In particular, we and others have recently shown that exosomes secreted by human tumor cells of various origins are able to induce apoptosis in activated T cells, through the expression of death ligands, inhibit NK functions and promote the generation of myeloid-derived suppressor cells from normal monocytes. These data, together with the reproducible evidence that exosomes of likely tumor origin can be abundantly found in plasma and neoplastic effusions of cancer patients support a role of tumor exosomes in molding host microenvironment to allow tumor growth and progression. The selection of these recombination sites defines modules that consist of a core sequence flanked by two 4 nt sequences. These modules can be amplified by PCR with primers designed to add flanking BsaI sites on each side of the modules , and cloned in an intermediate cloning vector and sequenced. A restriction-ligation performed on a mix containing all intermediate plasmids , the recipient acceptor vector, BsaI enzyme and ligase is expected to allow assembly of a library of shuffled genes. This is because each module is compatible and can be ligated only to a module belonging to the next consecutive set of homologous modules, or to the acceptor vector for the first and last modules, and because each module from a set of homologous modules can be ligated with equal probability to each module of a contiguous set. In addition, because of the restriction-ligation, only the desired assembled products are expected to accumulate since all other ligation products will contain BsaI sites and should therefore be immediately redigested with BsaI.The estimated annual cost of sarcopenia-related health issues to the US health care system is more than 18 billion dollars annually. Increased expression of VEGFR-1 by HIF-1a has been well established, but the Gefitinib consequence of activation of this signalling pathway and its role in stimulating VEGFR-2 expression is not as well characterized. VEGFR-1 is expressed in two forms. These include the full-length membrane bound receptor, capable of transducing a cellular signal, and a soluble receptor, capable of sequestering ligand or dimerizing with the full-length receptor to prevent signal transduction. It is known that the extracellular portion and the soluble fraction of VEGFR-1 have a 10-fold higher affinity for the VEGF ligand with limited or no detectable autophosphorylation activity. Furthermore, VEGFR-2 is phosphorylated approximately 10-fold more efficiently upon ligand binding. Seetharam et al. demonstrated a high binding affinity of VEGFR-1 to VEGF without generating a mitogenic response in transfected NIH3T3 fibroblasts that overexpressed VEGFR-1. We have shown here that inserts from nine separate plasmids can be easily and efficiently assembled and cloned in an acceptor vector in one step and one tube. The efficiency of this protocol comes from the fact that the only stable product issued from the restriction-ligation are the desired product ; these products are formed continuously with each cycle.