AmpTec GmbH

Hamburg, Germany

AmpTec GmbH

Hamburg, Germany
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Agency: European Commission | Branch: FP7 | Program: CP-FP-SICA | Phase: HEALTH.2011.2.3.3-2 | Award Amount: 8.37M | Year: 2012

WHO estimates that one of the main consequences of global warming will be an increased burden of vector-borne diseases. Among these, dengue appears to be particularly problematic, with tens of millions of cases of dengue fever estimated to occur annually, including up to 500,000 cases of the life-threatening dengue hemorrhagic fever/dengue shock syndrome. In recent years, the global burden of dengue disease has been rising dramatically and this prolific increase has been connected to societal changes such as population growth, urbanization and the transport of infected hosts and vectors. In addition, rising temperatures and global climate change may lead to the expansion of the range of major mosquito vectors, extension of the transmission season in areas with currently circulating dengue virus and increase in the mosquito spp. vectorial capacity. Active surveillance to detect in-coming dengue virus (DENV) in regions at the limits of DENV circulation are an important initial step in the prevention of dengue epidemics in Europe. Asymptomatic infections likely play a crucial role in the initial invasion process and DENV transmission and, although hitherto ignored, must be addressed. Using retrospective and prospective data from Asia, the main objectives of the program are (1) to identify key factors determining dengue transmission, outcome of infection and epidemics; (2) the development of novel diagnostic tools to detect asymptomatic infections. We will estimate the risk of DENV spreading to uninfected areas, especially in Southern Europe where susceptible vector exists. The major tools generated will be predictive models that enable specific interventions to reduce epidemic probability and diagnostic methods for surveillance. Inherent in this approach is the belief that improved surveillance and diagnosis of the asymptomatic dengue carriers will contribute to effective intervention, especially during early stages of pathogen invasion into a nave region.

Schroder W.,University of Tübingen | Bernhardt J.,University of Greifswald | Marincola G.,University of Tübingen | Marincola G.,University of Würzburg | And 5 more authors.
BMC Genomics | Year: 2014

Background: It has been shown previously that aminocoumarin antibiotics such as novobiocin lead to immediate downregulation of recA expression and thereby inhibit the SOS response, mutation frequency and recombination capacity in Staphylococcus aureus. Aminocoumarins function by inhibiting the ATPase activity of DNA gyrase subunit B with a severe impact on DNA supercoiling.Results: Here, we have analysed the global impact of the DNA relaxing agent novobiocin on gene expression in S. aureus. Using a novobiocin-resistant mutant, it became evident that the change in recA expression is due to gyrase inhibition. Microarray analysis and northern blot hybridisation revealed that the expression levels of a distinct set of genes were increased (e.g., recF-gyrB-gyrA, the rib operon and the ure operon) or decreased (e.g., arlRS, recA, lukA, hlgC and fnbA) by novobiocin. The two-component ArlRS system was previously found to decrease the level of supercoiling in S. aureus. Thus, downregulation of arlRS might partially compensate for the relaxing effect of novobiocin. Global analysis and gene mapping of supercoiling-sensitive genes did not provide any indication that they are clustered in the genome. Promoter fusion assays confirmed that the responsiveness of a given gene is intrinsic to the promoter region but independent of the chromosomal location.Conclusions: The results indicate that the molecular properties of a given promoter, rather than the chromosomal topology, dictate the responsiveness to changes in supercoiling in the pathogen Staphylococcus aureus. © 2014 Schröder et al.; licensee BioMed Central Ltd.

Bruder J.,Ludwig Maximilians University of Munich | Bruder J.,Max Planck Institute of Neurobiology | Siewert K.,Ludwig Maximilians University of Munich | Siewert K.,Max Planck Institute of Neurobiology | And 10 more authors.
Journal of Biological Chemistry | Year: 2012

In polymyositis and inclusion body myositis, muscle fibers are surrounded and invaded by CD8-positive cytotoxic T cells expressing the αβ-T cell receptor (αβ-TCR) for antigen. In a rare variant of myositis, muscle fibers are similarly attacked by CD8-negative T cells expressing the γδ-TCR (γδ-T cell-mediated myositis). We investigated the antigen specificity of a human γδ-TCR previously identified in an autoimmune tissue lesion of γδ-T cell-mediated myositis. We show that this Vγ1.3Vδ2-TCR, termed M88, recognizes various proteins from different species. Several of these proteins belong to the translational apparatus, including some bacterial and human aminoacyl-tRNA synthetases (AA-RS). Specifically, M88 recognizes histidyl-tRNA synthetase, an antigen known to be also targeted by autoantibodies called anti-Jo-1. The M88 target epitope is strictly conformational, independent of post-translational modification, and exposed on the surface of the respective antigenic protein. Extensive mutagenesis of the translation initiation factor-1 from Escherichia coli (EcIF1), which served as a paradigm antigen with known structure, showed that a short α-helical loop around amino acids 39 to 42 of EcIF1 is a major part of the M88 epitope. Mutagenesis of M88 showed that the complementarity determining regions 3 of both γδ-TCR chains contribute to antigen recognition. M88 is the only known example of a molecularly characterized γδ-TCR expressed by autoaggressive T cells in tissue. The observation that AA-RS are targeted by a γδ-T cell and by autoantibodies reveals an unexpected link between T cell and antibody responses in autoimmune myositis. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

Quabius E.S.,University of Kiel | Krupp G.,AmpTec GmbH
New Biotechnology | Year: 2015

Availability of high quality synthetic mRNAs (syn-mRNAs) has enabled progress in their applications. Important structural features and quality requirements are discussed. Developments in the application of mRNA-mediated manipulation of cells are presented (i) mRNA-directed expression of antigens in dendritic cells for vaccination projects in oncogenesis, infectious disease and allergy prevention; (ii) reprogramming of human fibroblasts to induced pluripotent stem cells with their subsequent differentiation to the desired cell type; (iii) applications in gene therapy. © 2014 Elsevier B.V.

Thompson N.,Ecole Polytechnique Federale de Lausanne | Gesina E.,Ecole Polytechnique Federale de Lausanne | Scheinert P.,AmpTec GmbH | Bucher P.,Ecole Polytechnique Federale de Lausanne | And 2 more authors.
Molecular and Cellular Biology | Year: 2012

Pancreas development is initiated by the specification and expansion of a small group of endodermal cells. Several transcription factors are crucial for progenitor maintenance and expansion, but their interactions and the downstream targets mediating their activity are poorly understood. Among those factors, PTF1a, a basic helix-loop-helix (bHLH) transcription factor which controls pancreas exocrine cell differentiation, maintenance, and functionality, is also needed for the early specification of pancreas progenitors. We used RNA profiling and chromatin immunoprecipitation (ChIP) sequencing to identify a set of targets in pancreas progenitors. We demonstrate that Mnx1, a gene that is absolutely required in pancreas progenitors, is a major direct target of PTF1a and is regulated by a distant enhancer element. Pdx1, Nkx6.1, and Onecut1 are also direct PTF1a targets whose expression is promoted by PTF1a. These proteins, most of which were previously shown to be necessary for pancreas bud maintenance or formation, form a transcription factor network that allows the maintenance of pancreas progenitors. In addition, we identify Bmp7, Nr5a2, RhoV, and P2rx1 as new targets of PTF1a in pancreas progenitors. © 2012, American Society for Microbiology.

Amptec Gmbh | Date: 2015-02-10

The present invention provides methods for the detection of RNA in a sample, comprising (a) mixing a specific amount of a rod-shaped virus-like particle with a sample, wherein the rod-shaped virus-like particle comprises a ribonucleic acid molecule and a viral coat, wherein: (i) the ribonucleic acid molecule comprises an origin-of-assembly sequence of a rod-shaped RNA virus and a heterologous sequence; and (ii) the viral coat comprises at least one type of coat protein of the rod-shaped RNA virus; (b) isolating RNA from the sample; and (c) detecting RNA comprising the heterologous sequence.

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