Activation of pluripotency genes in human fibroblast cells by a novel mRNA based approach

Background: Several methods have been used to induce somatic cells to re-enter the pluripotent state. Viral transduction of reprogramming genes yields higher efficiency but involves random insertions of viral sequences into the human genome. Although induced pluripotent stem (iPS) cells can be obtained with the removable PiggyBac transposon system or an episomal system, both approaches still use DNA constructs so that resulting cell lines need to be thoroughly analyzed to confirm they are free of harmful genetic modification. Thus a method to change cell fate without using DNA will be very useful in regenerative medicine. Methodology/Principal Findings: In this study, we synthesized mRNAs encoding OCT4, SOX2, cMYC, KLF4 and SV40 large T (LT) and electroporated them into human fibroblast cells. Upon transfection, fibroblasts expressed these factors at levels comparable to, or higher than those in human embryonic stem (ES) cells. Ectopically expressed OCT4 localized to the cell nucleus within 4 hours after mRNA introduction. Transfecting fibroblasts with a mixture of mRNAs encoding all five factors significantly increased the expression of endogenous OCT4, NANOG, DNMT3 beta, REX1 and SALL4. When such transfected fibroblasts were also exposed to several small molecules (valproic acid, BIX01294 and 5'-aza-2'-deoxycytidine) and cultured in human embryonic stem cell (ES) medium they formed small aggregates positive for alkaline phosphatase activity and OCT4 protein within 30 days. Conclusion/Significance: Our results demonstrate that mRNA transfection can be a useful approach to precisely control the protein expression level and short-term expression of reprogramming factors is sufficient to activate pluripotency genes in differentiated cells

Three String Junction and N=4 Dyon Spectrum

The exact spectrum of dyons in a class of N=4 supersymmetric string theories gives us information about dyon spectrum in N=4 supersymmetric gauge theories. This in turn can be translated into prediction about the BPS spectrum of three string junctions on a configuration of three parallel D3-branes. We show that this prediction agrees with the known spectrum of three string junction in different domains in the moduli space separated by walls of marginal stability.Comment: LaTeX file, 14 page

Probing metal ion binding and conformational properties of the colicin E9 endonuclease by electrospray ionization time-of-flight mass spectrometry

Nano-electrospray ionization time-of-flight mass spectrometry (ESI-MS) was used to study the conformational consequences of metal ion binding to the colicin E9 endonuclease (E9 DNase) by taking advantage of the unique capability of ESI-MS to allow simultaneous assessment of conformational heterogeneity and metal ion binding. Alterations of charge state distributions on metal ion binding/release were correlated with spectral changes observed in far- and near-UV circular dichroism (CD) and intrinsic tryptophan fluorescence. In addition, hydrogen/deuterium (H/D) exchange experiments were used to probe structural integrity. The present study shows that ESI-MS is sensitive to changes of the thermodynamic stability of E9 DNase as a result of metal ion binding/release in a manner consistent with that deduced from proteolysis and calorimetric experiments. Interestingly, acid-induced release of the metal ion from the E9 DNase causes dramatic conformational instability associated with a loss of fixed tertiary structure, but secondary structure is retained. Furthermore, ESI-MS enabled the direct observation of the noncovalent protein complex of E9 DNase bound to its cognate immunity protein Im9 in the presence and absence of Zn2+. Gas-phase dissociation experiments of the deuterium-labeled binary and ternary complexes revealed that metal ion binding, not Im9, results in a dramatic exchange protection of E9 DNase in the complex. In addition, our metal ion binding studies and gas-phase dissociation experiments of the ternary E9 DNase-Zn2+-Im9 complex have provided further evidence that electrostatic interactions govern the gas phase ion stability

String-mediated electroweak baryogenesis: a critical analysis

We study the scenario of electroweak baryogenesis mediated by nonsuperconducting cosmic strings. This idea relies upon electroweak symmetry being restored in a region around the core of the topological defect so that, within this region, the rate of baryon number violation is enhanced. We compute numerically how effectively baryon number is violated along a cosmic string, at an epoch when the baryon number violation rate elsewhere is negligible. We show that B-violation along nonsuperconducting strings is quite inefficient. When proper accounting is taken of the velocity dependence of the baryon number production by strings, it proves too small to explain the observed abundance by at least ten orders of magnitude, whether the strings are in the friction dominated or the scaling regime.We study the scenario of electroweak baryogenesis mediated by nonsuperconducting cosmic strings. This idea relies upon electroweak symmetry being restored in a region around the core of the topological defect so that, within this region, the rate of baryon number violation is enhanced. We compute numerically how effectively baryon number is violated along a cosmic string, at an epoch when the baryon number violation rate elsewhere is negligible. We show that B-violation along nonsuperconducting strings is quite inefficient. When proper accounting is taken of the velocity dependence of the baryon number production by strings, it proves too small to explain the observed abundance by at least ten orders of magnitude, whether the strings are in the friction dominated or the scaling regime

Non-Critical Liouville String Escapes Constraints on Generic Models of Quantum Gravity

It has recently been pointed out that generic models of quantum gravity must contend with severe phenomenological constraints imposed by gravitational Cerenkov radiation, neutrino oscillations and the cosmic microwave background radiation. We show how the non-critical Liouville-string model of quantum gravity we have proposed escapes these constraints. It gives energetic particles subluminal velocities, obviating the danger of gravitational Cerenkov radiation. The effect on neutrino propagation is naturally flavour-independent, obviating any impact on oscillation phenomenology. Deviations from the expected black-body spectrum and the effects of time delays and stochastic fluctuations in the propagation of cosmic microwave background photons are negligible, as are their effects on observable spectral lines from high-redshift astrophysical objects.Comment: 15 pages LaTeX, 2 eps figures include

Leak Rate Quantification Method for Gas Pressure Seals with Controlled Pressure Differential

An enhancement to the pressure decay leak rate method with mass point analysis solved deficiencies in the standard method. By adding a control system, a constant gas pressure differential across the test article was maintained. As a result, the desired pressure condition was met at the onset of the test, and the mass leak rate and measurement uncertainty were computed in real-time. The data acquisition and control system were programmed to automatically stop when specified criteria were met. Typically, the test was stopped when a specified level of measurement uncertainty was attained. Using silicone O-ring test articles, the new method was compared with the standard method that permitted the downstream pressure to be non-constant atmospheric pressure. The two methods recorded comparable leak rates, but the new method recorded leak rates with significantly lower measurement uncertainty, statistical variance, and test duration. Utilizing this new method in leak rate quantification, projects will reduce cost and schedule, improve test results, and ease interpretation between data sets

Dying Dyons Don't Count

The dyonic 1/4-BPS states in 4D string theory with N=4 spacetime supersymmetry are counted by a Siegel modular form. The pole structure of the modular form leads to a contour dependence in the counting formula obscuring its duality invariance. We exhibit the relation between this ambiguity and the (dis-)appearance of bound states of 1/2-BPS configurations. Using this insight we propose a precise moduli-dependent contour prescription for the counting formula. We then show that the degeneracies are duality-invariant and are correctly adjusted at the walls of marginal stability to account for the (dis-)appearance of the two-centered bound states. Especially, for large black holes none of these bound states exists at the attractor point and none of these ambiguous poles contributes to the counting formula. Using this fact we also propose a second, moduli-independent contour which counts the "immortal dyons" that are stable everywhere.Comment: 27 pages, 2 figures; one minus sign correcte

Supra-nutritional vitamin E supplementation for 28 days before slaughter maximises muscle vitamin E concentration in finisher pigs

A 4 × 3 factorial experiment (n= 8 pigs per treatment combination) was conducted with 96 female Landrace × Large White pigs to examine the required level of dietary vitamin E and optimum feeding duration before slaughter to maximise muscle vitamin E content in the Longissimus thoracis et lumborum (LTL) muscle. The respective factors were four dietary levels of vitamin E (supplemented as dl-α-tocopheryl acetate; 35, 300, 500, and 700 IU/kg) and three feeding durations (14, 28 and 42 days before slaughter). Vitamin E concentration in the LTL was maximised at 6 mg/kg, which was achieved by feeding a 700 IU vitamin E diet for 28 days before slaughter (P < 0.001). There was no further increase in the vitamin E content of the LTL by feeding the high vitamin E diet more than 28 days before slaughter

On walls of marginal stability in N=2 string theories

We study the properties of walls of marginal stability for BPS decays in a class of N=2 theories. These theories arise in N=2 string compactifications obtained as freely acting orbifolds of N=4 theories, such theories include the STU model and the FHSV model. The cross sections of these walls for a generic decay in the axion-dilaton plane reduce to lines or circles. From the continuity properties of walls of marginal stability we show that central charges of BPS states do not vanish in the interior of the moduli space. Given a charge vector of a BPS state corresponding to a large black hole in these theories, we show that all walls of marginal stability intersect at the same point in the lower half of the axion-dilaton plane. We isolate a class of decays whose walls of marginal stability always lie in a region bounded by walls formed by decays to small black holes. This enables us to isolate a region in moduli space for which no decays occur within this class. We then study entropy enigma decays for such models and show that for generic values of the moduli, that is when moduli are of order one compared to the charges, entropy enigma decays do not occur in these models.Comment: 40 pages, 2 figure

Aerobic Capacity Following Long Duration International Spaces Station (ISS) Missions: Preliminary Results

Maximum oxygen uptake (VO2max) is reduced immediately following space flights lasting 6%. WRmax also decreased on R+1/2 compared to preflight (Pre: 245+/-69, R+1/2: 210+/-45 W). On R+10, VO2max was 2.86+/-0.62 L(dot)/min, with 2 subjects still demonstrating a loss of > 6% from preflight. WRmax on R+10 was 240+/-49 W. HRmax did not change from pre to post-flight. Conclusions: These preliminary results, from the first 5 of 12 planned subjects of an ongoing ISS study, suggest that the majority of astronauts will experience a decrease in VO2max after long-duration space-flight. Interestingly, the two astronauts with the highest preflight VO2max had the greatest loss on R+1/2, and the astronaut with the lowest preflight VO2max increased by 13%. Thus, maintenance of VO2max may be more difficult in astronauts who have a high aerobic capacity, perhaps requiring more intense in-flight exercise countermeasure prescriptions