Overexpression of connexin 43 using a retroviral vector improves electrical coupling of skeletal myoblasts with cardiac myocytes in vitro.

BACKGROUND: Organ transplantation is presently often the only available option to repair a damaged heart. As heart donors are scarce, engineering of cardiac grafts from autologous skeletal myoblasts is a promising novel therapeutic strategy. The functionality of skeletal muscle cells in the heart milieu is, however, limited because of their inability to integrate electrically and mechanically into the myocardium. Therefore, in pursuit of improved cardiac integration of skeletal muscle grafts we sought to modify primary skeletal myoblasts by overexpression of the main gap-junctional protein connexin 43 and to study electrical coupling of connexin 43 overexpressing myoblasts to cardiac myocytes in vitro. METHODS: To create an efficient means for overexpression of connexin 43 in skeletal myoblasts we constructed a bicistronic retroviral vector MLV-CX43-EGFP expressing the human connexin 43 cDNA and the marker EGFP gene. This vector was employed to transduce primary rat skeletal myoblasts in optimised conditions involving a concomitant use of the retrovirus immobilising protein RetroNectin and the polycation transduction enhancer Transfectam. The EGFP-positive transduced cells were then enriched by flow cytometry. RESULTS: More than four-fold overexpression of connexin 43 in the transduced skeletal myoblasts, compared with non-transduced cells, was shown by Western blotting. Functionality of the overexpressed connexin 43 was demonstrated by microinjection of a fluorescent dye showing enhanced gap-junctional intercellular transfer in connexin 43 transduced myoblasts compared with transfer in non-transduced myoblasts. Rat cardiac myocytes were cultured in multielectrode array culture dishes together with connexin 43/EGFP transduced skeletal myoblasts, control non-transduced skeletal myoblasts or alone. Extracellular field action potential activation rates in the co-cultures of connexin 43 transduced skeletal myoblasts with cardiac myocytes were significantly higher than in the co-cultures of non-transduced skeletal myoblasts with cardiac myocytes and similar to the rates in pure cultures of cardiac myocytes. CONCLUSION: The observed elevated field action potential activation rate in the co-cultures of cardiac myocytes with connexin 43 transduced skeletal myoblasts indicates enhanced cell-to-cell electrical coupling due to overexpression of connexin 43 in skeletal myoblasts. This study suggests that retroviral connexin 43 transduction can be employed to augment engineering of the electrocompetent cardiac grafts from patients own skeletal myoblasts

Weak-localization corrections to the conductivity of double quantum wells

The weak-localization contribution \delta\sigma(B) to the conductivity of a tunnel-coupled double-layer electron system is evaluated and its behavior in weak magnetic fields B perpendicular or parallel to the layers is examined. In a perpendicular field B, \delta \sigma(B) increases and remains dependent on tunneling as long as the magnetic field is smaller than \hbar/e D \tau_t, where D is the in-plane diffusion coefficient and \tau_t the interlayer tunneling time. If \tau_t is smaller than the inelastic scattering time, a parallel magnetic field also leads to a considerable increase of the concuctivity starting with a B**2 law and saturating at fields higher than \hbar/e Z (D \tau_t)**(1/2), where Z is the interlayer distance. In the limit of coherent tunneling, when \tau_t is comparable to elastic scattering time, \delta \sigma(B) differs from that of a single-layer system due to ensuing modifications of the diffusion coefficient. A possibility to probe the weak-localization effect in double-layer systems by the dependence of the conductivity on the gate-controlled level splitting is discussed.Comment: Text 18 pages in Latex/Revtex format, 4 Postscript figures. J. Phys.: CM,in pres

Numerical Investigation of Second Mode Attenuation over Carbon/Carbon Surfaces on a Sharp Slender Cone

We have carried out axisymmetric numerical simulations of a spatially developing hypersonic boundary layer over a sharp 7$^{\circ{}}$-half-angle cone at $M_\infty=7.5$ inspired by the experimental investigations by Wagner (2015). Simulations are first performed with impermeable (or solid) walls with a one-time broadband pulse excitation applied upstream to determine the most convectively-amplified frequencies resulting in the range 260kHz -- 400kHz, consistent with experimental observations of second-mode instability waves. Subsequently, we introduce harmonic disturbances via continuous periodic suction and blowing at 270kHz and 350kHz. For each of these forcing frequencies complex impedance boundary conditions (IBC), modeling the acoustic response of two different carbon/carbon (C/C) ultrasonically absorptive porous surfaces, are applied at the wall. The IBCs are derived as an output of a pore-scale aeroacoustic analysis -- the inverse Helmholtz Solver (iHS) -- which is able to return the broadband real and imaginary components of the surface-averaged impedance. The introduction of the IBCs in all cases leads to a significant attenuation of the harmonically-forced second-mode wave. In particular, we observe a higher attenuation rate of the introduced waves with frequency of 350kHz in comparison with 270kHz, and, along with the iHS impedance results, we establish that the C/C surfaces absorb acoustic energy more effectively at higher frequencies.Comment: AIAA-SciTech 201

Solubility, aggregation and stability of Amphotericin B drug in pure organic solvents: Thermodynamic analysis and solid form characterization.

The solubility of amphotericin B (AmB) has been studied between temperatures of 298-343 K in a range of pure organic solvents including methanol, ethanol, isopropanol, butanol, acetone, tetrahydrofuran, and 1,4-dioxane. The initial solid form used in solubility determinations has been characterized by a series of techniques including powder X-ray diffraction (PXRD), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) and Raman spectroscopy, thermogravimetric analysis and differential scanning calorimetry. Pure AmB melts at 444.5 K with an associated enthalpy of fusion of 177.2 kJ/mol. The equilibrated solids in suspension have been characterized by SEM and PXRD and additionally, the equilibrated saturated solutions have been characterized by in-situ UV-vis to probe the saturation and the aggregation state of AmB. In the explored solvents, in terms of mass ratio, the solubility has been found to be remarkably low, decreasing in the order: ethanol > tetrahydrofuran > butanol ≃ 1,4-dioxane ≃ isopropanol > acetone. The relative order of solubility obtained in the alcohols correlates well with their polarity, revealing important interactions of the solvents' hydroxyl group with the polyol chain and the micosamine sugar carboxylic moiety of amphotericin B. Except in dioxane and isopropanol, the obtained Van't Hoff enthalpies of solution reveal an inverse proportionality to the experimental molar fraction solubility values obtained experimentally, indicating a larger energy requirement to solvate AmB molecules. Aggregation of AmB in the equilibrated solutions was not detected in any of the solvents studied yet HPLC analyses revealed that the API degraded in methanol during the equilibration time for the solubility determinations

A new array for the study of ultra high energy gamma-ray sources

The design and operation of a 32 x 1 10 to the 15th power sq m array of scintillation detectors for the detection of 10 to the 15th power eV cosmic rays is described with an expected angular resolution of 1 deg, thus improving the present signal/background ratio for gamma ray sources. Data are recorded on a hybrid CAMAC, an in-house system which uses a laser and Pockel-Cell arrangement to routinely calibrate the timing stability of the detectors

Physiotherapy intervention in Parkinson's disease: systematic review and meta-analysis

Objective To assess the effectiveness of physiotherapy compared with no intervention in patients with Parkinson’s disease. Design Systematic review and meta-analysis of randomised controlled trials. Data sources Literature databases, trial registries, journals, abstract books, and conference proceedings, and reference lists, searched up to the end of January 2012. Review methods Randomised controlled trials comparing physiotherapy with no intervention in patients with Parkinson’s disease were eligible. Two authors independently abstracted data from each trial. Standard meta-analysis methods were used to assess the effectiveness of physiotherapy compared with no intervention. Tests for heterogeneity were used to assess for differences in treatment effect across different physiotherapy interventions used. Outcome measures were gait, functional mobility and balance, falls, clinician rated impairment and disability measures, patient rated quality of life, adverse events, compliance, and economic analysis outcomes. Results 39 trials of 1827 participants met the inclusion criteria, of which 29 trials provided data for the meta-analyses. Significant benefit from physiotherapy was reported for nine of 18 outcomes assessed. Outcomes which may be clinically significant were speed (0.04 m/s, 95% confidence interval 0.02 to 0.06, P<0.001), Berg balance scale (3.71 points, 2.30 to 5.11, P<0.001), and scores on the unified Parkinson’s disease rating scale (total score −6.15 points, −8.57 to −3.73, P<0.001; activities of daily living subscore −1.36, −2.41 to −0.30, P=0.01; motor subscore −5.01, −6.30 to −3.72, P<0.001). Indirect comparisons of the different physiotherapy interventions found no evidence that the treatment effect differed across the interventions for any outcomes assessed, apart from motor subscores on the unified Parkinson’s disease rating scale (in which one trial was found to be the cause of the heterogeneity). Conclusions Physiotherapy has short term benefits in Parkinson’s disease. A wide range of physiotherapy techniques are currently used to treat Parkinson’s disease, with little difference in treatment effects. Large, well designed, randomised controlled trials with improved methodology and reporting are needed to assess the efficacy and cost effectiveness of physiotherapy for treating Parkinson’s disease in the longer term

Spatial analysis of malaria incidence at the village level in areas with unstable transmission in Ethiopia

<p>Abstract</p> <p>Background</p> <p>Malaria is the leading cause of morbidity and mortality in Ethiopia, accounting for over five million cases and thousands of deaths annually. The risks of morbidity and mortality associated with malaria are characterized by spatial and temporal variation across the country. This study examines the spatial and temporal patterns of malaria transmission at the local level and implements a risk mapping tool to aid in monitoring and disease control activities.</p> <p>Methods</p> <p>In this study, we examine the global and local patterns of malaria distribution in 543 villages in East Shoa, central Ethiopia using individual-level morbidity data collected from six laboratory and treatment centers between September 2002 and August 2006.</p> <p>Results</p> <p>Statistical analysis of malaria incidence by sex, age, and village through time reveal the presence of significant spatio-temporal variations. Poisson regression analysis shows a decrease in malaria incidence with increasing age. A significant difference in the malaria incidence density ratio (IDRs) is detected in males but not in females. A significant decrease in the malaria IDRs with increasing age is captured by a quadratic model. Local spatial statistics reveals clustering or hot spots within a 5 and 10 km distance of most villages in the study area. In addition, there are temporal variations in malaria incidence.</p> <p>Conclusion</p> <p>Malaria incidence varies according to gender and age, with males age 5 and above showing a statistically higher incidence. Significant local clustering of malaria incidence occurs between pairs of villages within 1–10 km distance lags. Malaria incidence was higher in 2002–2003 than in other periods of observation. Malaria hot spots are displayed as risk maps that are useful for monitoring and spatial targeting of prevention and control measures against the disease.</p