The rockefeller university

Cancer stem cells (CSCs) sustain tumor growth through their ability to self-renew and to generate differentiated progeny. These functions endow CSCs with the potential to initiate secondary tumors bearing characteristics similar to those of the parent. Recently the hair follicle stem cell marker CD34 was used to purify a CSC-like cell population from early skin tumors arising from treatment with 7,12-dimethylbenz[alpha] anthracene/12-o-tetradecanoylphorbol-13-acetate, which typically generates benign papillomas that occasionally progress to squamous cell carcinomas (SCCs). In the present study, we identify and characterize CSCs purified from malignant SCCs. We show that SCCs contain two highly tumorigenic CSC populations that differ in CD34 levels but are enriched for integrins and coexist at the SCC-stroma interface. Intriguingly, whether CD34(Io) or CD34(hi), alpha 6(hi)beta 1(hi) populations can initiate secondary tumors by serial limitdilution transplantation assays, but alpha 6(io)beta 1(io) populations cannot. Moreover, secondary tumors generated from a single CSC of either subtype contain both CD34(Io) and CD34(hi) alpha 6(hi)beta 1(hi)CSCs, indicating their nonhierarchical organization. Genomic profiling and hierarchical cluster analysis show that these two CSC subtypes share a molecular signature distinct from either the CD34(-) epidermal or the CD34hi hair follicle stem cell signature. Although closely related, alpha 6(hi)beta 1(hi)CD34(Io) and alpha 6(hi)beta 1(hi)CD34(hi) CSCs differ in cell-cycle gene expression and proliferation characteristics. Indeed, proliferation and expansion of alpha 6(hi)beta 1(hi)CD34(hi) CSCs is sensitive to whether they can initiate a TGF-beta receptor II-mediated response to counterbalance elevated focal adhesion kinase-mediated integrin signaling within the tumor. Overall, the coexistence and interconvertibility of CSCs with differing sensitivities to their microenvironment pose challenges and opportunities for SCC cancer therapies.

A search is performed for an excess of events, over the standard model expectations, with a photon, a lepton, and large missing transverse energy in pp collisions root s = 7TeV. Such events are expected in many new physics models, in particular a theory that is broken via a gauge-mediated mechanism, when the lightest and neutral gauginos are mass degenerate. The data sample used in this search to an integrated luminosity of 35 pb(-1) collected with the CMS detector at the No evidence of such an excess above the standard model backgrounds, dominated by production, is found. The results are presented as 95% confidence level upper limits on the cross section for a benchmark gauge-mediated scenario, and are then converted into exclusion limits on the squark, gluino, and wino masses.

The biochemical characterization of the bacterial transcription cycle has been greatly facilitated by the production and characterization of targeted RNA polymerase (RNAP) mutants. Traditionally, RNAP preparations containing mutant subunits have been produced by reconstitution of denatured RNAP subunits, a process that is undesirable for biophysical and structural studies. Although schemes that afford the production of in vivo-assembled, recombinant RNAP containing amino acid substitutions, insertions, or deletions in either the monomeric beta or beta' subunits have been developed, there is no such system for the production of in vivo-assembled, recombinant RNAP with mutations in the homodimeric alpha-subunits. Here, we demonstrate a strategy to generate in vivo-assembled, recombinant RNAP preparations free of the alpha C-terminal domain. Furthermore, we describe a modification of this approach that would permit the purification of in vivo-assembled, recombinant RNAP containing any alpha-subunit variant, including those variants that are lethal. Finally, we propose that these related approaches can be extended to generate in vivo-assembled, recombinant variants of other protein complexes containing homomultimers for biochemical, biophysical, and structural analyses.

Insomnia is a common problem for people with chronic pain. Cortical GABAergic neurons are part of the neurobiological substrate that underlies homeostatic sleep regulation. In the present study, we confirmed that sciatic nerve ligation caused thermal hyperalgesia and tactile allodynia in mice. In this experimental model for neuropathic pain, we found an increase in wakefulness and a decrease in non-rapid eye movement sleep under a neuropathic pain-like state. Under these conditions, membrane-bound GABA (c-aminobutyric acid) transporters (GATs) on activated glial fibrillary acidic protein-positive astrocytes were significantly increased in the cingulate cortex, and extracellular GABA levels in this area after depolarization were rapidly decreased by nerve injury. Furthermore, sleep disturbance induced by sciatic nerve ligation was improved by the intracingulate cortex injection of a GAT-3 inhibitor. These findings provide novel evidence that sciatic nerve ligation decreases extracellular-released GABA in the cingulate cortex of mice. These phenomena may, at least in part, explain the insomnia in patients with neuropathic pain. (C) 2011 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.

A study of event-shape observables in proton-antiproton collisions at root s = 1.96 TeV is presented. The data for this analysis were recorded by the CDF II Detector at the Tevatron Collider. The variables studied are the transverse thrust and thrust minor, both defined in the plane perpendicular to the beam direction. The observables are measured using energies from unclustered calorimeter cells. In addition to studies of the differential distributions, we present the dependence of event-shape mean values on the leading-jet transverse energy. Data are compared with PYTHIA Tune A and to resummed parton-level predictions that were matched to fixed-order results at next-to-leading-order (NLO) accuracy (NLO + NLL). Predictions from PYTHIA Tune A agree fairly well with the data. However, the underlying event contributes significantly to these observables, making it difficult to make direct comparisons to the NLO + NLL predictions, which do not account for the underlying event. To overcome this difficulty, we introduce a new observable, a weighted difference of the mean values of the thrust and thrust minor, which is less sensitive to the underlying event, allowing for a comparison with NLO + NLL. Both PYTHIA Tune A and the NLO + NLL calculations agree well within the 20% theoretical uncertainty with the data for this observable, indicating that perturbative QCD successfully describes shapes of the hadronic final states.

The ability to derive meaning from complex sensory input requires the integration of information over space and time, as well as cognitive mechanisms to shape that integration. We studied these processes in the primary visual cortex (V1), where neurons are thought to integrate visual inputs along contours defined by an association field (AF). We recorded extracellularly from single cells in macaque V1 to map the AF, by using an optimization algorithm to find the contours that maximally activated individual cells. We combined the algorithm with a delayed-match-to-sample task, to test how the optimal contours might be molded by the monkey's expectation for particular cue shapes. We found that V1 neurons were selective for complex shapes, a property previously ascribed to higher cortical areas. Furthermore, the shape selectivity was reprogrammed by perceptual task: Over the whole network, the optimal modes of geometric selectivity shifted between distinct subsets of the AF, alternately representing different stimulus features known to predominate in natural scenes. Our results suggest a general model of cortical function, whereby horizontal connections provide a broad domain of potential associations, and top-down inputs dynamically gate these linkages to task switch the function of a network.

The Y(1S), Y(2S), and Y(3S) production cross sections in proton-proton collisions at root s = 7 TeV are measured using a data sample collected with the CMS detector at the LHC, corresponding to an integrated luminosity of 3.1 +/- 0.3 pb(-1). Integrated over the rapidity range vertical bar y vertical bar < 2, we find the product of the Y(1S) production cross section and branching fraction to dimuons to be sigma(pp -> Y(1S)X) center dot B(Y(1S) -> mu(+)mu(-)) = 7.37 +/- 0.13(-0.42)(+0.61) +/- 0.81 nb, where the first uncertainty is statistical, the second is systematic, and the third is associated with the estimation of the integrated luminosity of the data sample. This cross section is obtained assuming unpolarized Y(1S) production. With the assumption of fully transverse or fully longitudinal production polarization, the measured cross section changes by about 20%. We also report the measurement of the Y(1S), Y(2S), and Y(3S) differential cross sections as a function of transverse momentum and rapidity.

The presence of multiple membrane-bound intracellular compartments is a major feature of eukaryotic cells. Many of the proteins required for formation and maintenance of these compartments share an evolutionary history. Here, we identify the SEA (Seh1-associated) protein complex in yeast that contains the nucleoporin Seh1 and Sec13, the latter subunit of both the nuclear pore complex and the COPII coating complex. The SEA complex also contains Npr2 and Npr3 proteins (upstream regulators of TORC1 kinase) and four previously uncharacterized proteins (Sea1-Sea4). Combined computational and biochemical approaches indicate that the SEA complex proteins possess structural characteristics similar to the membrane coating complexes COPI, COPII, the nuclear pore complex, and, in particular, the related Vps class C vesicle tethering complexes HOPS and CORVET. The SEA complex dynamically associates with the vacuole in vivo. Genetic assays indicate a role for the SEA complex in intracellular trafficking, amino acid biogenesis, and response to nitrogen starvation. These data demonstrate that the SEA complex is an additional member of a family of membrane coating and vesicle tethering assemblies, extending the repertoire of protocoatomer-related complexes. Molecular & Cellular Proteomics 10: 10.1074/mcp.M110.006478, 1-17, 2011.

The results of searches for new physics in events with two same-sign isolated leptons, hadronic jets, and missing transverse energy in the final state are presented. The searches use an integrated luminosity of 35 pb(-1) of pp collision data at a centre-of-mass energy of 7TeV collected by the CMS experiment at the LHC. The observed numbers of events agree with the standard model predictions, and no evidence for new physics is found. To facilitate the interpretation of our data in a broader range of new physics scenarios, information on our event selection, detector response, and efficiencies is provided.

Spindle checkpoint silencing is crucial for cell-cycle progression, but mechanisms underlying this process remain mysterious. Two papers, one in this issue of Developmental Cell (Meadows et al., 2011) and one in Current Biology (Rosenberg et al., 2011), begin to show how phosphatase PP(-)I -gamma connects chromosome-microtubule attachment with anaphase entry.