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

Effective mass and quantum lifetime in a Si/Si0.87Ge0.13/Si two-dimensional hole gas

Measurements of Shubnikov de Haas oscillations in the temperature range 0.3–2 K have been used to determine an effective mass of 0.23 m0 in a Si/Si0.87Ge0.13/Si two-dimensional hole gas. This value is in agreement with theoretical predictions and with that obtained from cyclotron resonance measurements. The ratio of the transport time to the quantum lifetime is found to be 0.8. It is concluded that the 4 K hole mobility of 11 000 cm2 V−1 s−1 at a carrier sheet density of 2.2×1011 cm−2 is limited by interface roughness and short-range interface charge scattering

Hole effective mass in remote doped Si/Si1−xGex quantum wells with 0.05x0.3

The effective masses in remote doped Si/Si1−xGex hole quantum wells with 0.05<=x<=0.3, have been determined from the temperature dependence of the Shubnikov–de Haas oscillations. The values are lower than previously observed by other workers, but still somewhat higher than the theoretical Gamma-point values for the ground-state heavy hole subband. The differences are attributed to finite carrier sheet densities and can be satisfactorily accounted for by nonparabolicity corrections

Effective mass and band nonparabolicity in remote doped Si/Si0.8Ge0.2 quantum wells

The effective masses in remote doped Si/Si0.8Ge0.2/Si quantum wells having sheet densities, Ns in the range 2 × 1011–1.1 × 1012 cm – 2 have been determined from the temperature dependencies of the Shubnikov–de Haas oscillations. The values obtained increase with magnetic field and Ns. This behavior is taken as evidence for the nonparabolicity of the valence band and accounts for the discrepancies in previously reported masses. Self-consistent band structure calculations for a triangular confinement of the carriers have also been carried out and provide confirmation of the increase in mass with Ns. Theory and experiment give extrapolated Gamma point effective masses of 0.21 and 0.20 of the free-electron mass, respectively

Hole effective mass in remote doped Si/Si1−xGex quantum wells with 0.05x0.3

The effective masses in remote doped Si/Si1−xGex hole quantum wells with 0.05<=x<=0.3, have been determined from the temperature dependence of the Shubnikov–de Haas oscillations. The values are lower than previously observed by other workers, but still somewhat higher than the theoretical Gamma-point values for the ground-state heavy hole subband. The differences are attributed to finite carrier sheet densities and can be satisfactorily accounted for by nonparabolicity corrections

How Physicians Draw Satisfaction and Overcome Barriers in their Practices: “It Sustains Me”

Objective Major reorganizations of medical practice today challenge physicians’ ability to deliver compassionate care. We sought to understand how physicians who completed an intensive faculty development program in medical humanism sustain their humanistic practices. Methods Program completers from 8 U.S. medical schools wrote reflections in answer to two open-ended questions addressing their personal motivations and the barriers that impeded their humanistic practice and teaching. Reflections were qualitatively analyzed using the constant comparative method. Results Sixty-eight physicians (74% response rate) submitted reflections. Motivating factors included: 1) identification with humanistic values; 2) providing care that they or their family would want; 3) connecting to patients; 4) passing on values through role modelling; 5) being in the moment. Inhibiting factors included: 1) time, 2) stress, 3) culture, and 4) episodic burnout. Conclusions Determination to live by one’s values, embedded within a strong professional identity, allowed study participants to alleviate, but not resolve, the barriers. Collaborative action to address organizational impediments was endorsed but found to be lacking. Practice implications Fostering fully mature professional development among physicians will require new skills and opportunities that reinforce time-honored values while simultaneously partnering with others to nurture, sustain and improve patient care by addressing system issues

Observation of an Excited Bc+ State

Using pp collision data corresponding to an integrated luminosity of 8.5 fb-1 recorded by the LHCb experiment at center-of-mass energies of s=7, 8, and 13 TeV, the observation of an excited Bc+ state in the Bc+π+π- invariant-mass spectrum is reported. The observed peak has a mass of 6841.2±0.6(stat)±0.1(syst)±0.8(Bc+) MeV/c2, where the last uncertainty is due to the limited knowledge of the Bc+ mass. It is consistent with expectations of the Bc∗(2S31)+ state reconstructed without the low-energy photon from the Bc∗(1S31)+→Bc+γ decay following Bc∗(2S31)+→Bc∗(1S31)+π+π-. A second state is seen with a global (local) statistical significance of 2.2σ (3.2σ) and a mass of 6872.1±1.3(stat)±0.1(syst)±0.8(Bc+) MeV/c2, and is consistent with the Bc(2S10)+ state. These mass measurements are the most precise to date

Factors affecting the accuracy of urine-based biomarkers of BSE

<p>Abstract</p> <p>Background</p> <p>Transmissible spongiform encephalopathy diseases are untreatable, uniformly fatal degenerative syndromes of the central nervous system that can be transmitted both within as well as between species. The bovine spongiform encephalopathy (BSE) epidemic and the emergence of a new human variant of Creutzfeldt-Jakob disease (vCJD), have profoundly influenced beef production processes as well as blood donation and surgical procedures. Simple, robust and cost effective diagnostic screening and surveillance tools are needed for both the preclinical and clinical stages of TSE disease in order to minimize both the economic costs and zoonotic risk of BSE and to further reduce the risk of secondary vCJD.</p> <p>Objective</p> <p>Urine is well suited as the matrix for an ante-mortem test for TSE diseases because it would permit non-invasive and repeated sampling. In this study urine samples collected from BSE infected and age matched control cattle were screened for the presence of individual proteins that exhibited disease specific changes in abundance in response to BSE infection that might form the basis of such an ante-mortem test.</p> <p>Results</p> <p>Two-dimensional differential gel electrophoresis (2D-DIGE) was used to identify proteins exhibiting differential abundance in two sets of cattle. The known set consisted of BSE infected steers and age matched controls throughout the course of the disease. The blinded unknown set was composed of BSE infected and control samples of both genders, a wide range of ages and two different breeds. Multivariate analyses of individual protein abundance data generated classifiers comprised of the proteins best able to discriminate between the samples based on disease state, breed, age and gender.</p> <p>Conclusion</p> <p>Despite the presence of confounding factors, the disease specific changes in abundance exhibited by a panel of urine proteins permitted the creation of classifiers able to discriminate between control and infected cattle with a high degree of accuracy.</p