Auxin-degron system identifies immediate mechanisms of OCT4.

The pluripotency factor OCT4 is essential for the maintenance of naive pluripotent stem cells in vitro and in vivo. However, the specific role of OCT4 in this process remains unknown. Here, we developed a rapid protein-level OCT4 depletion system that demonstrates that the immediate downstream response to loss of OCT4 is reduced expression of key pluripotency factors. Our data show a requirement for OCT4 for the efficient transcription of several key pluripotency factors and suggest that expression of trophectoderm markers is a subsequent event. In addition, we find that NANOG is able to bind to the genome in the absence of OCT4, and this binding is in fact enhanced. Globally, however, the active enhancer-associated histone mark H3K27ac is depleted. Our work establishes that, while OCT4 is required for the maintenance of the naive transcription factor network, at a normal embryonic stem cell levels it antagonizes this network through inhibition of NANOG binding

The role of the t-SNARE SNAP-25 in action potential-dependent calcium signaling and expression in GABAergic and glutamatergic neurons

Abstract Background The soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, comprised of SNAP-25, syntaxin 1A, and VAMP-2, has been shown to be responsible for action potential (AP)-dependent, calcium-triggered release of several neurotransmitters. However, this basic fusogenic protein complex may be further specialized to suit the requirements for different neurotransmitter systems, as exemplified by neurons and neuroendocrine cells. In this study, we investigate the effects of SNAP-25 ablation on spontaneous neuronal activity and the expression of functionally distinct isoforms of this t-SNARE in GABAergic and glutamatergic neurons of the adult brain. Results We found that neurons cultured from Snap25 homozygous null mutant (Snap25-/-) mice failed to develop synchronous network activity seen as spontaneous AP-dependent calcium oscillations and were unable to trigger glial transients following depolarization. Voltage-gated calcium channel (VGCC) mediated calcium transients evoked by depolarization, nevertheless, did not differ between soma of SNAP-25 deficient and control neurons. Furthermore, we observed that although the expression of SNAP-25 RNA transcripts varied among neuronal populations in adult brain, the relative ratio of the transcripts encoding alternatively spliced SNAP-25 variant isoforms was not different in GABAergic and glutamatergic neurons. Conclusion We propose that the SNAP-25b isoform is predominantly expressed by both mature glutamatergic and GABAergic neurons and serves as a fundamental component of SNARE complex used for fast synaptic communication in excitatory and inhibitory circuits required for brain function. Moreover, SNAP-25 is required for neurons to establish AP-evoked synchronous network activity, as measured by calcium transients, whereas the loss of this t-SNARE does not affect voltage-dependent calcium entry.</p

ZMYM2 inhibits NANOG-mediated reprogramming.

Background: NANOG is a homeodomain-containing transcription factor which forms one of the hubs in the pluripotency network and plays a key role in the reprogramming of somatic cells and epiblast stem cells to naïve pluripotency.  Studies have found that NANOG has many interacting partners and some of these were shown to play a role in its ability to mediate reprogramming. In this study, we set out to analyse the effect of NANOG interactors on the reprogramming process. Methods: Epiblast stem cells and somatic cells were reprogrammed to naïve pluripotency using MEK/ERK inhibitor PD0325901, GSK3β inhibitor CHIR99021 and Leukaemia Inhibitory Factor (together termed 2i Plus LIF). Zmym2 was knocked out using the CRISPR/Cas9 system or overexpressed using the PiggyBac system. Reprogramming was quantified after ZMYM2 deletion or overexpression, in diverse reprogramming systems. In addition, embryonic stem cell self renewal was quantified in differentiation assays after ZMYM2 removal or overexpression. Results: In this work, we identified ZMYM2/ZFP198, which physically associates with NANOG as a key negative regulator of NANOG-mediated reprogramming of both epiblast stem cells and somatic cells. In addition, ZMYM2 impairs the self renewal of embryonic stem cells and its overexpression promotes differentiation. Conclusions: We propose that ZMYM2 curtails NANOG's actions during the reprogramming of both somatic cells and epiblast stem cells and impedes embryonic stem cell self renewal, promoting differentiation.This study was supported by the Wellcome Trust through a Wellcome Trust Fellowship to J.C.R.S. [101861], Wellcome Trust Studentship to M.L. [079249], and a core funding grant jointly with the Medical Research Council (MRC) to the Wellcome-MRC Cambridge Stem Cell Institute [079249]

Absorption modes of Möbius strip resonators

This is the final version. Available on open access from Nature Research via the DOI in this record. In this work, the electromagnetic response of a mathematically interesting shape-a Möbius strip-is presented, along with a ring resonator for comparison. Both resonators consist of a central lossy dielectric layer bounded by perfectly conducting layers. For the case of the Möbius strips, the computational results show that there are a family of half-integer wavelength modes within the dielectric layer. These additional modes result in increased absorption, and a corresponding reduction in the radar cross section. Interestingly, rotational scans show that these modes can be excited over a large angular range. This investigation gives an understanding of the electromagnetic response of these structures, paving the way for future experiments on Möbius strip resonators.Engineering and Physical Sciences Research Council (EPSRC

Covert Images Using Surface Plasmon-Mediated Optical Polarization Conversion

This is the author accepted manuscript. The final version is available from Wiley via the DOI in this recordCovert optical signatures are a vital element in anticounterfeiting technologies. Plasmonic surfaces offer a means of manipulating the properties of light including the realization of colored pixels and images. In this work, concealed images with accurate color reproduction using plasmonic pixel arrays are demonstrated. The spectral and spatial control of optical polarization conversion is accomplished by tailoring the interaction of light with surface plasmons through the design and arrangement of surface nanostructures. The latent image is revealed using a polarization-sensitive optical system, which represents a means for the authentication of security features that can be created using these nanostructured devices. A red-green-blue color space is defined containing a wide gamut of chromaticities, enabling comprehensive full-color image capability. The device concept extends the functionality of a polarization-dependent plasmonic response to realize the encoding of a color image in covert form.This work was supported by the UK Engineering and Physical Sciences Research Council (EPSRC) Knowledge Transfer Account programme grant EP/H50012X/1, and by QinetiQ Ltd

To Go or Not to Go: A Proof of Concept Study Testing Food-Specific Inhibition Training for Women with Eating and Weight Disorders

This is the author accepted manuscript. The final version is available from Wiley via the DOI in this record.Inefficient food-specific inhibitory control is a potential mechanism that underlies binge eating in bulimia nervosa and binge eating disorder. Go/no-go training tools have been developed to increase inhibitory control over eating impulses. Using a within-subjects design, this study examined whether one session of food-specific go/no-go training, versus general inhibitory control training, modifies eating behaviour. The primary outcome measure was food consumption on a taste test following each training session. Women with bulimia nervosa and binge eating disorder had small non-significant reductions in high-calorie food consumption on the taste test following the food-specific compared with the general training. There were no effects on eating disorder symptomatic behaviour (i.e. binge eating/purging) in the 24 h post-training. The training task was found to be acceptable by the clinical groups. More research is needed with larger sample sizes to determine the effectiveness of this training approach for clinical populations.Robert Turton receives funding from the Medical Research Council (MRC) and the Psychiatry Research Trust (PRT) (Grant PCPTAAR). Dr. Bruno Palazzo Nazar has received funding from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and from the Federal University of Rio de Janeiro (UFRJ). Dr. Emilee Burgess received funding from the University of Alabama at Birmingham and Nova Scotia Health Authority. Dr. Colette Hirsch, Professor Janet Treasure OBE and Dr. Valentina Cardi, receive salary support from the National Institute for Health Research (NIHR), Mental Health Biomedical Research Centre at South London and Maudsley NHS Foundation Trust and King’s College London

Exit from Naive Pluripotency Induces a Transient X Chromosome Inactivation-like State in Males.

A hallmark of naive pluripotency is the presence of two active X chromosomes in females. It is not clear whether prevention of X chromosome inactivation (XCI) is mediated by gene networks that preserve the naive state. Here, we show that robust naive pluripotent stem cell (nPSC) self-renewal represses expression of Xist, the master regulator of XCI. We found that nPSCs accumulate Xist on the male X chromosome and on both female X chromosomes as they become NANOG negative at the onset of differentiation. This is accompanied by the appearance of a repressive chromatin signature and partial X-linked gene silencing, suggesting a transient and rapid XCI-like state in male nPSCs. In the embryo, Xist is transiently expressed in males and in females from both X chromosomes at the onset of naive epiblast differentiation. In conclusion, we propose that XCI initiation is gender independent and triggered by destabilization of naive identity, suggesting that gender-specific mechanisms follow, rather than precede, XCI initiation.This study was supported by a Wellcome Trust Fellowship (WT101861) to J.C.R.S., who is a Wellcome Trust Senior Research Fellow. E.J.S. is the recipient of a Ph.D. fellowship from the Portuguese Foundation for Sciences and Technology, FCT (SFRH/BD/52197/2013). H.T.S. and L.E.B. are recipients of an MRC Ph.D. studentship

Effective-periodicity effects in Fibonacci slot arrays (article)

This is the final version. Available from the American Physical Society via the DOI in this record. The data that support the findings of this study are available from the corresponding author upon reasonable request.The dataset associated with this article is available in ORE at: https://doi.org/10.24378/exe.3783In this Letter, the transmission properties of a nonperiodic array of slots arranged in the form of a Fibonacci sequence are investigated. By arranging the slots in this manner, an additional periodicity can be utilized, resulting in corresponding resonance features in the transmitted signal. By investigating the transmission response of a perforated metallic sheet over a broad frequency range (6–40 GHz), it is shown that this simple one-dimensional chain supports two periodicities, one due to the regular periodic separation and one due to average spacing—which is related to the golden ratio. This response replicates the resonant behavior of a two-dimensional periodic array with a single nonperiodic array also creating new families of diffraction lobes in the far-field region.Engineering and Physical Sciences Research CouncilEngineering and Physical Sciences Research CouncilMCIN/AEI/10.13039/50110001103

Efficient mm-wave photomodulation via coupled Fabry–Perot cavities

This is the author accepted manuscript. The final version is available from AIP Publishing via the DOI in this recordData availability: The data that support the findings of this study are available from the corresponding author upon reasonable request.An efficient mm-wave photomodulator is designed based on coupled Fabry–Perot modes in a low-lifetime silicon wafer and an adjacent cavity formed from a transparent reflector, such as indium tin oxide. The modulation of a reflected beam using this coupled-cavity design is increased by a factor of 7 compared with that from an isolated silicon wafer, while also introducing a degree of tunability and maintaining low angular dispersion. For the particular design built and tested, a modulation of 32% is achieved for an extremely low optical illumination of just 0.006W/cm2 and with a maximum operation rate of more than 3 kHz. The large increase in modulation, coupled with the flexibility of the design and the fact that all components can be industrially manufactured, makes this photomodulator a promising candidate for many communication, imaging, and sensing applications.Engineering and Physical Sciences Research Council (EPSRC)QinetiQ Ltd

OCT4 induces embryonic pluripotency via STAT3 signaling and metabolic mechanisms.

OCT4 is a fundamental component of the molecular circuitry governing pluripotency in vivo and in vitro. To determine how OCT4 establishes and protects the pluripotent lineage in the embryo, we used comparative single-cell transcriptomics and quantitative immunofluorescence on control and OCT4 null blastocyst inner cell masses at two developmental stages. Surprisingly, activation of most pluripotency-associated transcription factors in the early mouse embryo occurs independently of OCT4, with the exception of the JAK/STAT signaling machinery. Concurrently, OCT4 null inner cell masses ectopically activate a subset of trophectoderm-associated genes. Inspection of metabolic pathways implicates the regulation of rate-limiting glycolytic enzymes by OCT4, consistent with a role in sustaining glycolysis. Furthermore, up-regulation of the lysosomal pathway was specifically detected in OCT4 null embryos. This finding implicates a requirement for OCT4 in the production of normal trophectoderm. Collectively, our findings uncover regulation of cellular metabolism and biophysical properties as mechanisms by which OCT4 instructs pluripotency.This work was supported by the University of Cambridge, BBSRC project grant RG74277, BB/R018588/1 and MR/R017735/1 to HS and LB respectively, MRC PhD studentship for AK and a core support grant from the Wellcome Trust and MRC to the Wellcome Trust – Medical Research Council Cambridge Stem Cell Institute