@article{14628, abstract = {We introduce a compact, intuitive procedural graph representation for cellular metamaterials, which are small-scale, tileable structures that can be architected to exhibit many useful material properties. Because the structures’ “architectures” vary widely—with elements such as beams, thin shells, and solid bulks—it is difficult to explore them using existing representations. Generic approaches like voxel grids are versatile, but it is cumbersome to represent and edit individual structures; architecture-specific approaches address these issues, but are incompatible with one another. By contrast, our procedural graph succinctly represents the construction process for any structure using a simple skeleton annotated with spatially varying thickness. To express the highly constrained triply periodic minimal surfaces (TPMS) in this manner, we present the first fully automated version of the conjugate surface construction method, which allows novices to create complex TPMS from intuitive input. We demonstrate our representation’s expressiveness, accuracy, and compactness by constructing a wide range of established structures and hundreds of novel structures with diverse architectures and material properties. We also conduct a user study to verify our representation’s ease-of-use and ability to expand engineers’ capacity for exploration.}, author = {Makatura, Liane and Wang, Bohan and Chen, Yi-Lu and Deng, Bolei and Wojtan, Christopher J and Bickel, Bernd and Matusik, Wojciech}, issn = {1557-7368}, journal = {ACM Transactions on Graphics}, keywords = {Computer Graphics and Computer-Aided Design}, number = {5}, publisher = {Association for Computing Machinery}, title = {{Procedural metamaterials: A unified procedural graph for metamaterial design}}, doi = {10.1145/3605389}, volume = {42}, year = {2023}, } @phdthesis{14651, abstract = {For self-incompatibility (SI) to be stable in a population, theory predicts that sufficient inbreeding depression (ID) is required: the fitness of offspring from self-mated individuals must be low enough to prevent the spread of self-compatibility (SC). Reviews of natural plant populations have supported this theory, with SI species generally showing high levels of ID. However, there is thought to be an under-sampling of self-incompatible taxa in the current literature. In this thesis, I study inbreeding depression in the SI plant species Antirrhinum majus using both greenhouse crosses and a large collected field dataset. Additionally, the gametophytic S-locus of A. majus is highly heterozygous and polymorphic, thus making assembly and discovery of S-alleles very difficult. Here, 206 new alleles of the male component SLFs are presented, along with a phylogeny showing the high conservation with alleles from another Antirrhinum species. Lastly, selected sites within the protein structure of SLFs are investigated, with one site in particular highlighted as potentially being involved in the SI recognition mechanism.}, author = {Arathoon, Louise S}, issn = {2663-337X}, pages = {96}, publisher = {Institute of Science and Technology Austria}, title = {{Investigating inbreeding depression and the self-incompatibility locus of Antirrhinum majus}}, doi = {10.15479/at:ista:14651}, year = {2023}, } @inproceedings{14735, abstract = {Scaling blockchain protocols to perform on par with the expected needs of Web3.0 has been proven to be a challenging task with almost a decade of research. In the forefront of the current solution is the idea of separating the execution of the updates encoded in a block from the ordering of blocks. In order to achieve this, a new class of protocols called rollups has emerged. Rollups have as input a total ordering of valid and invalid transactions and as output a new valid state-transition. If we study rollups from a distributed computing perspective, we uncover that rollups take as input the output of a Byzantine Atomic Broadcast (BAB) protocol and convert it to a State Machine Replication (SMR) protocol. BAB and SMR, however, are considered equivalent as far as distributed computing is concerned and a solution to one can easily be retrofitted to solve the other simply by adding/removing an execution step before the validation of the input. This “easy” step of retrofitting an atomic broadcast solution to implement an SMR has, however, been overlooked in practice. In this paper, we formalize the problem and show that after BAB is solved, traditional impossibility results for consensus no longer apply towards an SMR. Leveraging this we propose a distributed execution protocol that allows reduced execution and storage cost per executor (O(log2n/n)) without relaxing the network assumptions of the underlying BAB protocol and providing censorship-resistance. Finally, we propose efficient non-interactive light client constructions that leverage our efficient execution protocols and do not require any synchrony assumptions or expensive ZK-proofs.}, author = {Stefo, Christos and Xiang, Zhuolun and Kokoris Kogias, Eleftherios}, booktitle = {27th International Conference on Financial Cryptography and Data Security}, isbn = {9783031477539}, issn = {0302-9743}, location = {Bol, Brac, Croatia}, pages = {3--20}, publisher = {Springer Nature}, title = {{Executing and proving over dirty ledgers}}, doi = {10.1007/978-3-031-47754-6_1}, volume = {13950}, year = {2023}, } @misc{15292, abstract = {We present a rigid body animation technique which prevents solids from interpenetrating, dissipates energy through friction, and propagates shocks through contacts. We employ the Alternating Direction Method of Multipliers (ADMM) to couple non-smooth Coulomb friction with impact propagation, allowing efficient and accurate non-smooth dynamics along with a correct transmission of impacts through assemblies of rigid bodies. We further extend our method to model adhesion, dynamic friction and lubricated contact.}, author = {Chen, Yi-Lu and Ly, Mickaël and Wojtan, Christopher J}, booktitle = {Proceedings of the ACM SIGGRAPH/Eurographics Symposium on Computer Animation}, location = {Los Angeles, CA, United States}, publisher = {ACM}, title = {{Unified treatment of contact, friction and shock-propagation in rigid body animation}}, doi = {10.1145/3606037.3606836}, year = {2023}, } @unpublished{20572, abstract = {We present an elementary non-recursive formula for the multivariate moments of the Dirichlet distribution on the standard simplex, in terms of the pattern inventory of the moments' exponents. We obtain analog formulas for the multivariate moments of the Dirichlet-Ferguson and Gamma measures. We further introduce a polychromatic analogue of Ewens sampling formula on colored integer partitions, discuss its relation with suitable extensions of Hoppe's urn model and of the Chinese restaurant process, and prove that it satisfies an adapted notion of consistency in the sense of Kingman.}, author = {Dello Schiavo, Lorenzo and Quattrocchi, Filippo}, booktitle = {arXiv}, keywords = {Dirichlet distribution, Ewens sampling formula, Hoppe urn model, colored partitions}, title = {{Multivariate Dirichlet moments and a polychromatic Ewens sampling formula}}, doi = {10.48550/arXiv.2309.11292}, year = {2023}, } @inproceedings{14863, author = {Polesello, Andrea and Muller, Caroline J and Pasquero, Claudia and Meroni, Agostino N.}, booktitle = {EGU General Assembly 2023}, location = {Vienna, Austria & Virtual}, publisher = {European Geosciences Union}, title = {{Intensification mechanisms of tropical cyclones}}, doi = {10.5194/egusphere-egu23-6157}, year = {2023}, } @phdthesis{12885, abstract = {High-performance semiconductors rely upon precise control of heat and charge transport. This can be achieved by precisely engineering defects in polycrystalline solids. There are multiple approaches to preparing such polycrystalline semiconductors, and the transformation of solution-processed colloidal nanoparticles is appealing because colloidal nanoparticles combine low cost with structural and compositional tunability along with rich surface chemistry. However, the multiple processes from nanoparticle synthesis to the final bulk nanocomposites are very complex. They involve nanoparticle purification, post-synthetic modifications, and finally consolidation (thermal treatments and densification). All these properties dictate the final material’s composition and microstructure, ultimately affecting its functional properties. This thesis explores the synthesis, surface chemistry and consolidation of colloidal semiconductor nanoparticles into dense solids. In particular, the transformations that take place during these processes, and their effect on the material’s transport properties are evaluated. }, author = {Calcabrini, Mariano}, isbn = {978-3-99078-028-2}, issn = {2663-337X}, pages = {82}, publisher = {Institute of Science and Technology Austria}, title = {{Nanoparticle-based semiconductor solids: From synthesis to consolidation}}, doi = {10.15479/at:ista:12885}, year = {2023}, } @inproceedings{14260, abstract = {This paper presents Lincheck, a new practical and user-friendly framework for testing concurrent algorithms on the Java Virtual Machine (JVM). Lincheck provides a simple and declarative way to write concurrent tests: instead of describing how to perform the test, users specify what to test by declaring all the operations to examine; the framework automatically handles the rest. As a result, tests written with Lincheck are concise and easy to understand. The framework automatically generates a set of concurrent scenarios, examines them using stress-testing or bounded model checking, and verifies that the results of each invocation are correct. Notably, if an error is detected via model checking, Lincheck provides an easy-to-follow trace to reproduce it, significantly simplifying the bug investigation. To the best of our knowledge, Lincheck is the first production-ready tool on the JVM that offers such a simple way of writing concurrent tests, without requiring special skills or expertise. We successfully integrated Lincheck in the development process of several large projects, such as Kotlin Coroutines, and identified new bugs in popular concurrency libraries, such as a race in Java’s standard ConcurrentLinkedDeque and a liveliness bug in Java’s AbstractQueuedSynchronizer framework, which is used in most of the synchronization primitives. We believe that Lincheck can significantly improve the quality and productivity of concurrent algorithms research and development and become the state-of-the-art tool for checking their correctness.}, author = {Koval, Nikita and Fedorov, Alexander and Sokolova, Maria and Tsitelov, Dmitry and Alistarh, Dan-Adrian}, booktitle = {35th International Conference on Computer Aided Verification}, isbn = {9783031377051}, issn = {1611-3349}, location = {Paris, France}, pages = {156--169}, publisher = {Springer Nature}, title = {{Lincheck: A practical framework for testing concurrent data structures on JVM}}, doi = {10.1007/978-3-031-37706-8_8}, volume = {13964}, year = {2023}, } @phdthesis{14547, abstract = {Superconductor-semiconductor heterostructures currently capture a significant amount of research interest and they serve as the physical platform in many proposals towards topological quantum computation. Despite being under extensive investigations, historically using transport techniques, the basic properties of the interface between the superconductor and the semiconductor remain to be understood. In this thesis, two separate studies on the Al-InAs heterostructures are reported with the first focusing on the physics of the material motivated by the emergence of a new phase, the Bogoliubov-Fermi surface. The second focuses on a technological application, a gate-tunable Josephson parametric amplifier. In the first study, we investigate the hypothesized unconventional nature of the induced superconductivity at the interface between the Al thin film and the InAs quantum well. We embed a two-dimensional Al-InAs hybrid system in a resonant microwave circuit allowing measurements of change in inductance. The behaviour of the resonance in a range of temperature and in-plane magnetic field has been studied and compared with the theory of conventional s-wave superconductor and a two-component theory that includes both contribution of the $s$-wave pairing in Al and the intraband $p \pm ip$ pairing in InAs. Measuring the temperature dependence of resonant frequency, no discrepancy is found between data and the conventional theory. We observe the breakdown of superconductivity due to an applied magnetic field which contradicts the conventional theory. In contrast, the data can be captured quantitatively by fitting to a two-component model. We find the evidence of the intraband $p \pm ip$ pairing in the InAs and the emergence of the Bogoliubov-Fermi surfaces due to magnetic field with the characteristic value $B^* = 0.33~\mathrm{T}$. From the fits, the sheet resistance of Al, the carrier density and mobility in InAs are determined. By systematically studying the anisotropy of the circuit response, we find weak anisotropy for $B < B^*$ and increasingly strong anisotropy for $B > B^*$ resulting in a pronounced two-lobe structure in polar plot of frequency versus field angle. Strong resemblance between the field dependence of dissipation and superfluid density hints at a hidden signature of the Bogoliubov-Fermi surface that is burried in the dissipation data. In the second study, we realize a parametric amplifier with a Josephson field effect transistor as the active element. The device's modest construction consists of a gated SNS weak link embedded at the center of a coplanar waveguide resonator. By applying a gate voltage, the resonant frequency is field-effect tunable over a range of 2 GHz. Modelling the JoFET minimally as a parallel RL circuit, the dissipation introduced by the JoFET can be quantitatively related to the gate voltage. We observed gate-tunable Kerr nonlinearity qualitatively in line with expectation. The JoFET amplifier has 20 dB of gain, 4 MHz of instantaneous bandwidth, and a 1dB compression point of -125.5 dBm when operated at a fixed resonant frequency. In general, the signal-to-noise ratio is improved by 5-7 dB when the JoFET amplifier is activated compared. The noise of the measurement chain and insertion loss of relevant circuit elements are calibrated to determine the expected and the real noise performance of the JoFET amplifier. As a quantification of the noise performance, the measured total input-referred noise of the JoFET amplifier is in good agreement with the estimated expectation which takes device loss into account. We found that the noise performance of the device reported in this document approaches one photon of total input-referred added noise which is the quantum limit imposed in nondegenerate parametric amplifier.}, author = {Phan, Duc T}, issn = {2663-337X}, keywords = {superconductor-semiconductor, superconductivity, Al, InAs, p-wave, superconductivity, JPA, microwave}, pages = {80}, publisher = {Institute of Science and Technology Austria}, title = {{Resonant microwave spectroscopy of Al-InAs}}, doi = {10.15479/14547}, year = {2023}, } @phdthesis{14641, abstract = {Mutation rates represent the net result of complex interactions among various cellular processes and can dramatically influence the evolutionary fate of microbial populations. However, many popular techniques used to study mutations are subject to the confounding effects of heredity and the subtleties of adaptation to selection, all of which make it difficult to observe any dynamic responses of mutation rates to fitness challenges. Furthermore, in spite of the ubiquity of quorum sensing systems across the bacterial domain and relevance for many physiological behaviors, the effects of such mechanisms on mutation rate and adaptation remain poorly understood. In the following work, I present the development of a microfluidic droplet-based method to measure single base-pair mutation rates in growing populations of the bacterium Escherichia coli. I use this method to observe a stress-induced increase in mutation rate that is mediated by luxS, a highly conserved bacterial quorum sensing component. I also show that the aforementioned increase in mutation rate, and its associated control by luxS, corresponds to a higher degree of adaptability under competitive environments.}, author = {Hennessey-Wesen, Mike}, issn = {2663-337X}, keywords = {microfluidics, miceobiology, mutations, quorum sensing}, pages = {104}, publisher = {Institute of Science and Technology Austria}, title = {{Adaptive mutation in E. coli modulated by luxS}}, doi = {10.15479/at:ista:14641}, year = {2023}, } @article{12675, abstract = {Aromatic side chains are important reporters of the plasticity of proteins, and often form important contacts in protein--protein interactions. By studying a pair of structurally homologous cross-β amyloid fibrils, HET-s and HELLF, with a specific isotope-labeling approach and magic-angle-spinning (MAS) NMR, we have characterized the dynamic behavior of Phe and Tyr aromatic rings to show that the hydrophobic amyloid core is rigid, without any sign of "breathing motions" over hundreds of milliseconds at least. Aromatic residues exposed at the fibril surface have a rigid ring axis but undergo ring flips, on a variety of time scales from ns to µs. Our approach provides direct insight into hydrophobic-core motions, enabling a better evaluation of the conformational heterogeneity generated from a NMR structural ensemble of such amyloid cross-β architecture.}, author = {Becker, Lea Marie and Berbon, Mélanie and Vallet, Alicia and Grelard, Axelle and Morvan, Estelle and Bardiaux, Benjamin and Lichtenecker, Roman and Ernst, Matthias and Loquet, Antoine and Schanda, Paul}, issn = {1521-3773}, journal = {Angewandte Chemie International Edition}, keywords = {General Chemistry, Catalysis}, number = {19}, publisher = {Wiley}, title = {{The rigid core and flexible surface of amyloid fibrils probed by Magic‐Angle Spinning NMR of aromatic residues}}, doi = {10.1002/anie.202219314}, volume = {62}, year = {2023}, } @unpublished{15039, abstract = {A crucial property for achieving secure, trustworthy and interpretable deep learning systems is their robustness: small changes to a system's inputs should not result in large changes to its outputs. Mathematically, this means one strives for networks with a small Lipschitz constant. Several recent works have focused on how to construct such Lipschitz networks, typically by imposing constraints on the weight matrices. In this work, we study an orthogonal aspect, namely the role of the activation function. We show that commonly used activation functions, such as MaxMin, as well as all piece-wise linear ones with two segments unnecessarily restrict the class of representable functions, even in the simplest one-dimensional setting. We furthermore introduce the new N-activation function that is provably more expressive than currently popular activation functions. We provide code at this https URL.}, author = {Prach, Bernd and Lampert, Christoph}, booktitle = {arXiv}, title = {{1-Lipschitz neural networks are more expressive with N-activations}}, doi = {10.48550/ARXIV.2311.06103}, year = {2023}, } @inproceedings{13221, abstract = {The safety-liveness dichotomy is a fundamental concept in formal languages which plays a key role in verification. Recently, this dichotomy has been lifted to quantitative properties, which are arbitrary functions from infinite words to partially-ordered domains. We look into harnessing the dichotomy for the specific classes of quantitative properties expressed by quantitative automata. These automata contain finitely many states and rational-valued transition weights, and their common value functions Inf, Sup, LimInf, LimSup, LimInfAvg, LimSupAvg, and DSum map infinite words into the totallyordered domain of real numbers. In this automata-theoretic setting, we establish a connection between quantitative safety and topological continuity and provide an alternative characterization of quantitative safety and liveness in terms of their boolean counterparts. For all common value functions, we show how the safety closure of a quantitative automaton can be constructed in PTime, and we provide PSpace-complete checks of whether a given quantitative automaton is safe or live, with the exception of LimInfAvg and LimSupAvg automata, for which the safety check is in ExpSpace. Moreover, for deterministic Sup, LimInf, and LimSup automata, we give PTime decompositions into safe and live automata. These decompositions enable the separation of techniques for safety and liveness verification for quantitative specifications.}, author = {Boker, Udi and Henzinger, Thomas A and Mazzocchi, Nicolas Adrien and Sarac, Naci E}, booktitle = {34th International Conference on Concurrency Theory}, isbn = {9783959772990}, issn = {1868-8969}, location = {Antwerp, Belgium}, publisher = {Schloss Dagstuhl - Leibniz-Zentrum für Informatik}, title = {{Safety and liveness of quantitative automata}}, doi = {10.4230/LIPIcs.CONCUR.2023.17}, volume = {279}, year = {2023}, } @article{12762, abstract = {Neurons in the brain are wired into adaptive networks that exhibit collective dynamics as diverse as scale-specific oscillations and scale-free neuronal avalanches. Although existing models account for oscillations and avalanches separately, they typically do not explain both phenomena, are too complex to analyze analytically or intractable to infer from data rigorously. Here we propose a feedback-driven Ising-like class of neural networks that captures avalanches and oscillations simultaneously and quantitatively. In the simplest yet fully microscopic model version, we can analytically compute the phase diagram and make direct contact with human brain resting-state activity recordings via tractable inference of the model’s two essential parameters. The inferred model quantitatively captures the dynamics over a broad range of scales, from single sensor oscillations to collective behaviors of extreme events and neuronal avalanches. Importantly, the inferred parameters indicate that the co-existence of scale-specific (oscillations) and scale-free (avalanches) dynamics occurs close to a non-equilibrium critical point at the onset of self-sustained oscillations.}, author = {Lombardi, Fabrizio and Pepic, Selver and Shriki, Oren and Tkačik, Gašper and De Martino, Daniele}, issn = {2662-8457}, journal = {Nature Computational Science}, pages = {254--263}, publisher = {Springer Nature}, title = {{Statistical modeling of adaptive neural networks explains co-existence of avalanches and oscillations in resting human brain}}, doi = {10.1038/s43588-023-00410-9}, volume = {3}, year = {2023}, } @inproceedings{13120, abstract = {We formalized general (i.e., type-0) grammars using the Lean 3 proof assistant. We defined basic notions of rewrite rules and of words derived by a grammar, and used grammars to show closure of the class of type-0 languages under four operations: union, reversal, concatenation, and the Kleene star. The literature mostly focuses on Turing machine arguments, which are possibly more difficult to formalize. For the Kleene star, we could not follow the literature and came up with our own grammar-based construction.}, author = {Dvorak, Martin and Blanchette, Jasmin}, booktitle = {14th International Conference on Interactive Theorem Proving}, isbn = {9783959772846}, issn = {1868-8969}, location = {Bialystok, Poland}, publisher = {Schloss Dagstuhl - Leibniz-Zentrum für Informatik}, title = {{Closure properties of general grammars - formally verified}}, doi = {10.4230/LIPIcs.ITP.2023.15}, volume = {268}, year = {2023}, } @article{13049, abstract = {We propose a computational design approach for covering a surface with individually addressable RGB LEDs, effectively forming a low-resolution surface screen. To achieve a low-cost and scalable approach, we propose creating designs from flat PCB panels bent in-place along the surface of a 3D printed core. Working with standard rigid PCBs enables the use of established PCB manufacturing services, allowing the fabrication of designs with several hundred LEDs. Our approach optimizes the PCB geometry for folding, and then jointly optimizes the LED packing, circuit and routing, solving a challenging layout problem under strict manufacturing requirements. Unlike paper, PCBs cannot bend beyond a certain point without breaking. Therefore, we introduce parametric cut patterns acting as hinges, designed to allow bending while remaining compact. To tackle the joint optimization of placement, circuit and routing, we propose a specialized algorithm that splits the global problem into one sub-problem per triangle, which is then individually solved. Our technique generates PCB blueprints in a completely automated way. After being fabricated by a PCB manufacturing service, the boards are bent and glued by the user onto the 3D printed support. We demonstrate our technique on a range of physical models and virtual examples, creating intricate surface light patterns from hundreds of LEDs.}, author = {Freire, Marco and Bhargava, Manas and Schreck, Camille and Hugron, Pierre-Alexandre and Bickel, Bernd and Lefebvre, Sylvain}, issn = {1557-7368}, journal = {Transactions on Graphics}, keywords = {PCB design and layout, Mesh geometry models}, location = {Los Angeles, CA, United States}, number = {4}, publisher = {Association for Computing Machinery}, title = {{PCBend: Light up your 3D shapes with foldable circuit boards}}, doi = {10.1145/3592411}, volume = {42}, year = {2023}, } @article{14343, abstract = {The total energy of an eigenstate in a composite quantum system tends to be distributed equally among its constituents. We identify the quantum fluctuation around this equipartition principle in the simplest disordered quantum system consisting of linear combinations of Wigner matrices. As our main ingredient, we prove the Eigenstate Thermalisation Hypothesis and Gaussian fluctuation for general quadratic forms of the bulk eigenvectors of Wigner matrices with an arbitrary deformation.}, author = {Cipolloni, Giorgio and Erdös, László and Henheik, Sven Joscha and Kolupaiev, Oleksii}, issn = {2050-5094}, journal = {Forum of Mathematics, Sigma}, publisher = {Cambridge University Press}, title = {{Gaussian fluctuations in the equipartition principle for Wigner matrices}}, doi = {10.1017/fms.2023.70}, volume = {11}, year = {2023}, } @article{14421, abstract = {Only recently has it been possible to construct a self-adjoint Hamiltonian that involves the creation of Dirac particles at a point source in 3d space. Its definition makes use of an interior-boundary condition. Here, we develop for this Hamiltonian a corresponding theory of the Bohmian configuration. That is, we (non-rigorously) construct a Markov jump process $(Q_t)_{t\in\mathbb{R}}$ in the configuration space of a variable number of particles that is $|\psi_t|^2$-distributed at every time t and follows Bohmian trajectories between the jumps. The jumps correspond to particle creation or annihilation events and occur either to or from a configuration with a particle located at the source. The process is the natural analog of Bell's jump process, and a central piece in its construction is the determination of the rate of particle creation. The construction requires an analysis of the asymptotic behavior of the Bohmian trajectories near the source. We find that the particle reaches the source with radial speed 0, but orbits around the source infinitely many times in finite time before absorption (or after emission).}, author = {Henheik, Sven Joscha and Tumulka, Roderich}, issn = {1751-8121}, journal = {Journal of Physics A: Mathematical and Theoretical}, number = {44}, publisher = {IOP Publishing}, title = {{Creation rate of Dirac particles at a point source}}, doi = {10.1088/1751-8121/acfe62}, volume = {56}, year = {2023}, } @phdthesis{12491, abstract = {The extracellular matrix (ECM) is a hydrated and complex three-dimensional network consisting of proteins, polysaccharides, and water. It provides structural scaffolding for the cells embedded within it and is essential in regulating numerous physiological processes, including cell migration and proliferation, wound healing, and stem cell fate. Despite extensive study, detailed structural knowledge of ECM components in physiologically relevant conditions is still rudimentary. This is due to methodological limitations in specimen preparation protocols which are incompatible with keeping large samples, such as the ECM, in their native state for subsequent imaging. Conventional electron microscopy (EM) techniques rely on fixation, dehydration, contrasting, and sectioning. This results in the alteration of a highly hydrated environment and the potential introduction of artifacts. Other structural biology techniques, such as nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography, allow high-resolution analysis of protein structures but only work on homogenous and purified samples, hence lacking contextual information. Currently, no approach exists for the ultrastructural and structural study of extracellular components under native conditions in a physiological, 3D environment. In this thesis, I have developed a workflow that allows for the ultrastructural analysis of the ECM in near-native conditions at molecular resolution. The developments I introduced include implementing a novel specimen preparation workflow for cell-derived matrices (CDMs) to render them compatible with ion-beam milling and subsequent high-resolution cryo-electron tomography (ET). To this end, I have established protocols to generate CDMs grown over several weeks on EM grids that are compatible with downstream cryo-EM sample preparation and imaging techniques. Characterization of these ECMs confirmed that they contain essential ECM components such as collagen I, collagen VI, and fibronectin I in high abundance and hence represent a bona fide biologically-relevant sample. I successfully optimized vitrification of these specimens by testing various vitrification techniques and cryoprotectants. In order to obtain high-resolution molecular insights into the ultrastructure and organization of CDMs, I established cryo-focused ion beam scanning electron microscopy (FIBSEM) on these challenging and complex specimens. I explored different approaches for the creation of thin cryo-lamellae by FIB milling and succeeded in optimizing the cryo-lift-out technique, resulting in high-quality lamellae of approximately 200 nm thickness. High-resolution Cryo-ET of these lamellae revealed for the first time the architecture of native CDM in the context of matrix-secreting cells. This allowed for the in situ visualization of fibrillar matrix proteins such as collagen, laying the foundation for future structural and ultrastructural characterization of these proteins in their near-native environment. In summary, in this thesis, I present a novel workflow that combines state-of-the-art cryo-EM specimen preparation and imaging technologies to permit characterization of the ECM, an important tissue component in higher organisms. This innovative and highly versatile workflow will enable addressing far-reaching questions on ECM architecture, composition, and reciprocal ECM-cell interactions.}, author = {Zens, Bettina}, isbn = {978-3-99078-027-5}, issn = {2663-337X}, keywords = {cryo-EM, cryo-ET, FIB milling, method development, FIBSEM, extracellular matrix, ECM, cell-derived matrices, CDMs, cell culture, high pressure freezing, HPF, structural biology, tomography, collagen}, pages = {187}, publisher = {Institute of Science and Technology Austria}, title = {{Ultrastructural characterization of natively preserved extracellular matrix by cryo-electron tomography}}, doi = {10.15479/at:ista:12491}, year = {2023}, } @phdthesis{13984, abstract = {Social insects fight disease using their individual immune systems and the cooperative sanitary behaviors of colony members. These social defenses are well explored against externally-infecting pathogens, but little is known about defense strategies against internally-infecting pathogens, such as viruses. Viruses are ubiquitous and in the last decades it has become evident that also many ant species harbor viruses. We present one of the first studies addressing transmission dynamics and collective disease defenses against viruses in ants on a mechanistic level. I successfully established an experimental ant host – viral pathogen system as a model for the defense strategies used by social insects against internal pathogen infections, as outlined in the third chapter. In particular, we studied how garden ants (Lasius neglectus) defend themselves and their colonies against the generalist insect virus CrPV (cricket paralysis virus). We chose microinjections of virus directly into the ants’ hemolymph because it allowed us to use a defined exposure dose. Here we show that this is a good model system, as the virus is replicating and thus infecting the host. The ants mount a clear individual immune response against the viral infection, which is characterized by a specific siRNA pattern, namely siRNAs mapping against the viral genome with a peak of 21 and 22 bp long fragments. The onset of this immune response is consistent with the timeline of viral replication that starts already within two days post injection. The disease manifests in decreased survival over a course of two to three weeks. Regarding group living, we find that infected ants show a strong individual immune response, but that their course of disease is little affected by nestmate presence, as described in chapter four. Hence, we do not find social immunity in the context of viral infections in ants. Nestmates, however, can contract the virus. Using Drosophila S2R+ cells in culture, we showed that 94 % of the nestmates contract active virus within four days of social contact to an infected individual. Virus is transmitted in low doses, thus not causing disease transmission within the colony. While virus can be transmitted during short direct contacts, we also assume transmission from deceased ants and show that the nestmates’ immune system gets activated after contracting a low viral dose. We find considerable potential for indirect transmission via the nest space. Virus is shed to the nest, where it stays viable for one week and is also picked up by other ants. Apart from that, we want to underline the potential of ant poison as antiviral agent. We determined that ant poison successfully inactivates CrPV in vitro. However, we found no evidence for effective poison use to sanitize the nest space. On the other hand, local application of ant poison by oral poison uptake, which is part of the ants prophylactic behavioral repertoire, probably contributes to keeping the gut of each individual sanitized. We hypothesize that oral poison uptake might be the reason why we did not find viable virus in the trophallactic fluid. The fifth chapter encompasses preliminary data on potential social immunization. However, our experiments do not confirm an actual survival benefit for the nestmates upon pathogen challenge under the given experimental settings. Nevertheless, we do not want to rule out the possibility for nestmate immunization, but rather emphasize that considering different experimental timelines and viral doses would provide a multitude of options for follow-up experiments. In conclusion, we find that prophylactic individual behaviors, such as oral poison uptake, might play a role in preventing viral disease transmission. Compared to colony defense against external pathogens, internal pathogen infections require a stronger component of individual physiological immunity than behavioral social immunity, yet could still lead to collective protection.}, author = {Franschitz, Anna}, isbn = {978-3-99078-034-3}, issn = {2663-337X}, pages = {89}, publisher = {Institute of Science and Technology Austria}, title = {{Individual and social immunity against viral infections in ants}}, doi = {10.15479/at:ista:13984}, year = {2023}, }