Template for Developing Guidelines for the Evaluation of the Clinical Efficacy of Psychophysiological Interventions

An essential function of both the Association for Applied Psychophysiology and Biofeedback (AAPB) and the Society for Neuronal Regulation (SNR) is the systematic evaluation of psychophysiological interventions that have been developed for the treatment of medical and psychiatric disorders. In order to address scientific concerns regarding the efficacy of specific clinical applications of biofeedback, these two societies formed and Efficacy Task Force. The process to be used in the assessment of treatment efficacy, specificity and clinical utility is presented in the form of a template that will serve as the foundation for a series of scientific reviews and practice guidlines to be published by both societies

GENE SILENCING. Epigenetic silencing by the HUSH complex mediates position-effect variegation in human cells.

Forward genetic screens in Drosophila melanogaster for modifiers of position-effect variegation have revealed the basis of much of our understanding of heterochromatin. We took an analogous approach to identify genes required for epigenetic repression in human cells. A nonlethal forward genetic screen in near-haploid KBM7 cells identified the HUSH (human silencing hub) complex, comprising three poorly characterized proteins, TASOR, MPP8, and periphilin; this complex is absent from Drosophila but is conserved from fish to humans. Loss of HUSH components resulted in decreased H3K9me3 both at endogenous genomic loci and at retroviruses integrated into heterochromatin. Our results suggest that the HUSH complex is recruited to genomic loci rich in H3K9me3, where subsequent recruitment of the methyltransferase SETDB1 is required for further H3K9me3 deposition to maintain transcriptional silencing.This work was supported by a Wellcome Trust Principal Research Fellowship to P.J.L. (084957/Z/08/Z) and studentship to I.A.T., an MRC Centenary Award to R.T.T., and the Cambridge Biomedical Research Centre (UK). The CIMR is in receipt of a Wellcome Trust Strategic Award.This is the author accepted manuscript. The final version is available from AAAS via http://dx.doi.org/10.1126/science.aaa722

A separable domain of the p150 subunit of human Chromatin Assembly Factor-1 promotes protein and chromosome associations with nucleoli

Chromatin Assembly Factor-1 (CAF-1) is a three-subunit protein complex conserved throughout eukaryotes that deposits histones during DNA synthesis. Here, we present a novel role for the human p150 subunit in regulating nucleolar macromolecular interactions. Acute depletion of p150 causes redistribution of multiple nucleolar proteins and reduces nucleolar association with several repetitive element-containing loci. Notably, a point mutation in a SUMO-interacting motif (SIM) within p150 abolishes nucleolar associations, whereas PCNA or HP1 interaction sites within p150 are not required for these interactions. Additionally, acute depletion of SUMO-2 or the SUMO E2 ligase Ubc9 reduces alpha-satellite DNA association with nucleoli. The nucleolar functions of p150 are separable from its interactions with the other subunits of the CAF-1 complex, because an N-terminal fragment of p150 (p150N) that cannot interact with other CAF-1 subunits is sufficient for maintaining nucleolar chromosome and protein associations. Therefore, these data define novel functions for a separable domain of the p150 protein, regulating protein and DNA interactions at the nucleolus

Promiscuous Targeting of Cellular Proteins by Vpr Drives Systems-Level Proteomic Remodeling in HIV-1 Infection.

HIV-1 encodes four "accessory proteins" (Vif, Vpr, Vpu, and Nef), dispensable for viral replication in vitro but essential for viral pathogenesis in vivo. Well characterized cellular targets have been associated with Vif, Vpu, and Nef, which counteract host restriction and promote viral replication. Conversely, although several substrates of Vpr have been described, their biological significance remains unclear. Here, we use complementary unbiased mass spectrometry-based approaches to demonstrate that Vpr is both necessary and sufficient for the DCAF1/DDB1/CUL4 E3 ubiquitin ligase-mediated degradation of at least 38 cellular proteins, causing systems-level changes to the cellular proteome. We therefore propose that promiscuous targeting of multiple host factors underpins complex Vpr-dependent cellular phenotypes and validate this in the case of G2/M cell cycle arrest. Our model explains how Vpr modulates so many cell biological processes and why the functional consequences of previously described Vpr targets, identified and studied in isolation, have proved elusive.This work was supported by the Wellcome Trust (PRF 210688/Z/18/Z to PJL), the MRC (CSF MR/P008801/1 to NJM), NHSBT (WPA15-02 to NJM), the NIHR Cambridge BRC, and a Wellcome Trust Strategic Award to the CIMR

Functional proteomic atlas of HIV infection in primary human CD4+ T cells.

Viruses manipulate host cells to enhance their replication, and the identification of cellular factors targeted by viruses has led to key insights into both viral pathogenesis and cell biology. In this study, we develop an HIV reporter virus (HIV-AFMACS) displaying a streptavidin-binding affinity tag at the surface of infected cells, allowing facile one-step selection with streptavidin-conjugated magnetic beads. We use this system to obtain pure populations of HIV-infected primary human CD4+ T cells for detailed proteomic analysis, and quantitate approximately 9000 proteins across multiple donors on a dynamic background of T cell activation. Amongst 650 HIV-dependent changes (q < 0.05), we describe novel Vif-dependent targets FMR1 and DPH7, and 192 proteins not identified and/or regulated in T cell lines, such as ARID5A and PTPN22. We therefore provide a high-coverage functional proteomic atlas of HIV infection, and a mechanistic account of host factors subverted by the virus in its natural target cell

Variable Sodium Absorption in a Low-Extinction Type Ia Supernova

Recent observations have revealed that some Type Ia supernovae exhibit narrow, time-variable Na I D absorption features. The origin of the absorbing material is controversial, but it may suggest the presence of circumstellar gas in the progenitor system prior to the explosion, with significant implications for the nature of the supernova progenitors. We present the third detection of such variable absorption, based on six epochs of high-resolution spectroscopy of the Type Ia supernova SN 2007le from Keck and the HET. The data span ~3 months, from 5 days before maximum light to 90 days after maximum. We find that one component of the Na D absorption lines strengthened significantly with time, indicating a total column density increase of ~2.5 x 10^12 cm^-2. The changes are most prominent after maximum light rather than at earlier times when the UV flux from the SN peaks. As with SN 2006X, we detect no change in the Ca II H&K lines over the same time period, rendering line-of-sight effects improbable and suggesting a circumstellar origin for the absorbing material. Unlike the previous two SNe exhibiting variable absorption, SN 2007le is not highly reddened (E_B-V = 0.27 mag), also pointing toward circumstellar rather than interstellar absorption. Photoionization models show that the data are consistent with a dense (10^7 cm^-3) cloud or clouds of gas located ~0.1 pc from the explosion. These results broadly support the single-degenerate scenario previously proposed to explain the variable absorption, with mass loss from a nondegenerate companion star responsible for providing the circumstellar gas. We also present tentative evidence for narrow Halpha emission associated with the SN, which will require followup observations at late times to confirm. [abridged]Comment: 16 pages, 10 figures (8 in color), 5 tables. Accepted for publication in Ap

NGC 2770 - a supernova Ib factory?

NGC 2770 has been the host of three supernovae of Type Ib during the last 10 years, SN 1999eh, SN 2007uy and SN 2008D. SN 2008D attracted special attention due to the serendipitous discovery of an associated X-ray transient. In this paper, we study the properties of NGC 2770 and specifically the three SN sites to investigate whether this galaxy is in any way peculiar to cause a high frequency of SNe Ib. We model the global SED of the galaxy from broadband data and derive a star-formation and SN rate comparable to the values of the Milky Way. We further study the galaxy using longslit spectroscopy covering the major axis and the three SN sites. From the spectroscopic study we find subsolar metallicities for the SN sites, a high extinction and a moderate star-formation rate. In a high resolution spectrum, we also detect diffuse interstellar bands in the line-of-sight towards SN 2008. A comparison of NGC 2770 to the global properties of a galaxy sample with high SN occurance (at least 3 SN in the last 100 years) suggests that NGC 2770 is not particularly destined to produce such an enhancement of observed SNe observed. Its properties are also very different from gamma-ray burst host galaxies. Statistical considerations on SN Ib detection rates give a probability of ~1.5% to find a galaxy with three Ib SNe detected in 10 years. The high number of rare Ib SNe in this galaxy is therefore likely to be a coincidence rather than special properties of the galaxy itself. NGC 2770 has a small irregular companion, NGC 2770B, which is highly starforming, has a very low mass and one of the lowest metallicities detected in the nearby universe as derived from longslit spectroscopy. In the most metal poor part, we even detect Wolf-Rayet features, against the current models of WR stars which require high metallicities.Comment: 15 pages, 10 figures, submitted to Ap

Microrheology with an anisotropic optical trap

Microrheology with optical tweezers (MOT) measurements are usually performed using optical traps that are close to isotropic across the plane being imaged, but little is known about what happens when this is not the case. In this work, we investigate the effect of anisotropic optical traps on microrheology measurements. This is an interesting problem from a fundamental physics perspective, but it also has practical ramifications because in 3D all optical traps are anisotropic due to the difference in the intensity distribution of the trapping laser along axes parallel and perpendicular to the direction of beam propagation. We find that attempting viscosity measurements with highly anisotropic optical traps will return spurious results, unless the axis with maximum variance in bead position is identified. However, for anisotropic traps with two axes of symmetry such as traps with an elliptical cross section, the analytical approach introduced in this work allows us to explore a wider range of time scales than those accessible with symmetric traps. We have also identified a threshold level of anisotropy in optical trap strength of ~ 30 %, below which conventional methods using a single arbitrary axis can still be used to extract valuable microrheological results. We envisage that the outcomes of this study will have important practical ramifications on how all MOT measurements should be conducted and analysed in future applications

Stabilizing bubble and droplet interfaces using dipeptide hydrogels

Hydrophobic dipeptide molecules can be used to create interfacial films covering bubbles and droplets made from a range of oils. At high pH, the dipeptide molecules form micelles which transform into a hydrogel of fibres in response to the addition of salt. We characterize the properties of the hydrogel for two different salt (MgSO4) concentrations and then we use these gels to stabilize interfaces. Under high shear, the hydrogel is disrupted and will reform around bubbles or droplets. Here, we reveal that at low dipeptide concentration, the gel is too weak to prevent ripening of the bubbles; this then reduces the long-term stability of the foam. Under the same conditions, emulsions prepared from some oils are highly stable. We examine the wetting properties of the oil droplets at a hydrogel surface as a guide to the resulting emulsions