The intestinal microbiome has a central role in the host's health. In pig production, newborn and weaned piglets are particularly affected by diarrheal diseases of multifactorial origin, which can often only be controlled with antibiotics. However, in line with the “One Health” approach, the aim is to reduce the use of antibiotics in animal husbandry. Targeted modulation of the intestinal microbiome could enable new preventive measures, with the mother sow playing a key role in the initial microbial colonization of newborn piglets and intestinal colonization up to weaning. This approach was investigated by the “OptiBiom” project by analyzing comprehensive microbiome and farm data and combined with targeted microbiome modulation and vaccines to develop new anti-infection strategies. For this purpose, fecal samples were collected from sows before and after birth and their piglets before and after weaning from 20 commercial farms. This thesis analyzes data on the fecal microbiome, antibiotic resistance genes, and bacterial virulence factors. In addition, the use of antibiotics in the corresponding production cycles was documented and evaluated for each farm. The hypothesis that the fecal microbiota composition and the occurrence of resistance and virulence genes differ significantly between the farms and age groups of the animals was investigated. In addition, correlations were examined between farm differences and the sow-piglet relationship, as well as between antibiotic use and the occurrence of bacterial genes. In the first publication, 802 DNA extracts from fecal samples were sequenced using Illumina NextSeq and analyzed for diversity and microbial composition. Detected genera were compared between the different age groups of the animals, and the differences could be visualized using non-metric multidimensional scaling and hierarchical clustering. Data from sows and their piglets were pooled to investigate the family effect on farm differences. In the second publication, quantitative real-time PCR was used to detect resistance genes of relevant antibiotics (sulfonamides, trimethoprim, colistin, cefotaxime) and an integron responsible for spreading resistance genes (Int1). In addition, the samples were examined for relevant pathogens associated with diarrhea in piglets (Clostridioides difficile, Clostridium perfringens, Salmonella, Escherichia/Shigella/Hafnia, Escherichia coli). The farms were analyzed for cluster formation based on their antibiotic use. Latent class analysis was then used to identify the genes relevant for cluster formation in each animal group. Adapted regression models were used to test correlations with the antibiotics administered. The phyla Bacillota (formerly Firmucutes) and Bacteroidota (formerly Bacteroidetes) dominate all samples with varying relative abundances depending on the production phase. Parturition (sows) and weaning (piglets) led to significant changes in the fecal microbiota composition. After birth, the relative abundance of non-dominant genera, in particular, decreased significantly, while previously dominant genera continued to predominate. In piglets, on the other hand, originally non-dominant genera became predominant after weaning, while previously dominant genera from the suckling phase decreased. The microbiome of suckling piglets was the most uniform and showed the least diversity. Combining the microbiome data of sows and their piglets revealed a distinctive cluster formation for two farms. The use of antibiotics on each farm led to different clusters for sows, suckling piglets, and weaners. The greatest diversity of antibiotics used was found in weaned piglets, including amoxicillin, lincomycin and enrofloxacin, which were associated with antibiotic resistance genes (mcr1, dfrA1, blaCTX-M). However, direct effects of a particular antibiotic on the associated resistance gene were rare, similarly between antibiotics and virulence genes. However, enrofloxacin and florfenicol favored the occurrence of C. difficile in sows. The Int1 gene was ubiquitously present and could be associated with the prevalence of F4 fimbriae in E. coli, while other E. coli fimbrial genes were not shown to be affected by antibiotics. The vertical transmission of the microbiota from sows to their offspring could be an important factor in the microbial individuality of farms and provide an approach for individual intervention strategies to get piglets through the vulnerable weaning phase in good health. Nevertheless, future studies must include other influencing factors such as the farm environment, detailed health data, and feeding. Extensive databases and integrating artificial intelligence and machine learning can be helpful here. The long-term goal should be indicators for defining a target microbiota that enables the highest possible disease resilience and productivity. It is essential to develop modulation strategies without the use of antibiotics.

Spin systems are known for their purely interacting nature, causing them to lack a canonical perturbative limit, which usually helps to understand the fundamental physics of a system. A method that has proven effective for the description of strongly correlated systems is Functional Renormalization Group (FRG), which can be thought of as an alternative to the path integral formalism as an approach to many-body quantum mechanics. The basic concept is to cure infrared divergences by introducing a cutoff to correlation functions. Continuously lowering the infrared cutoff and hence allowing the system to occupy lower-lying energy states is called the FRG flow and is formulated in terms of differential equations. The solutions to these equations are functions which describe effective interactions between particles and hence contain information about particle correlations and thermodynamic quantities.

Formulating FRG in terms of spins turns out to be a subtle endeavor, though, as the spin algebra relations are relatively complicated compared to, for example, the canonical anti-commutation relations of fermions. In order to use the advantages of FRG for spin systems, one can map spins onto fermionic operators, which leads to pseudo-fermion FRG (PFFRG), developed by Reuther and Wölfle.[198] Alter- natively, one can express spins in terms of Majorana operators, leading to pseudo- Majorana FRG (PMFRG), which has been developed by Niggemann, Sbierski, and Reuther.[163] PMFRG is particularly powerful at finite temperatures, where useful properties of Majorana operators render the method more accurate than PFFRG. In contrast to many other methods, PFFRG and PMFRG are applicable to any spin system, even frustrated ones.

In this work, PMFRG is being generalized and applied to highly frustrated spin systems. In particular, spin representations in terms of Majorana operators for spins with arbitrary large spin magnitude S are being investigated and classified thoroughly, which closes a gap in the literature about the second quantization of spin operators. Moreover, it is presented how to generalize PMFRG for Heisenberg models with full SU(2) symmetry to XXZ models exhibiting only a U(1) symmetry. This opens up a wide range of interesting applications for PMFRG. An example is the XXZ model on the pyrochlore lattice, which is known to exhibit conventional long-range order but also exotic spin liquid ground states in the so-called spin ice phase. PMFRG is applied to this model, and the results are being compared to experiments. Furthermore, the entire phase diagram of the XXZ model is being mapped out. Also, the capability of PMFRG to reproduce low-energy field theory predictions and to determine critical exponents is being tested.

Exultet iam angelica turba coelorum! – Frohlocket, ihr Chöre der Engel! – mit diesen Worten setzt das Osterlob ein; sein Eingangswort Exultet gab den Manuskripten, auf denen das Lob als zentraler Bestandteil der Osterliturgie notiert war, seinen Namen. Die zwischen dem 10. und 14. Jahrhundert vor allem in Apulien und Kampanien entstandenen Exultet-Rollen kombinieren Schrift, Bild und musikalische Notation in einzigartiger Weise.

Das Buch untersucht erstmals materielle und mediale Besonderheiten der Rollen aus interdisziplinärer Perspektive, indem es nicht nur die Objekte selbst, sondern auch ihre Nutzung und Wahrnehmung in der beneventanischen Liturgie in den Blick nimmt. Es wird gezeigt, inwiefern die Rollen visuell, auditiv und olfaktorisch zur Konstitution liturgischer und sozialer Räume beitrugen.

Transform coding methods play a fundamental role in image and video coding technologies like the Versatile Video Coding (VVC) standard. Typically, the employed transforms are linear maps with strong energy compaction capabilities. Therefore, efficient quantization and entropy coding methods can be designed for transmitting and storing the transform coefficients. In recent years, there have been considerable efforts to design coding-efficient transforms from learning-based methods. As for video compression, additional bitrate savings are achieved by optimizing linear block transforms with respect to the different intra prediction modes. In contrast, end-to-end optimized image codecs have been obtained from deep-learning experiments. Learned codecs like the JPEG AI coding standard rely on using non-linear, neural networks as forward and inverse transform. Remarkably, JPEG AI is reported to have superior compression efficiency relative to conventional still image coding. Since the transforms in learned image compression are non-linear, it is not clear if rate-distortion optimization methods designed for linear blocks transforms are well-suited. Thus, this thesis studies the impact of different signal-dependent encoder optimizations on the quantization when a learned image codec is used. As a main result, an algorithm for rate-distortion optimized scalar quantization is developed which achieves bitrate savings between 1 % and 7 %. Furthermore, it has been shown that a rate-constrained vector quantizer improves the coding efficiency on a similar scale. Its design has similarities with the trellis-coded quantization stage in VVC. Thus, since rate-constrained quantization is shown to be effective when applied to non-linear transforms, different non-linear transform coding tools for block-based video compression are developed. These tools employ neural networks which are obtained from a data-driven optimization. The first tool, a non-linear coefficient prediction, uses reconstructed coefficients and the reference samples from the block boundary for predicting low-frequency coefficients. Therefore, only the difference between the predicted value and the original coefficient is quantized and coded. The second tool, a non-linear transform offset, is applied after reconstructing all coefficients and also depends on the reference samples as input. The offset is added before the synthesis transform and has been trained to improve the reconstruction quality. A combination of both methods yields coding gains between 1.0 % and 2.8 % over VVC in All-Intra configuration. Finally, non-linear transforms and intra modes are obtained from an end-to-end training method. The learned transforms do not depend on the reference samples. The training goal is to minimize the expected rate-distortion cost by using an approximation of the transform coefficients’ bitrate. The average All-intra bitrate savings of the learned transforms and intra modes are 0.9 % against VVC.

Autonomous Driving (AD) has advanced significantly in recent years, yet widespread deployment remains limited. One of the most enduring challenges in Autonomous Vehicle (AV) development is planning a safe, comfortable, and efficient motion in complex, real-world environments. This thesis addresses motion planning across three distinct domains: urban shuttles, passenger vehicles, and truck-trailer systems. It contributes practical insights and novel approaches toward scalable autonomous mobility. The first part of this work presents an integrated motion planning framework for the Continental Urban Mobility Experience (CUbE) driverless shuttle. Extensive real-world testing over several years highlights the system’s robustness and underscores the importance of long-term validation in urban settings. Key innovations include a multi-layered planning stack and a data-driven motion forecasting approach that enhances interaction with human traffic participants. The second part investigates the behavior of human drivers in understructured traffic environments. Those are areas that fall between well-defined road systems and fully unstructured spaces. A novel analysis framework is introduced for mining patterns from naturalistic trajectory datasets, enabling AVs to better blend into human traffic and navigate ambiguous scenarios with improved predictability and safety. The final part of the thesis explores Deep Reinforcement Learning (DRL) for planning and controlling complex truck-trailer maneuvers. A DRL-based approach is developed and evaluated in simulated environments, demonstrating the method’s potential to handle the nonlinear dynamics of articulated vehicles. These contributions advance the field of motion planning by combining theoretical insights, system-level integration, and empirical evaluation. They offer pathways for improving AV behavior across diverse platforms and use cases, ultimately supporting the broader adoption of AD technologies.

This dissertation examines Luri oral traditions in Kohgiluyeh and Boyer-Ahmad, focusing on role in shaping historical memory and identity through narratives of Alexander the Great. Using a comparative and interdisciplinary approach, it traces the evolution of Alexander’s legend across Greek, Byzantine, Syriac, and Persian sources, demonstrating that oral traditions are not passive reflections of history but active historiographical forces that preserve, reinterpret, and resist dominant narratives. By analyzing onomastics (naming practices), toponyms (place names), and gendered representations, the study examines how Luri oral traditions function as sites of cultural resistance, challenging Persian nationalist historiography. It explores how Reżā Šāh’s policies marginalized regional oral traditions, promoting the Šāhnāme while sidelining Niẓāmī’s Iskandarnāme, yet Lur oral performers continued to preserve alternative narratives, maintaining distinct historical perspectives. Through ethnographic fieldwork and an analysis of oral storytelling techniques, the study categorizes Luri oral narratives into Mas̩al (proverbs), Muqawwm (chants), Tims̩āl (literary adaptations and genealogies), Matīl (fables), and Iʿtiqāt (beliefs), demonstrating how these traditions actively shape and transmit historical meaning. The findings underscore the urgency of preserving regional oral traditions, as their erosion threatens to erase alternative historiographies and counter-histories essential to understanding Iran’s diverse historical landscape. This study contributes to oral history, historiography, and cultural memory studies, offering insights into how oral traditions function as mechanisms of historical negotiation, cultural resistance, and identity formation beyond Iran.

The Paradoxes of Intimacy explores the formal and narrative possibilities and limitations of writing about intimate human relationships in the second decade of the twenty-first century. In an age marked by globalization, growing social and environmental insecurities, and intensification of capitalist markets (Jeffrey Nealon), the ways we bond with each other have come under increasing scrutiny in scientific research -not least for its links to problems of alienation, solipsism, and social disintegration. Recent sociological research on intimacy has attested to a crisis in the subject describing contemporary forms of close relationships as dramatically cooling (Eva Illouz), unbinding (Zygmunt Baumann), and even eroding (Byung- Chul Han). Meanwhile, the emergence of new forms of intimate bonding, such as same-sex relationships, the #Metoo social movement’s public exposure of private harm, the rise of emotion-aware technologies, and the sentimentalization of capitalist markets have radically transformed the ways intimacies are created and experienced both in actual life and in artful practices. While the subject has been extensively studied in sociological and philosophical disciplines, it is still underexplored in literary scholarship, where intimacy is often used interchangeably with love to refer to either romantic or erotic relationships. Performing a cross-generic analysis of Rachel Cusk’s The Outline Trilogy, Margaret Atwood’s The Heart Goes Last, and Marilynne Robinson’s Lila, The Paradoxes of Intimacy argues that contemporary women’s fiction reimagines the aesthetics of intimacy by moving beyond the exhausted structures of romantic and confessional narratives. Investigating the emergent forms of literary intimacies as they are problematized and re-invented in postmillennial women’s fiction, it critically addresses the question of how literature navigates the liminal spaces of (textual and narrative) encounters and temporalities (of imagined futures and old forms) to dismantle the fictional representation of intimacy as a private matter. While literary intimacies are traditionally predicated on an individualized point of view in which the interiority of the narrator or a character is rendered through introspection, contemporary women’s fiction uses interior monologues and indirect style not to deliver psychological depth, but to reveal the sociality, artificiality, and paradoxicality of connection. In tracing these formal and thematic transformations, the dissertation contends that contemporary women’s fiction does not seek to restore intimacy or propose new modes of authentic bonding. Rather, it reflects and aestheticizes the pervasive disconnection that defines the postmillennial world. Cusk’s autofictional narratives turn intimacy into a performative repetition of stories that reveal the emptiness of self-disclosure; Atwood’s dystopian satire renders love as an apparatus of emotional and economic control; and Robinson’s Lila gestures toward a fragile fleeting alternative that neither resolves nor escapes this crisis. In this sense, The Paradoxes of Intimacy argues that intimacy in contemporary fiction becomes not a site of resolution, but a literary form of crisis and paradox -a mode that exposes the limits of relation, the failure of reciprocity, and the persistent yearning for connection in an age increasingly hostile to the intimate.

Die vorliegende Arbeit befasst sich mit der automatisierten Erkennung und Positionskontrolle von vier spezifischen Arten von Therapiehilfen (Endotrachealtubus bzw. Trachealkanüle, verschiedene Arten von Kathetern, Thoraxdrainagen und Magensonden) in Röntgenthorax Bildern unter Verwendung von tiefen neuronalen Netzwerken. Durch den Einsatz von Segmentationsmodellen und dem Self-Aware-Mechanismus soll die röntgenbasierte Positionskontrolle dieser Therapiehilfen, die in der Intensiv- und Notfallmedizin eine zentrale Rolle spielen, optimiert und automatisiert werden. Dabei werden die Herausforderungen der Positionierung in der klinischen Praxis adressiert, die aufgrund der kritischen Bedeutung der korrekten Positionierung für die Patientensicherheit und Therapieeffizienz von hoher Relevanz sind. Zunächst wurden drei verschiedene methodische Ansätze zur Segmentierung von Therapiehilfen untersucht, die sich in der Komplexität und Spezialisierung der verwendeten Modelle unterscheiden. Dabei zeigte sich, dass der Ansatz mit spezialisierten Modellen für jede Art von Therapiehilfe (Ansatz II) die besten Ergebnisse liefert. Die Evaluierung der Modelle basierte auf den Klassifikationsmetriken AUROC, Sensitivität, Spezifität, Genauigkeit und balancierte Genauigkeit sowie den Segmentierungsmetriken Dice und Dice+, letztere wurde speziell für diese Arbeit entwickelt. Insbesondere die SegResNet-Architektur zeichnete sich durch eine überlegene Segmentationsleistung aus. Ein weiterer Schwerpunkt war die Entwicklung und Evaluierung des Self-Aware-Mechanismus (SA), der in fast allen Szenarien zu einer Verbesserung der Segmentationsleistung führte. Dieser Mechanismus ermöglicht es dem Modell, seine eigenen Vorhersagen zu überprüfen und zu verfeinern, was insbesondere bei der Endpunktverfeinerung und dem Auffüllen oder Verbinden unvollständiger Segmente von Vorteil ist. Die Untersuchung des Einflusses von Trainingsdatenmenge und Bildauflösung auf die Segmentationsleistung ergab, dass sowohl eine größere Datenmenge als auch eine höhere Bildauflösung zu besseren Ergebnissen führen, wobei die Verbesserungen mit zunehmender Datenmenge bzw. Auflösung einem abnehmenden Grenznutzen folgen. Das Ergebnis der Arbeit ist, dass durch den Einsatz von Deep Learning und insbesondere des Self Aware-Mechanismus eine deutliche Leistungssteigerung in der automatisierten Detektion und Positionskontrolle von Therapiehilfen im Röntgenthorax erreicht werden kann. Diese Erkenntnisse bieten wertvolle Ansatzpunkte für die Weiterentwicklung automatisierter Systeme in der medizinischen Bildverarbeitung mit dem Potenzial, die Effizienz und Genauigkeit von automatisierten Kontrollsystemen in der klinischen Praxis signifikant zu verbessern.

Die Habilitationsschrift untersucht neue Aspekte der Einteilung, Diagnostik und Therapie funktioneller und traumatischer Schulterinstabilitäten. Besonders die komplexe funktionelle Schulterinstabilität (FSI) wurde neu charakterisiert und klassifiziert, woraus ein innovativer konservativer Therapieansatz entstand: der Schulter-Schrittmacher. Dieser zeigte in Studien deutliche Vorteile gegenüber der alleinigen Physiotherapie. Zudem wurden moderne Ruhigstellungskonzepte und operative Verfahren verglichen. Die Abduktions-Außenrotationsruhigstellung erwies sich als wirksame Option nach Erstluxation, während operative Stabilisierung niedrigere Rezidivraten bot. Neue operative Techniken, darunter die Bankart-Plus-Methode und der Latarjet-Eingriff nach Voroperationen, wurden hinsichtlich Wirksamkeit und Komplikationen bewertet. Trotz Fortschritten bleiben Fragen zur funktionellen Stabilität, zu Indikationsgrenzen knöcherner Augmentation und zur optimalen Nachbehandlung offen.

Chimeric Antigen Receptor (CAR-) T cell therapy represents one of the newest, most promising treatments for cancer. Here, the patient’s own T lymphocytes are genetically modified with a cancer-specific CAR. CAR T cell therapy has shown significant clinical success, especially for the treatment of hematologic cancers by targeting the B cell antigen CD19. Nevertheless, selective pressure on the cancer cells frequently causes suppression or complete loss of the target antigen, leading to cancer relapse with fewer treatment options. This phenomenon, known as antigen escape, renders CAR-T cells ineffective representing a major clinical challenge. Consequently, there is an urgent need for targets that are less prone to antigen escape. In this thesis, a novel type of CAR-T cells is described, employing cysteine-engineered receptors that interact with altered extracellular redox states of B cell non-Hodgkin lymphoma (B-NHL). First, the redox-reactive nanobody CB2 was used for the generation of CB2-CAR-T cells, interacting directly with B-NHL redox states via a non-canonical cysteine in the complementarity-determining region (CDR) 3. The activation and specific cytotoxicity of CB2-CAR-T cells were verified in vitro against different subtypes of B-NHL, including antigen escape models. Next, the universality of the approach was confirmed by cysteine-engineering of a non-cancer-related nanobody-CAR. A single amino acid substitution with cysteine was sufficient to redirect these CAR-T cells to specifically target B-NHL, rendering cysteine-engineered CAR-T cells (or CysCAR-T cells) a universal strategy to enable antigen-independent targeting. Additionally, it was shown that cysteine engineering of state-of-the-art CD19-CAR-T cells enables co-targeting of both CD19-positive and -negative B-NHL. No systemic toxicity associated with these bifunctional CysCD19-CAR-T cells was observed in mice. Furthermore, T cells expressing the cysteine-engineered CAR delayed tumor growth and prolonged survival of mice engrafted with CD19-positive and -negative lymphoma. These findings introduce a novel type of bifunctional CAR-T cells that simultaneously target conventional antigens and altered redox states, potentially reducing the risk of antigen escape. It was shown that those CysCAR-T cells act through an antigen-independent mechanism and that cysteine engineering can be applied to CARs of diverse specificities. Altered redox states have been described for various cancers, including breast cancer and leukemia. Hence, these results indicate a broad therapeutic scope for CysCAR-T cells in preventing antigen escape associated with conventional targets.

Current treatment options for Clostridioides difficile infections of the gut are limited by reliance on antibiotics, resulting in high recurrence and mortality. Since the secreted toxin TcdB is the source of pathogenesis, a more effective treatment route may target TcdB directly. Allosteric control of the cysteine protease domain (CPD) of TcdB offers a promising intervention point for deactivating the toxin. Here, we apply computational methods to uncover the allosteric mechanism of TcdB CPD and thereby advance the larger rational drug design project. Free energy calculations feature prominently in our toolkit. We use umbrella sampling to sample the large conformational transition of TcdB CPD and reproduce the allosteric effect on free energy surfaces. We also use free energy perturbation calculations to distinguish binding affinities among highly complex protonation states of the allosteric modulator phytate (IP6). Based on these and other computations, we construct a detailed allosteric mechanism of in the form of a switchable interaction network. This mechanism is thoroughly validated by both computations and experiments. We additionally contribute to the characterization of IP6 analogues with thiophosphate substitutions in the form of docking studies. Finally, we uncover the structural origins of the net loss of one proton on IP6–TcdB CPD complexation. Together, this computational treatment provides detailed structural and mechanistic insight on an important ligand-protein system.

Sensory processing and sleep are essential behaviors critical for survival from animal models to humans. The presynaptic active zone (AZ) plays a central role in both processes, yet how molecular diversity at the AZ contributes to functional variability and behavioral specificity remains unclear. In the Drosophila brain, distinct nanoscopic coupling distances between Ca²⁺ channels and the priming factors Unc13A and Unc13B underlie diverse modes of short-term plasticity (STP), which are essential for sensory decoding. Unc13A drives fast phasic transmission, while Unc13B mediates adaptive dynamics such as sensory prediction error coding. However, how this balance is maintained—and spatially controlled—at individual synapses had remained unresolved. Our recent study identified a previously unrecognized AZ protein, Blobby, encoded by the Drosophila gene CG42795. Blobby is a BRP-interacting factor homologous to human TBC1D30, a large Rab GTPase-activating protein (Rab-GAP). At the developing third-instar larval neuromuscular junction (NMJ), Blobby localizes adjacent to BRP scaffold and arrives at the newly forming AZ later than BRP. Loss of Blobby in blobbyNull mutant leads to ectopic accumulation of large BRP aggregates and a reduction in evoked synaptic transmission, which is likely due to a decrease in both release probability and number of release sites.Here, in my thesis, by combining biochemistry, genetics, diverse behavioral approaches, living imaging and super resolution microscopy, I provide a comprehensive characterization of Blobby in regulating nanoscopic localization of BRP and Unc13 proteins in the adult brain, and in its critical function in odor sensation and sleep regulation. The blobbyNull mutant is surprisingly homozygous adult-viable with severely reduced Mendelian ratio and the adult flies appeared healthy at young age. However, they suffered from a strong decrease of lifespan and hypersensitivity to oxidative stress. Furthermore, the blobbyNull mutant flies exhibit a marked loss of olfactory-driven behaviors in response to both appetitive and aversive odors. By mapping through the Drosophila olfactory circuit through a targeted knockout strategy, I show that Blobby is required in the excitatory projection neurons (PN) and the mushroom body (MB) Kenyon cells (KC), but not in the olfactory receptor neurons. Importantly, at the presynapse of PN in Calyx, loss of blobby either only in PN or throughout the whole brain causes nanoscopic redistribution of Unc13B toward AZ center and increases its overall levels, without affecting the localization of the tight-coupling Unc13A. Two-photon calciumimaging at PN::KC synapses reveal that PN-specific blobby knockout resulted in a drastic reduction in odor-evoked postsynaptic calcium response. Strikingly, knocking down Unc13B in PN was sufficient to rescue the olfactory sensitivity to both blobbyNull mutant and PN-specific blobby knockout flies. These data suggest Blobby is a bona fide regulator for sensory processing, most likely through the fine-tuning of synaptic transmission in the olfactory circuit via a redistribution of Unc13B. As sleep and sensory processing are closely coupled, I further show that loss of Blobby had very limited effects on the sleep patterns. In contrast, Unc13B mutant flies exhibited a strong increase in both daytime and nighttime sleep, with shorter sleep latency and more consolidated sleep. Remarkably, the sleep phenotypes of Unc13B mutant flies were fully rescued by loss of Blobby, indicating that the interaction between Blobby and Unc13B extends from the coding of sensory information to the maintenance of sleep. In conclusion, Blobby is an essential AZ protein in tuning synaptic transmission via nanoscopic localization and balancing of Unc13 isoforms. The interactive regulatory relationship between Blobby and Unc13B in sleep and odor sensation highlights the role of synapse and synaptic plasticity in gating, filtering and integrating behavioral relevant information. By identifying novel regulators such as Blobby in regulating core AZ scaffolds and release factors, we will provide better molecular understandings in bridging nanometer-scale vesicle priming to organism-scale control of sleep and sensory behavior.

The transcription machinery in the phylum spirochetes is poorly characterized at the molecular level yet is of significant evolutionary and medical importance. Pathogenic spirochetes can easily move through the mammalian tissues, penetrate blood vessels, cross the blood-brain barrier, and cause serious diseases, such as Lyme disease, relapsing fever, and syphilis. At the same time, many non-pathogenic spirochete species exist. Spirochetal RNA polymerases (RNAPs) are naturally resistant to rifampicin (Rif), the best-known transcription inhibitor in clinical use. Spirochetes evolved independently from other bacterial phyla and are not closely related to the well-established model organisms Escherichia coli (Eco) and Bacillus subtilis (Bsu), suggesting that the regulation of transcription in spirochetes includes distinct and novel strategies. Transcription is the first step in the highly regulated process of gene expression. It is divided into three phases, initiation, elongation and termination, that determines the start and the end of the transcription unit. To initiate transcription, RNAP, together with a Sigma factor (holoenzyme), recognizes promoter motifs on the DNA template and starts RNA synthesis. Many regulatory factors are associated with RNAP during initiation and modulate its activity, including CarD, GreA and DksA. Sequence alignments identified additional or distinct domains of some of these transcription factors in spirochetes compared to most other bacteria phyla, suggesting that they may act through different molecular mechanisms. To investigate transcription mechanisms regulated by CarD, GreA, and DksA, we reconstituted initiation complexes with each initiation factor and determined their structures using cryo-electron microscopy (cryo-EM). Here, I present the cryo-EM structures of Spirochaeta africana (Sfc) RNAP open promoter complex (RPo) in the presence or absence of CarD. Sfc RNAP, together with Sfc σ 70 , binds to the promoter DNA in an open conformation in which the duplex DNA is unwounded and the transcription bubble is formed. The structures suggest that Sfc RPo is unstable and could be stabilized by Sfc CarD via binding to the upstream template DNA (tDNA) of the transcription bubble. I also present the cryo-EM structure of the Sfc RPo in complex with GreA, indicating that Sfc GreA, like Sfc CarD, can stabilize the open promoter complex. This stabilization is achieved through the CarD-like domain, confirming the role of Sfc GreA as an initiation factor. During transcription elongation, transcription factors NusG and NusA are recruited by RNAP in most operons, while LoaP comes specifically in certain operons, promoting the RNA chain extended correctly. In addition, Sfc NusA has two additional acidic disordered loops on the AR1 and AR2 domains compared to Eco NusA. To investigate the functions of Sfc NusA and LoaP, we reconstituted elongation complexes with NusG/NusA and LoaP/NusA, and determined their structures using cryo-EM. I present the cryo-EM structures of Sfc RNAP in complex with NusG/NusA and LoaP/NusA, respectively. Although Sfc NusA does not change much the conformation of RNAP in the presence of NusG, it cooperates with LoaP to push nucleic acids away from RNAP, ensuring that the correct transcription elongation factor is recruited into the appropriate operon. The structures provide essential insights into the overall architecture of Sfc RNAP, the initiation and elongation complexes, and form the basis for further functional and structural analyses of spirochete-specific transcription factors and their regulation.

Die Prognose von Patientinnen und Patienten mit AGÖ/M ist trotz moderner Therapiestrategien schlecht. Die Stratifikation der Therapie erfolgt aktuell in Abhängigkeit vom Nachweis der prognostischen Faktoren Lymphknoten- und Fernmetastasierung. In dieser Arbeit wurden wei-tere Biomarker im Hinblick auf ihr prognostisches Potential für AGÖ/M untersucht. Originalarbeit 1 konnte an einer Kohorte von 129 Magenfrühkarzinomen UICC I ohne Lymphknotenmetastasen eine Reduktion des 5-Jahresüberleben bei Vorliegen einer venösen Infiltration um 36,7% und bei Vorliegen einer Tumormorphologie eines medullären/hepatoiden Subtyps um 23,1% nachweisen. In einem Panel von 30 theoretisch möglichen immunhistoche-mischen Markern konnte keine prognostische Relevanz der untersuchten Marker für dieses Tumorstadium festgestellt werden. Nach Erweiterung der Kohorte auf 393 Patientinnen und Patienten über alle Tumorstadien er-folgte in der Originalarbeit 2 die Analyse der Prävalenz und des prognostischen Potentials des für eine Therapie mit Zolbetuximab prädiktiven Biomarkers Claudin 18.2. Die Analyse zeigte eine Prävalenz von 21,8% Claudin 18.2 moderat exprimierender und 17,1% hochexprimieren-der Tumore ohne signifikante Assoziationen zu bestimmten Subgruppen und ohne Einfluss auf das Überleben. Die erhobenen Daten zeigen keinen prognostischen Effekt von Claudin 18.2. Die positiven Daten der Phase I- und II-Studien zu Zolbetuximab sind deshalb wahrscheinlich auf den direkten zytotoxischen Effekt des Antikörpers zurückzuführen. Originalarbeit 3 untersuchte den prognostischen Effekt von tumorinfiltrierenden Lymphozyten (TIL) in der beschriebenen AGÖ/M-Kohorte. Die multivariate Analyse zeigte, dass TIL mit einer HR 0,326 (KI95% 0,124 – 0,858; p=0,023) ein unabhängiger prognostischer Marker für AGÖ/Ms sind. Die Subgruppenanalyse ergab, dass dieser Effekt nur für frühe Tumorstadien (UICC I und II) nachweisbar ist und nur zutrifft, wenn PD-1 nicht auf den TIL exprimiert wird. Der prognostisch günstige Effekt von TIL im lokalisierten Tumorstadium kann somit durch die Expression von PD-1 aufgehoben werden. Den prädiktiven Wert von TIL für den Einsatz von Checkpoint-Inhibitoren muss prospektiv untersucht werden. Originalarbeit 4 analysiert die Rolle des Wnt-Signal-Zielproteins S100A4 auf die Prognose von AGÖ/M und die Möglichkeit einer pharmakologischen Inhibition. In unserer Kohorte zeig-te sich S100A4 negativ prognostisch (HR 1,366; 95% KI 1.010 – 1,847; p=0,043). Der stärkste Effekt für diesen Marker konnte in Niedrigrisikosituationen wie Tumorstadium UICC I/II, N0, L0 oder V0 Status nachgewiesen werden. In vitro-Analysen an Magenkarzinomzelllinien zeig-ten einen signifikanten Effekt des Proteins auf die Motilität der Zellen, die durch den Einsatz von Niclosamid inhibiert werden konnten. S100A4 ist daher einerseits ein vielversprechender Biomarker bei AGÖ/M und birgt andererseits das Potential als therapeutische Zielstruktur un-tersucht zu werden. Originalarbeit 5 untersucht die Rolle des HEG/MET-Signalweg-Regulators MACC1 als prognostischen Biomarker und Metastaseninduktor bei AGÖ/M. MACC1 ist in der untersuch-ten Kohorte ein unabhängig negativ prognostischer Maker. Besondere klinische Relevanz dürfte der Befund haben, dass Patientinnen und Patienten mit morphologischen Niedrigrisikokonstel-lationen (L0 und V0) bei Vorliegen einer MACC1-Überexpression eine deutlich schlechtere Prognose haben. Sowohl in Zelllinien als auch im Tiermodell zeigte sich, dass MACC1 einen signifikanten Einfluss auf die Motilität der Zellen und die Metastasierung ausübt, der durch Ein-satz von Selumetinib inhibiert werden konnte. Auch MACC1 stellt sich in dieser Analyse als ein prognostischer Marker dar, der als therapeutische Zielstruktur weiter untersucht werden sollte. Zusammenfassend konnten in den vorliegenden Arbeiten mehrere Biomarker für AGÖ/M iden-tifiziert werden, die vor allem in lokalisierten Tumorstadien eine hohe prognostische Relevanz zeigten und möglicherweise eine Risikostratifizierung erlauben könnten, um damit die Prognose von Hochrisikogruppen in lokalisierten Tumorstadien durch die Erweiterung der Therapie zu verbessern. Darüber hinaus könnte der Nachweis von hoher Infiltration von TIL in Kombinati-on mit niedriger PD-1-Expression die Prädiktion für die Therapie mit CPI verbessern, und die Weiterentwicklung von S100A4- und MACC1-Inhibitoren neue Zielstrukturen für die Therapie von AGÖ/M darstellen.

Im Rahmen dieser Habilitationsarbeit wurden mikrobielle Infektionen durch molekulare Bildgebung (Fluoreszenz in situ Hybridisierung in Kombination mit PCR und Sequenzierung – FISHseq) erforscht. Bei FISHseq handelt es sich um eine mikroskopische Methode, die die Vorteile von Molekularbiologie, Pathologie und Mikrobiologie miteinander vereint. Mit Hilfe von FISHseq gelingt der Nachweis auch von schwer-kultivierbaren oder Biofilm-assoziierten Erregern, die mit den Routine-Methoden verpasst würden. Darüber hinaus kann FISHseq durch die Bildgebung die Einordnung zweifelhafter diagnostischer Ergebnisse erleichtern: einerseits die Interpretation von seltenen, und daher fraglichen Erregern, andererseits aber auch die Relevanz von Spezies der Standortflora, die ebenfalls schwer einzuschätzen sein kann. Zudem können bildgebende Verfahren das Verständnis pathogener Prozesse verbessern und neue diagnostische Ansätze ermöglichen. FISHseq ermöglicht Aussagen nicht nur zur Identität der Pathogene in einer klinischen Probe, sondern auch zu deren Lokalisation, Formation (Biofilm-Staging), sowie zum Aktivitätszustand der Mikroorganismen basierend auf deren Ribosomengehalt. Von einer reinen Forschungsanwendung gelang die Translation der Anwendung der Methode in den klinisch-diagnostischen Routineeinsatz bei bestimmten Fragestellungen. Bei der Infektiösen Endokarditis ist FISHseq seit 2023 Teil der diagnostischen ISCVID Duke Kriterien und der ESC Guidelines für das Management der Infektiösen Endokarditis. Die Methode kann zudem nun überall dort, wo die üblichen mikrobiologischen Nachweisverfahren versagen, in der klinischen Mikrobiologie eingesetzt werden, um so eine bessere Patientenversorgung zu ermöglichen.

Das Ziel dieser Studie war es, die Effektivität des TAP-Blocks im Vergleich zur alleinigen intravenösen Opioidtherapie bei offenen Prostatektomien zu zeigen. 66 Patienten, die sich einer offenen retropubischen radikalen Prostatektomie am Charité Campus Mitte unterzogen, wurden in drei Gruppen unterteilt. Alle Patienten erhielten neben einer Allgemeinanästhesie für den Eingriff eine patientenkontrollierte Morphinpumpe für die postoperative Schmerztherapie. 22 Patienten in der Kontrollgruppe bekamen kein zusätzliches Regionalanästhesieverfahren (PCA-Gruppe). 22 Patienten erhielten präoperativ zusätzlich einen lateralen beidseitigen ultraschall-gestützten TAP-Block mit je 30 ml Ropivacain 0,375 % (TAP- Gruppe). In einer zweiten Kontrollgruppe wurde bei wiederum 22 Patienten präoperativ zusätzlich ein PDK mit einer kontinuierlichen Laufrate von Ropivacain 0,2 % und Sufentanil 1 µg/ml angelegt. Die Ergebnisse dieser Gruppe sind nicht in die vorliegende Arbeit eingeflossen. In den ersten 24 Stunden postoperativ wurde auf die Gabe zusätzlicher Analgetika verzichtet. Die vorliegende Arbeit stellt eine Subgruppenanalyse dar, die ausschließlich die Gruppen PCA und TAP vergleicht. Der primäre Endpunkt war der postoperative Ruheschmerz im Operationsgebiet, gemessen anhand der Numerischen Rating Skala (NRS). Sekundär wurden der postoperative Schmerz bei Bewegung, der Morphinbedarf, die Gesamtzufriedenheit mit der Schmerztherapie und das Auftreten von Nebenwirkungen wie PONV, Pruritus und vesikale Tenesmen verglichen. Die Messungen der Endpunkte erfolgten zu den Zeitpunkten 1 h, 2 h, 8 h, 24 h und 72 h postoperativ. Für den statistischen Vergleich des primären Endpunktes wurde der Mann-Whitney-U-Test verwendet. Bei den Patienten der TAP-Gruppe waren die NRS-Scores in Ruhe im Vergleich den Patienten der PCA-Gruppe zu den Zeitpunkten 1 h (1 [0;3] vs. 4 [2;6], p < 0,01), 2 h (1 [0;2] vs. 3 [2;5], p < 0,01) und 8 h (0 [0;2] vs. 2 [1;4], p < 0,01) signifikant niedriger. Sie hatten ebenfalls niedrigere NRS-Scores zu diesen Zeitpunkten bei postoperativen Schmerzen in Bewegung. Der Morphinverbrauch unterschied sich zu keinem Zeitpunkt. Die Gesamtzufriedenheit und das Auftreten von Nebenwirkungen unterschieden sich nicht signifikant. Zusammenfassend hat der beidseitige TAP-Block bei offenen Prostatektomien in der frühen postoperativen Phase einen analgetischen Vorteil gegenüber der alleinigen postoperativen Schmerzbehandlung mit Morphin.

Leichte Schädel-Hirn-Traumata (SHTs) wurden lange Zeit als folgenlose Bagatellverletzungen betrachtet. Forschungsergebnisse der letzten zwei Jahrzehnte liefern jedoch Hinweise, dass wiederholte leichte SHTs einen kumulativen Effekt mit anhaltenden funktionellen und neurometabolischen Veränderungen haben können und das Risiko für neurodegenerative und psychische Erkrankungen erhöhen. Im Fokus dieser Arbeit steht die Untersuchung chronischer Effekte wiederholter leichter SHTs auf die motorkortikale Exzitabilität, Inhibition und synaptische Plastizität sowie deren Modulierbarkeit durch anodale Gleichstromstimulation (atDCS). Zur Beantwortung der Forschungsfrage wurde ein randomisiertes, einfach-verblindetes, placebokontrolliertes Crossover-Design gewählt. Das Studienkollektiv umfasste 16 Personen mit zwei oder mehr leichten SHTs, die mindestens sechs Monate zurücklagen, sowie 21 gesunde Kontrollpersonen. An zwei Untersuchungsterminen erfolgte in randomisierter Reihenfolge eine 20-minütige anodale tDCS mit einer Stromstärke von 1 mA und eine Scheinstimulation. Parameter der motorkortikalen Exzitabilität und Inhibition wurden durch transkranielle Magnetstimulation (TMS) bestimmt. Zur Induktion LTP-ähnlicher synaptischer Plastizität im Motorcortex diente gepaarte assoziative Stimulation (PAS). Beide Versuchsgruppen unterschieden sich weder in den soziodemografischen Merkmalen noch im Ausgangsniveau der motorkortikalen Exzitabilität und Inhibition. Obwohl im Gesamtkollektiv nach anodaler Stimulation Hinweise auf eine gesteigerte LTP-ähnliche synaptische Plastizität vorlagen, ließen sich keine statistisch signifikanten Gruppen- oder Stimulationseffekte auf die motorkortikale Exzitabilität, Inhibition und synaptische Plastizität nachweisen. Die neurophysiologischen Parameter zeigten insgesamt eine hohe intra- und interindividuelle Variabilität. Diese sowie der heterogene Charakter leichter SHTs und die kleine Stichprobengröße könnten das Fehlen statistisch signifikanter Effekte erklären. Zudem ist nicht auszuschließen, dass in der SHT-Gruppe in der Zeit seit dem letzten SHT bereits eine weitgehende Normalisierung stattgefunden hat. Auch wenn diese Arbeit ältere Ergebnisse nur teilweise reproduzieren konnte, steht sie im Einklang mit neueren Publikationen, die eine hohe Variabilität und einen substanziellen Anteil an Non-Respondern bei nichtinvasiven Stimulationsverfahren berichten.

Einleitung Endometriose (EM) gehört zu den häufigsten gynäkologischen Erkrankungen. Bei hohen Infertilitätsraten ist Kinderwunsch für viele Betroffene ein zentrales Thema. Zunehmend wird beobachtet, dass eine EM auch Auswirkungen auf den Verlauf der Schwangerschaft und Geburt haben kann und Plazentationsstörungen und vermehrte Blutungen zu begünstigen scheint. Aktuell gelten Schwangere mit EM jedoch nicht als Risikoschwangere. Etwa 2% aller peripartalen Hämorrhagien (PPH) werden durch Plazenta praevia (PP), Placenta-accreta-Spektrum (PAS), Vasa praevia (VP) und Insertio velamentosa (IV) verursacht. In der Regel können sie sonographisch diagnostiziert und Komplikationen antizipiert werden. In einer prospektiven Studie untersuchten wir den möglichen Zusammenhang von EM mit Störungen der Plazentation und Nabelschnurinsertion. Methoden In der prospektiven Beobachtung begleiteten wir 82 Schwangere mit und 82 ohne EM. Primär untersuchten wir, ob PP, PAS, IV oder VP auftraten. Sekundär erhoben wir das Auftreten schwerer PPH und den Geburtsmodus, sowie den Beratungsbedarf bei EM. Die Kontrollen wurden nach Alter und Konzeptionsmodus gematcht. Ergebnisse In der Gruppe der Schwangeren mit EM beobachteten wir vermehrt Störungen der Plazenta und Nabelschnur, darunter PP, PAS, IV und VP, verglichen mit dem Auftreten einer PP in der Gruppe der gesunden Schwangeren (10% vs. 1%). Schwangere mit EM gebaren seltener spontan als Teilnehmende der Kontrollgruppe (43% vs. 67%). Dementsprechend erhöht war die Rate an Sectiones (40% vs. 26%) und vaginal-operativen Geburten (17% vs. 7%), sowie an Geburten mit einer schweren PPH (13% vs. 1%) in der EM-Gruppe. Von den befragten Teilnehmenden mit EM wurde ein Großteil nicht zu EM in der Schwangerschaft beraten (60%) und die Mehrheit hätte sich mehr Beratung zu dem Thema gewünscht (76%). Diskussion Bei Schwangeren mit EM beobachteten wir vermehrt Störungen der Plazenta oder Nabelschnur, es kam häufiger zu schweren PPH und seltener zur spontanen Geburt als in der Kontrollgruppe. Die Ergebnisse legen einen Zusammenhang von EM mit Plazentations- und Nabelschnurauffälligkeiten nahe. Diese sind Risikofaktoren für Schwangerschaft und Geburt und können die Behandlung der Schwangeren sowie eine Sectio caesarea verkomplizieren. Aufgrund der heterogenen Diagnose und Ausprägung der Erkrankung ist unklar, ob dies für alle oder nur für bestimmte Subformen der EM gilt. In jedem Fall sollten die Plazentation und Nabelschnurinsertion sorgfältig untersucht und eine Risikoberechnung für PPH vorgenommen werden. Ideale Bedingungen für den besonderen Beratungsbedarf und die individuelle Betreuung von Schwangeren mit EM finden sich in einem Perinatalzentrum.

Introduction: Preclinical curricula in medical education are increasingly overflowing with other basic sciences, and while anatomy remains a central subject, its instructional time has seen a steady decrease. Educators are concerned about declining levels of anatomical knowledge, and it has become clear: the instruction of anatomy must adapt to contemporary teaching and curricula changes. Cadaveric dissection prevails as the main instructional method in most universities, but how to make this method more efficient? Many agree to see great potential in integrating radiographic imaging to increase interest, and deeper understanding of anatomy. While the current body of research has mostly focused on medical students, this study has focused on studying the effects of this integrated teaching approach on the performance and motivation of dentistry students. Methods: The third preclinical semester was divided into a control and an intervention group. A curriculum was conceptualized and was delivered to students of the intervention group in parallel with their dissection course. The curriculum was written utilizing the backwards design method by Wiggins and McTighe. To analyse the courses’ effect on student motivation, a survey was designed and handed out at the end of the course to the intervention group. To assess the effects on student performance, an assessment form was created utilizing three levels of Bloom’s taxonomy and delivered at the end of the course to both the intervention and control group for comparison. Key descriptive statistics were applied to analyse responses in the student survey and hypothesis testing followed, with an independent samples two paired t-test to assess significant changes in performance after attending the integrated approach. Results: Students reported high levels of satisfaction and thought it to be altogether advantageous for their education. They recognized the integrated approach for an increased level of knowledge in gross anatomy and radiology. Though not statistically significant, a positive impact on student performance was observed, especially in lower domains of cognitive demand. Conclusions: An integrated approach of teaching anatomy with radiology is a resourceful and cost-effective way to address the gap created by the shortage of instruction time, while improving students' knowledge of anatomy and boosting their interest levels.

This dissertation explores the development of a novel three-dimensional (3D) bone marrow (BM) culture system, marking a significant advancement in the ex vivo survival of human plasma cells (PCs). By employing primary femoral BM tissue and a collagen-hyaluronic acid (Col-HA) hydrogel, this study established a more physiologically relevant environment than traditional cell culture models. A specifically designed Col-HA hydrogel, which reproduces some BM biomechanical properties, was the essential method implemented to preserve primary tissue integrity in vitro. Together with a PCs-tailored culture medium, our investigation demonstrated the obtained 3D BM culture system’s ability to sustain key survival factors and replicate the complex BM microenvironment, thus supporting the long-term viability and antibody production of PCs, which is crucial for studying haematological conditions and developing new treatments. The additional integration of a dynamic microphysiological system (MPS) aimed to further explore a more realistic simulation of the BM niche, providing valuable insights into cell behaviour and interactions within the hydrogel matrix. The present work also highlights the potential for future advancements in the outlined 3D BM model. Ongoing and upcoming research will focus on further optimizing the system through vascularization, which is critical for nutrient supply and waste removal, thereby enhancing cell survival and function. Single-cell analyses, such as scRNA CITE-seq, will be essential for detailed profiling of the in-tissue composition, marker expression, and interactions of different cell subpopulations. These analyses will provide deeper insights into the BM microenvironment’s complexity and inform the development of advanced therapeutic strategies. One of the significant aims of this study was the establishment of a testing platform to integrate chimeric antigen receptor (CAR) T cells into the 3D BM model. To this end, a collaborative project presented here achieved efficient and safe generation of CAR T cells via CRISPR-Cas9, offering a promising alternative to traditional viral transduction methods. TRAC locus insertion of CAR transgenes into T cells might pave the way for the creation of an effective therapeutic off-the-shelf strategy. Preliminary observations indicate that TRAC-replaced CAR T cells interact effectively with the Col-HA hydrogel in a 3D conformation. In conclusion, this work underscores the potential of the 3D BM culture system in advancing our understanding of BM biology and improving the development of targeted therapies, such as CAR T cell treatments. By connecting in vitro models with in vivo conditions, this research lays the foundation for more precise and efficient therapeutic approaches, with the ultimate goal of advancing clinical applications and enhancing patient outcomes.