellung der originalen Traktat-Zuordnung deutlich macht: hierfür wird die geringere Traktat- Zählung der Kf auf die 14 Traktate der lateinischen Lf erweitert und diese erstmalig jeweils den originalen plotinischen Haupt-Fragestellungen und Theoremen zugeordnet, die in den drei Fassungen noch erkennbar sind. Für die Bestimmung des Verhältnisses von Lf und Kf dienen die judäo-arabischen Fragmente, die der Orientalist Andrei Borisov in den zwanziger Jahren des 20ten Jh.s entdeckt hat, als wichtige Textzeugen. Die Parallelisierung zeigt, dass alle Themen der Traktate in der ThA auf den Haupt-Fragestellungen bzw. Theoremen der 31sten Schrift und der anderen verwendeten Schriften Plotins basieren. Die Kf und Lf hatten somit ursprünglich die gleiche Traktat-Einteilung. Die frühere durchdachte Komposition und Thematik der ThA werden auf diese Weise auch in der Kf für die Forschung sichtbar gemacht. Inhaltlich können durch die judäo-arabischen Fragmente und die lateinische Lf originale (verlorene) arabische Passagen der Lf rekonstruiert werden und umgekehrt kann die erhaltene arabische Kf für verlorene judäo-arabische Fragmente als Vorlage dienen [Kap. 2]. Die Darstellung von Plotins in der ThA verwendeten Schriften und seines Unterrichts geben Hinweise auf die Intention des Verfassers der ThA [Kap. 3]. Die erstmalige deutsche Übersetzung und Textanalyse der in der lateinischen Lf bearbeiteten 31sten Schrift mit Vergleichen zu den anderen Fassungen zeigen deren Funktion, Komposition und Themen in der ThA. Ferner bestätigt die Analyse, dass die Lf die ältere ist und nicht die Kf, wie es im aktuellen Konsens heißt, und sie legt dar, inwieweit diese Fassungen übereinstimmen. Die ThA zeigt auf der Basis der 31sten Schrift [Kap. 4–7], wie jeder Mensch dank seines Intellekts die innere Schönheit sowohl der Sinneswelt und seines Selbst als auch der Himmels- welt durch aktive geistige Kontemplation erfassen kann und auf diese Weise die menschliche Seele die Schönheit der geistigen Welt, in der sich die Ideen bzw. die geistige Urform befinden. Ihrem Anspruch nach ist die ThA eine Universal-Theologie, da es für jeden Menschen möglich ist zu seinem geistigen Ursprung zurückzukehren. Die ThA endet mit der Genesis, dem in Erscheinung tretenden Logos Gottes. Hier stimmen Ziel und Anfang überein. Auf Grund der herausgearbeiteten und analysierten Themen können neue Hinweise auf die Intention und Identität eines möglichen Verfassers der ThA gewonnen werden [Kap. 8]. Dieser kannte Plotins zentrale Lehren wie die der Ideen bzw. Universalien in der geistigen Welt und hat die Logos-Lehre weiterentwickelt. Die herausgearbeiteten Themen der 31sten Schrift Plotins in der ThA sind: Schönheit und Glanz der sinnlichen und geistigen Welt und des Geistes (nous, ‘aql, intellectus), Emanation (perilampsis), Lehre der Seele (psychê, nafs, anima), Lehre des Intellekts – (innere und äu- ßere) Kontemplation, Logos-Lehre – Hervortreten (prohodos, ẖurūǧ, proventus / processio) – Erkenne- Dich-Selbst – Umkehr und Rückkehr (epistrophê, ruǧū‛, reversio) – Aufstieg (anhodos, ṣu‛ūḏ, ascensio) – Erleuchtung (ellampsis, nūr / iḍā’a, illuminatio) – Intuition – Wiederherstellung (apokatastasis), die Lehre der Ideen im Geist, Demiurg – Erstes Licht – Schöpfer – Hen, Intellectus agens et possibilis, Übereinstimmung von Anfang und Ziel (archê, ‛illa fā‛ila, principium und telos, ‛illa tamām, finis), Schöpfung (genesis, ẖalq, creatio), Universal-Theologie.

Das Thema der Arbeit “Strafrechtliche Verantwortlichkeit im Zusammenhang mit autonomen Fahrzeugen” ist weltweit bedeutend, dennoch werden nur einzelne Fragen dieses Themas in der Rechtswissenschaft, besonders im Strafrecht bzw. Nebenstrafrecht, behandelt. Anhand der besonders aktuellen Probleme ist es von Relevanz, strafrechtliche Theorie und die Verantwortung im Unfall in Verbindung zu setzen. Die Arbeit behandelt strafrechtliche Fragestellungen im Zusammenhang mit automatisierten bzw. autonomen Fahrzeugen. Sie widmet sich der Überlegung, ob die bestehenden strafrechtlichen Theorien und die Gesetze auch diesbezüglich angewendet werden können. In diesem Zusammenhang werden viele Fragen aufgeworfen, u.a. welche Strafbarkeit in verschiedenen kritischen Konstellationen vorliegen könnte, wie die Verantwortlichkeit zwischen Verkehrsteilnehmern verteilt wird, ob der Nutzer eines automatisierten bzw. autonomen Fahrzeugs dem Fahrzeug bzw. Fahrsystem vertrauen darf und welche Sorgfaltspflichten für Hersteller zu erwarten sind. Es ist noch zu fragen, ob das automatisierte bzw. autonome Fahrsystem bestraft werden kann und wie die Programmierung eines Algorithmus dogmatisch einzuordnen ist. Durch diese Überprüfung wird erkennbar, welche Lücken bei der Anwendung der geltenden Gesetze und strafrechtlichen Theorien noch vorfindbar sind. Es muss herausgefunden werden, welche Maßnahmen ergriffen werden können, um Unfälle zu vermeiden, die bei der Nutzung eines automatisierten bzw. autonomen Fahrsystems verursacht werden und in welche Richtung sich das Strafrecht richten sollte. Außerdem ist vorzustellen, wie die Gesetze in anderen Ländern autonome Fahrzeuge und Unternehmensverantwortung regulieren und welche Maßnahmen gegen Verletzung der Sorgfaltspflichten von Automobilunternehmern ergriffen werden. Das Ziel dieser Arbeit ist somit, unter Bezugnahme der Rolle des Strafrechts in der Risikogesellschaft und der präventiven Funktion von Strafe für die Verstärkung der Sorgfaltspflichten von Automobilunternehmern, konstruktive Gespräche zur strafrechtlichen Regulierung autonomer Fahrzeuge zu führen und befriedigende strafrechtliche Maßnahmen herauszuarbeiten und ggf. vorzuschlagen.

In the context of rapid urbanisation and growing mental health concerns, there has been an increasing focus on understanding how indoor environments, where humans spend the majority of their time, influence psychological well-being and mental health. It is now widely suggested that certain environmental features, such as geometry and forms, significantly shape individuals’ affective and behavioural responses. Notably, a recurrent phenomenon in research highlights a seemingly consistent preference for curvature over angularity across various stimulus categories including simple lines and shapes as well as more complex everyday objects. This preference is believed to also extend to the built environment. Generally, the underlying reasons for this phenomenon are still debated. Some views attribute the preference effects to a threat elicited by angles while others ascribe them to a pleasant intrinsic effect of curves. Yet, the limited body of available research exploring architecture and interior design settings often relies on unrealistic or poorly matched stimuli, mainly presented as images, and focuses on the aesthetic dimension, neglecting other psychological domains. These limitations raise questions about the robustness and implications of the reported effects, especially given the variability in findings across some studies, underscoring the need for further investigation. This dissertation comprises three publications aimed at systematically investigating the causal effects of short-term exposure to angular versus curved interior designs on affect, cognition, and behaviour, employing both virtual reality (VR) and images as presentation modes. Paper I (Tawil et al., 2021) introduces a novel VR paradigm to assess the psychological impact of interior environments with angular versus curved features across various domains, including affective and spatial experience, momentary affect, cognitive performance, and perceived restorativeness. The study used two pairs of photo-realistic three-dimensional living spaces, exclusively differing in form (angular versus curved) and style (modern versus classic). Out of 33 variables assessed, only two revealed differences, favouring rooms with angular features on the “order” and “novelty” dimensions. Further Bayesian analysis supported the results from the initial frequentist approach, challenging the preference for curvature and suggesting that psychological responses to angularity versus curvature in close-to-reality architectural settings may entail greater complexity. Paper II (Tawil et al., 2022) focuses on the effects of interior environments with angular versus curved features on aesthetic preference (i.e., liking and beauty) and stress ratings (i.e., restfulness and stressfulness), using 20 images derived from the virtual rooms used in Paper I, controlled for low-level image features. Additionally, style and sex effects were examined to account for potential interactions. Results indicated that participants reported higher aesthetic preference and lower stress in response to images of curved compared to angular interiors. Notably, effects on stress responses were consistent, whereas aesthetic preference for curves was context- and sex-dependent, being limited to modern style and the female subgroup. These findings suggest that the impact of angular versus curved interiors appears to extend more widely to psychological and potentially physiological stress responses than to aesthetic evaluations that may be influenced by person-specific preferences, perhaps based on differential experiences and cognitive processes. Paper III (Tawil et al., 2023) examines the preference for curves more implicitly, employing a battery of four reaction time paradigms that particularly focus on approach-avoidance behaviour, aligning with the different accounts regarding the source of the effect. The study aimed to capture attentional, motoric, as well as associative-semantic and -motoric biases towards angular versus curved interior designs. Results confirmed implicit effects with two paradigms, establishing links with approach-avoidance tendencies using experimental tasks and according reaction times. Participants semantically associated curvature with approach and angularity with avoidance, with stronger effects in women. Moreover, biases in motoric representations were identified, with participants exhibiting faster approach and slower avoidance responses to curvature, while no differences in responses to angularity were observed. These effects were intensified for interiors with the modern style. The findings may hint at (partially) automatic responses to curvature in interior design settings. In conclusion, this thesis presents empirical evidence supporting the potential positive effects of interior spaces with curved features, particularly when presented as images, rather than in a dynamic spatial experience (i.e., VR). It underscores significant impacts on explicit (i.e., self- reported aesthetic preference and stress) as well as implicit responses (i.e., semantic and motoric representations relating to approach-avoidance tendencies). Furthermore, it highlights the importance of accounting for contextual (e.g., style) and person-specific (e.g., participant sex) factors when exploring the effects of angular versus curved designs on human psychology and physiology. Ultimately, this research contributes to the expanding fields of environmental and architectural psychology. Its findings hold promise for evidence-based design strategies aimed at considering affective and behavioural human responses. This may be particularly relevant to spaces intended to promote mental health and well-being, but also everyday environments, fostering a more informed approach to architecture and interior design.

Huntingtin (HTT) is a highly conserved and ubiquitously expressed, multi-functional protein. Mutations within exon 1 of the Huntingtin gene (HTT) are translated into a pathogenic expansion (>36 glutamines) of the polyglutamine (polyQ) tract, giving rise to an aggressive neurodegenerative disease: Huntington’s disease (HD). Aggregates are well established as a key hallmark which are linked to the pathogenesis of HD. A small, truncated mutant fragment of HTT (mHTTex1), has been identified as a component of neuronal aggregates in HD patient brains, and its presence recapitulates HD-like symptoms in model systems. However, the molecular mechanisms dictating mHTTex1 aggregation and toxicity remain elusive.

To this end, a novel Drosophila Melanogaster (D. melanogaster) model was developed to investigate mHTTex1 aggregates. Transgenic fly strains were generated that pan-neuronally co-express HTTex1 mNeongreen (HTTex1-mNG) and HTTex1 mScarlet-I (HTTex1-mSc-I) fusion proteins with 17, 52, or 75 glutamine repeats (HTTex1Q17-mNG/-mSc-I, HTTex1Q52- mNG/-mSc-I, and HTTex1Q75-mNG/-mSc-I) which enabled detection of mHTTex1 aggregation in situ by measuring Forster Resonance Energy Transfer (FRET). Through the neuronal expression of this FRET-based HTTex1 aggregation biosensor, the localisation of HTTex1 within the fly brain was tracked. Co-expression of pathogenic mHTTex1 fusion proteins (HTTex1Q52-mNG/-mSc-I and HTTex1Q75-mNG/-mSc-I) resulted in the formation of SDS stable mHTTex1 aggregates , which correlated with reduced lifespan and mobility in flies. A novel FRET-based fluorescence-activated cell sorting method (FACS) was established which enabled quantitative readout of mHTTex1 aggregates within live cells derived from fly brains.

To interrogate the impact of mHTTex1 aggregates on the proteome, label free quantitative proteomics (LC-MS) analysis of both immunoprecipitated (IP) mHTTex1 aggregates from fly head lysates and whole fly brain samples was performed. Immunoprecipitates and whole brain lysates from flies expressing mHTTex1 with both pathogenic and non-pathogenic polyQ tracts were compared. The results obtained revealed that mHTTex1 aggregates significantly associate with proteins involved in intercellular transport and lead to a global increase of proteins associated with the endomembrane system.

The findings of this work establish mHTTex1 aggregates as profound disrupters of the neuronal proteome. RNAi knockdown of the key upregulated protein sff resulted in significant toxicity, highlighting a potential protective role increased sff protein levels may play against HTTex1-induced toxicity.

This work is concerned with the ultrafast magnetization dynamics found in rareearth and transition metals. In the first part, we have investigated the magnetization dynamics of 60 nm Gd(0001) grown on W(110) in XMCD in reflection. Our analysis shows that the ultrafast timescale of demagnetization, which was found by previous authors in 10 nm thick Gd films [2, 20] can be attributed to effects at the Gd/W interface, which we interpret as magnon driven spin currents. In the second part of the work, we have applied both XRMR and XMCD in reflection to study the magnetic structure of the synthetic ferrimagnet Fe/Gd grown on W(110). We find clear signs of a twisted magnetization state, as proposed by Camley and Tilley [10, 11] in which both layers form a domain-wall-like structure with the magnetic moments being aligned almost perpendicular to the field at the Fe/Gd interface and tilting stepwise towards field-aligned throughout both layers. We further find very diverse magnetization dynamics in XMCD in reflection, ranging from an increase of the detected magnetization component on a slow ps-timescale to all-optical switching of only the Gd component within less than 1 ps. The differences are caused by the temperature dependent orientation of the magnetization in the twisted state. As it can likewise be manipulated by the external magnetic field strength, synthetic ferrimagnets offer a great amount of controllability of their magnetization dynamics.

The present dissertation discusses the origins and development of anticommunist cooperation between the Brazilian government under Getúlio Vargas and the Third Reich. The analysis identifies that this cooperation underwent three distinct phases between the years 1933 and 1938. The initial phase, covering 1933 and 1934, was marked by the independent implementation of anticommunist policies, with Brazilian authorities drawing inspiration from the Third Reich for the conception of their own authoritarian project. The second phase, spanning 1935 to 1937, represents the peak of anticommunist collaboration between the two regimes, catalyzed by the communist uprisings in Brazil in November 1935. During this period, anticommunism became part of the bilateral diplomatic agenda, leading to cooperation in counterintelligence operations, law enforcement coordination, and the formulation and distribution of anticommunist propaganda. The third phase, from 1937 to 1942, saw the decline of the German-Brazilian anticommunist partnership, eventually leading the former allies to fight on opposite sides during World War II.

Cell-free protein synthesis can circumvent the challenge of producing membrane proteins and toxic proteins in living cells. Its open environment permits the addition of specific components to enhance protein solubility and introduce novel chemical reactants, such as non-canonical amino acids, for site-specific protein modifications. Eukaryotic cell-free systems are particularly advantageous for producing complex proteins, as they support post-translational modifications and optimal protein folding conditions. However, cultivating mammalian cells, like the commonly used Chinese hamster ovary (CHO) cells for pharmaceutical protein production, is costly. This doctoral thesis aims to develop a cost-effective cell-free system for producing difficult-to-express proteins. To achieve this, an optimal gene location for stable CHO cell line development and subsequent preparation of translationally active cell lysate was identified. The efficiency of cell-free transcription was enhanced by incorporating T7 RNA polymerase into CHO cells, while translation initiation was improved by using CHO cells expressing a mutant of the α subunit of eukaryotic initiation factor 2. Additionally, straightforward orthogonal cell-free translation was achieved by introducing orthogonal aminoacyl tRNA synthetases into CHO cells, facilitating the cell-free production of site-specifically modified membrane proteins. Furthermore, the identified locus in the CHO genome was utilized to create a tetracycline-inducible expression system, enabling both cell-free and inducible cell-based production of challenging proteins at desired time points and cell densities. Another goal of this research was to develop a yeast-based cell-free system using Pichia pastoris. This system combines the benefits of low-cost production, a eukaryotic environment, and the opportunity to synthesize difficult-to-express proteins. Moreover, it could be demonstrated that statistical Design of Experiments can be performed in a short time and on a small scale to identify optimal reaction parameters, facilitating the rapid adjustment of required protein synthesis conditions. In summary, the established methods streamlined induced cell-based expression and cell-free protein synthesis by minimizing the need for external supplements, improving protein production, and reducing costs.

Hypocalcämie ist eine häufig auftretende Stoffwechselstörung der Milchkuh im Zeitraum bis 48 Stunden nach der Abkalbung. Die subklinische Hypocalcämie wird in diesem Zusammenhang häufig als „gateway disease“ bezeichnet, da sie zu zahlreichen Folgeerkrankungen prädisponiert und so zu erheblichen wirtschaftlichen Einbußen und einer Beeinträchtigung der Tiergesundheit führt. Seit den 1970er-Jahren wird bei mehrkalbigen Milchkühen ca. fünf bis sieben Tage vor der erwarteten Kalbung Cholecalciferol gespritzt. Ziel ist es, die Mechanismen der Calciumhomöostase bereits vor der Kalbung zu aktivieren und die Tiere so besser auf das bei Einsetzen der Milchproduktion entstehende Calciumdefizit vorzubereiten (Gürtler et al. 1977). In der vorliegenden Studie wurde der Einfluss einer Cholecalciferol-Injektion a.p. auf die Serumdynamik von Calcium, Phosphor und Magnesium nach der Kalbung sowie auf die Milchleistung und verschiedene Reproduktionsparameter in der Folgelaktation untersucht. Dazu wurden 446 pluripare Milchkühe, die in der Trockenstehphase eine Ration mit positiver DCAB erhielten, von 11 Betrieben in Nordwest-Deutschland sieben Tage vor dem Kalbedatum mit einer intramuskulären Injektion von 10 Mio. I.E. Cholecalciferol oder einem Placebo behandelt. Alle Tiere wurden fünf bis zehn Tage nach der Kalbung auf Anzeichen einer Metritis untersucht und der Kalbeverlauf sowie das Auftreten von Zwillingen, Totgeburten, klinischer Hypocalcämie und Retentio secundinarum dokumentiert. Bei einer Teilstrichprobe von 95 Tieren wurden an Tag 1, 2 und 3 p.p. jeweils eine Blutprobe entnommen und auf die Konzentration von Calcium, Phosphor und Magnesium untersucht. Des Weiteren wurde bei 38 Tieren zusätzlich zwei Tage nach der Injektion eine weitere Blutprobe entnommen und der Gehalt an Progesteron und Östradiol bestimmt. Die Blutprobensätze von 30 Tieren, bei denen zu allen vier Zeitpunkten Blut gewonnen wurde, wurden zudem auf die Konzentrationen von Cholecalciferol, 25-Hydroxycholecalciferol (25OHD3), 1,25-Dihydroxycholecalciferol (1,25(OH)2D3) und 24,25-Dihydroxycholecalciferol (24,25(OH)2D3) untersucht, um die Metabolisierung des Cholecalciferols im Organismus nachvollziehen zu können. Nach Ende des Beobachtungszeitraums (Mai 2022 bis September 2023) wurden die gesammelten Milchleistungsdaten und Reproduktionsparameter ausgewertet. Die Injektion von 10 Mio. I.E. Cholecalciferol sieben Tage vor der Kalbung hatte keinen Einfluss auf die Milchleistung in der Folgelaktation (P = 0,67). Die Tiere der Behandlungsgruppe hatten tendenziell höhere Serum-Ca-Werte in den ersten drei Tagen p.p (P = 0,08). Es konnten außer einer erhöhten Inzidenz von Retentio secundinarum in der Behandlungsgruppe (Kontrolle: 7,9%, Behandlung: 14,8%, P = 0,02) keine Auswirkungen der Cholecalciferol-Injektion auf die Inzidenzen von Totgeburten, klinischer Hypocalcämie und Metritis sowie die Milchleistungsdaten und Reproduktionsparameter beobachtet werden. Die Analyse der Konzentrationen von Progesteron und Östradiol vor der Kalbung ergab keine Unterschiede zwischen den beiden Gruppen (P = 0,81). Die Hypothese aus einer ähnlich aufgebauten Studie von Venjakob et al. (2022) mit Fütterung einer negativen DCAB in der Vorbereiterration, dass die Cholecalciferol-Injektion den Steroidhormonmetabolismus dahingehend beeinflusst hätte, dass es zu einem frühzeitigen Abbau von Progesteron zu Östradiol und damit zu einer vorzeitigen Initiierung der Abkalbung gekommen sein könnte, konnte anhand dieser Ergebnisse nicht bestätigt werden. Die Untersuchung der Blutproben auf den Gehalt von Cholecalciferol, 25OHD3, 1,25(OH)2D3 und 24,25(OH)2D3 ergab einen etwa 100-fachen Anstieg der Cholecalciferol-Konzentration auf 464,0 ng/ml zwei Tage nach der Injektion und einen vierfachen Anstieg der 25OHD3-Konzentration auf bis zu 205,5 ng/ml in den ersten drei Tagen p.p. in der Behandlungsgruppe. Die 1,25(OH)2D3-Konzentration im Serum der mit Cholecalciferol behandelten Tiere erreichte fünf Tage vor der Kalbung deutlich höhere Werte (89,2 pg/ml) als bei den mit einem Placebo behandelten Tieren (34,9 pg/ml). Nach der Kalbung zeigte jedoch die 1,25(OH)2D3-Konzentration in der Kontrollgruppe einen stärkeren Anstieg (Behandlung: 122,9 pg/ml; Kontrolle: 166,0 pg/mL an Tag zwei p.p.), sodass der 1,25(OH)2D3-Wert an Tag drei p.p. in der Kontrollgruppe signifikant höher lag (Behandlung: 101,6 pg/ml; Kontrolle: 155,6 pg/ml; P < 0,05). Diese Ergebnisse zeigen, dass die Wirkung der Cholecalciferol-Injektion auf den Serum-Ca-Spiegel p.p. in der vorliegenden Untersuchung vermutlich durch 25OHD3 vermittelt wurde. Insgesamt zeigen die Resultate dieser Untersuchung, dass die Kombination einer Injektion von 10 Mio. I.E. Cholecalciferol ca. sieben Tage vor der erwarteten Kalbung und der Fütterung einer kationenreichen Ration in der Trockenstehphase zu deutlich geringeren Auswirkungen auf die Serumdynamik von Ca und Pi als die Kombination mit einer Vorbereiterration mit negativer DCAB führte. Es konnten abgesehen von einer höheren Inzidenz von Retentio secundinarum in der Behandlungsgruppe dennoch keine negativen Effekte auf die Inzidenz von Metritis sowie die Milchleistung und Güstzeit in der Folgelaktation nachgewiesen werden.

Mourning as Melancholia—Works of Grief in Contemporary Literature and Theory is a study that observes how experiences of loss and bereavement are portrayed in contemporary texts. My dissertation’s argument is based on the assumption that the portrayal of mourning does no longer comply with Sigmund Freud’s psychoanalytical agenda. It does, to be more precise, no longer follow the assumption that the mourner has to ‘work’ through loss in order to overcome it and achieve a former state of wellbeing and functionality. Recently published autobiographical, fictional, and theoretical accounts of mourning thus challenge established assumptions about what grief is and how it affects us. This is why this study reassesses the notion of the ‘grief work’ (‘Trauerarbeit’) that Freud developed in his canonical “Mourning and Melancholia.” In Freud’s essay, mourning is cast as a necessary process that enables the mourner to externalize his or her grief. Melancholia is on the contrary framed as a pathological condition because here, the melancholic

identifies with its enigmatic sense of loss and thus holds onto and incorporates the lost object. My study proposes that the narrators and protagonists, whom I observe, can be described as melancholic mourners, who continue to identify with their relation to the loved person without, however, being able to fully comprehend or explain it. As a consequence, they remain inconsolable, vulnerable, and impaired. The essentially bereaved melancholic figures that populate contemporary texts do therefore not only present grief as incomprehensible and potentially interminable, they also strongly identify with the vulnerability that the experience of loss has exposed them to. In doing so, they refuse to tell stories that coherently incorporate all of their essential episodes, but instead insist on the meaninglessness of loss. This is why this study has made it its task to ask how a life story that no longer ‘makes sense’ because it revolves around the stubbornly enigmatic void of loss can be written, read, and understood today. The first chapter retraces the social history of the conceptualization of grief. It demonstrates that the experience of grief is dependent upon the cultural context from which it emanates. The fact that grief was once clad in public rituals but is today perceived as a private feeling already indicates how impressionable the concept of grief is. In order to highlight the dynamics that shaped our idea of what it means to mourn, Freud’s psychoanalytical theorization of the ‘work of mourning’ will be reassessed and aligned with Max Weber’s critique of a paradigmatically American ‘work ethic.’ Recent sociological approaches that address the social construction and determination of emotions will factor into a discussion that focuses on the interceding functions that texts about mourning can assume today.

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After chapter one discusses Joan Didion’s The Year of Magical Thinking (2005) and Meghan O’Rourke’s The Long Goodbye (2011) as paradigmatic examples for the growing genre of the ‘grief memoir,’ chapter two focuses on Dave Eggers A Heartbreaking Work of Staggering Genius (2000). In Eggers’ book, the traumatic experience of parental death is circumvented rather than rendered. Yet it is precisely the young narrator’s inability to articulate his grief and inexplicable sense of loss that both motivates and disables his story. This raises the complex question of how to tell an experience that one cannot and does not want to integrate into one’s life story, for fear of ‘making sense’ of the essential meaninglessness that melancholic mourning incorporates. Chapter three retraces Roland Barthes’s concern with the lacerating pain of emotional

suffering. It observes how the critic’s perspective changes after his mother’s death. A close- reading of Barthes’s posthumously published Mourning Diary (2010) and late Camera

Lucida (1980) highlight that Barthes insisted as much on the meaninglessness of his mother’s death as on the intractable uniqueness of her being, the latter of which he intended to capture in an envisioned, but never realized literary text called Vita Nova. His failure to render his mother’s essential being without imbuing her death with conventional meaning had a grave impact on Barthes’s worldview: it essentially changed his perspective on the relation between the writing subject and the written text. Siri Hustvedt’s works are the focal point of the fourth chapter. Her narratives exhibit characters that are defined by their affective ties and unconscious desires. It is through the experience of grief that the design of their relationally constituted identities is brought to light. Despite the fact that Hustvedt’s stories are deeply steeped in psychoanalytic theory and rely heavily on Freud’s assertions, they are told from the point of view of narrators, who grapple with a melancholic form of grief that arises from their complex, often ambivalent and always persistently captivating relationship to the deceased.

Gender has proven to be a relevant factor in schools, particularly given the finding that girls tend to outperform boys. Empirical findings relating to not just gender but also constructs associated with gender, such as gender stereotypes and gender role self-concepts, can offer insight into academic, social, and personal functioning of children and adolescents. In regarding not academic achievement but rather a variety of variables associated with the educational system, such as perceived likability and competence, attributions of success, and self-esteem and achievement motivation, the present works expands upon the existing literature. Past work has examined gender in relation to these constructs, but research on how gender stereotypes and gender role self-concept relate to perceived likability and competence, attributions of success, and self-esteem constructs and achievement motivation is less common. As the school environment features a variety of unique roles, the perspectives of educators, peers, and students themselves are taken into account in the present works. In manuscript 1, we gathered gender stereotypes about young children held by German adults in a first study consisting of N = 397 participants. This information allowed the construction of vignettes in which gender stereotypical and nonstereotypical three-year-old boys and girls were portrayed. In our main study, pre-service pedagogical educators (N = 414) indicated their liking, perceived competence, creativity, self-esteem, prosocial behavior, internalizing and externalizing problems of hypothetical children using a 2 x 2 between participants design (target gender: boy, girl; target stereotyped behavior: masculine, feminine). We hypothesized that counterstereotypical children, particularly feminine boys, would receive the lowest ratings in likability. Further, we hypothesized that ratings of competence would differ between stereotypical and nonstereotypical children and expected the latter children to receive positive ratings in terms of creativity and self-esteem. The analyses revealed no significant difference in liking but showed that masculine girls received better ratings in terms of competence, creativity, and self-esteem compared to feminine girls. This stands in contrast with past literature, which shows adults displaying negative biases towards feminine boys (Coyle et al., 2016; Sullivan et al., 2018; Thomas & Blakemore, 2013). The backlash for feminine boys, seen in the past literature, and our findings, showing benefits for masculine girls, highlight the social standing of masculinity (Feinman, 1981). In manuscript 2, we investigated a peer perspective on how gender stereotyped classroom behavior could impact causal attributions of achievements, based on work by Kessels and Heyder (2020) and the paradox of praise (Meyer et al., 1979; Möller, 2005). Study 2 was an experimental vignette study in which high-achieving students displaying prosocial or nondescript behavior were described. N = 324 9th grade students participated in the 2 x 2 mixed design study, where target gender (girl, boy) was varied between participants and vignette (prosocial, nondescript) was varied within participants. Participants were asked to indicate (1) whether the hypothetical students would receive positive reactions from the teacher, as well as (2) how the students’ high achievements could be attributed, and (3) how they would rate targets on social and gender-related outcomes, such as perceived intelligence, masculinity, and femininity. We hypothesized that prosocial students would be perceived as receiving more positive teacher reactions. We further expected prosocial students to receive attributions of greater effort and lower ability, which should be mediated by the expected teacher reaction (Möller, 2005; Weiner & Kukla, 1970). Prosocial students were perceived as more effortful and less able than nondescript students, but the hypothesized teacher reactions did not mediate this association. The findings highlight that social classroom behavior, here feminine stereotyped prosocial behavior, can impact how academic achievements are attributed by peers. The belief that prosocial students are less able than nondescript peers is particularly relevant for female students. Manuscript 3, a cross-sectional study, established the relation between self-esteem constructs and achievement motivation with gender role self-concept. N = 355 9th grade students provided information on their gender role self-concept, global self-esteem, academic contingent self-esteem, hope for success, and fear of failure. We hypothesized positive relations between instrumentality and global self-esteem and hope for success, while expecting a negative relation with instrumentality and academic contingent self-esteem and fear of failure. Importantly, gender differential relations were hypothesized, as past work has shown that the association between a masculine gender role self-concept and self-esteem measures can be stronger in female individuals (Cate & Sugawara, 1986; Hirokawa & Dohi, 2007; Whitley, 1988). Structural equation modeling showed a benefit of an instrumental orientation, which was associated with greater self-esteem and hope for success, and lower academic contingent self-esteem and fear of failure. The association between instrumentality and global self-esteem was stronger for girls, for whom this association also indirectly related to lower fear of failure, highlighting how an instrumental gender role self-concept can particularly benefit girls. Our findings that instrumentality can benefit students expand the literature on the feminization of school, which has presented masculinity in school as mostly negative (Lyng, 2009). The results of all three empirical works highlight a pattern of benefits of masculinity and the disadvantages of femininity from the perspective of educators, peers, and students themselves. The discussion of the present dissertation thus outlines how gender can be understood as a status characteristic (J. Berger et al., 1972; Gerber, 2009), with maleness and masculinity perceived as holding more status than femaleness and femininity. This is seemingly in conflict with past work on the feminization of school, presenting the better fit between femininity and school (Heyder & Kessels, 2013). We discuss some practical and theoretical limitations of the present work and outline future research directions. The implications of the work focus on the feminization of school and on the question of whether reducing gender salience in schools could be advantageous, ultimately highlighting current trends in society regarding focus on gender.

This thesis advances the field of bioprinting and tissue modeling by addressing the limitations of current processing protocols for dECM-based bioresins. It explores effective enzymes for thermogelling dECM pre-gels, investigates the grafting of different methacryloylated groups on dECM for vat 3D bioprinting, and uses 3D printing to fabricate advanced dynamic culture systems that apply mechanical shear stress on epithelial cells to develop advanced in vitro tissue models. Precise control over the physical, mechanical, and biochemical properties of dECM hydrogels after solubilization and gelation is crucial for their application. While animal-derived pepsin is traditionally used for solubilization of dECM, in our first study, we investigated various nonanimal-based alternatives, including plant-derived papain, bacterial-based collagenase, and α- amylase. Although α-amylase-digested dECM exhibited the highest content of growth factors and cytokines, both α-amylase and collagenase treatments disrupted collagen self-assembly of dECM digests and compromised the protein structural integrity. Moreover, their insufficient digestion efficiency resulted in significantly reduced yields of dECM digests (∼ 7 – 10 %). In contrast, papain preserved the physical gel formation capacity of digested dECM and demonstrated comparable yields (∼79-86 %), thermo-gelation kinetics, and bulk mechanical properties to pepsin, along with increased amounts of retained growth factors and superior cell adhesion. As a conclusion, papain was found to be a cost-effective and reproducible alternative to pepsin for producing liver-derived dECM hydrogels, eliminating the risk for zoonotic disease transmission. The second study covered functionalization of papain-digested liver dECM with glycidyl methacrylate (dECM-GMA) and methacrylic anhydride (dECM-MA) into photoactive bioresins for vat photopolymerization. Both chemical modifications yielded a comparable degree of functionalization (DOF ∼ 81.5 – 83.5 %) without disrupting the dECM structure. Interestingly, dECM-GMA showed enhanced solubility and higher shear viscosity compared to dECM-MA due to the additional hydroxyl groups and the increased hydrogen bonding. When formulated into resins, both modifications yielded a similar degree of photocrosslinking (30 – 60 %, depending on the condition), enabling efficient on-demand crosslinking and good printing fidelity. The bulk stiffness of 3D printed dECM-GMA hydrogels was slightly softer (∼ 0.2 – 1.2 kPa) compared to dECM-MA (∼ 0.5 – 2 kPa), both being within the range of healthy human liver tissue (∼ 0.15 – 5.9 kPa), making them suitable candidates for bioprinting of HepaRG cells. dECM-GMA hydrogels exhibited enhanced hydrophilicity and cell compatibility and faster biodegradation compared to dECM-MA gels, highlighting the critical role of chemical functionalization in optimizing bioresins for vat bioprinting. In the third study, the papain digestion protocol established in the first study was applied to porcine intestinal dSIS, achieving characteristics similar to liver dECM and thereby demonstrating its versatility. Functionalized with methacrylic anhydride (MA) for vat 3D printing, dSIS produced biomimetic dSIS-MA hydrogels with tunable stiffness (3.7 ± 0.2 kPa), matching healthy human small intestinal tissue (∼ 1.3 – 4.0 kPa). Moreover, the functionalization enhanced biostability of the hydrogels when seeded with intestinal epithelial cells, essential for advanced intestinal in vitro tissue models. To enable dynamic cell culture conditions, a biocompatible 3D-printed millifluidic tissue chamber was designed to host cellular dSIS-MA hydrogel scaffolds while maintaining a laminar flow profile, as confirmed by computational fluid dynamics simulations. The millifluidic system enables advanced intestinal models with tunable stiffness and fluid shear stresses, offering a scalable and cost-effective solution for in vitro tissue modeling. As demonstrated by culturing HT29-MTX monolayers under low physiological shear stress (∼ 0.01 dyne/cm2), the system induced 3D tissue reorganization and enhanced differentiation and mucus production, particularly MUC5AC, when compared to static culture conditions as evidenced by immunofluorescent proteins staining. Experiments with human organoid-derived ileum ISCs showed that physiological shear stress combined with biomimetic hydrogel scaffolds decreased proliferation and stemness of the cells while promoting their multi-lineage differentiation. Changes in alkaline phosphatase (ALP) activity and secreted mucus demonstrated functional cell differentiation into enterocyte and goblet cell lineages. The studies in this thesis covered the critical steps in dECM bioprinting, from enzyme-based solubilization of dECM through their chemical functionalization for vat (bio)printing until the integration of advanced fluidic systems for tissue modeling. The studies in this thesis covered the critical steps in dECM bioprinting, from enzyme-based solubilization of dECM through their chemical functionalization for vat (bio)printing until the integration of advanced fluidic systems for tissue modeling. Each step advances the understanding and development of biomimical materials and strategies, enhancing the efficiency and functionality of bioengineered tissues. This paves the way for future applications in regenerative medicine and tissue engineering.

The growing popularity of anti-establishment parties and politicians among voters in Western democracies since the mid-2010s has led to a number of explanations. One widely accepted approach, both in journalistic and academic circles, is the theory that the "left behind" voters are primarily responsible for this development. Despite the popularity of this theory, it has two central problems: firstly, a clear definition of what “left behind” means and secondly, a lack of empirical testing. This dissertation addresses these problems using the 2016 U.S. presidential election as an example. Based on an examination of media and academic discourse, an empirically testable definition of the theory is distilled, operationalized, and then empirically tested at the county level through two regression models. The results of the analysis show that the chosen operationalization of the theory can explain approximately two-thirds of the variance in the vote share for Donald Trump. In order to understand the proportion of unexplained variance, counties in which Trump's share of the vote as predicted by the statistical models deviates significantly from the actual vote are then examined in descriptive analysis in exploratory form. It becomes clear that the reasons for the strong deviations lie less in the selection of the chosen variables, but probably in their lack of weighting, as well as in other factors such as voter turnout and historical loyalty to a party. The dissertation places this empirical research in a broader context by viewing Trump's election as a symptom of a transitional phase between two political paradigms. Both by adopting this perspective and through the empirical aspects of the work, this dissertation contributes to a better understanding of the origins of Trump's electoral success and opens up new ways to address them.

Directional proton transport across the lipid bilayer of a biological membrane is a process of fundamental importance in nature. Due to the protons’ small size, the study of proton transfer in biological macromolecules such as proteins, is a challenging task. One of the few techniques, that allows one to observe protonation reactions is infrared (IR) difference spectroscopy. Its chemical sensitivity sheds insight into the changes in protonation state of single moieties within proteins but is at the same time sensitive to all other molecular alterations accompanying proton transfer. IR difference spectroscopy is therefore a powerful tool to study the functional mechanism of proton-translocating proteins. A very intriguing question is how proteins that act as membrane-bound proton pumps achieve vectorial proton transport. The first part of this thesis aims to address this question by the spectroscopic investigation of a novel microbial rhodopsin acting as an inward proton pump. I characterized the sequence of protonation reactions with a focus on the cytoplasmic half channel of the protein. Here, deprotonation of an aspartic acid located on the cytoplasmic side is correlated to the deprotonation of the distantly located retinal Schiff base. The sequence of these protonation events has important implications on the functional mechanism of inward proton transport, which is compared to the mechanism of a prototypical outward proton pump. The structural changes associated with inward proton transfer are not only localized within the protein itself but are also transmitted to the surrounding lipid bilayer. Photoactivation of this microbial rhodopsin is sensed by the surrounding lipid molecules by two different processes occurring on a fast (< 100 ns) and a slow time scale (> μs). While time-resolved IR spectroscopy is an established tool to monitor light-induced absorption changes of reversible processes, it has found only limited application in the study of irreversible and light-insensitive reactions. In the second part of this thesis, I showcase two novel IR absorption techniques based on quantum cascade lasers (QCL) to study protein reactions. Both techniques were used to monitor protein conformational changes as well as protonation dynamics by single-shot experiments, i.e. without the need to average multiple acquisitions. After a thorough comparison of both techniques, I applied them to the study of the irreversible activation process of the G-protein-coupled receptor rhodopsin. This approach demonstrates that QCL-based IR spectroscopy is a powerful tool for the spectroscopic study of irreversible and, in the future, also light-insensitive (protein) reactions.

The use of cannabidiol (CBD) products is becoming increasingly popular among animal owners and veterinarians as an alternative treatment for stress, anxiety or pain in horses. In equestrian sports, all cannabinoids are banned due to their potentially psychotropic effects. However, there are only a few studies on the detection times of CBD concentrations in blood or urine, and the actual effectiveness in horses. The aim of this study was to determine the pharmacokinetic properties of CBD after oral administration in healthy horses and to analyse stress parameters, including behavioural observations, heart rate and cortisol levels. Study products were two pastes for oral administration, one with CBD as active ingredient and one without active ingredient. Paste administration was blinded. In the first study part (dose escalation study), the pastes were administered in escalating trials as single doses (0.2 mg CBD/kg, 1 mg CBD/kg, 3 mg CBD/kg) to a treatment and a control group. In the second part of the study (multiple dose study), both pastes were administered twice daily for 15 days (treatment group: 3 mg CBD/kg). For the pharmacokinetic analysis, blood and urine samples were taken daily during both study parts. After day 15 of the multiple dose study, additional samples were collected for two weeks to analyse the elimination phase. Concentrations of CBD, CBD metabolites and other cannabinoids were determined using gas chromatography/tandem mass spectrometry. Pharmacokinetic parameters were assessed using two approaches: Non-compartmental analysis and population pharmacokinetic analysis using a nonlinear mixed-effects model. During the elimination phase, the ratio between the steady-state concentrations of CBD in urine to serum (Rss) was calculated. In the dose escalation study, behavioural parameters were assessed using photographs to evaluate the horses' facial expressions on a specifically developed scale, which was based on existing scales (FaceSed and Horse Grimace Scale). To identify potential sedation, the horses' reactions to acoustic and visual stimuli were video recorded. The evaluation of the photos and videos was conducted in a blinded manner. In addition, the heart rate was continuously recorded via heart rate sensors throughout the study with subsequent analysis of heart rate (HR) and heart rate variability (HRV). In the multiple dose study, facial expressions and the depth of sedation were analysed daily following the same protocol as in the dose escalation study. In addition, blood and saliva samples were daily collected and analysed for cortisol levels using liquid chromatography/tandem mass spectrometry. The behavioural observations and cortisol levels were compared between the groups. A novel object test and a trailer test were performed prior to study start. Both tests were repeated on study day 13. Assessment included reactivity, movement patterns such as gait changes and behavioural characteristics. Heart rate was recorded during the tests and analysed using HR and HRV parameters. Blood and saliva samples were obtained before and after the tests for cortisol analysis. The CBD paste was well-tolerated and no side effects were observed. The non-compartmental analysis showed a maximum serum concentration of 12.2 ng/ml after single administration of CBD (3 mg/kg). The population pharmacokinetic analysis showed that a three-compartment model with zero-order absorption most accurately describes the pharmacokinetic properties of CBD. High volumes of distribution into peripheral compartments and high concentrations of the metabolite 7-carboxy-CBD were identified. In the multiple dose study, the mean maximum serum concentration was 38.4 ng/mL. The terminal half-life was 161.3 hours in serum and Rss was 4.5. In the dose escalation study, analysis of behavioural parameters, HR and HRV showed no consistently significant differences between the treatment and control group. During the multiple dose study, daily behavioural observations and cortisol levels also did not differ between treatment and control group. When analysing reactivity, movement patterns, HR, HRV and cortisol levels during the novel object test and the trailer test, no consistently significant differences were observed between groups. This study was the first to investigate pharmacokinetic parameters combined with the effect of CBD on behaviour and stress after regular oral administration of CBD in horses over two weeks. The pharmacokinetic analysis showed an extensive metabolism of CBD with a high distribution into peripheral tissues and a long elimination phase. The results of the behavioural assessments provided no reliable evidence for a stress-reducing or sedative effect of CBD in horses after regular oral administration at a dose of 3 mg/kg twice daily. The main limitation of this study is the small sample size. Further investigation of the potential stress-reducing effects of CBD in conjunction with pharmacokinetic analysis is essential to determine relevant CBD concentrations for medication control at equestrian sport events. Subsequent studies may consider administering higher CBD doses, such as 10 mg CBD/kg, and specifically explore the effect on horses known to exhibit signs of nervousness and are easily stressed.

Dynamical systems on networks, which can be seen as a special type of agent-based model, are widely used to model systems that consist of many interacting entities called agents. In this framework the nodes in the network represent the agents, the edges represent the relations between them, and the state of each agent evolves over time in dependence on the states of its neighbors, typically in a stochastic manner. Although the state evolution of each single agent is often dictated by simple rules and mechanisms, the overall collective or emergent behavior of the system, which is the result of many individual interactions, can be incredibly hard to anticipate and understand. The investigation of this collective behavior is the main focus of this thesis. Even though the collective behavior is difficult to predict, it is typically much less complex than the vast number of degrees of freedom would allow for and instead (approximately) follows some low-dimensional dynamics. The understanding of collective behavior hence consists of two steps. Firstly, a projection that maps the high-dimensional microscopic state, containing the state of each agent, to a low-dimensional macroscopic state, containing only the essential aggregated information to describe the collective behavior, has to be found. This projection, which filters out unnecessary degrees of freedom and quickly decaying processes of the original system, is called a collective variable (CV). Secondly, the reduced macroscopic system that dictates the evolution of the macroscopic state has to be derived. If the choice of CV was appropriate, this macroscopic system is able to reproduce the low-dimensional projection of the original model, i.e., the collective behavior. Akin to the law of large numbers, the aggregated random actions of many agents sometimes lead to an approximately deterministic macroscopic dynamics, which is referred to as a concentration effect. This thesis mainly considers Markovian discrete-state dynamics on (random) networks and addresses efficient simulation, discovery of CVs and macroscopic dynamics, and the occurrence of concentration effects. In this setting the shares of each discrete state in the system, or in certain subsystems, constitute an important choice of CVs. Conditions that guarantee the convergence of the dynamics of the shares to a deterministic mean-field ordinary differential equation in the large population limit are proved. These conditions enable the derivation of parameter bounds for popular random graph models, e.g., Erdős-Rényi random graphs, the stochastic block model, and random regular graphs, that ensure convergence to the mean-field limit, which is demonstrated for a continuous-time noisy voter model (CNVM). For systems that do not exhibit convergence to the mean-field limit and for which the simple state shares are not appropriate CVs because they lack essential state information, a data-driven method for algorithmically learning good and interpretable CVs from model simulations is presented. This method permits to assess the quality of the learned CVs and to infer their relation to topological features of the network. In combination with established techniques for learning dynamics from data, an automatic evaluation of the collective behavior can be achieved. This is demonstrated for the CNVM on scale-free networks.

Im Zentrum dieser Arbeit steht die Frage: Welche Rolle spielt in China städtischer öffentlicher Raum als soziales Ordnungsmedium? Im Mittelpunkt stehen seine soziale Produktion, In- und Exklusionsmechanismen sowie die Beziehung zwischen staatlichen Idealen und alltäglicher Aneignung in Chinas Reformperiode. Die soziale Produktion öffentlichen Raums als Ordnungsmedium wird hauptsächlich am Beispiel reziproker Handlungsabläufe von staatlichen Akteuren und „Vagabunden“ (Straßenhändler inkl. mobile Wahrsager und Bettler) als unerwünschte Nutzergruppe herausgearbeitet. In Anlehnung an Henri Lefèbvre, Martina Löw u.a. definiere ich für diese Arbeit die soziale Produktion des Raums als kontinuierlichen Prozess der Aneignung/Appropriation durch die (Neu)Ordnung physischer Objekte (inklusive des eigenen Körpers) einerseits und/oder sozialer Bedeutung andererseits. Urbaner öffentlicher Raum meint den „nicht-eingehausten“ Stadtraum, wo die Chance auf ein größeres Publikum und auf Anonymität als dominierendem Merkmal besteht, wie z.B. populäre Straßen, Plätze, Parks, Promenaden etc. Den lokalen Kontext dieser Untersuchung bietet dabei das südchinesische Guangzhou, wo ich von 2010 bis 2014 lebte und meine Feldforschung abschloss. Als Forschungsfragen stellen sich: Welche öffentlichen Räume werden vom „Staat“ und den „Vagabunden“ okkupiert, reguliert, gebaut? Welche strukturellen Mechanismen und diskursiven Motivationen stecken dahinter? Wie okkupieren und formen die untersuchten Nutzergruppen den öffentlichen Raum um? Wie interagieren dabei die jeweiligen Parteien? Welche Konsequenzen hat die Appropriation öffentlichen Raums für die städtische In- und Exklusion der „Vagabunden“ als marginalisierte Nutzer? Welchen Einfluss hat die Reform- und Öffnungspolitik auf die Appropriation der untersuchten Nutzergruppen? In welchem urbanen und historischen Entwicklungskontext steht die heutige soziale Produktion von Guangzhous öffentlichem Raum? Ergebnisse: 1) Entwicklung von Städtebau, -governance und -gesellschaft entlang der Dimension des städtischen öffentlichen Raums in China: Hier sind insbesondere nationalistische Bewegungen der Modernisierung, Zivilisierung und „Spektakularisierung“ des öffentlichen Raums von Bedeutung. Angelehnt an Debord meint Letzteres die (Ab)Lenkung bürgerlicher Aufmerksamkeit und die Herrschaft durch das „Image“. Dabei nimmt öffentlicher Raum eine doppelte Rolle, nach innen und außen, ein. Er fungiert als Medium der staatlichen Darstellung und der Erziehung der Stadtbewohner. Die politische Konnotation und Funktion öffentlichen Raums im städtischen China kommt deutlich zum Vorschein. Im Alltag entwickelt sich die Offenheit bzw. Schließung des öffentlichen Raums entlang der Zonierung und Rhythmisierung öffentlicher Ordnung. Es kommt zu einer Fragmentierung des öffentlichen Raums entlang der Logik wirtschaftlichen Profits und politischen Prestiges. Er entwickelt sich als „Inseln des Spektakels“ die zeitlich und räumlich expandierenden. Das Spektakularisierung des städtischen Raums dehnt sich in den Alltag aus. 2) Prozesse der Exklusion, Strategien und Taktiken der Inklusion marginalisierter Migranten in Guangzhou: Die Frage der städtischen Teilhabe ist eine Frage des Raums. Die Auseinandersetzungen zeigen zwar eine Dominanz, aber keine „Alleinherrschaft“ der staatlichen Akteure. Am Beispiel der Straßenhändler arbeite ich die „Taktiken der Präsenz“ heraus, die zeigen, wie sie sich an die staatliche Raumlogik anpassen, ihr aus dem Weg gehen und sie herausfordern. Mobile Wahrsager verdeutlichen, dass Zugang zum öffentlichen Raum mehr bedeutet als ein bloßes „Sich-im-Raum-befinden“. Sie eignen sich den öffentlichen Raum via Performance an, die auf das umgebende Publikum ausgerichtet ist. Sie zeigen, nicht jede Aktion bedeutet Kontestation, denn sie orientieren sich am historisch gewachsenen Raumerbe, an der Raumkultur, d.h. am sozial akzeptierten Verhalten an den Tempeln, Konsum- und Unterhaltungsgebieten. Bettler zeigen, wie beide Aspekte – staatliche Raumlogik und Raumkultur/-erbe – zusammen wirken. Im Ergebnis verschaffen sie sich Zugang zum öffentlichen Raum durch die Kombination von Entertainment und Darstellung der Authentizität ihres Leids. Die Exklusion der „Vagabunden“ aus dem öffentlichen Raum ist nicht absolut, sondern durch Barrieren der Regulation gekennzeichnet. Um diese zu überwinden, bedarf es vonseiten der „Vagabunden“ wachsender Investitionen – in Mobilität, Flexibilität und Performances. Das heißt, ohne Ressourcen haben diese städtischen Armen kaum Zutritt zum öffentlichen Raum. Dabei ist ihre Rolle für die soziale Produktion nicht marginal: Sie beeinflussen das Vorgehen der staatlichen Akteure, tragen zur Vielfalt des öffentlichen Raums, aber auch zu seiner „Spektakularisierung“ bei. 3) Desiderate des Konzepts des öffentlichen Raums und Lehren aus der chinesischen Empirie: Die soziale Produktion öffentlichen Raums hält eine neue Perspektive bereit, die einige Annahmen zum öffentlichen Raum infrage stellt. Die chinesische Empirie verweist darüber hinaus auf die „Tücken“ des theoretischen Konzepts hin. Dazu gehören vor allem und dessen starke normative Konnotation sowie die Vielfältigkeit der sich überlappenden Perspektiven. Darüber hinaus ergeben sich folgende Lehren, die den „westlichen“ diskursdominanten Annahmen zum öffentlichen Raum entgegenstehen: Öffentlicher Raum ist nicht physisch-räumlich determiniert. Er ist weder räumlich noch zeitlich fix. Öffentlicher Raum ist nicht offen und zugänglich für „Alle“, er war es noch nie – weder in Europa noch in China. Er lässt sich nicht per se über den Besitz in öffentlicher Hand definieren. Daher ist auch die Privatisierung und/oder Kommerzialisierung an sich auch nicht ein Zeichen des Verlusts öffentlichen Raums. Öffentlicher Raum ist ein wichtiges Ordnungsmedium, der die qualitative Dimension von Verstädterung prägt. Die konkreten Öffnungs- und Schließungsmechanismen strukturieren die städtische Gesellschaft. Deshalb steht der öffentliche Raum im Zentrum von Urbanisierung und Urbanität.