Comparative transcriptomics enlarges the toolkit of known developmental genes in mollusks

Data used for the phylogenetic analysis of Hox and ParaHox genes, including the respective GenBank accession numbers. (DOC 31 kb

Constitutive and inducible co-expression systems for non-viral osteoinductive gene therapy

Tissue regenerative gene therapy requires expression strategies that deliver therapeutic effective amounts of transgenes. As physiological expression patterns are more complex than high-level expression of a singular therapeutic gene, we aimed at constitutive or inducible co-expression of 2 transgenes simultaneously. Co-expression of human bone morphogenetic protein 2 and 7 (BMP2/7) from constitutively expressing and doxycycline inducible plasmids was evaluated in vitro in C2C12 cells with osteocalcin reporter gene assays and standard assays for osteogenic differentiation. The constitutive systems were additionally tested in an in vivo pilot for ectopic bone formation after repeated naked DNA injection to murine muscle tissue. Inductor controlled differentiation was demonstrated in vitro for inducible co-expression. Both co-expression systems, inducible and constitutive, achieved significantly better osteogenic differentiation than single factor expression. The potency of the constitutive co-expression systems was dependent on relative expression cassette topology. In vivo, ectopic bone formation was demonstrated in 6/13 animals (46 % bone formation efficacy) at days 14 and 28 in hind limb muscles as proven by in vivo μCT and histological evaluation. In vitro findings demonstrated that the devised single vector BMP2/7 co-expression strategy mediates superior osteoinduction, can be applied in an inductor controlled fashion and that its efficiency is dependent on expression cassette topology. In vivo results indicate that co-expression of BMP2/7 applied by non-viral naked DNA gene transfer effectively mediates bone formation without the application of biomaterials, cells or recombinant growth factors, offering a promising alternative to current treatment strategies with potential for clinical translation in the future

Measurement Of Quasiparticle Transport In Aluminum Films Using Tungsten Transition-Edge Sensors

We report new experimental studies to understand the physics of phonon sensors which utilize quasiparticle diffusion in thin aluminum films into tungsten transition-edge-sensors (TESs) operated at 35 mK. We show that basic TES physics and a simple physical model of the overlap region between the W and Al films in our devices enables us to accurately reproduce the experimentally observed pulse shapes from x-rays absorbed in the Al films. We further estimate quasiparticle loss in Al films using a simple diffusion equation approach.Comment: 5 pages, 6 figures, PRA

Expression, purification and in vitro biological activity from human recombinant BMP-2 produced by a novel approach

Bone morphogenetic proteins have promoted great biomedical interest due to their ability in inducing new bone formation when used as powerful osteoinductive components of several late-stage bone grafting products. Recombinant human bone morphogenetic protein-2 (rhBMP-2) is obtained from mammalian cell expressing systems in low amounts or from bacteria inclusion bodies after timeconsuming refolding methods. Thus, there is a need to establish novel approaches for producing rhBMP-2 in high yields by simple and cheap procedures.Portuguese Foundation for Science and Technolog y, FCT (PhD Grant to PC Bessa , SFRH/BD/17049/2004). This work was also partially supported by the European STREP HIPPOCRATES (NMP3 - CT - 2003 - 505758) and carried out under the scope of European NoE EXPERTISSUES (NMP3 - CT - 2004 - 500283).info:eu-repo/semantics/publishedVersio

Expression, purification and in vitro biological activity from human recombinant BMP-2 produced by a novel approach

Bone tissue engineering has been an increasing field of research during the last years. The ideal approach for a regenerative application would consist in the use of cells from the patient, scaffolding materials and differentiation growth factors. Bone morphogenetic protein-2 (BMP-2) is one such growth factors with a strong ability to induce new bone and cartilage formation and has been used as a powerful osteoinductive component of several late-stage tissue engineering products for bone grafting. In this work, we aimed at obtaining high yields of human recombinant BMP-2 in a stable, pure and biologically active form by use of a new bacteria expression system that circumvents the disadvantages of conventional recombinant protein preparation methods and to perform a study of the stability conditions and functionality of these peptides in vitro in human mesenchymal stem cells and C2C12 murine cell line.Portuguese Foundation for Science and Technology, FCT (PhD Grant to PC Bessa, to PC Bessa, SFRH/BD/17049/2004 SFRH/BD/17049/2004 ). This work was ). This work was also partially supported by the European STREP HIPPOCRATES (NMP3 also partially supported by the European STREP HIPPOCRATES (NMP3--CTCT--2003 2003--505758) and carried out under the scope of 505758) and carried out under the scope of European NoE EXPERTISSUES (NMP3 European NoE EXPERTISSUES (NMP3--CTCT- -2004 2004 --500283). 500283info:eu-repo/semantics/publishedVersio

A novel system for producing human recombinant BMP-2 and study of the growth factor stabilizing conditions

Bone tissue engineering has been an increasing field of research during the last years. The ideal approach for a regenerative application would consist in the use of cells from the patient, scaffolding materials and differentiation growth factors. Bone morphogenetic protein-2 (BMP-2) is one such growth factors with a strong ability to induce new bone and cartilage formation and has been used as a powerful osteoinductive component of several late-stage tissue engineering products for bone grafting. In this work, we aimed at obtaining high yields of human recombinant BMP-2 in a stable, pure and biologically active form by use of a new bacteria expression system that circumvents the disadvantages of conventional recombinant protein preparation methods and to perform a study of the stability conditions and the functionality of these peptides in vitro in human mesenchymal stem cells and C2C12 murine cell line.Portuguese Foundation for Science and Technology (PhD Grant to PC Bessa, SFRH/BD/17049/2004). This work was also partially supported by the European STREP HIPPOCRATES (NMP3-CT-2003-505758) and carried out under the scope of European NoE EXPERTISSUES (NMP3-CT-2004- 500283).info:eu-repo/semantics/publishedVersio