Inhibitors of the 5-lipoxygenase arachidonic acid pathway induce ATP release and ATP-dependent organic cation transport in macrophages

AbstractWe have previously described that arachidonic acid (AA)-5-lipoxygenase (5-LO) metabolism inhibitors such as NDGA and MK886, inhibit cell death by apoptosis, but not by necrosis, induced by extracellular ATP (ATPe) binding to P2X7 receptors in macrophages. ATPe binding to P2X7 also induces large cationic and anionic organic molecules uptake in these cells, a process that involves at least two distinct transport mechanisms: one for cations and another for anions. Here we show that inhibitors of the AA-5-LO pathway do not inhibit P2X7 receptors, as judged by the maintenance of the ATPe-induced uptake of fluorescent anionic dyes. In addition, we describe two new transport phenomena induced by these inhibitors in macrophages: a cation-selective uptake of fluorescent dyes and the release of ATP. The cation uptake requires secreted ATPe, but, differently from the P2X7/ATPe-induced phenomena, it is also present in macrophages derived from mice deficient in the P2X7 gene. Inhibitors of phospholipase A2 and of the AA-cyclooxygenase pathway did not induce the cation uptake. The uptake of non-organic cations was investigated by measuring the free intracellular Ca2+ concentration ([Ca2+]i) by Fura-2 fluorescence. NDGA, but not MK886, induced an increase in [Ca2+]i. Chelating Ca2+ ions in the extracellular medium suppressed the intracellular Ca2+ signal without interfering in the uptake of cationic dyes. We conclude that inhibitors of the AA-5-LO pathway do not block P2X7 receptors, trigger the release of ATP, and induce an ATP-dependent uptake of organic cations by a Ca2+- and P2X7-independent transport mechanism in macrophages

Measurement of the W gamma Production Cross Section in Proton-Proton Collisions at root s=13 TeV and Constraints on Effective Field Theory Coefficients

A fiducial cross section for W gamma production in proton-proton collisions is measured at a center-of-mass energy of 13 TeV in 137 fb(-1) of data collected using the CMS detector at the LHC. The W -> e nu and mu nu decay modes are used in a maximum-likelihood fit to the lepton-photon invariant mass distribution to extract the combined cross section. The measured cross section is compared with theoretical expectations at next-to-leading order in quantum chromodynamics. In addition, 95% confidence level intervals are reported for anomalous triple-gauge couplings within the framework of effective field theory.Peer reviewe

Performance of the CMS muon trigger system in proton-proton collisions at √s = 13 TeV

The muon trigger system of the CMS experiment uses a combination of hardware and software to identify events containing a muon. During Run 2 (covering 2015-2018) the LHC achieved instantaneous luminosities as high as 2 × 10 cm s while delivering proton-proton collisions at √s = 13 TeV. The challenge for the trigger system of the CMS experiment is to reduce the registered event rate from about 40 MHz to about 1 kHz. Significant improvements important for the success of the CMS physics program have been made to the muon trigger system via improved muon reconstruction and identification algorithms since the end of Run 1 and throughout the Run 2 data-taking period. The new algorithms maintain the acceptance of the muon triggers at the same or even lower rate throughout the data-taking period despite the increasing number of additional proton-proton interactions in each LHC bunch crossing. In this paper, the algorithms used in 2015 and 2016 and their improvements throughout 2017 and 2018 are described. Measurements of the CMS muon trigger performance for this data-taking period are presented, including efficiencies, transverse momentum resolution, trigger rates, and the purity of the selected muon sample. This paper focuses on the single- and double-muon triggers with the lowest sustainable transverse momentum thresholds used by CMS. The efficiency is measured in a transverse momentum range from 8 to several hundred GeV

Search for long-lived particles decaying to jets with displaced vertices in proton-proton collisions at root s=13 Te V

A search is presented for long-lived particles produced in pairs in proton-proton collisions at the LHC operating at a center-of-mass energy of 13 TeV. The data were collected with the CMS detector during the period from 2015 through 2018, and correspond to a total integrated luminosity of 140 fb(-1). This search targets pairs of long-lived particles with mean proper decay lengths between 0.1 and 100 mm, each of which decays into at least two quarks that hadronize to jets, resulting in a final state with two displaced vertices. No significant excess of events with two displaced vertices is observed. In the context of R-parity violating supersymmetry models, the pair production of long-lived neutralinos, gluinos, and top squarks is excluded at 95% confidence level for cross sections larger than 0.08 fb, masses between 800 and 3000 GeV, and mean proper decay lengths between 1 and 25 mm.Peer reviewe

Search for top squark production in fully hadronic final states in proton-proton collisions at root s=13 TeV

A search for production of the supersymmetric partners of the top quark, top squarks, is presented. The search is based on proton-proton collision events containing multiple jets, no leptons, and large transverse momentum imbalance. The data were collected with the CMS detector at the CERN LHC at a center-of-mass energy of 13 TeV, and correspond to an integrated luminosity of 137 fb(-1). The targeted signal production scenarios are direct and gluino-mediated top squark production, including scenarios in which the top squark and neutralino masses are nearly degenerate. The search utilizes novel algorithms based on deep neural networks that identify hadronically decaying top quarks and W bosons, which are expected in many of the targeted signal models. No statistically significant excess of events is observed relative to the expectation from the standard model, and limits on the top squark production cross section are obtained in the context of simplified supersymmetric models for various production and decay modes. Exclusion limits as high as 1310 GeVare established at the 95% confidence level on the mass of the top squark for direct top squark production models, and as high as 2260 GeV on the mass of the gluino for gluino-mediated top squark production models. These results represent a significant improvement over the results of previous searches for supersymmetry by CMS in the same final state.Peer reviewe

Search for a heavy resonance decaying to a top quark and a w boson at √s = 13 tev in the fully hadronic final state

A search for a heavy resonance decaying to a top quark and a W boson in the fully hadronic final state is presented. The analysis is performed using data from proton-proton collisions at a center-of-mass energy of 13 TeV, corresponding to an integrated luminosity of 137 fb−1 recorded by the CMS experiment at the LHC. The search is focused on heavy resonances, where the decay products of each top quark or W boson are expected to be reconstructed as a single, large-radius jet with a distinct substructure. The production of an excited bottom quark, b*, is used as a benchmark when setting limits on the cross section for a heavy resonance decaying to a top quark and a W boson. The hypotheses of b* quarks with left-handed, right-handed, and vector-like chiralities are excluded at 95% confidence level for masses below 2.6, 2.8, and 3.1 TeV, respectively. These are the most stringent limits on the b* quark mass to date, extending the previous best limits by almost a factor of two

Measurements of production cross sections of the Higgs boson in the four-lepton final state in proton–proton collisions at √s=13Te

Production cross sections of the Higgs boson are measured in the H → Z Z → 4 ℓ (ℓ=e,μ) decay channel. A data sample of proton–proton collisions at a center-of-mass energy of 13Te, collected by the CMS detector at the LHC and corresponding to an integrated luminosity of 137fb-1 is used. The signal strength modifier μ, defined as the ratio of the Higgs boson production rate in the 4 ℓ channel to the standard model (SM) expectation, is measured to be μ=0.94±0.07(stat)-0.08+0.09(syst) at a fixed value of mH=125.38Ge. The signal strength modifiers for the individual Higgs boson production modes are also reported. The inclusive fiducial cross section for the H → 4 ℓ process is measured to be 2.84-0.22+0.23(stat)-0.21+0.26(syst)fb, which is compatible with the SM prediction of 2.84±0.15fb for the same fiducial region. Differential cross sections as a function of the transverse momentum and rapidity of the Higgs boson, the number of associated jets, and the transverse momentum of the leading associated jet are measured. A new set of cross section measurements in mutually exclusive categories targeted to identify production mechanisms and kinematical features of the events is presented. The results are in agreement with the SM predictions.STFC, Marie-Curie program and the European Research Council and Horizon 2020 Gran

Identification of εPKC targets during cardiac ischemic injury

Submitted by Sandra Infurna ([email protected]) on 2017-05-02T12:44:55Z No. of bitstreams: 1 andre_teixeira_etal_IOC_2012.pdf: 1862111 bytes, checksum: a1e9d15c61bac74b6d078f3163f11e37 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2017-05-02T12:56:37Z (GMT) No. of bitstreams: 1 andre_teixeira_etal_IOC_2012.pdf: 1862111 bytes, checksum: a1e9d15c61bac74b6d078f3163f11e37 (MD5)Made available in DSpace on 2017-05-02T12:56:37Z (GMT). No. of bitstreams: 1 andre_teixeira_etal_IOC_2012.pdf: 1862111 bytes, checksum: a1e9d15c61bac74b6d078f3163f11e37 (MD5) Previous issue date: 2012Stanford University School of Medicine. Department of Chemical and Systems Biology. São Paulo, SP, Brasil.Instituto do Coração. São Paulo, SP, Brasil.Stanford University School of Medicine. Department of Chemical and Systems Biology. São Paulo, SP, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ. Brasil.Instituto do Coração. São Paulo, SP, Brasil.tanford University School of Medicine. Department of Chemical and Systems Biology. São Paulo, SP, Brasil.Universidade de São Paulo. Instituto de Química. Departamento de Bioquímica. São Paulo, SP, Brasil.Background—Activation of ε protein kinase C (εPKC) protects hearts from ischemic injury. However, some of the mechanism(s) of εPKC mediated cardioprotection are still unclear. Identification of εPKC targets may aid to elucidate εPKC–mediated cardioprotective mechanisms. Previous studies, using a combination of εPKC transgenic mice and difference in gel electrophoresis (DIGE), identified a number of proteins involved in glucose metabolism, whose expression was modified by εPKC. These studies, were accompanied by metabolomic analysis, and suggested that increased glucose oxidation may be responsible for the cardioprotective effect of εPKC. However, whether these εPKC-mediated alterations were due to differences in protein expression or phosphorylation was not determined

Modulation of Mouse Embryonic Stem Cell Proliferation and Neural Differentiation by the P2X7 Receptor

<div><p>Background</p><p>Novel developmental functions have been attributed to the P2X7 receptor (P2X7R) including proliferation stimulation and neural differentiation. Mouse embryonic stem cells (ESC), induced with retinoic acid to neural differentiation, closely assemble processes occurring during neuroectodermal development of the early embryo.</p><p>Principal Findings</p><p>P2X7R expression together with the pluripotency marker Oct-4 was highest in undifferentiated ESC. In undifferentiated cells, the P2X7R agonist Bz-ATP accelerated cell cycle entry, which was blocked by the specific P2X7R inhibitor KN-62. ESC induced to neural differentiation with retinoic acid, reduced Oct-4 and P2X7R expression. P2X7R receptor-promoted intracellular calcium fluxes were obtained at lower Bz-ATP ligand concentrations in undifferentiated and in neural-differentiated cells compared to other studies. The presence of KN-62 led to increased number of cells expressing SSEA-1, Dcx and β3-tubulin, as well as the number of SSEA-1 and β3-tubulin-double-positive cells confirming that onset of neuroectodermal differentiation and neuronal fate determination depends on suppression of P2X7R activity. Moreover, an increase in the number of Ki-67 positive cells in conditions of P2X7R inhibition indicates rescue of progenitors into the cell cycle, augmenting the number of neuroblasts and consequently neurogenesis.</p><p>Conclusions</p><p>In embryonic cells, P2X7R expression and activity is upregulated, maintaining proliferation, while upon induction to neural differentiation P2X7 receptor expression and activity needs to be suppressed.</p></div

Immunofluorescence studies of P2X7 receptor and differentiation-stage specific proteins in undifferentiated and differentiated ESC.

<p>P2X7R expression and co-localization with differentiation-stage specific proteins was determined by confocal microscopy and immunofluorescence assays as described in Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096281#s2" target="_blank">Methods</a>. (<b>A</b>). Left panel: Co-localization between SSEA-1 (stage-specific embryonic antigen-1) and P2X7R immunofluorescence in undifferentiated ES cells. Co-localization of the protein is shown by Z-stack analysis. Right panel: Double-immunostaining for nestin (neural stem and precursor marker) and P2X7R expression in cells induced to differentiation for 8 days. (<b>B</b>). Left panel: Immunostaining for Oct-4 (pluripotency marker) in undifferentiated cells. Middle panel: Immunostaining for Oct-4 and SSEA-1 in undifferentiated cells. Right panel: Staining pattern for the neuron-specific marker β-tubulin in neural-differentiated cells. Cell nuclei were visualized by DAPI staining. Scale bar, 50 µm.</p