Using the Surveyor nuclease assay and flow cytometric analysis, we confirmed ZFN-induced cleavage of the target sequence and the disappearance of EGFP fluorescence expression in ZFN-treated cells. In addition, sequence analysis revealed that ZFN-induced mutations such as base substitution, deletion, or insertion were generated in the ZFN cleavage site of EGFP-expression negative cells that were cloned from ZFN-treated cells, thereby showing it was possible

to disrupt (i.e., knock out) the function of the EGFP gene in porcine somatic cells. To our knowledge, this study provides the first evidence that the ZFN-KO system can be applied to pigs. These findings may open a new avenue to the creation of gene KO pigs using ZFN-treated cells and somatic cell nuclear transfer. (C) 2010 Elsevier Inc. All rights reserved.”
“Limited number of hematopoietic stem cells in umbilical cord blood (UCB) presents a problem when using UCB for stem PF-04929113 ic50 cell transplantation. Improving their homing capacity could reduce the need for high initial cell numbers find more during transplantation procedures. Although it is evident that protein kinase B (PKB/c-Akt) plays an important role in regulation of migration of various cell types, a role for PKB in regulation of migration and homing of human hematopoietic stem and progenitor cells remains to be determined. PKB activity was found to be required for induction

of adhesion compound screening assay to bone marrow-derived stromal cells and detrimental for migration of UCB-derived CD34(+) hematopoietic progenitors. In addition, PKB activity was found to positively regulate integrin expression. CD34(+) hematopoietic progenitors, and their capacity to form colonies in vitro, were not affected by transient inhibition of PKB. Finally, transplantation of beta 2-microglobulin(-/-) nonobese diabetic/severe combined immunodeficient mice with CD34(+) cells ectopically expressing constitutively active PKB resulted

in reduced migration to the bone marrow, whereas inhibition of PKB activity resulted in an induction in bone marrow homing and engraftment. These results indicate that transient inhibition of PKB activity may provide a means for ex vivo stem cell manipulation to improve bone marrow transplantation regimes. (Blood. 2010; 116(13): 2373-2384)”
“Background: A spectrum of myopathic manifestations has been recognized as associated with lipid lowering drug therapy (LLT), but their effect on quality of life and physical functioning is uncertain. We conducted a prospective cohort study in which physical functioning was the dependent variable, in patients with and without exposure to LLT.\n\nMethods: Consecutive patients attending a risk reduction clinic were invited to participate in a questionnaire study which included demographic data, muscular symptoms, the SF-36 Physical Function Score (PF), and the modified Health Assessment Questionnaire (mHAQ). Laboratory and co-morbidity data was recorded.