Polysaccharide films built by simultaneous or alternate spray: a rapid way to engineer biomaterial surfaces.

We investigated polysaccharide films obtained by simultaneous and alternate spraying of a chitosan (CHI) solution as polycation and hyaluronic acid (HA), alginate (ALG), and chondroitin sulfate (CS) solutions as polyanions. For simultaneous spraying, the film thickness increases linearly with the cumulative spraying time and passes through a maximum for polyanion/CHI molar charge ratios lying between 0.6 and 1.2. The size of polyanion/CHI complexes formed in solution was compared with the simultaneously sprayed film growth rate as a function of the polyanion/CHI molar charge ratio. A good correlation was found. This suggests the importance of polyanion/polycation complexation in the simultaneous spraying process. Depending on the system, the film topography is either liquid-like or granular. Film biocompatibility was evaluated using human gingival fibroblasts. A small or no difference is observed in cell viability and adhesion between the two deposition processes. The CHI/HA system appears to be the best for cell adhesion inducing the clustering of CD44, a cell surface HA receptor, at the membrane of cells. Simultaneous or alternate spraying of CHI/HA appears thus to be a convenient and fast procedure for biomaterial surface modifications.journal articleresearch support, non-u.s. gov't2012 Jun 052012 05 23importe

Comparative Study of Hematopoietic Differentiation between Human Embryonic Stem Cell Lines

Directed differentiation of human embryonic stem cells (hESCs) into any desired cell type has been hailed as a therapeutic promise to cure many human diseases. However, substantial roadblocks still exist for in vitro differentiation of hESCs into distinct cell types, including T lymphocytes. Here we examined the hematopoietic differentiation potential of six different hESC lines. We compare their ability to develop into CD34+ or CD34+CD45+ hematopoietic precursor populations under several differentiation conditions. Comparison of lymphoid potential of hESC derived- and fetal tissue derived-hematopoietic precursors was also made. We found diverse hematopoietic potential between hESC lines depending on the culture or passage conditions. In contrast to fetal-derived hematopoietic precursors, none of the CD34+ precursors differentiated from hESCs were able to develop further into T cells. These data underscore the difficulties in the current strategy of hESC forward differentiation and highlight distinct differences between CD34+ hematopoietic precursors generated in vitro versus in vivo

Molecular-ion-beam-assisted deposition of stable SiOF films

Recently we reported on low refractive index fluorinated silica (SiOF) films obtained by a Molecular Ion Beam Assisted Deposition (MIBAD) process, using a fluorocarbon precursor in an End Hall ion source during evaporation of silica grains. These films were unstable due to moisture absorption and a subsequent chemical reaction, leading to an irreversible change in composition and increase of refractive index.In this paper we investigate an inert gas enhanced MIBAD process, involving the simultaneous use of a fluorocarbon precursor and an inert gas in the ion gun. Ion beam composition was analyzed by energy selective ion mass spectrometry. Addition of inert gas to the fluorocarbon precursor not only increases the total ion current, but also enhances the dissociation of the fluorocarbon precursor into smaller fragments.The films were characterized by ellipsometry, IR transmission spectroscopy and in situ and ex situ stress measurements.Time evolution of film properties was followed for several weeks. For a given fluorocarbon precursor flow rate, refractive index measured directly after deposition increases with increasing inert gas flow rate, while refractive index change with time is reduced. Fluorine content in the films is maximum for low inert gas flow rates, but decreases for higher flow rates to reach approximately the same level as for pure fluorocarbon ion beam. The observed refractive indexstabilization for high inert gas flow rates can be explained by a change in film structure rather than film composition, i.e.by reduced film porosity due to high inert gas ion bombardment.With carefully chosen deposition conditions, stable films with low refractive index can be obtained

The “Dispensable” Portion of RAG2 Is Necessary for Efficient V-to-DJ Rearrangement during B and T Cell Development

AbstractPrevious in vitro studies defined the minimal regions of RAG1 and RAG2 essential for V(D)J recombination. In order to characterize the role of the C-terminal “dispensable” portion of RAG2, we generated core-RAG2 knock-in mice. We found that the core-RAG2-containing recombinase complex is selectively defective in catalyzing V-to-DJ rearrangement at the IgH and TCRβ loci, resulting in partial developmental blocks in B and T lymphopoiesis. Analysis of recombination intermediates showed defects at the cleavage phase of the reaction. We also observed a reduction in overall recombinase activity in core-RAG2-expressing thymocytes, leading us to suggest that the interaction of a defective recombinase with RSS sequences unique to VH and Vβ gene segments may underlie the specific V-to-DJ rearrangement defect in core-RAG2 mice

Os compostos fenólicos são mais influenciados pelos fatores clima e cultivar quando comparados aos perfis metabólicos de aminoácidos e RMN 1H.

o HPLC é um método analítico com excelente sensibilidade que permite analisar diferentes metabólitos a partir de extratos vegetais ou animais. Ele é usado com sucesso para a determinação de compostos fenólicos e aminoácidos de uvas e vinhos. Estes dois grupos de compostos são importantes para os padrões de qualidade no meio enológico. A concentração dos compostos relacionados pode variar fortemente em relação às condições do meio