Polyethylenimine-mediated gene delivery into human bone marrow mesenchymal stem cells from patients

Transplantation of mesenchymal stem cells (MSCs) derived from adult bone marrow has been proposed as a potential therapeutic approach for post-infarction left ventricular (LV) dysfunction. However, age-related functional decline of stem cells has restricted their clinical benefits after transplantation into the infarcted myocardium. The limitations imposed on patient cells could be addressed by genetic modification of stem cells. This study was designed to improve our understanding of genetic modification of human bone marrow derived mesenchymal stem cells (hMSCs) by polyethylenimine (PEI, branched with Mw 25 kD), one of non-viral vectors that show promise in stem cell genetic modification, in the context of cardiac regeneration for patients. We optimized the PEI-mediated reporter gene transfection into hMSCs, evaluated whether transfection efficiency is associated with gender or age of the cell donors, analysed the influence of cell cycle on transfection and investigated the transfer of therapeutic vascular endothelial growth factor gene (VEGF). hMSCs were isolated from patients with cardiovascular disease aged from 41 to 85 years. Optimization of gene delivery to hMSCs was carried out based on the particle size of the PEI/DNA complexes, N/P ratio of complexes, DNA dosage and cell viability. The highest efficiency with the cell viability near 60% was achieved at N/P ratio 2 and 6.0 μg DNA/cm 2. The average transfection efficiency for all tested samples, middle-age group (<65 years), old-age group (>65 years), female group and male group was 4.32%, 3.85%, 4.52%, 4.14% and 4.38%, respectively. The transfection efficiency did not show any correlation either with the age or the gender of the donors. Statistically, there were two subpopulations in the donors; and transfection efficiency in each subpopulation was linearly related to the cell percentage in S phase. No significant phenotypic differences were observed between these two subpopulations. Furthermore, PEI-mediated therapeutic gene VEGF transfer could significantly enhance the expression level.DFG/SFB/Transregio 37BMBF/0313191German Helmholtz AssociationDFG/0402710Ministry of Education/0312138 AMinistry of Economy (Mecklenburg-West Pommerania)/V220-630-08-TFMVF/S-035Marie Curie International Research Staff Exchange Scheme (IRSES, FP7-PEOPLE-2009-IRSES)Reference and Translation Center for Cardiac Stem Cell Therapy (RTC

Организация работ по локализации и ликвидации аварийного разлива нефти и нефтепродуктов на водной поверхности

Объектом исследования являются методы локализации и ликвидации разливов нефти на водной поверхности. Цель работы – провести анализ существующих методов и тактических мероприятий по локализации и ликвидации разливов нефти на водной поверхности. В процессе исследования проводился анализ имеющихся данных, на основе которых были выделение наиболее эффективные методы локализации и ликвидации разливов нефти на водной поверхности. Проведен расчет объема вытекшей нефти и количества сорбента, требуемого для ликвидации разлива. В результате исследования было выявлено, что наиболее эффективным способом локализации разливов на водной поверхности являются боны с различными механизмами их установки, а наиболее эффективными методами ликвидации – использование скиммеров и сорбентов.Объектом исследования являются методы локализации и ликвидации разливов нефти на водной поверхности. Цель работы – провести анализ существующих методов и тактических мероприятий по локализации и ликвидации разливов нефти на водной поверхности. В процессе исследования проводился анализ имеющихся данных, на основе которых были выделение наиболее эффективные методы локализации и ликвидации разливов нефти на водной поверхности. Проведен расчет объема вытекшей нефти и количества сорбента, требуемого для ликвидации разлива. В результате исследования было выявлено, что наиболее эффективным способом локализации разливов на водной поверхности являются боны с различными механизмами их установки, а наиболее эффективными методами ликвидации – использование скиммеров и сорбентов

Identification of novel biomarkers in chronic immune thrombocytopenia (ITP) by microarray-based serum protein profiling

The pathological mechanisms underlying the development of immune thrombocytopenia (ITP) are unclear and its diagnosis remains a process of exclusion. Currently, there are no known specific biomarkers for ITP to support differential diagnosis and treatment decisions. Profiling of serum proteins may be valuable for identifying such biomarkers. Sera from 46 patients with primary chronic ITP and 34 healthy blood donors were analysed using a microarray of 755 antibodies. We identified 161 differentially expressed proteins. In addition to oncoproteins and tumour-suppressor proteins, including apoptosis regulator BCL2, breast cancer type 1 susceptibility protein (BRCA1), Fanconi anaemia complementation group C (FANCC) and vascular endothelial growth factor A (VEGFA), we detected six anti-nuclear autoantibodies in a subset of ITP patients: anti-PCNA, anti-SmD, anti-Ro/SSA60, anti-Ro/SSA52, anti-La/SSB and anti-RNPC antibodies. This finding may provide a rational explanation for the association of ITP with malignancies and other autoimmune diseases. While RUNX1mRNA expression in the peripheral blood mononuclear cells (PBMC) of patients was significantly downregulated, an accumulation of RUNX1 protein was observed in the platelets of ITP patients. This may indicate dysregulation of RUNX1 expression in PBMC and megakaryocytes and may lead to an imbalanced immune response and impaired thrombopoiesis. In conclusion, we provide novel insights into the pathogenic mechanisms of ITP that warrant further exploration

Coronary artery surgery: cardiotomy suction or cell salvage?

Coronary artery bypass grafting (CABG) today results in what may be regarded as acceptable levels of blood loss with many institutions avoiding allogeneic red cell transfusion in over 60% of their patients. The majority of cardiac surgeons employ cardiotomy suction to preserve autologous blood during on-pump coronary artery bypass surgery; however the use of cardiotomy suction is associated with a more pronounced systemic inflammatory response and a resulting coagulopathy as well as exacerbating the microembolic load. This leads to a tendency to increased blood loss, transfusion requirement and organ dysfunction. Conversely, the avoidance of cardiotomy suction in coronary artery bypass surgery is not associated with an increased transfusion requirement. There is therefore no indication for the routine use of cardiotomy suction in on-pump coronary artery surgery

Does the administration of the dose of perfluorocarbon when starting partial liquid ventilation affect haemodynamics and cerebral blood flow?

Background and aims: The disaster at the Nuclear Power Plant in Chernobyl in April 1986 resulted in the exposure of a large number of people to ionizing radiation that varied substantially, creating a new situation for epidemiology. We aim to demonstrate lasting genetic or reproductive detrimental health effects after Chernobyl in moderately and highly affected European countries.   Methods: Time trend analyses and spatial-temporal analyses based on logistic regression.    Results: Long-term dose dependent impacts of radioactive fallout after Chernobyl on stillbirths, birth defects, and the human sex ratio at birth have been found. For example, from nearly all published data concerning Down’s syndrome, long term increases after Chernobyl may be seen. Significant ecological relative risks for stillbirths and birth defects are in the range of 1.005 to 1.020 per kBq/m2 137Cs. A relative risk coefficient of 1.010 per kBq/m2 137Cs translates to a preliminary relative risk coefficient of 1.60 per mSv/a. German district by district data imply a sex odds ratio of 1.015 per mSv/a. Furthermore, there are striking jumps in the sex ratio in 1987 in several central and eastern European countries.   Conclusion: The effects of ionizing radiation at doses below 10 mSv are little understood. Recent epidemiologic evidence suggests that there is harm at doses below 1 mSv or that the established dose concept is invalid

Endothelial NOS is required for SDF-1alpha/CXCR4-mediated peripheral endothelial adhesion of c-kit+ bone marrow stem cells

In the era of intravascular approaches for regenerative cell therapy, the underlying mechanisms of stem cell migration to non-marrow tissue have not been clarified. We hypothesized that next to a local inflammatory response implying adhesion molecule expression, endothelial nitric oxide synthase (eNOS)-dependent signaling is required for stromal- cell-derived factor-1 alpha (SDF-1alpha)-induced adhesion of c-kit+ cells to the vascular endothelium. SDF-1alpha/tumor necrosis factor-alpha (TNF-alpha)-induced c-kit+-cell shape change and migration capacity was studied in vitro using immunohistochemistry and Boyden chamber assays. In vivo interaction of c-kit+ cells from bone marrow with the endothelium in response to SDF-1alpha/TNF-alpha stimulation was visualized in the cremaster muscle microcirculation of wild-type (WT) and eNOS (-/-) mice using intravital fluorescence microscopy. In addition, NOS activity was inhibited with N-nitro-L-arginine-methylester-hydrochloride in WT mice. To reveal c-kit+-specific adhesion behavior, endogenous leukocytes (EL) and c-kit+ cells from peripheral blood served as control. Moreover, intercellular adhesion molecule-1 (ICAM-1) and CXCR4 were blocked systemically to determine their role in inflammation-related c-kit+-cell adhesion. In vitro, SDF-1alpha enhanced c-kit+-cell migration. In vivo, SDF-1alpha alone triggered endothelial rolling-not firm adherence-of c-kit+ cells in WT mice. While TNF-alpha alone had little effect on adhesion of c-kit+ cells, it induced maximum endothelial EL adherence. However, after combined treatment with SDF-1alpha+TNF-alpha, endothelial adhesion of c-kit+ cells increased independent of their origin, while EL adhesion was not further incremented. Systemic treatment with anti-ICAM-1 and anti-CXCR4-monoclonal antibody completely abolished endothelial c-kit+-cell adhesion. In N-nitro-L-arginine-methylester-hydrochloride-treated WT mice as well as in eNOS (-/-) mice, firm endothelial adhesion of c-kit+ cells was entirely abrogated, while EL adhesion was significantly increased. The chemokine SDF-1alpha mediates firm adhesion c-kit+ cells only in the presence of TNF-alpha stimulation via an ICAM-1- and CXCR4-dependent mechanism. The presence of eNOS appears to be a crucial and specific factor for firm c-kit+-cell adhesion to the vascular endothelium

Gene delivery to the heart by magnetic nanobeads

10.1016/j.jmmm.2006.11.201Journal of Magnetism and Magnetic Materials3111 SPEC. ISS.336-341JMMM