Targeted expression profiling reveals distinct stages of early canine fibroblast reprogramming are regulated by 2-oxoglutarate hydroxylases

Background: Ectopic expression of a defined set of transcription factors allows the reprogramming of mammalian somatic cells to pluripotency. Despite continuous progress in primate and rodent reprogramming, limited attention has been paid to cell reprogramming in domestic and companion species. Previous studies attempting to reprogram canine cells have mostly assessed a small number of presumptive canine induced pluripotent stem cell (iPSC) lines for generic pluripotency attributes. However, why canine cell reprogramming remains extremely inefficient is poorly understood. Methods: To better characterize the initial steps of pluripotency induction in canine somatic cells, we optimized an experimental system where canine fetal fibroblasts (cFFs) are transduced with the Yamanaka reprogramming factors by Sendai virus vectors. We use quantitative PCR arrays to measure the expression of 80 target genes at various stages of canine cell reprogramming. We ask how cFF reprogramming is influenced by small molecules affecting the epigenomic modification 5-hydroxymethylcytosine, specifically L-ascorbic acid and retinoic acid (AA/RA). Results: We found that the expression and catalytic output of a class of 2-oxoglutarate-dependent (2-OG) hydroxylases, known as ten-eleven translocation (TET) enzymes, can be modulated in canine cells treated with AA/RA. We further show that AA/RA treatment induces TET1 expression and facilitates early canine reprogramming, evidenced by upregulation of epithelial and pluripotency markers. Using a chemical inhibitor of 2-OG hydroxylases, we demonstrate that 2-OG hydroxylase activity regulates the expression of a subset of genes involved in mesenchymal-to-epithelial transition (MET) and pluripotency in early canine reprogramming. We identify a set of transcription factors depleted in maturing reprogramming intermediates compared to pluripotent canine embryonic stem cells. Conclusions: Our findings highlight 2-OG hydroxylases have evolutionarily conserved and divergent functions regulating the early reprogramming of canine somatic cells and show reprogramming conditions can be rationally optimized for the generation of maturing canine iPSC

Optical properties of Au colloids self-organized into rings via copolymer templates

The investigation of the Localized Surface Plasmon Resonance for plasmonic nanoparticles has opened new perspectives for optical nanosensors. Today, an issue in plasmonics is the development of large scale and low cost devices. We focus here on the Langmuir-Blodgett technique to self-organize gold nanoparticles (~ 7 nm) into rings (~ 60 nm) via polystyrene-b-polymethylmethacrylate templates. In particular, we investigated the optical properties of organized gold nanoparticle rings over large areas and report experimental evidence for plasmon resonances of both individual nanoparticles and collective modes. This paves the way for designing devices with multiple resonances in the visible-Infra-red spectrum and developing optical sensors

Amounts of selected coagulation factors in pre-and post-mortem follicular fluid are similar and do not correlate with molecular mass

Abstract This study was designed to evaluate the amounts of coagulation factors and to determine whether the protein profile in pre-ovulatory ovarian follicular fluid aspirated from ovaries collected from mares at slaughter are representative of that in follicular fluid collected from live animals. The proteins evaluated included, (i) albumin, ceruloplasmin and fibronectin, (ii) the procoagulant plasma proteins, Factor V (FV), Factor VII (FVII), Factor X (FX) and prothrombin, and (iii) the anticoagulant plasma proteins, antithrombin and ␣2-macroglobulin. The amounts of the individual proteins were similar in both types of follicular fluid. There was no correlation between the activity of FV, FVII, FX or prothrombin in follicular fluid and their molecular size although a correlation was found for the other proteins. These results suggest that the procoagulant proteins in follicular fluid are not likely derived from plasma. The total protein content of follicular fluid samples collected from both sources was similar and the results determined with the Biuret, Lowry and Biorad methods were also not significantly different (P>0.05)

Antimicrobial peptide capsids of de novo design

The spread of bacterial resistance to antibiotics poses the need for antimicrobial discovery. With traditional search paradigms being exhausted, approaches that are altogether different from antibiotics may offer promising and creative solutions. Here, we introduce a de novo peptide topology that-by emulating the virus architecture-assembles into discrete antimicrobial capsids. Using the combination of high-resolution and real-time imaging, we demonstrate that these artificial capsids assemble as 20-nm hollow shells that attack bacterial membranes and upon landing on phospholipid bilayers instantaneously (seconds) convert into rapidly expanding pores causing membrane lysis (minutes). The designed capsids show broad antimicrobial activities, thus executing one primary function-they destroy bacteria on contact

Separation of instrumental and astrophysical foregrounds for mapping CMB anisotropies

We address the extended problem of component separation for CMB applications when a mixture of both astrophysical and instrumental components are present in the observations, and show how standard methods can be adapted to handle this more complex inversion problem.Comment: 12 pages, 5 figures, submitted to MNRA

Brane inflation and the WMAP data: a Bayesian analysis

The Wilkinson Microwave Anisotropy Probe (WMAP) constraints on string inspired ''brane inflation'' are investigated. Here, the inflaton field is interpreted as the distance between two branes placed in a flux-enriched background geometry and has a Dirac-Born-Infeld (DBI) kinetic term. Our method relies on an exact numerical integration of the inflationary power spectra coupled to a Markov-Chain Monte-Carlo exploration of the parameter space. This analysis is valid for any perturbative value of the string coupling constant and of the string length, and includes a phenomenological modelling of the reheating era to describe the post-inflationary evolution. It is found that the data favour a scenario where inflation stops by violation of the slow-roll conditions well before brane annihilation, rather than by tachyonic instability. Concerning the background geometry, it is established that log(v) > -10 at 95% confidence level (CL), where "v" is the dimensionless ratio of the five-dimensional sub-manifold at the base of the six-dimensional warped conifold geometry to the volume of the unit five-sphere. The reheating energy scale remains poorly constrained, Treh > 20 GeV at 95% CL, for an extreme equation of state (wreh ~ -1/3) only. Assuming the string length is known, the favoured values of the string coupling and of the Ramond-Ramond total background charge appear to be correlated. Finally, the stochastic regime (without and with volume effects) is studied using a perturbative treatment of the Langevin equation. The validity of such an approximate scheme is discussed and shown to be too limited for a full characterisation of the quantum effects.Comment: 65 pages, 15 figures, uses iopart. Shortened version, updated references. Matches publication up to appendix B kept on the arXi

Nanoscale imaging reveals laterally expanding antimicrobial pores in lipid bilayers

Antimicrobial peptides are postulated to disrupt microbial phospholipid membranes. The prevailing molecular model is based on the formation of stable or transient pores although the direct observation of the fundamental processes is lacking. By combining rational peptide design with topographical (atomic force microscopy) and chemical (nanoscale secondary ion mass spectrometry) imaging on the same samples, we show that pores formed by antimicrobial peptides in supported lipid bilayers are not necessarily limited to a particular diameter, nor they are transient, but can expand laterally at the nano-to-micrometer scale to the point of complete membrane disintegration. The results offer a mechanistic basis for membrane poration as a generic physicochemical process of cooperative and continuous peptide recruitment in the available phospholipid matrix