Structural and energetic characteristics of the heparin-binding site in antithrombotic protein C.
AuthorsFriedrich U, Blom AM, Dahlbäck B, Villoutreix BO,
JournalJ Biol Chem
PubMed ID11316800
'Human activated protein C (APC) is a key component of a natural anticoagulant system that regulates blood coagulation. In vivo, the catalytic activity of APC is regulated by two serpins, alpha1-antitrypsin and the protein C inhibitor (PCI), the inhibition by the latter being stimulated by heparin. We have identified a ... More
FRET measurements of intracellular cAMP concentrations and cAMP analog permeability in intact cells.
'Real-time measurements of second messengers in living cells, such as cAMP, are usually performed by ratiometric fluorescence resonance energy transfer (FRET) imaging. However, correct calibration of FRET ratios, accurate calculations of absolute cAMP levels and actual permeabilities of different cAMP analogs have been challenging. Here we present a protocol that ... More
A critical role for choline kinase-alpha in the aggressiveness of bladder carcinomas.
AuthorsHernando E, Sarmentero-Estrada J, Koppie T, Belda-Iniesta C, Ramírez de Molina V, Cejas P, Ozu C, Le C, Sánchez JJ, González-Barón M, Koutcher J, Cordón-Cardó C, Bochner BH, Lacal JC, Ramírez de Molina A,
JournalOncogene
PubMed ID19448670
'Bladder cancer is one of the most common causes of death in industrialized countries. New tumor markers and therapeutic approaches are still needed to improve the management of bladder cancer patients. Choline kinase-alpha (ChoKalpha) is a metabolic enzyme that has a role in cell proliferation and transformation. Inhibitors of ChoKalpha ... More
Delivery of proteins into living cells by reversible membrane permeabilization with streptolysin-O.
AuthorsWalev I, Bhakdi SC, Hofmann F, Djonder N, Valeva A, Aktories K, Bhakdi S,
JournalProc Natl Acad Sci U S A
PubMed ID11248053
'The pore-forming toxin streptolysin O (SLO) can be used to reversibly permeabilize adherent and nonadherent cells, allowing delivery of molecules with up to 100 kDa mass to the cytosol. Using FITC-labeled albumin, 10(5)-10(6) molecules were estimated to be entrapped per cell. Repair of toxin lesions depended on Ca(2+)-calmodulin and on ... More
Fabrication of synthetic polymer coatings and their use in feeder-free culture of human embryonic stem cells.
The culture of human embryonic stem (hES) cells in defined and xenogeneic-free conditions will contribute substantially to future biotechnological and medical applications. To achieve this goal, we developed the first fully defined synthetic polymer coating poly[2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl)ammonium hydroxide] (PMEDSAH) that sustains long-term growth of hES cells in different culture media. ... More
Molecular rearrangements of the extracellular vestibule in NMDAR channels during gating.
Authors Sobolevsky Alexander I; Beck Christine; Wollmuth Lonnie P;
JournalNeuron
PubMed ID11779481
Many N-methyl-D-aspartate receptor (NMDAR) channel blockers that have therapeutic potential can be trapped in the closed state. Using a combination of the substituted cysteine accessibility method and open channel blockers, we found that the M3 segment forms the core of the extracellular vestibule, including a deep site for trapping blockers. ... More
Ascorbic-acid transporter Slc23a1 is essential for vitamin C transport into the brain and for perinatal survival.
Authors Sotiriou Sotiria; Gispert Suzana; Cheng Jun; Wang Yaohui; Chen Amy; Hoogstraten-Miller Shelley; Miller Georgina F; Kwon Oran; Levine Mark; Guttentag Susan H; Nussbaum Robert L;
JournalNat Med
PubMed ID11984580
The only proven requirement for ascorbic acid (vitamin C) is in preventing scurvy, presumably because it is a cofactor for hydroxylases required for post-translational modifications that stabilize collagen. We have created mice deficient in the mouse ortholog (solute carrier family 23 member 1 or Slc23a1) of a rat ascorbic-acid transporter, ... More
Statin prevents tissue factor expression in human endothelial cells: role of Rho/Rho-kinase and Akt pathways.
Authors Eto Masato; Kozai Toshiyuki; Cosentino Francesco; Joch Hana; Lüscher Thomas F;
JournalCirculation
PubMed ID11956113
BACKGROUND: Tissue factor plays a pivotal role in thrombus formation in acute coronary syndromes. However, the regulatory mechanisms underlying tissue factor expression are poorly understood. Statins are effective in patients with acute coronary syndromes. Hence, the aim of this study was to clarify in human endothelial cells the signaling pathways ... More
Malignant effusions and immunogenic tumour-derived exosomes.
BACKGROUND: Exosomes derived from tumours are small vesicles released in vitro by tumour cell lines in culture supernatants. To assess the role of these exosomes in vivo, we examined malignant effusions for their presence. We also investigated whether these exosomes could induce production of tumour-specific T cells when pulsed with ... More
Reduced interleukin-18 levels in BAL specimens from patients with asthma compared to patients with sarcoidosis and healthy control subjects.
Authors Ho Ling-Pei; Davis Margaret; Denison Alan; Wood Fraser T; Greening Andrew P;
JournalChest
PubMed ID12006423
STUDY OBJECTIVES: To investigate whether differing airway interleukin (IL)-18 levels may be implicated in the pathogenesis of asthma and sarcoidosis. SETTING: University teaching hospital. Patients and methods: IL-18 levels were measured in BAL fluid and in the supernatant of lipopolysaccharide (LPS)-stimulated alveolar macrophages obtained by BAL from 15 patients with ... More