Cytological profiling: providing more haystacks for chemists' needles

Conventional high-throughput 'chemical genetic' screening seeks to identify small-molecule inhibitors of a specific protein or pathway. A recent study describes how unbiased screening of cellular morphology, followed by affinity purification to identify targets of compounds with interesting effects, can lead to the identification of novel inhibitors

An engineered cardiac reporter cell line identifies human embryonic stem cell-derived myocardial precursors.

Unlike some organs, the heart is unable to repair itself after injury. Human embryonic stem cells (hESCs) grow and divide indefinitely while maintaining the potential to develop into many tissues of the body. As such, they provide an unprecedented opportunity to treat human diseases characterized by tissue loss. We have identified early myocardial precursors derived from hESCs (hMPs) using an α-myosin heavy chain (αMHC)-GFP reporter line. We have demonstrated by immunocytochemistry and quantitative real-time PCR (qPCR) that reporter activation is restricted to hESC-derived cardiomyocytes (CMs) differentiated in vitro, and that hMPs give rise exclusively to muscle in an in vivo teratoma formation assay. We also demonstrate that the reporter does not interfere with hESC genomic stability. Importantly, we show that hMPs give rise to atrial, ventricular and specialized conduction CM subtypes by qPCR and microelectrode array analysis. Expression profiling of hMPs over the course of differentiation implicate Wnt and transforming growth factor-β signaling pathways in CM development. The identification of hMPs using this αMHC-GFP reporter line will provide important insight into the pathways regulating human myocardial development, and may provide a novel therapeutic reagent for the treatment of cardiac disease

Inflation from Susy quantum cosmology

We propose a realization of inverted hybrid inflation scenario in the context of n=2 supersymmetric quantum cosmology. The spectrum of density fluctuations is calculated in the de Sitter regimen as a function of the gravitino and the Planck mass, and explicit forms for the wave function of the universe are found in the WKB regimen for a FRW closed and flat universes.Comment: 9 pages, one figure, to appear in Phys. Rev.

Pharmacologic Inhibition of the Anaphase-Promoting Complex Induces A Spindle Checkpoint-Dependent Mitotic Arrest in the Absence of Spindle Damage

SummaryMicrotubule inhibitors are important cancer drugs that induce mitotic arrest by activating the spindle assembly checkpoint (SAC), which, in turn, inhibits the ubiquitin ligase activity of the anaphase-promoting complex (APC). Here, we report a small molecule, tosyl-L-arginine methyl ester (TAME), which binds to the APC and prevents its activation by Cdc20 and Cdh1. A prodrug of TAME arrests cells in metaphase without perturbing the spindle, but nonetheless the arrest is dependent on the SAC. Metaphase arrest induced by a proteasome inhibitor is also SAC dependent, suggesting that APC-dependent proteolysis is required to inactivate the SAC. We propose that mutual antagonism between the APC and the SAC yields a positive feedback loop that amplifies the ability of TAME to induce mitotic arrest

Ubistatins Inhibit Proteasome-Dependent Degradation by Binding the Ubiquitin Chain

To identify previously unknown small molecules that inhibit cell cycle machinery, we performed a chemical genetic screen in Xenopus extracts. One class of inhibitors, termed ubistatins, blocked cell cycle progression by inhibiting cyclin B proteolysis and inhibited degradation of ubiquitinated Sic1 by purified proteasomes. Ubistatins blocked the binding of ubiquitinated substrates to the proteasome by targeting the ubiquitin-ubiquitin interface of Lys^(48)-linked chains. The same interface is recognized by ubiquitin-chain receptors of the proteasome, indicating that ubistatins act by disrupting a critical protein-protein interaction in the ubiquitin-proteasome system

Five Dimensional Rotating Black Hole in a Uniform Magnetic Field. The Gyromagnetic Ratio

In four dimensional general relativity, the fact that a Killing vector in a vacuum spacetime serves as a vector potential for a test Maxwell field provides one with an elegant way of describing the behaviour of electromagnetic fields near a rotating Kerr black hole immersed in a uniform magnetic field. We use a similar approach to examine the case of a five dimensional rotating black hole placed in a uniform magnetic field of configuration with bi-azimuthal symmetry, that is aligned with the angular momenta of the Myers-Perry spacetime. Assuming that the black hole may also possess a small electric charge we construct the 5-vector potential of the electromagnetic field in the Myers-Perry metric using its three commuting Killing vector fields. We show that, like its four dimensional counterparts, the five dimensional Myers-Perry black hole rotating in a uniform magnetic field produces an inductive potential difference between the event horizon and an infinitely distant surface. This potential difference is determined by a superposition of two independent Coulomb fields consistent with the two angular momenta of the black hole and two nonvanishing components of the magnetic field. We also show that a weakly charged rotating black hole in five dimensions possesses two independent magnetic dipole moments specified in terms of its electric charge, mass, and angular momentum parameters. We prove that a five dimensional weakly charged Myers-Perry black hole must have the value of the gyromagnetic ratio g=3.Comment: 23 pages, REVTEX, v2: Minor changes, v3: Minor change

Hierarchical Neutrino Mass Matrices, CP violation and Leptogenesis

In this work we study examples of hierarchical neutrino mass matrices inspired by family symmetries, compatible with experiments on neutrino oscillations, and for which there is a connection among the low energy CP violation phase associated to neutrino oscillations, the phases appearing in the amplitude of neutrinoless double beta decay, and the phases relevant for leptogenesis. In particular, we determine the predictions from a texture based on an underlying SU(3) family symmetry together with a GUT symmetry, and a strong hierarchy for the masses of the heavy right handed Majorana masses. We also give some examples of inverted hierarchies of neutrino masses, which may be motivated in the context of U(1) family symmetries.Comment: 34 pages. Replaced with published version -typos, corrections and references adde

Ubiquitin Chains Are Remodeled at the Proteasome by Opposing Ubiquitin Ligase and Deubiquitinating Activities

SummaryThe ubiquitin ligase Hul5 was recently identified as a component of the proteasome, a multisubunit protease that degrades ubiquitin-protein conjugates. We report here a proteasome-dependent conjugating activity of Hul5 that endows proteasomes with the capacity to extend ubiquitin chains. hul5 mutants show reduced degradation of multiple proteasome substrates in vivo, suggesting that the polyubiquitin signal that targets substrates to the proteasome can be productively amplified at the proteasome. However, the products of Hul5 conjugation are subject to disassembly by a proteasome-bound deubiquitinating enzyme, Ubp6. A hul5 null mutation suppresses a ubp6 null mutation, suggesting that a balance of chain-extending and chain-trimming activities is required for proper proteasome function. As the association of Hul5 with proteasomes was found to be strongly stabilized by Ubp6, these enzymes may be situated in proximity to one another. We propose that through dynamic remodeling of ubiquitin chains, proteasomes actively regulate substrate commitment to degradation