Quantification of lentiviral vector copy numbers in individual hematopoietic colony-forming cells shows vector dose-dependent effects on the frequency and level of transduction

Lentiviral vectors are effective tools for gene transfer and integrate variable numbers of proviral DNA copies in variable proportions of cells. The levels of transduction of a cellular population may therefore depend upon experimental parameters affecting the frequency and/or the distribution of vector integration events in this population. Such analysis would require measuring vector copy numbers (VCN) in individual cells. To evaluate the transduction of hematopoietic progenitor cells at the single-cell level, we measured VCN in individual colony-forming cell (CFC) units, using an adapted quantitative PCR (Q-PCR) method. The feasibility, reproducibility and sensitivity of this approach were tested with characterized cell lines carrying known numbers of vector integration. The method was validated by correlating data in CFC with gene expression or with calculated values, and was found to slightly underestimate VCN. In spite of this, such Q-PCR on CFC was useful to compare transduction levels with different infection protocols and different vectors. Increasing the vector concentration and re-iterating the infection were two different strategies that improved transduction by increasing the frequency of transduced progenitor cells. Repeated infection also augmented the number of integrated copies and the magnitude of this effect seemed to depend on the vector preparation. Thus, the distribution of VCN in hematopoietic colonies may depend upon experimental conditions including features of vectors. This should be carefully evaluated in the context of ex vivo hematopoietic gene therapy studies

A comprehensive review of techniques for biofunctionalization of titanium

A number of surface modification techniques using immobilization of biofunctional molecules of Titanium (Ti) for dental implants as well as surface properties of Ti and Ti alloys have been developed. The method using passive surface oxide film on titanium takes advantage of the fact that the surface film on Ti consists mainly of amorphous or low-crystalline and non-stoichiometric TiO2. In another method, the reconstruction of passive films, calcium phosphate naturally forms on Ti and its alloys, which is characteristic of Ti. A third method uses the surface active hydroxyl group. The oxide surface immediately reacts with water molecules and hydroxyl groups are formed. The hydroxyl groups dissociate in aqueous solutions and show acidic and basic properties. Several additional methods are also possible, including surface modification techniques, immobilization of poly(ethylene glycol), and immobilization of biomolecules such as bone morphogenetic protein, peptide, collagen, hydrogel, and gelatin

Potential for a Circular Autopoietic Economy on Canavese Territory

The aim of this gigamap is to describe and summarize a student work carried out during the semester course Open Systems Design at Politecnico Torino. The map is one of the outputs from an analysis of the Canavese, Piedmont region and subsequently an in-depth study of the relationships and “flows” with certain “currencies” between some selected economic and public actors. Canavese is a historical-geographical area located in the province of Turin, Italy; it’s full of great natural and geological spots, with a strong enogastronomic identity, historical re-enactments, a well-known manufacture sector like the ceramic of Castellamonte, as well as the leading high-tech branch like Arduino. Through the approach deriving from systemic design, it was possible to identify some strategies for the creation of new networks aimed at transforming the economy and production processes from linear to circular approach. Five types of “circular flows” were assessed and “designed into” the current linear system, thus proposing an improved economy based on circularity: flows of matter (energy/CO2, water, materials), economic flows, and social flows. The Gigamap will be presented to stakeholders in a public hearing and supposedly be used to illustrate and incubate a circular economy that is more resilient and more regenerative. Reading the map The reading should start at the top left, where the abstract is located, next, there is an outline that shows the connections between the five topics investigated. Subsequently, there is a timeline with some fundamental events in the history of the Canavese. In the center there is the circularity map with the main cities, focusing on new opportunities of circular flows between the 15 analysed economic actors. At the bottom, some general data and on the right a brief description of the role and territorial qualities of the companies. Lastly, there is a summary of the emerging properties

Endosseous implant treated with phosphonic acid-group containing compounds

Endosseous implant to be applied to a human or animal bone, said implant having a smooth or rough surface texture and wherein said surface has been treated with at least one organic compound carrying at least one phosphonic acid group or a salt thereof and method for producing said implant