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

Impacts of climate change on coastal habitats, relevant to the coastal and marine environment around the UK

Coastal habitats are at risk from both direct (temperature, rainfall), and indirect (sea-level rise, coastal erosion) impacts due to a changing climate. Beyond the environmental impacts and ensuing habitat loss, the changing climate will have a significant societal impact to coastal communities ranging from health to livelihoods, as well as the loss of important ecosystem services such as coastal defence – particularly relevant with predicted increase in storminess. Vegetated coastal ecosystems sequester carbon – another ‘ecosystem service’ that could be disrupted due to climate change. There has been considerable recent attention to the potential role these habitats could play in climate mitigation, and also in transferring carbon across the land–sea interface. To understand the relative importance of these habitats within the global carbon cycle, coastal habitats need to be accounted for in national greenhouse gas inventories, and a true multidisciplinary catchment-to-coast approach to research is required. Management options exist that can reduce the immediate impacts of climate change, such as managed realignment and sediment recharge. Fixed landward coastal defences are becoming unsustainable and creating ‘coastal squeeze’, highlighting the need to work with natural processes to recreate more-natural shorelines where possible

Renormalization scale uncertainty in tne DIS 2+1 jet cross-section

The deep inelastic scattering 2+1 jet cross- section is a useful observable for precision tests of QCD, e.g. measuring the strong coupling constant alpha(s). A consistent analysis requires a good understanding of the theoretical uncertainties and one of the most fundamental ones in QCD is due to the renormalization scheme and scale ambiguity. Different methods, which have been proposed to resolve the scale ambiguity, are applied to the 2+1 jet cross-section and the uncertainty is estimated. It is shown that the uncertainty can be made smaller by choosing the jet definition in a suitable way.Comment: 24 pages, uuencoded compressed tar file, DESY 94-082, TSL-ISV-94-009

Evolution of active and polar photospheric magnetic fields during the rise of Cycle 24 compared to previous cycles

The evolution of the photospheric magnetic field during the declining phase and minimum of Cycle 23 and the recent rise of Cycle 24 are compared with the behavior during previous cycles. We used longitudinal full-disk magnetograms from the NSO's three magnetographs at Kitt Peak, the Synoptic Optical Long-term Investigations of the Sun (SOLIS) Vector Spectro-Magnetograph (VSM), the Spectromagnetograph and the 512-Channel Magnetograph instruments, and longitudinal full-disk magnetograms from the Mt. Wilson 150-foot tower. We analyzed 37 years of observations from these two observatories that have been observing daily, weather permitting, since 1974, offering an opportunity to study the evolving relationship between the active region and polar fields in some detail over several solar cycles. It is found that the annual averages of a proxy for the active region poloidal magnetic field strength, the magnetic field strength of the high-latitude poleward streams, and the time derivative of the polar field strength are all well correlated in each hemisphere. These results are based on statistically significant cyclical patterns in the active region fields and are consistent with the Babcock-Leighton phenomenological model for the solar activity cycle. There was more hemispheric asymmetry in the activity level, as measured by total and maximum active region flux, during late Cycle 23 (after around 2004), when the southern hemisphere was more active, and Cycle 24 up to the present, when the northern hemisphere has been more active, than at any other time since 1974. The active region net proxy poloidal fields effectively disappeared in both hemispheres around 2004, and the polar fields did not become significantly stronger after this time. We see evidence that the process of Cycle 24 field reversal has begun at both poles.Comment: Accepted for publication in Solar Physic

One-Loop Matching of the Heavy-Light A_0 and V_0 Currents with NRQCD Heavy and Improved Naive Light Quarks

One-loop matching of heavy-light currents is carried out for a highly improved lattice action, including the effects of dimension 4 O(1/M) and O(a) operators. We use the NRQCD action for heavy quarks, the Asqtad improved naive action for light quarks, and the Symanzik improved glue action. As part of the matching procedure we also present results for the NRQCD self energy and for massless Asqtad quark wavefunction renormalization with improved glue.Comment: 25 pages, 3 eps-figure

Determination of the Strong Coupling \boldmath{\as} from hadronic Event Shapes and NNLO QCD predictions using JADE Data

Event Shape Data from $e^+e^-$ annihilation into hadrons collected by the JADE experiment at centre-of-mass energies between 14 GeV and 44 GeV are used to determine the strong coupling $\alpha_S$. QCD predictions complete to next-to-next-to-leading order (NNLO), alternatively combined with resummed next-to-leading-log-approximation (NNLO+NLLA) calculations, are used. The combined value from six different event shape observables at the six JADE centre-of-mass energies using the NNLO calculations is $\alpha_S(M_Z)$= 0.1210 +/- 0.0007(stat.) +/- 0.0021(expt.) +/- 0.0044(had.) +/- 0.0036(theo.) and with the NNLO+NLLA calculations the combined value is $\alpha_S$= 0.1172 +/- 0.0006(stat.) +/- 0.0020(expt.) +/- 0.0035(had.) +/- 0.0030(theo.) . The stability of the NNLO and NNLO+NLLA results with respect to missing higher order contributions, studied by variations of the renormalisation scale, is improved compared to previous results obtained with NLO+NLLA or with NLO predictions only. The observed energy dependence of $\alpha_S$ agrees with the QCD prediction of asymptotic freedom and excludes absence of running with 99% confidence level.Comment: 9 pages, EPHJA style, 4 figures, corresponds to published version with JADE author lis

The perseverance of Pacioli's goods inventory accounting system

This paper details sources of the 'undoubtedly strange' (Yamey, 1994a, p.119) system of goods inventory records described in Pacioli’s 1494 bookkeeping treatise and traces the longevity and widespread use of this early perpetual inventory recording (EPIR) system in English language texts. By doing so and contrasting this system with the bookkeeping treatment of modern texts, it is shown that the EPIR system persisted as the dominant form of goods inventory accounting for between 400 and 500 years and that the reasons for its demise are worthy of further consideration and research

A rapid review of variation in the use of dental general anaesthetics in children

Introduction: The use of dental general anaesthetics (DGAs) remains a cause for concern due to additional strains placed on health services. There are numerous factors influencing the prevalence and use of DGAs, and understanding these is an important first step in addressing the issue. Aim: Conduct a rapid review of current peer-reviewed and grey literature on the variation in the use of DGAs in children. Methods: Electronic searching using Medline via Ovid covering DGA articles from 1998 onwards, written in English. Publication types included primary and secondary sources from peer-reviewed journals and reports, as well as grey literature. Results: From 935 results, 171 articles were included in the final review. Themes emerging from the literature included discussions of DGA variation, variations in standards of service provision by health services, and the socio-demographic and geographical characteristics of children. Prominent socio-demographic and geographical characteristics included age, other health conditions, ethnic and cultural background, socioeconomic status and deprivation, and geographical location. Conclusions: This review identified numerous variations in the patterns associated with DGA provision and uptake at both a health service and individual level. The findings demonstrate the complicated and multifaceted nature of DGA practices worldwide

DNA-PKcs modulates progenitor cell proliferation and fibroblast senescence in idiopathic pulmonary fibrosis

Published online: 29 August 2019BACKGROUND: Recent studies have highlighted the contribution of senescent mesenchymal and epithelial cells in Idiopathic Pulmonary Fibrosis (IPF), but little is known regarding the molecular mechanisms that regulate the accumulation of senescent cells in this disease. Therefore, we addressed the hypothesis that the loss of DNA repair mechanisms mediated by DNA protein kinase catalytic subunit (DNA-PKcs) in IPF, promoted the accumulation of mesenchymal progenitors and progeny, and the expression of senescent markers by these cell types. METHODS: Surgical lung biopsy samples and lung fibroblasts were obtained from patients exhibiting slowly, rapidly or unknown progressing IPF and lung samples lacking any evidence of fibrotic disease (i.e. normal; NL). The expression of DNA-Pkcs in lung tissue was assessed by quantitative immunohistochemical analysis. Chronic inhibition of DNA-PKcs kinase activity was mimicked using a highly specific small molecule inhibitor, Nu7441. Proteins involved in DNA repair (stage-specific embryonic antigen (SSEA)-4+ cells) were determined by quantitative Ingenuity Pathway Analysis of transcriptomic datasets (GSE103488). Lastly, the loss of DNA-PKc was modeled in a humanized model of pulmonary fibrosis in NSG SCID mice genetically deficient in PRKDC (the transcript for DNA-PKcs) and treated with Nu7441. RESULTS: DNA-PKcs expression was significantly reduced in IPF lung tissues. Chronic inhibition of DNA-PKcs by Nu7441 promoted the proliferation of SSEA4+ mesenchymal progenitor cells and a significant increase in the expression of senescence-associated markers in cultured lung fibroblasts. Importantly, mesenchymal progenitor cells and their fibroblast progeny derived from IPF patients showed a loss of transcripts encoding for DNA damage response and DNA repair components. Further, there was a significant reduction in transcripts encoding for PRKDC (the transcript for DNA-PKcs) in SSEA4+ mesenchymal progenitor cells from IPF patients compared with normal lung donors. In SCID mice lacking DNA-PKcs activity receiving IPF lung explant cells, treatment with Nu7441 promoted the expansion of progenitor cells, which was observed as a mass of SSEA4+ CgA+ expressing cells. CONCLUSIONS: Together, our results show that the loss of DNA-PKcs promotes the expansion of SSEA4+ mesenchymal progenitors, and the senescence of their mesenchymal progeny.David M. Habiel, Miriam S. Hohmann, Milena S. Espindola, Ana Lucia Coelho, Isabelle Jones, Heather Jones, Richard Carnibella, Isaac Pinar, Freda Werdiger and Cory M. Hogaboa

'Caught Between a Rock and a Hard Place': Anti-discrimination Legislation in the Liberal State and the Fate of the Australian Disability Discrimination Act

This article offers a critical analysis of some of the practical implications for disabled people of the Disability Discrimination Act of 1992. Specifically, it raises questions about politics and the role of the law as an instrument of social change?taking greater account of the interests of disabled people?on the one hand, and of the reliance of the social model of disability on a strategy based upon legal rights on the other. The article also suggests that the constraining effects of Australia's constitutional protections of rights and its federal system of government hinder the mildly progressive elements of the Disability Discrimination Act. To illustrate this, the paper employs empirical evidence to suggest that these effects have been exacerbated by the passage of the Human Rights Legislation Amendment Act in 1999