Generation of mouse-induced pluripotent stem cells by transient expression of a single nonviral polycistronic vector

Induced pluripotent stem (iPS) cells have generated keen interestdue to their potential use in regenerative medicine. They havebeen obtained from various cell types of both mice and humans byexogenous delivery of different combinations of Oct4, Sox2, Klf4,c-Myc, Nanog, and Lin28. The delivery of these transcription factorshas mostly entailed the use of integrating viral vectors (retrovirusesor lentiviruses), carrying the risk of both insertional mutagenesisand oncogenesis due to misexpression of these exogenousfactors. Therefore, obtaining iPS cells that do not carry integratedtransgene sequences is an important prerequisite for their eventualtherapeutic use. Here we report the generation of iPS cell linesfrom mouse embryonic fibroblasts with no evidence of integrationof the reprogramming vector in their genome, achieved by nucleofectionof a polycistronic construct coexpressing Oct4, Sox2, Klf4,and c-Myc

A protocol describing the genetic correction of somatic human cells and subsequent generation of IPS cell

The generation of patient-specific induced pluripotent stem cells (iPSCPSCPSCs) offers unprecedented opportunities for modeling and treating human disease. In combination with gene therapy, the iPSCPSCPSC technology can be used to generate disease-free progenitor cells of potential interest for autologous cell therapy. We explain a protocol for the reproducible generation of genetically corrected iPSCPSCPSCs starting from the skin biopsies of Fanconi anemia patients using retroviral transduction with OCT4, SOX2 and KLF4. Before reprogramming, the fibroblasts and/or keratinocytes of the patients are genetically corrected with lentiviruses expressing FANCA. The same approach may be used for other diseases susceptible to gene therapy correction. Genetically corrected, characterized lines of patient-specific iPSCPSCPSCs can be obtained in 4–5 months.

Generation of induced pluripotent stem cells from human cord blood cells with only two factors:OCT4 and SOX2

Induced pluripotent stem cells (iPSC ) provide an invaluable resource for regenerative medicine as they allow the generationof patient-specific progenitors with potential value for cell therapy. However, in many instances, an off-the-shelf approach isdesirable, such as for cell therapy of acute conditions or when the patient’s somatic cells are altered as a consequence of a chronicdisease or aging. Cord blood (CB) stem cells appear ideally suited for this purpose as they are young cells expected to carryminimal somatic mutations and possess the immunological immaturity of newborn cells; additionally, several hundred thousandimmunotyped CB units are readily available through a worldwide network of CB banks. Here we present a detailed protocol for thederivation of CB stem cells and how they can be reprogrammed to pluripotency by retroviral transduction with only two factors(OCT 4 and SO X2) in 2 weeks and without the need for additional chemical compounds.

Generation of cardiomyocytes from new human embrionic stem cell lines derived from poor-quanlity blastocysts

Human embryonic stem (hES) cells represent a potential source for cell replacement therapy of many degenerative diseases. Most frequently, hES cell lines are derived from surplus embryos from assisted reproduction cycles, independent of their quality or morphology. Here, we show that hES cell lines can be obtained from poor-quality blastocysts with the same efficiency as that obtained from good- or intermediate-quality blastocysts. Furthermore, we show that the self-renewal, pluripotency, and differentiation ability of hES cell lines derived from either source are comparable. Finally, we present a simple and reproducible embryoid body-based protocol for the differentiation of hES cells into functional cardiomyocytes. The five new hES cell lines derived here should widen the spectrum of available resources for investigating the biology of hES cells and advancing toward efficient strategies of regenerative medicine.

Semantic web adoption: online tools for web evaluation and metadata extraction

This work briefly analyses the difficulties to adopt the Semantic Web, and in particular proposes systems to know the present level of migration to the different technologies that make up the Semantic Web. It focuses on the presentation and description of two tools, DigiDocSpider and DigiDocMetaEdit, designed with the aim of verifYing, evaluating, and promoting its implementation.

Women of the beat generation : la traducción de una obra poética coral

Comentario a la traducción por encargo del libro WOTBG, centrado en las muestras de obra poética de las distintas autoras y la necesidad de conservar sus similitudes y sus diferéncias.

The illusion of control, the structures of prizes and the demand for football pools in Spain

Using data for all the fixtures for the seasons from 1972-73 to 2002-03, we estimate a dynamic model of demand for football pools in Spain paying attention to whether their main economic explanatory variable is the effective price of a ticket or the jackpot. Additionally, we evaluate the importance of the composition of the list of games in terms of whether First Division matches are included or not. Results show that the jackpot model is preferred to the effective price model, having important implications in terms of how the structure of the game should be changed in order to increase demand.

Notes for a new guide to Keynes (I): Wages, aggregate demand, and employment

I revisit the General Theory's discussion of the role of wages inemployment determination through the lens of the New Keynesianmodel. The analysis points to the key role played by the monetarypolicy rule in shaping the link between wages and employment, andin determining the welfare impact of enhanced wage flexibility. I showthat the latter is not always welfare improving.

Discrete devaluations and multiple equilibria in a first generation model of currency crises

The first generation models of currency crises have often been criticized because they predict that, in the absence of very large triggering shocks, currency attacks should be predictable and lead to small devaluations. This paper shows that these features of first generation models are not robust to the inclusion of private information. In particular, this paper analyzes a generalization of the Krugman-Flood-Garber (KFG) model, which relaxes the assumption that all consumers are perfectly informed about the level of fundamentals. In this environment, the KFG equilibrium of zero devaluation is only one of many possible equilibria. In all the other equilibria, the lack of perfect information delays the attack on the currency past the point at which the shadow exchange rate equals the peg, giving rise to unpredictable and discrete devaluations.

On money as a medium of exchange when goods vary by supply and demand

Models of the exchange process based on search theory can be usedto analyze the features of objects that make them more or less likely toemerge as ``money'' in equilibrium. These models illustrate the trade--offbetween endogenous acceptability (an equilibrium property) and intrinsiccharacteristics of goods, such as storability, recognizability, etc. Inthis paper, we look at how the relative supply and demand for various goodsaffect their likelihood of becoming money. Intuitively, goods in highdemand and/or low supply are more likely to appear as commodity money,subject to the qualification that which object ends up circulating as amedium of exchange depends at least partly on convention. Welfare propertiesare discussed.