Contacts with Diasporas and Diaspora Organisations as a Key to a Successful Migrant Integration Policy in the EU

We analyse European Commission and European Parliament documents - directives, communications, conclusions, recommendations - and best practices for EU member states' international projects focusing on migrant integration in both the EU states and the countries of origin. Special emphasis is placed on the role of diasporas and the efforts taken by the EU to involve them in the integration process. We stress the need for a new supranational EU immigration and integration policy, in view of new migration trends and the so-called migrant crisis. The study shows that the EU integration policy is directed towards both migrants and host countries and the countries of origin. Since the early 2010s, the involvement of various diaspora organisations in the implementation of the EU migrant integration policy has significantly increased. An analysis of completed projects shows that diasporas have a potential of becoming a key actor in the EU integration policy. We suggest expanding the list of the objectives of the national integration policy of the EU countries. This may be achieved by boosting efforts to reduce the gap in the socio-economic development of the host countries and the countries of origin, particularly, by promoting multilateral cooperation with diaspora organisations

Mitochondrial Electron Transport Chain in Heavy Metal-Induced Neurotoxicity: Effects of Cadmium, Mercury, and Copper

To clarify the role of mitochondrial electron transport chain (mtETC) in heavy-metal-induced neurotoxicity, we studied action of Cd2+, Hg2+, and Cu2+ on cell viability, intracellular reactive oxygen species formation, respiratory function, and mitochondrial membrane potential of rat cell line PC12. As found, the metals produced, although in a different way, dose- and time-dependent changes of all these parameters. Importantly, Cd2+ beginning from 10 [mu]M and already at short incubation time (3 h) significantly inhibited the FCCP-uncoupled cell respiration; besides, practically the complete inhibition of the respiration was reached after 3 h incubation with 50 [mu]M Hg2+ or 500 [mu]M Cd2+, whereas even after 48 h exposure with 500 [mu]M Cu2+, only a 50% inhibition of the respiration occurred. Against the Cd2+-induced cell injury, not only different antioxidants and mitochondrial permeability transition pore inhibitors were protective but also such mtETC effectors as FCCP and stigmatellin (complex III inhibitor). However, all mtETC effectors used did not protect against the Hg2+- or Cu2+-induced cell damage. Notably, stigmatellin was shown to be one of the strongest protectors against the Cd2+-induced cell damage, producing a 15–20% increase in the cell viability. The mechanisms of the mtETC involvement in the heavy-metal-induced mitochondrial membrane permeabilization and cell death are discussed

Mitochondrial mechanisms by which gasotransmitters (HS, NO and CO) protect cardiovascular system against hypoxia

Over past few years, there has been a dramatic increase in studying physiological mechanisms of the activity of various signaling low-molecular molecules that directly or indirectly initiate adaptive changes in the cardiovascular system cells (CVSC) to hypoxia. These molecules include biologically active endogenous gases or gasotransmitters (HS, NO and CO) that influence on many cellular processes, including mitochondrial biogenesis, oxidative phosphorylation, K/Ca exchange, contractility of cardiomyocytes (CM) and vascular smooth muscle cells (VSMC) under conditions of oxygen deficiency. The present review focuses on the mechanistic role of the gasotransmitters (NO, HS, CO) in cardioprotection. The structural components of these mechanisms involve mitochondrial enzyme complexes and redox signal proteins, K and Ca channels, and mitochondrial permeability transition pore (MPTP) that have been considered as the final molecular targets of mechanisms underlying antioxidant and mild mitochondrial uncoupling effects, preconditioning, vasodilatation and adaptation to hypoxia. In this article, we have reviewed recent findings on the gasotransmitters and proposed a unifying model of mitochondrial mechanisms of cardioprotection

Atrial fibrillation is associated with increased susceptibility of mitochondria to permeability transition pore opening

Background/significance: Although atrial fibrillation (AF) is known to result in progressive electrical, contractile and structural remodeling of the atria with gradual cell loss and replacement fibrosis, the molecular basis for the progressive structural alterations is not fully elucidated. Since mitochondria play an essential role in regulation of cell death through the opening of mitochondrial permeability transition pore (mPTP), we hypothesized that the susceptibility of mitochondria from patients with AF to PTP opening is increased contributing to the substrate that promote progression of AF to long-lasting forms. Purpose: The aim of the study was to characterize the sensitivity of mitochondria to Ca2+-induced mPTP opening in human atria from patients with and without AF. Methods: Freshly removed left atrial appendage tissue from patients undergoing cardiac bypass surgery with (AF) and without history of AF (nAF) was used for mitochondrial isolation. The sensitivity of mitochondria towards Ca2+-induced mPTP opening was assessed by exposing the isolated mitochondria to sequential additions of 10 μM of Ca2+ and monitoring simultaneously abrupt mitochondrial Ca2+ release (Fluo-5N fluorescence), mitochondrial depolarization (Safranin O fluorescence), and swelling of mitochondrial matrix (decrease in light scattering). Differences in the expression of proteins participating in mPTP formation, including voltage-dependent anion channel (VDAC), adenine nucleotide translocase (ANT), and phosphate carrier protein (PCP) were determined by Western blotting in isolated mitochondria. The protein expression level was expressed in arbitrary units normalized to mitochondrial protein. Results: The sensitivity of mitochondria to mPTP opening was increased in patients with AF compared to nAF (41±12 vs. 66±8 μM Ca2+). Cyclosporine A, an inhibitor of mPTP, increased tolerance of the mitochondria to Ca2+ loading (74±15 vs. 93±9) and reduced mPTP opening in both group of patients. Increased sensitivity towards mPTP opening in patients with AF was associated with a 2 fold downregulation of the expression of PCP (217±66 vs. 437.6±18) and 2 fold increase in expression of VDAC (428±45 vs. 197±19). A trend towards increased level of expression of ANT was observed in those with AF (900±13 vs. 774±29). Conclusion: The increased sensitivity of mitochondria from AF patients to mPTP opening can be partly explained by selective downregulation of PCP protein, putative component of mPTP with upregulation of regulatory VDAC and ANT proteins

A novel high throughput approach for quantification of cell density

Background: Current approach to cell counting using hemacytometer is limited by requirement for high cell concentration and is prone to error. In biological experiments using cells from human cardiac tissues with limited number of cells, this approach results in large variation in cell counts. Here, we demonstrate the utility of a novel approach using a 96-well microplate that accurately provides the density of cells as low as 15,000 cells/cm2, which fulfills an unmet need in experiments with limited cell availability. Purpose: To develop and test the accuracy of a high-throughput 96-well microplate assay in assessing the cell density in comparison to existing methods. Methods: NIH/3T3 fibroblasts were cultured and differentiated and grown to different cell density. Cell number obtained using hemacytometer was compared to the total fluorescence of propidium iodide, binding to the nuclei of cells permeabilized with Triton X-100 (0.25%), and assessed using multiplate reader. In addition, the total activity of lactate dehydrogenase, an intracellular enzyme, was used to assess the total volume of cytoplasm released from permeabilized cells. Furthermore, the ratio of live/ dead cells was determined by propidium iodide-positive cells and lactate dehydrogenase activity before and after permeabilization in each well of the 96-well plate. Results:There was a linear relationship between increasing intensity of propidium iodide fluorescence with the density of the cells in the 96-well microplate (ranging from 5,000 to 100,000 cells/cm2). Similarly, linear relationship was observed between the intensity of propidium iodide fluorescence and cellular lactate dehydrogenase activity in corresponding wells. At low cell density ( Conclusion: Proposed propidium iodide and lactate dehydrogenase assays are useful tools for quantification of cell number in high-throughput manner with greater accuracy at low cell density, higher reproducibility and overall time saving. This assay is especially useful in experiments using limited cell number such as cells isolated from the human heart

Correlation of potential noninvasive biomarkers of extracellular matrix remodeling with postoperative heart failure - a preliminary study

Background: Postoperative heart failure (PHF) is a major factor that prolongs hospital stay and contributes to increased cost and morbidity after surgery. Clinical predictors of PHF have been identified but lack specificity and predictive accuracy; therefore, identifying the candidates at risk for PHF remains difficult, thus, necessitating further investigation. Since cardiac fibrosis in the elderly contributes to abnormal cardiac contractility, elevated markers of extracellular matrix turnover can be used to identify those at risk for PHF. Purpose: To identify biomarkers in those at risk for PHF. Methods: Serum biomarkers of collagen synthesis (C-terminal propeptide type 1 of procollagen [PICP] and N-terminal propeptide of type III procollagen [PIIINP]), collagen degradation (C-terminal telopeptide of collagen type 1 [CITP]), and extracellular matrix remodeling (matrix metalloprotease-1 [MMP-1] and tissue inhibitor of metalloproteases-1 [TIMP-1]) were determined by ELISA in preoperative blood samples collected from patients with no prior history of heart failure who were undergoing cardiac surgery and correlated to PHF. Results: Of 45 patients enrolled (mean age 69 ± 11 years, 77% male), 11 (24%, mean age 66 ± 10 years) developed PHF requiring additional inotropic support (dobutamine) and management for heart failure. Overall, there were no significant differences in baseline demographics and comorbidities between those who did or did not develop PHF. Ventricular function was preserved, with no significant differences in left ventricular ejection fraction (60 ± 11% vs. 50 ± 16%; P=0.11) or atrial and ventricular dimensions and function between the two groups. Mean PICP levels showed elevated trend in PHF (661 ± 375 ng/ ml vs. 609 ± 410 ng/ml in the non-PHF group; P=0.07); however, PIIINP levels were not significantly different between the two groups (134 ± 72 in PHF vs. 157 ± 73; P=0.47). However, levels of CITP were significantly lower in PHF patients (6 ± 3 ng/ml vs. 9 ± 7 ng/ml; P=0.03). The overall ratio of PICP/CITP was higher in patients with PHF (119 vs. 108; P=0.7). The MMP-1, MMP-2 and TIMP-1 levels were not significantly different between the two groups. Conclusion: The preliminary data obtained from 45 patients demonstrated a trend toward higher PICP levels indicative of collagen synthesis in those at risk for PHF but was not statistically significant. This is likely due to the small sample size and the heterogeneity of the patients, indicating a larger number of heterogeneous patients are needed to demonstrate the prognostic significance of serum biomarkers of extracellular matrix remodeling

Prolonged post-differentiation culture influences the expression and biophysics of Na+ and Ca2+ channels in induced pluripotent stem cell-derived ventricular-like cardiomyocytes

Several studies have been reported in various domains from induction methods to utilities of somatic cell pluripotent reprogramming. However, one of the major struggles facing the research field of induced pluripotent stem cell (iPSC)-derived target cells is the lack of consistency in observations. This could be due to variety of reasons including varied culture periods post-differentiation. The cardiomyocytes (CMs) derived from iPSCs are commonly studied and proposed to be utilized in the comprehensive in vitro proarrhythmia initiative for drug safety screening. As the influence of varied culture periods on the electrophysiological properties of iPSC-CMs is not clearly known, using whole-cell patch clamp technique, we compared two groups of differentiated ventricular-like iPSC-CMs that are cultured for 10 to 15 days (D10-15) and more than 30 days (≥ D30) both under current and voltage clamps. The prolonged culture imparts increased excitability with high-frequency spontaneous action potentials, robust increase in the magnitude of peak Na+ current density, relatively shallow inactivation kinetics of Na+ channels, faster recovery from inactivation, and augmented Ca2+ current density. Quantitative real-time PCR studies of α-subunit transcripts showed enhanced mRNA expression of SCN1A, SCN5A Na+ channel subtypes, and CACNA1C, CACNA1G, and CACNA1I Ca2+ channel subtypes, in ≥ D30 group. Conclusively, the prolonged culture of differentiated iPSC-CMs affects the excitability, single-cell electrophysiological properties, and ion channel expressions. Therefore, following standard periods of culture across research studies while utilizing ventricular-like iPSC-CMs for in vitro health/disease modeling to study cellular functional mechanisms or test high-throughput drugs\u27 efficacy and toxicity becomes crucial