Analysis of knockout/knockin mice that express a mutant FasL lacking the intracellular domain

Fas ligand (FasL; CD178; CD95L) is a type II transmembrane protein belonging to the tumour necrosis factor family; its binding to the Fas receptor (CD95; APO-1) triggers apoptosis in the receptor-bearing cell. Signalling through this pathway plays a pivotal role during the immune response and in immune system homeostasis. Similar to other TNF family members, the intracellular domain has been reported to transmit signals to the inside of the FasL-bearing cell (reverse signalling). Recently, we identified the proteases ADAM10 and SPPL2a as molecules important for the processing of FasL. Protease cleavage releases the intracellular domain, which then is able to translocate to the nucleus and to repress reporter gene activity. To study the physiological importance of FasL reverse signalling in vivo, we established knockout/knockin mice with a FasL deletion mutant that lacks the intracellular portion (FasLDeltaIntra). Co-culture experiments confirmed that the truncated FasL protein is still capable of inducing apoptosis in Fas-sensitive cells. Preliminary immune histochemistry data suggest that, in contrast to published data, the absence of the intracellular FasL domain does not alter the intracellular FasL localization in activated T cells. We are currently investigating signalling and proliferative capacities of T cells derived from homozygous FasLDeltaIntra mice to validate a co-stimulatory role of FasL reverse signalling

Law and Finance in Transition Economies

This paper offers the first comprehensive analysis of legal change in the protection of shareholder and creditor rights in transition economies and its impact on the propensity of firms to raise external finance. Following La Porta et al. (1998), the paper constructs an expanded set of legal indices to capture a range of potential conflicts between different stakeholders of the firm. It supplements the analysis of the law on the books with an analysis of the effectiveness of legal institutions. Our main finding is that the effectiveness of legal institutions has a much stronger impact on external finance than does the law on the books, despite legal change that has substantially improved shareholder and creditor rights. This finding supports the proposition that legal transplants and extensive legal reforms are not sufficient for the evolution of effective legal and market institutions.shareholder and creditor rights, legal effectiveness, external finance, transition

Efficient Explicit Time Stepping of High Order Discontinuous Galerkin Schemes for Waves

This work presents algorithms for the efficient implementation of discontinuous Galerkin methods with explicit time stepping for acoustic wave propagation on unstructured meshes of quadrilaterals or hexahedra. A crucial step towards efficiency is to evaluate operators in a matrix-free way with sum-factorization kernels. The method allows for general curved geometries and variable coefficients. Temporal discretization is carried out by low-storage explicit Runge-Kutta schemes and the arbitrary derivative (ADER) method. For ADER, we propose a flexible basis change approach that combines cheap face integrals with cell evaluation using collocated nodes and quadrature points. Additionally, a degree reduction for the optimized cell evaluation is presented to decrease the computational cost when evaluating higher order spatial derivatives as required in ADER time stepping. We analyze and compare the performance of state-of-the-art Runge-Kutta schemes and ADER time stepping with the proposed optimizations. ADER involves fewer operations and additionally reaches higher throughput by higher arithmetic intensities and hence decreases the required computational time significantly. Comparison of Runge-Kutta and ADER at their respective CFL stability limit renders ADER especially beneficial for higher orders when the Butcher barrier implies an overproportional amount of stages. Moreover, vector updates in explicit Runge--Kutta schemes are shown to take a substantial amount of the computational time due to their memory intensity

Symptoms in Swiss adolescents in relation to exposure from fixed site transmitters : a prospective cohort study

There is public concern regarding potential health effects of radiofrequency electromagnetic fields (RF-EMF) emitted by fixed site transmitters. We therefore investigated whether self-reported general well-being in adolescents is affected by RF-EMF exposure from mobile phone base stations (downlink) and broadcast transmitters (TV and radio).; In a prospective cohort study, 439 study participants aged 12-17 years, completed questionnaires about their self-reported well-being and possible confounding factors at baseline and one year later. Exposure from fixed site transmitters at home and school was calculated by using a geospatial propagation model. Data were analysed using a mixed-logistic cross-sectional model of a combined dataset consisting of baseline and follow-up data and a longitudinal approach where we investigated whether exposure at baseline (cohort analysis) or changes in exposure between baseline and follow-up (change analysis) were related to a new onset of a symptom between baseline and follow-up. All analyses were adjusted for relevant confounders.; Mean exposure (median; 75(th)) for broadcast transmitters, downlink and total exposure at baseline were 1.9 μW/m(2) (1.0 μW/m(2); 2.8 μW/m(2)), 14.4 μW/m(2) (3.8 μW/m(2); 11.0 μW/m(2)) and 16.3 μW/m(2) (5.8 μW/m(2); 13.4 μW/m(2)), respectively. In cross-sectional analyses no associations were observed between any symptom and RF-EMF exposure from fixed site transmitters. In the cohort and change analyses only a few significant associations were observed including an increased OR for tiredness (2.94, 95%CI: 1.43 to 6.05) for participants in the top 25(th) percentile of total RF-EMF exposure from fixed site transmitters at baseline, in comparison to participants exposed below the median and a decreased OR for exhaustibility (0.50, 95%CI: 0.27 to 0.93) for participants with an exposure increase between baseline and follow-up.; In this cohort study, using a geospatial propagation model, RF-EMF exposure from fixed site transmitters was not consistently associated with self-reported symptoms in Swiss adolescents. The few observed associations have to be interpreted with caution and might represent chance findings

Farm restructuring and agricultural recovery in Kazakhstan's grain region: An update

Against the rising global concern of how to achieve sustainable output expansion in food, we document the main outcomes of post-Soviet agricultural recovery and restructuring in the Kazakhstan grain region. Together with an expansion of cropland area and increasing capital input, real agricultural value added has almost doubled within the recent decade. Privatisation legislation has allowed private ownership of land. However, access to state land and capital continues to be strongly regulated, and private lenders even turn away from agriculture. There are now three dominant groups of agricultural producers in the region: large agricultural enterprises and smaller individual farms mostly engaged in grain, and tiny household economies focusing on vegetable and live-stock. While agricultural enterprises have been growing more persistently than individual farms in recent years, average land productivity of both farm types is practically identical and wheat yields are even higher in individual farms. Both vertically and horizontally integrated agroholdings have emerged among the agricultural enterprises and have brought outside investment and management to the region. With stable employment in agriculture, nominal consumption spending of rural households has tripled over the last decade and has risen much faster than the costs of living. While North Kazakhstan looks much like a success story, constrained factor markets are likely to dampen further growth. The Kazakh government should improve the legal conditions for a functioning land rental market, avoid driving commercial lenders out of the market, and make sure that future access to qualified labour in agriculture is warranted. --agricultural productivity,agricultural transition,farm organisation,Kazakhstan

Vaccinia virus modulates the host cell cycle to promote infection

Vaccinia Virus (VACV) is well-known as the vaccine used for the eradication of smallpox. It serves as the model orthopox virus and has gained further clinical significance as an oncolytic virus. As a member of the poxvirus family, VACV is a double-stranded DNA virus that replicates exclusively in the cytoplasm of infected cells. Early research suggested that VACV alters the host cell cycle and inhibits cellular DNA synthesis. Later, VACV was described to modulate key cell cycle regulators during late timepoints of infection. However, the relevance of this cell cycle subversion to VACV replication and how it is achieved remains undefined. In this PhD project, I combined state of the art techniques with classical assays to determine the (viral) effector proteins, their mode of action, and the contribution of the host cell cycle to productive VACV infection. Using recombinant VACV strains, RNAi, biochemistry, and super-resolution microscopy, I demonstrate that VACV early gene expression inhibits cell proliferation after viral entry. Concurrently, the cellular CDK inhibitor p21 is upregulated, while the tumour suppressor p53 is targeted for degradation by the viral kinase B1 and/or its paralog pseudokinase B12. The second wave of viral gene expression shifts the cell cycle from G1 to S/G2/M, while still inhibiting cell proliferation. Additionally, the viral kinase F10 was shown to be necessary and sufficient to cause degradation of p21, and for activation of the cellular DNA damage response (DDR), a process known to be essential for viral DNA replication. By probing these cellular pathways with a small molecule inhibitor library I defined their requirement for the viral life cycle. Screening for defects in viral late gene expression, I found inhibition of Aurora Kinases, selected CDKs, ATR and Chk1/2 interferes with infection. Collectively, I demonstrate that VACV modulates cell cycle checkpoints and identify the viral kinases B1 and F10 as potential temporal controllers of the host cell cycle that serve to promote productive viral replication

Development of a New Generation of Metal-Based Anticancer Drugs

Kinases are a large family of transferase enzymes, of which at least 538 different members are present in the human body.[11] By catalysing the transfer of the γ-phosphate group of ATP onto a specific side chain of their substrate, they mediate most cellular signal transductions in the body, thereby regulating various critical cellular activities. These include proliferation, survival, apoptosis, metabolism, transcription, differentiation, and a wide array of other cellular processes.[9,10] It is no surprise that the kinome has been extensively investigated as a family of potential drug targets[12] and the development of kinase inhibitors has therefore become an essential part of biological and medical research. Small molecule kinase inhibitors have been studied to treat various human diseases including cardiovascular diseases,[23] autoimmune disorders such as rheumatoid arthritis,[24] neurodegenerative conditions like Alzheimer’s disease, diabetes or liver disorders.[25] Originally based on unselective natural product inhibitors like staurosporine (51), research has led to almost 30 FDA approved inhibitors by the middle of 2016.[26] Most of them were developed to treat oncological conditions, which are still a major focus of kinase inhibitor research today. Over the last decade the MEGGERS group has established highly potent and selective kinase inhibitors using inert metal centres as unique structural templates, thus mimicking and enhancing the globular shape of the non-selective inhibitor staurosporine (51)