Development of osteogenic cell sheets for bone tissue engineering applications

The use of scaffolds in combination with osteogenic cells has been the gold standard in bone tissue engineering strategies. These strategies have, however, in many cases failed to produce the desired results due to issues such as the immunogenicity of the biomaterials used and cell necrosis at the bulk of the scaffold related to deficient oxygen and nutrients diffusion. Here, we originally propose the use of cell sheet (CS) engineering as a possible way to overcome some of these obstacles. Osteogenic CSs were fabricated by culturing rat bone marrow stromal cells in thermoresponsive culture dishes. The CSs were recovered from the dishes using a low-temperature treatment and then were implanted subcutaneously in nude mice. New bone formation was verified from day 7 post-transplantation using X-ray, microcomputed tomography, and histological analysis. The presence of a vascularized marrow was also verified in the newly formed bone after 6 weeks of transplantation. Further, osteocytes were found in this newly formed tissue, supporting the conclusion that mature bone was formed after ectopically transplanting osteogenic CSs. These results therefore confirm the great potentiality of CS engineering to be used in bone tissue engineering applicationsThis study was partially supported by Formation of Innovation Center for Fusion of Advanced Technologies in the Special Coordination Funds for Promoting Science and Technology "Cell Sheet Tissue Engineering Center (CSTEC)" and the Global COE program, Multidisciplinary Education and Research Center for Regenerative Medicine (MER-CREM), from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.Financial support to R. P. Pirraco by the Portuguese Foundation for Science and Technology (FCT) through the PhD grant SFRH/BD/44893/2008 is also acknowledged

Sazonalidade e qualidade do sêmen fresco e criopreservado de touros Pantaneiro e Nelore.

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Community profiling and gene expression of fungal assimilatory nitrate reductases in agricultural soil

Although fungi contribute significantly to the microbial biomass in terrestrial ecosystems, little is known about their contribution to biogeochemical nitrogen cycles. Agricultural soils usually contain comparably high amounts of inorganic nitrogen, mainly in the form of nitrate. Many studies focused on bacterial and archaeal turnover of nitrate by nitrification, denitrification and assimilation, whereas the fungal role remained largely neglected. To enable research on the fungal contribution to the biogeochemical nitrogen cycle tools for monitoring the presence and expression of fungal assimilatory nitrate reductase genes were developed. To the ∼100 currently available fungal full-length gene sequences, another 109 partial sequences were added by amplification from individual culture isolates, representing all major orders occurring in agricultural soils. The extended database led to the discovery of new horizontal gene transfer events within the fungal kingdom. The newly developed PCR primers were used to study gene pools and gene expression of fungal nitrate reductases in agricultural soils. The availability of the extended database allowed affiliation of many sequences to known species, genera or families. Energy supply by a carbon source seems to be the major regulator of nitrate reductase gene expression for fungi in agricultural soils, which is in good agreement with the high energy demand of complete reduction of nitrate to ammonium

Roles of silver nanoclusters in surface-enhanced Raman spectroscopy

The cause for the huge enhancement factors of surface-enhanced Raman spectroscopy (SERS) by the addition of small silver nanoclusters is theoretically investigated by focusing on the difference between resonance Raman activity and surface plasmon effects. First, the resonance and off-resonance Raman spectra are calculated using the incident light wavenumbers of the low-lying charge transfer excitations for the surface (S) and vertex (V) complexes of the pyridine molecule attaching to three small silver nanoclusters: Ag-5, Ag-10, and Ag-20. As a result, it is found that the incident radiation dramatically increases the resonance Raman activities with the enhancement factors up to 10(12). This indicates that the resonance Raman effects are dominant in the enhancement factors of SERS, at least when to use small silver clusters. It is also found that the resonance Raman spectra significantly depend on the adsorption sites given in S or V complexes, and on the inclusion or exclusion of the long-range correction for density functional theory, irrespective of the size of the silver clusters. The electromagnetic field enhancement effects called surface plasmon effects are also examined for the Ag-20 cluster to confirm this conclusion. Consequently, the enhancement in the electric field is roughly evaluated as less than one for the static polarizability of this small cluster. It is, therefore, concluded that the resonance Raman activity effect is dominant in the huge SERS enhancement factors for, at least, small silver nanoclusters

Introduction process of n-methylglucamino groups for binding antimony (III) to a polymer brush

Polymer brushes containing epoxy groups were grafted onto a porous hollow-fiber membrane by radiation-induced graft polymerization of glycidyl methacrylate. N-Methylglucamino (NMG) groups for binding antimony(III) were introduced to the epoxy groups of the polymer brush using a range of reaction times. To determine the process of the introduction of NMG groups to the polymer brush, the water permeability through the membrane, the swelling ratio of the membrane, and the antimony(III) adsorptivity of the membrane were determined for different NMG conversion values of 0 - 76%. The water permeability and the swelling ratio indicated that the NMG groups reacted with the epoxy groups of the polymer brush on the outside of the base of the polymer first, and then with those inside the base of the polymer matrix. The molar ratio between the NMG groups and the antimony(III) adsorbed on the polymer brush was always 0.5, demonstrating that the NMG groups were introduced from the tail end of the polymer brush to the fixed end due to diffusion and reaction of the NMG groups along the polymer brush