Survival of patients with poor prognosis or relapsed haematopoietic malignancies can be markedly improved by allogeneic haematopoietic stem cell transplantation (HSCT). HSCT reconstitutes the immune and haematopoietic systems after myeloablative conditioning and inhibits the recurrence of the malignancy by a graft-versus-leukaemia (GVL) response mediated by donor T cells. However, significant post-transplant complications such as graft-versus-host disease (GVHD) continue to plague the event-free survival of this curative procedure. GVHD is facilitated by donor T cells that recognise histocompatibility antigens on host antigen presenting cells (APC), such as dendritic cells (DC). Current treatment options for GVHD are focused on these T cells. However, these treatments result in an increased incidence of infection, graft rejection and relapse. A novel means of immunosuppression in GVHD is the use of multi-potent, mesenchymal stromal cells (MSC). MSC are non-immunogenic cells that actively suppress T cell function in vitro, and can resolve steroid-refractory GVHD in the clinic. Despite their use in the clinic, there is a paucity of pre-clinical data. Our aim was to investigate the in vivo efficacy of MSC to control GVHD while maintaining the beneficial GVL effect, and to begin to understand the mechanism by which MSC exert their immunosuppressive effects. We isolated and characterised MSC from murine bone/bone marrow and demonstrated that they suppressed T cell proliferation in vitro, even at low ratios of 1 MSC per 100 T cells. This was true of both donor-derived MSC, and MSC derived from unrelated donors (third party). Importantly, we observed that MSC significantly reduced T cell production of the pro-inflammatory cytokines TNFα and IFNγ in culture supernatants and that IFNγ plays a key role in the ability of MSC to suppress T cell proliferation. In vivo, we examined the effects of donor-derived MSC on GVHD severity and onset in two myeloablative murine models of HSCT. A major histocompatibility complex (MHC)-mismatched donor-recipient pair combination was used as a proof–of-principle model [UBI-GFP/BL6 (H-2b)àBALB/c (H-2d)], and an MHC-matched, minor histocompatibility antigen (miHA) mismatched donor-recipient pair combination was used to mimic MHC-matched sibling transplantation [UBI-GFP/BL6 (H-2b)àBALB.B (H-2b)]. We examined a number of variables related to MSC infusion including timing, dose and route of injection. We found that early post transplant infusion of MSC by the intraperitoneal injection was most effective at delaying death from GVHD, compared to pre-transplant infusion or intravenous injection. Furthermore, we found that the dose of MSC was critical, as infusion of too few MSC was ineffective and infusion of too many MSC exacerbated the development of GVHD. Taken together, these results suggest that timing, dose and route of injection are all important factors to be considered to ensure successful therapeutic outcome. To investigate the in vivo mechanism of action, we conducted timed sacrifice experiments in the MHC-mismatched model to determine if MSC altered cytokine secretion and cellular effectors, such as DC, known to play a key role in GVHD. Despite the fact that MSC given post-HSCT enter an environment full of activated DC and IFNγ levels, by day 3 and 6 post infusion, these activated DC and IFNγ levels are decreased compared to controls or mice infused with MSC pre-transplant (p<0.05). This confirmed our in vitro data that IFNγ played an important role in MSC-mediated immunosuppression. In addition, when we removed a major source of IFNγ production in vivo by administering the T cell depleting antibody KT3 to mice with or without MSC, we found that although T cell depletion prolonged survival, MSC were unable to further enhance this effect. This was also true when MSC were used in combination with the conventional immunosuppressant cyclosporine. Finally, we examined whether the infusion of MSC would compromise the GVL effect. We found that whilst MSC could delay the onset of GVHD, in our model they did not alter the anti-tumour effects of the donor T cells. Overall, we have shown that MSC can delay but not prevent death from GVHD when administered at an appropriate time and dose and that IFNγ is required for MSC-mediated immunosuppression in our model. These data suggest that patients undergoing HSCT should be monitored for IFNγ, and administered MSC when high levels are reached. Whilst MSC may be a promising therapy for patients with severe GVHD, we highlight that further investigation is warranted before MSC are accepted for widespread use in the clinic. The risks and benefits for transplant recipients should be carefully considered before utilising MSC to treat or prevent GVHD.

Poly (glycerol-sebacate) (PGS) is a synthetic bioelastomer with a covalently crosslinked, three-dimensional network of random coils with hydroxyl groups attached to its backbone. This biodegradable polymer is biocompatible (in vitro and in vivo), tough, elastic, inexpensive, and flexible, and finds potential applications in tissue engineering and regenerative medicine. Due to the slow rate of step-growth polymerisation, the synthesis of PGS prepolymer requires 24-48 h. A batch and a continuous process, if developed, could address the inherent deficiencies (eg. long residence time, venting) associated with the large-scale synthesis of such bioelastomers. However, in order to assess whether this particular system may be adapted to continuous processes, such as reactive extrusion, studies on kinetics of controlled condensation reactions are of vital importance. FT-Raman spectroscopy was used to study the kinetics of the step-growth reactions between glycerol (G) and sebacic acid (SA) at three molar ratios (G:SA= 0.6,0.8,1.0) and three temperatures (120, 130, 140 ˚C). The rate curves followed first-order kinetics with respect to sebacic acid concentration in the kinetics regime. An increase in the molar ratio (G : SA) of the reactants decreased the average functionality of the system and the crosslinking density, resulting in the lowering of the activation energy and pre-exponential factor. The average functionality of the system had a profound effect on the crosslinking density, mechanical properties, and the reaction kinetics of the system. Three different PGS oligomers and films (PGS 0.6, PGS 0.8, PGS 1.0) were thoroughly characterised using Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), wide angle X-ray scattering (WAXS), differential scanning calorimetry (DSC), and contact angle measurements. FTIR spectra of PGS oligomers confirmed the formation of ester bonds (1740 cm -1). Quantification of various functional groups in PGS films using XPS was in agreement with the theoretical values of the proposed structure. WAXS results indicated that PGS system with a higher average functionality possesses a higher degree of crystallinity. Crystallisation exotherms and melting endotherms of PGS systems revealed that the average functionality influences the density of crosslinking, degree of crystallinity, and the network structure of bioelastomers. Contact angle studies confirmed that an increase in the average functionality of PGS system increases hydrophilicity, and the surface treatment through aminolysis further increases the hydrophilicity of the films. Batch studies were performed on a Brabender Plasticorder®. The samples collected over a reaction period of 5 h were characterised using Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The number-average molecular weight (Mn) and the weight-average molecular weight (Mw) of the oligoesters were determined using matrix-assisted laser desroption/ionization time-of-flight spectroscopy (MALDI-TOF) and compared with the corresponding values from the benchtop synthesis. It was found that due to higher shear-mixing and better orientation of functional groups, the degree of polymerisation at any stage of the reaction was higher in the Brabender than in the benchtop process. The gel-point of the reaction was determined from the crossover point of storage and loss moduli, and the reaction rate constant was calculated using the torque vs time data of the rheometer. The kinetics rate constant and the extent of the reaction in the Brabender were found to be higher than the corresponding values obtained from the conventional benchtop process by a factor of 2. PGS was found to be thermo-mouldable and adaptable to high-shear mixing, and hence is a better candidate for making thermoplastic elastomers using reactive extrusion. The challenges and possibilities in scaling up a batch process to a continuous process were investigated. The use of a wiped film reactor or a disk reactor along with reactive extrusion and batch-mixing (as a post-extrusion operation) is a commercially viable method to synthesise PGS oligomers. Such a continuous process will boost the production of bioelastomers for tissue engineering application by addressing the constraints in step-growth polymerisation. Finally, the effect of PGS substrate stiffness and surface treatment (aminolysis, hydrolysis, layer-by-layer deposition) on the morphology and lineage of mesenchymal stem cells – which have a capacity to differentiate themselves into cartilage, adipose, tendon, and muscle tissues – was analysed using fluorescence microscopy and DNA and protein assays. Stiffness of the PGS surface and the method of treatment influenced the cell attachment and spreading on different surfaces. However, cells did not differentiate into definite phenotypes at the end of 14 d time-point, indicating that higher time-points are needed to be considered to study the effect of matrix stiffness and surface treatment on cell attachment and phenotype differentiation.

Generating samples from multivariate distributions efficiently is an important task in Monte Carlo integration and many other stochastic simulation problems. Markov chain Monte Carlo has been shown to be very efficient compared to "conventional methods", especially when many dimensions are involved. In this article we propose a Hit-and-Run sampler in combination with the Ratio-of-Uniforms method. We show that it is well suited for an algorithm to generate points from quite arbitrary distributions, which include all log-concave distributions. The algorithm works automatically in the sense that only the mode (or an approximation of it) and an oracle is required, i.e., a subroutine that returns the value of the density function at any point x. We show that the number of evaluations of the density increases slowly with dimension. (author's abstract) / Series: Preprint Series / Department of Applied Statistics and Data Processing

We use inequalities to design short universal algorithms that can be used to generate random variates from large classes of univariate continuous or discrete distributions (including all log-concave distributions). The expected time is uniformly bounded over all these distributions. The algorithms can be implemented in a few lines of high level language code. In opposition to other black-box algorithms hardly any setup step is required and thus it is superior in the changing parameter case. (author's abstract) / Series: Preprint Series / Department of Applied Statistics and Data Processing

In the last years multilevel preconditioners like BPX became more and more popular for solving second-order elliptic finite element discretizations by iterative methods. P. Oswald has adapted these methods for discretizations of the fourth order biharmonic problem by rectangular conforming Bogner-Fox-Schmidt elements and nonconforming Adini elements and has derived optimal estimates for the condition numbers of the preconditioned linear systems. In this paper we generalize the results from Oswald to the construction of BPX and Multilevel Diagonal Scaling (MDS-BPX) preconditioners for the elasticity problem of thin smooth shells of arbitrary forms where we use Koiter's equations of equilibrium for an homogeneous and isotropic thin shell, clamped on a part of its boundary and loaded by a resultant on its middle surface. We use the two discretizations mentioned above and the preconditioned conjugate gradient method as iterative method. The parallelization concept is based on a non-overlapping domain decomposition data structure. We describe the implementations of the multilevel preconditioners. Finally, we show numerical results for some classes of shells like plates, cylinders, and hyperboloids.

In this paper we describe a version of transformed density rejection that requires less uniform random numbers. Random variates below the squeeze are generated by inversion. For the expensive part between squeeze and density an algorithm that uses a coverering with triangles is introduced. (author's abstract) / Series: Preprint Series / Department of Applied Statistics and Data Processing

We use inequalities to design short universal algorithms that can be used to generate random variates from large classes of univariate continuous or discrete distributions (including all log-concave distributions). The expected time is uniformly bounded over all these distributions for a particular generator. The algorithms can be implemented in a few lines of high level language code. (author's abstract) / Series: Preprint Series / Department of Applied Statistics and Data Processing

Let G be a connected graph with n vertices and let x=(x1, ..., xn) be a real vector. A positive (negative) sign graph of the vector x is a maximal connected subgraph of G on vertices xi>0 (xi<0). For an eigenvalue of a generalized Laplacian of a tree: We characterize the maximal number of sign graphs of an eigenvector. We give an O(n2) time algorithm to find an eigenvector with maximum number of sign graphs and we show that finding an eigenvector with minimum number of sign graphs is an NP-complete problem. (author's abstract) / Series: Preprint Series / Department of Applied Statistics and Data Processing

Der Elektrik/Elektronik- und der IT-Anteil steigt derzeit in den aktuellen Premium -Fahrzeugen stetig an. Durch den Verbau von immer mehr (hochvernetzten) Steuergeräten im Fahrzeug wird versucht, dem Wunsch der Kunden nach mehr Funktionalität, Sicherheit, etc. gerecht zu werden. Aufgrund der Komplexität und der großen Entwicklungssprünge sind jedoch die neuen Steuergeräte meistens nicht mehr kompatibel mit den Vorgänger-Steuergeräten. Hier ist ein großes Einsparpotenzial vorhanden und dies ist auch der Ansatzpunkt des Forschungsthemas ”CompA“ (Compatibility Analysis of Electronic Control Units), das in diesem Dokument beschrieben wird. Im Rahmen dieses Forschungsthemas wird eine Methode definiert, mit der zwei Steuergeräte auf Rückwärtskompatibilität untersucht werden können. Der Ansatz baut auf drei Schwerpunkten auf: • Definition eines Spezifikationsansatzes zur hinreichenden Beschreibung von Steuergeräten auf Basis eines XML-Schemas. Es werden hierbei sowohl die statischen als auch die dynamischen Eigenschaften abgebildet. Dieser neuartige Spezifikationsansatz bildet die Basis für den nächsten Schwerpunkt. • Definition einer Methode zur Analyse von Rückwärtskompatibilität von Steuergeräten auf Basis eines XML-Schemas. Die Rückwärtskompatibilität zweier Steuergeräte bzw. Systeme wird auf Basis der zugehörigen XML-Dokumente analysiert. Kern der Vergleichsmethode ist hierbei ein effizientes Mapping der XML-Dokumente und ein Experten-Regelwerk. • Definition einer Methode zur Kompatibilitätsanalyse von Message Sequence Charts (MSC). MSCs werden eingesetzt, um dynamisches Verhalten an der Schnittstelle von Steuergeräten zu beschreiben. In diesem Dokument wird ein Ansatz definiert, mit dem MSCs zueinander auf Rückwärtskompatibilität geprüft werden können. Der Vergleich erfolgt auf Basis deterministischer Automaten. Des Weiteren wird im vorliegenden Dokument ein Konzept für eine graphische Benutzeroberfläche (GUI) vorgestellt, die zur Spezifikation von Steuergeräten geeignet ist und sich adaptiv unterschiedlichen Schemata anpasst. Alle vorgestellten Konzepte wurden in einem Software-Tool implementiert und die Gültigkeit an mehreren Beispielen validiert.

Perfect slice sampling is a method to turn Markov Chain Monte Carlo (MCMC) samplers into exact generators for independent random variates. The originally proposed method is rather slow and thus several improvements have been suggested. However, two of them are erroneous. In this article we give a short introduction to perfect slice sampling, point out incorrect methods, and give a new improved version of the original algorithm. (author's abstract) / Series: Preprint Series / Department of Applied Statistics and Data Processing