cDNA cloning and expression of a hamster α-thrombin receptor coupled to Ca2+ mobilization

AbstractThe serine protease α-thrombin (thrombin) potently stimulates G-protein-coupled signaling pathways and DNA synthesis in CCL39 hamster lung fibroblasts. To clone a thrombin receptor cDNA, selective amplification of mRNA sequences displaying homology to the transmembrane domains of G-protein-coupled receptor genes was performed by polymerase chain reaction. Using reverse transcribed poly(A)+ RNA from CCL39 cells and degenerate primers corresponding to conserved regions of several phospholipase C-coupled receptors, three novel putative receptor sequences were identified. One corresponds to an mRNA transcript of 3.4 kb in CCL39 cells and a relatively abundant cDNA. Microinjection of RNA transcribed in vitro from this cDNA in Xenopus oocytes leads to the expression of a functional thrombin receptor. The hamster thrombin receptor consists of 427 amino acid residues with 8 hydrophobic domains, including one at the extreme N-terminus that is likely to represent a signal peptide. A thrombin consensus cleavage site is present in the N-terminal extracellular region of the receptor sequence followed by a negatively charged cluster of residues present in a number of proteins that interact with the anion-binding exosite of thrombin

HIV-1 drug-resistance patterns among patients on failing treatment in a large number of European countries

Background: Information about patterns of HIV-1 drug resistance among treatment-exposed patients is crucial for the development of novel effective drugs. Currently no system exists that monitors patterns of resistance in patients failing therapy. Methods: The study included 1,988 HIV-1 sequences from patients experiencing therapy failure collected between 2000 and 2004 in 15 European countries. Genotypic resistance was interpreted using the ANRS algorithm. Phenotypic resistance was predicted using the Virco geno- to phenotype system. Results: 80.7% of the sequences included at least one drug-resistance mutation. Mutations were found for NRTIs (73.5%), NNRTIs (48.5%), and protease inhibitors (35.8%). Ninety percent of sequences with genotypic resistance harbored M184V, M41L, K103N, D67N, and/or T215Y. Among NRTIs, resistance was most frequently predicted for lamivudine. About half of all sequences had reduced susceptibility for NNRTIs. Resistance to most boosted protease inhibitors was found in &lt; 25%. No sequence had resistance to all currently available drugs. Conclusion: Levels of resistance among patients with therapy failure were high. The patterns of resistance reflect resistance to drugs available for a longer time. Fully suppressive regimens can be designed even for the most mutated HIV because boosted protease inhibitors have remained active against most circulating viruses and new drug classes have become available.</p

CITY AUTOMATED TRANSPORT SYSTEM (CATS): THE LEGACY OF AN INNOVATIVE EUROPEAN PROJECT

CATS is a collaborative European project promoting driverless vehicles that ended in December 2014. This contribution explains how the project evolved, including the handling of unexpected events and concentrating on lessons learned. The constructor and vehicle had to be changed for economic reasons in the middle of the project timeline. A second constructor went bankrupt, although access to his vehicles could be secured. For security and legal reasons, part of the final demonstration was relocated at short notice to the EPFL campus in Lausanne, Switzerland, where around 1600 people were transported during 16 days of vehicle operation. Reactions to the driverless vehicle concept were overwhelmingly positive. Implications for the acceptability of driverless vehicles in Europe and elsewhere are discussed

BioMoon: a concept for a mission to advance space life sciences and astrobiology on the Moon

As humans advance their presence in space and seek to improve the quality of life on Earth, a variety of science questions in support of these two objectives can be answered using the Moon. In this paper, we present a concept for an integrated mission focused on answering fundamental and applied biological questions on the Moon: BioMoon. The mission was designed to investigate the effects of the lunar radiation, gravity, and regolith on biological systems ranging from biomolecules to systems with complex trophic interactions, spanning a range of model organisms. Using common analytical systems and data processing, BioMoon represents a systems-level integrated life sciences mission. It would provide fundamental insights into biological responses to the lunar environment, as well as applied knowledge for In-Situ Resource Utilisation (ISRU), closed-loop life support system development, planetary protection and human health care. The mission was conceived to test biotechnology and sensor technology for lunar and terrestrial application and provide education and outreach opportunities. Although BioMoon was considered in the context of the European Space Agency’s Argonaut (European Large Logistics Lander) concept, the mission design provides a template for any integrated life sciences experimental suite on the Moon and other celestial bodies, implemented either robotically or by human explorers.CSC acknowledges support from the Science and Technology Facilities Council (Grants ST/V000586/1 and ST/Y001788/1).Discover Spac

Emissions Trading, CDM, JI, and More - The Climate Strategy of the EU

The objective of this paper is to assess the likely allocation effects of the current cli-mate protection strategy as it is laid out in the National Allocation Plans (NAPs) for the European Emissions Trading Scheme (ETS). The multi-regional, multi-sectoral CGE-model DART is used to simulate the effects of the current policies in the year 2012 when the Kyoto targets need to be met. Different scenarios are simulated in or-der to highlight the effects of the grandfathering of permits to energy-intensive instal-lations, the use of the project-based mechanisms (CDM and JI), and the restriction imposed by the supplementarity criterion

Aspects génétiques et biochimiques de la régulation de la transcription chez la bactérie E. coli et ses phages

SCOPUS: re.jinfo:eu-repo/semantics/publishe

Etude génétique et biochimique de la régulation de la transcription

Doctorat en Sciencesinfo:eu-repo/semantics/nonPublishe

Etude génétique et biochimique de la régulation de la transcription

Doctorat en Sciencesinfo:eu-repo/semantics/nonPublishe

Differential action of trypsin on the β′ subunit of DNA-dependent RNA polymerase from E. coli

SCOPUS: ar.jinfo:eu-repo/semantics/publishe