Efficient Symmetry Reduction and the Use of State Symmetries for Symbolic Model Checking

One technique to reduce the state-space explosion problem in temporal logic model checking is symmetry reduction. The combination of symmetry reduction and symbolic model checking by using BDDs suffered a long time from the prohibitively large BDD for the orbit relation. Dynamic symmetry reduction calculates representatives of equivalence classes of states dynamically and thus avoids the construction of the orbit relation. In this paper, we present a new efficient model checking algorithm based on dynamic symmetry reduction. Our experiments show that the algorithm is very fast and allows the verification of larger systems. We additionally implemented the use of state symmetries for symbolic symmetry reduction. To our knowledge we are the first who investigated state symmetries in combination with BDD based symbolic model checking

Lipid-Iron Nanoparticle with a Cell Stress Release Mechanism Combined with a Local Alternating Magnetic Field Enables Site-Activated Drug Release

Simple Summary A novel active release system magnetic sphingomyelin-containing liposome encapsulated with indocyanine green, fluorescent marker, or the anticancer drug cisplatin was evaluated. The liposomal sphingomyelin is a target for the sphingomyelinase enzyme, which is released by stressed cells. Thus, sphingomyelin containing liposomes behave as a sensitizer for biological stress situations. In addition, the liposomes were engineered by adding paramagnetic beads to act as a receiver of outside given magnetic energy. The enzymatic activity towards liposomes and destruction caused by the applied magnetic field caused the release of the content from the liposomes. By using these novel liposomes, we could improve the drug release feature of liposomes. The improved targeting and drug-release were shown in vitro and the orthotopic tongue cancer model in mice optical imaging. The increased delivery of cisplatin prolonged the survival of the targeted delivery group versus free cisplatin. Most available cancer chemotherapies are based on systemically administered small organic molecules, and only a tiny fraction of the drug reaches the disease site. The approach causes significant side effects and limits the outcome of the therapy. Targeted drug delivery provides an alternative to improve the situation. However, due to the poor release characteristics of the delivery systems, limitations remain. This report presents a new approach to address the challenges using two fundamentally different mechanisms to trigger the release from the liposomal carrier. We use an endogenous disease marker, an enzyme, combined with an externally applied magnetic field, to open the delivery system at the correct time only in the disease site. This site-activated release system is a novel two-switch nanomachine that can be regulated by a cell stress-induced enzyme at the cellular level and be remotely controlled using an applied magnetic field. We tested the concept using sphingomyelin-containing liposomes encapsulated with indocyanine green, fluorescent marker, or the anticancer drug cisplatin. We engineered the liposomes by adding paramagnetic beads to act as a receiver of outside magnetic energy. The developed multifunctional liposomes were characterized in vitro in leakage studies and cell internalization studies. The release system was further studied in vivo in imaging and therapy trials using a squamous cell carcinoma tumor in the mouse as a disease model. In vitro studies showed an increased release of loaded material when stress-related enzyme and magnetic field was applied to the carrier liposomes. The theranostic liposomes were found in tumors, and the improved therapeutic effect was shown in the survival studies.Peer reviewe

Improving BDD Based Symbolic Model Checking with Isomorphism Exploiting Transition Relations

Symbolic model checking by using BDDs has greatly improved the applicability of model checking. Nevertheless, BDD based symbolic model checking can still be very memory and time consuming. One main reason is the complex transition relation of systems. Sometimes, it is even not possible to generate the transition relation, due to its exhaustive memory requirements. To diminish this problem, the use of partitioned transition relations has been proposed. However, there are still systems which can not be verified at all. Furthermore, if the granularity of the partitions is too fine, the time required for verification may increase. In this paper we target the symbolic verification of asynchronous concurrent systems. For such systems we present an approach which uses similarities in the transition relation to get further memory reductions and runtime improvements. By applying our approach, even the verification of systems with an previously intractable transition relation becomes feasible.Comment: In Proceedings GandALF 2011, arXiv:1106.081

Symbolic BDD-based Model Checking of Asynchronous Concurrent Systems

Today, information and communication systems are ubiquitous and consist very often of several interacting and communicating components. One reason is the widespread use of multi-core processors and the increasing amount of concurrent software for the efficient usage of multi-core processors. Also, the dissemination of distributed emergent technologies like sensor networks or the internet of things is growing. Additionally, a lot of internet protocols are client-server architectures with clients which execute computations in parallel and servers that can handle requests of several clients in parallel. Systems which consist of several interacting and communicating components are often very complex and due to their complexity also prone to errors. Errors in systems can have dramatic consequenses, especially in safety-critical areas where human life can be endangered by incorrect system behavior. Hence, it is inevitable to have methods that ensure the proper functioning of such systems. This thesis aims on improving the verifiability of asynchronous concurrent systems using symbolic model checking based on Binary Decision Diagrams (BDDs). An asynchronous concurrent system is a system that consists of several components, from which only one component can execute a transition at a time. Model checking is a formal verification technique. For a given system description and a set of desired properties, the validity of the properties for the system is decided in model checking automatically by software tools called model checkers. The main problem of model checking is the state-space explosion problem. One approach to reduce this problem is the use of symbolic model checking. There, system states and transitions are not stored explicitely as in explicit model checking. Instead, in symbolic model checking sets of states and sets of transitions are stored and also manipulated together. The data structure which is used in this thesis to store those sets are BDDs. BDD-based symbolic model checking has already been used successful in industry for several times. Nevertheless, BDD-based symbolic model checking still suffers from the state-space explosion problem and further improvements are necessary to improve its applicability. Central operations in BDD-based symbolic model checking are the computation of successor and predecessor states of a given set of states. Those computations are called image computations. They are applied repeatedly in BDD-based symbolic model checking to decide the validity of properties for a given system description. Hence, their efficient execution is crucial for the memory and runtime requirements of a model checker. In an image computation a BDD for a set of transitions and a BDD for a set of states are combined to compute a set of successor or predecessor states. Often, also the size of the BDDs to represent the transition relation is critical for the successful use of model checking. To further improve the applicability of symbolic model checking, we present in this thesis new data structures to store the transition relation of asynchronous concurrent systems. Additionally, we present new image computation algorithms. Both can lead to large runtime and memory reductions for BDD-based symbolic model checking. Asynchronous concurrent systems often contain symmetries. A technique to exploit those symmetries to diminish the state-space explosion problem is symmetry reduction. In this thesis we also present a new efficient algorithm for symmetry reduction in BDD-based symbolic model checking.In unserem Alltag kommen wir heute ständig mit Systemen der Informations- und Kommunikationstechnik in Kontakt. Diese bestehen häufig aus mehreren interagierenden und kommunizierenden Komponenten, wie zum Beispiel nebenläufige Software zur effizienten Nutzung von Mehrkernprozessoren oder Sensornetzwerke. Systeme, die aus mehreren interagierenden und kommunizierenden Komponenten bestehen sind häufig komplex und dadurch sehr fehleranfällig. Daher ist es wichtig zuverlässige Methoden, die helfen die korrekte Funktionsweise solcher Systeme sicherzustellen, zu besitzen. Im Rahmen dieser Doktorarbeit wurden neue Methoden zur Verbesserung der Verifizierbarkeit von asynchronen nebenläufigen Systemen durch Anwendung der symbolischen Modellprüfung mit binären Entscheidungsdiagrammen (BDDs) entwickelt. Ein asynchrones nebenläufiges System besteht aus mehreren Komponenten, von denen zu einem Zeitpunkt jeweils nur eine Komponente Transitionen ausführen kann. Die Modellprüfung ist eine Technik zur formalen Verifikation, bei der die Gültigkeit einer Menge von zu prüfenden Eigenschaften für eine gegebene Systembeschreibung automatisch durch Softwarewerkzeuge, die Modellprüfer genannt werden, entschieden wird. Das Hauptproblem der symbolischen Modellprüfung ist das Problem der Zustandsraumexplosion und es sind weitere Verbesserungen notwendig, um die symbolische Modellprüfung häufiger erfolgreich durchführen zu können. Bei der BDD-basierten symbolischen Modellprüfung werden Mengen von Systemzuständen und Mengen von Transitionen jeweils durch BDDs repräsentiert. Zentrale Operationen bei ihr sind die Berechnung von Nachfolger- und Vorgängerzuständen von gegebenen Zustandsmengen, welche Bildberechnungen genannt werden. Um die Gültigkeit von Eigenschaften für eine gegebene Systembeschreibung zu überprüfen, werden wiederholt Bildberechnungen durchgeführt. Daher ist ihre effiziente Berechnung entscheidend für eine geringe Laufzeit und einen niedrigen Speicherbedarf der Modellprüfung. In einer Bildberechnung werden ein BDD zur Repräsentation einer Menge von Transitionen und ein BDD für eine Menge von Zuständen kombiniert, um eine Menge von Nachfolger- oder Vorgängerzuständen zu berechnen. Oft ist auch die Größe von BDDs zur Repräsentation der Transitionsrelation von Systemen entscheidend für die erfolgreiche Anwendbarkeit der Modellprüfung. In der vorliegenden Arbeit werden neue Datenstrukturen zur Repräsentation der Transitionsrelation von asynchronen nebenläufigen Systemen bei der BDD-basierten symbolischen Modellprüfung vorgestellt. Zusätzlich werden neue Algorithmen zur Durchführung von Bildberechnungen präsentiert. Beides kann zu großen Reduktionen der Laufzeit und des Speicherbedarfs führen. Asynchrone nebenläufige Systeme besitzen häufig Symmetrien. Eine Technik zur Reduktion des Problems der Zustandsraumexplosion ist die Symmetriereduktion. In dieser Arbeit wird ebenfalls ein neuer effizienter Algorithmus zur Symmetriereduktion bei der symbolischen Modellprüfung mit BDDs aufgeführt

Structure Formation of Metallopolymer-Grafted Block Copolymers

Microphase separation drives the structure formation in block copolymers. Here, functional metallopolymer-grafted diblock copolymers consisting of polystyrene-<i>block</i>-polyisoprene (PS-<i>b</i>-PI) as polymer backbone featuring low molar mass poly­ferrocenyl­dimethyl­silane (PFS) and polyvinyl­ferrocene (PVFc) are synthesized via an iterative anionic grafting-to polymerization strategy. PS-<i>b</i>-PI block copolymers having about 30 mol % 1,2-polyisoprene moieties are subjected to platinum-catalyzed hydrosilylation reaction for the introduction of chlorosilane groups. The Si–Cl moieties are shown to efficiently react with the active metallopolymers yielding block-selective metallopolymer-grafted copolymers with 34 vol % PVFc and 43 vol % PFS as evidenced by ¹H NMR spectroscopy as well as size exclusion chromatography. The microphase separation of the functional metallopolymer-grafted block copolymers is evidenced via TEM measurements revealing fascinating morphologies. The structure formation of the PVFc-grafted block copolymers is studied in more detail by TEM, small-angle X-ray scattering, wide-angle X-ray scattering, and atomic force microscopy measurements evidencing a lamellar morphology featuring a spherical substructure for the PVFc segments inside the polyisoprene lamellae

Multimodal [GdO]+[ICG]− Nanoparticles for Optical, Photoacoustic, and Magnetic Resonance Imaging

Multimodal contrast agents with high biocompatibility and biodegradability, as well as low material complexity, are in great demand for clinical diagnostics at different scales of resolution and/or for translating preclinical diagnosis into intraoperative imaging. Multimodality, however, often results in multicomponent and multistructured materials with complexity becoming a severe restriction for synthesis, approval, and use in routine clinical practice. Here, we present sulfonate-based saline [GdO]+[ICG]− (ICG, indocyanine green) inorganic-organic hybrid nanoparticles (IOH-NPs with an inorganic [GdO]+ cation and an organic [ICG]− anion) as a novel, multimodality contrast agent for optical, photoacoustic, and magnetic resonance imaging (OI, PAI, MRI). [GdO]+[ICG]− IOH-NPs have a plain composition based on clinically used constituents and are prepared as an insoluble saline compound in water. The high [ICG]− content (81 wt %) ensures intense near-infrared emission (780-840 nm) and a strong photoacoustic signal. First, in vitro studies demonstrate longer detectability and greater emission intensity for [GdO]+[ICG]− IOH-NP suspensions than for ICG solutions, as well as a reduced toxicity compared to that of Gd-DTPA, a standard MRI contrast agent. Conceptual in vivo studies confirm the utility of the [GdO]+[ICG]− IOH-NPs for optical and magnetic resonance imaging with a T1 relaxivity better than that of Gd-DTPA. Taken together, [GdO]+[ICG]− represents a new compound and nanomaterial that can be highly interesting as a multimodal contrast agent

Targeting and Modulation of Liver Myeloid Immune Cells by Hard-Shell Microbubbles

Poly n-butylcyanoacrylate (PBCA)-based hard-shell microbubbles (MB) have manifold biomedical applications, including targeted drug delivery or contrast agents for ultrasound (US)-based liver imaging. MB and their fragments accumulate in phagocytes, especially in the liver, but it is unclear if MB affect the function of these immune cells. Herein, it is shown that human primary monocytes internalize different PBCA-MB by phagocytosis, which transiently inhibits monocyte migration in vertical chemotaxis assays and renders monocytes susceptible to cytotoxic effects of MB during US-guided destruction. Conversely, human macrophage viability and function, including cytokine release and polarization, remain unaffected after MB uptake. After intravenous injection in mice, MB predominantly accumulate in liver, especially in hepatic phagocytes (monocytes and Kupffer cells). Despite efficiently targeting myeloid immune cells in liver, MB or MB after US-elicited burst do not cause overt hepatotoxicity or inflammation. Furthermore, MB application with or without US-guided burst does not aggravate the course of experimental liver injury in mice or the inflammatory response to liver injury in vivo. In conclusion, PBCA-MB have immunomodulatory effects on primary human myeloid cells in vitro, but do not provoke hepatotoxicity, inflammation or altered response to liver injury in vivo, suggesting the safety of these MB for diagnostic and therapeutic purposes

Correlation between FMISO-PET based hypoxia in the primary tumour and in lymph node metastases in locally advanced HNSCC patients

Purpose: This secondary analysis of the prospective study on repeat [18F]fluoromisonidazole (FMISO)-PET in patients with locally advanced head and neck squamous cell carcinoma (HNSCC) assessed the correlation of hypoxia in the primary tumour and lymph node metastases (LN) prior to and during primary radiochemotherapy. Methods: This analysis included forty-five LN-positive HNSCC patients having undergone FMISO-PET/CTs at baseline, and at week 1, 2 and 5 of radiochemotherapy. The quantitative FMISO-PET/CT parameters maximum standardised uptake value (SUVmax, corrected for partial volume effect) and peak tumour-to-background ratio (TBRpeak) were estimated in the primary tumour as well as in index and large LN, respectively. Statistical analysis was performed using the Spearman correlation coefficient ρ. Results: In 15 patients with large LN (FDG-PET positive volume >5 ml), there was a significant correlation between the hypoxia measured in the primary tumour and the large LN at three out of four time-points using the TBRpeak (baseline: ρ = 0.57, p = 0.006; week 2: ρ = 0.64, p = 0.003 and week 5: ρ = 0.68, p = 0.001). For the entire cohort (N = 45) only assessed prior to the treatment, there was a statistically significant, though weak correlation between FMISO-SUVmax of the primary tumour and the index LN (ρ = 0.36, p = 0.015). Conclusions: We observed a significant correlation between FMISO-based hypoxia in the primary tumour and large lymph node(s) in advanced stage HNSCC patients. However, since most patients only had relatively small hypoxic lymph node metastases, a comprehensive assessment of the primary tumour and lymph node hypoxia is essential. Keywords: Hypoxia, FMISO, PET, Primary tumour, Lymph node metastases, Locally advanced HNSCC, Radiochemotherp

Multimodal [GdO]⁺[ICG]⁻ Nanoparticles for Optical, Photoacoustic, and Magnetic Resonance Imaging

Multimodal contrast agents with high biocompatibility and biodegradability, as well as low material complexity, are in great demand for clinical diagnostics at different scales of resolution and/or for translating preclinical diagnosis into intraoperative imaging. Multimodality, however, often results in multicomponent and multistructured materials with complexity becoming a severe restriction for synthesis, approval, and use in routine clinical practice. Here, we present sulfonate-based saline [GdO]⁺[ICG]⁻ (ICG, indocyanine green) inorganic-organic hybrid nanoparticles (IOH-NPs with an inorganic [GdO]⁺ cation and an organic [ICG]⁻ anion) as a novel, multimodality contrast agent for optical, photoacoustic, and magnetic resonance imaging (OI, PAI, MRI). [GdO]⁺[ICG]⁻ IOH-NPs have a plain composition based on clinically used constituents and are prepared as an insoluble saline compound in water. The high [ICG]⁻ content (81 wt %) ensures intense near-infrared emission (780–840 nm) and a strong photoacoustic signal. First, <i>in vitro</i> studies demonstrate longer detectability and greater emission intensity for [GdO]⁺[ICG]⁻ IOH-NP suspensions than for ICG solutions, as well as a reduced toxicity compared to that of Gd-DTPA, a standard MRI contrast agent. Conceptual <i>in vivo</i> studies confirm the utility of the [GdO]⁺[ICG]⁻ IOH-NPs for optical and magnetic resonance imaging with a <i>T</i>₁ relaxivity better than that of Gd-DTPA. Taken together, [GdO]⁺[ICG]⁻ represents a new compound and nanomaterial that can be highly interesting as a multimodal contrast agent