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

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

Effect of two-weeks endurance training wearing additional clothing in a temperate outdoor environment on performance and physiology in the heat

This investigation assessed performance, physiological and perceptual responses to wearing additional clothing during endurance training for two-weeks in temperate environments, to determine if this approach could be used as a practical, alternative, heat acclimation strategy for athletes. Fifteen trained male triathletes assigned to performance-matched groups completed a two-week unsupervised endurance cycling and running program in either (i) shorts and a short sleeve top (CON; n = 8) or (ii) additional clothing of full-length pants, a “winter” jacket and gloves made from nylon, polyurethane and polyester (AC; n = 7). Participants completed three separate (i.e. familiarisation, pre-program and post-program), identical, pre-loaded cycling time-trials (20 min at 180 W followed by a 40 min self-paced time trial) in 32.5 ± 0.1°C and 55 ± 6% RH. Core and skin temperatures, heart rate, sweat rate, perceived exertion, thermal sensation and thermal comfort were measured across the pre-loaded time trials, and heart rate and thermal sensation were measured across the training program. All of the participants recorded in their diaries that they completed all of the programmed training sessions in the required attire. Mean thermal sensation was most likely hotter in AC (5.5 ± 0.4 AU) compared to CON (4.4 ± 0.4 AU; ES = 1.61, ± 0.68) during the training sessions. However, follow up tests revealed no physiological or perceptual signs of heat acclimation, and the change in time-trial performance from pre-post between groups was trivial (CON: −3.5 ± 12.0 W, AC: −4.1 ± 9.6 W; difference = -0.7%, ± 5.4%). Training in additional clothing for two-weeks in a temperate environment was not an effective heat acclimation strategy for triathletes

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

Pre-exercise carbohydrate or protein ingestion influences substrate oxidation but not performance or hunger compared with cycling in the fasted state

Nutritional intake can influence exercise metabolism and performance, but there is a lack of research comparing protein-rich pre-exercise meals with endurance exercise performed both in the fasted state and following a carbohydrate-rich breakfast. The purpose of this study was to determine the effects of three pre-exercise nutrition strategies on metabolism and exercise capacity during cycling. On three occasions, seventeen trained male cyclists (VO2peak 62.2 ± 5.8 mL·kg−1·min−1, 31.2 ± 12.4 years, 74.8 ± 9.6 kg) performed twenty minutes of submaximal cycling (4 × 5 min stages at 60%, 80%, and 100% of ventilatory threshold (VT), and 20% of the difference between power at the VT and peak power), followed by 3 × 3 min intervals at 80% peak aerobic power and 3 × 3 min intervals at maximal effort, 30 min after consuming a carbohydrate-rich meal (CARB; 1 g/kg CHO), a protein-rich meal (PROTEIN; 0.45 g/kg protein + 0.24 g/kg fat), or water (FASTED), in a randomized and counter-balanced order. Fat oxidation was lower for CARB compared with FASTED at and below the VT, and compared with PROTEIN at 60% VT. There were no differences between trials for average power during high-intensity intervals (367 ± 51 W, p = 0.516). Oxidative stress (F2-Isoprostanes), perceived exertion, and hunger were not different between trials. Overall, exercising in the overnight-fasted state increased fat oxidation during submaximal exercise compared with exercise following a CHO-rich breakfast, and pre-exercise protein ingestion allowed similarly high levels of fat oxidation. There were no differences in perceived exertion, hunger, or performance, and we provide novel data showing no influence of pre-exercise nutrition ingestion on exercise-induced oxidative stress