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neutrophil substrate

P Nyssen, A Mouithys-Mickalad, G Minguet, E Sauvage, J Wouters, T Franck, M Hoebeke
Morphine is an opioid alkaloid commonly used in clinical practice for its analgesic properties. The phenolic hydroxyl group of that phenanthrene derivative is pivotal for binding to opioid receptors but it may also be responsible for the antioxidant behavior of morphine reported in several in vitro experiments. In this study, we assessed the effect of morphine on myeloperoxidase (MPO), a hemic enzyme from azurophilic granules of polymorphonuclear neutrophils involved in the production of cytotoxic and microbicidal reactive oxidants during inflammatory response...
July 10, 2018: Biochimica et Biophysica Acta
Shiliang Feng, Lüwen Zhou, Yan Zhang, Shouqin Lü, Mian Long
Directional neutrophil migration during human immune responses is a highly coordinated process regulated by both biochemical and biomechanical environments. In this paper, we developed an integrative mathematical model of neutrophil migration using a lattice Boltzmann-particle method built in-house to solve the moving boundary problem with spatiotemporal regulation of biochemical components. The mechanical features of the cell cortex are modeled by a series of spring-connected nodes representing discrete cell-substrate adhesive sites...
July 2, 2018: Biomechanics and Modeling in Mechanobiology
Rao Fu, Soichiro Tajima, Kimitaka Suetsugu, Hiroyuki Watanabe, Nobuaki Egashira, Satohiro Masuda
Calcineurin inhibitors (CNIs), such as cyclosporine A and tacrolimus, are widely used immunosuppressive agents for the prevention of post-transplantation rejection and have improved 1-year graft survival rates by up to 90%. However, CNIs can induce severe reactions, such as acute or chronic allograft nephropathy, hypertension, and neurotoxicity. Because CNIs have varied bioavailabilities, narrow therapeutic ranges, and individual propensities for toxic effects, therapeutic drug monitoring is necessary for all CNIs...
June 27, 2018: Acta Pharmacologica Sinica
Mieke Gouwy, Mieke De Buck, Sara Abouelasrar Salama, Jennifer Vandooren, Sofie Knoops, Noëmie Pörtner, Lotte Vanbrabant, Nele Berghmans, Ghislain Opdenakker, Paul Proost, Jo Van Damme, Sofie Struyf
Serum amyloid A1 (SAA1) is a prototypic acute phase protein, induced to extremely high levels by physical insults, including inflammation and infection. Human SAA and its NH2 -terminal part have been studied extensively in the context of amyloidosis. By contrast, little is known about COOH-terminal fragments of SAA. Intact SAA1 chemoattracts leukocytes via the G protein-coupled receptor formyl peptide receptor like 1/formyl peptide receptor 2 (FPR2). In addition to direct leukocyte activation, SAA1 induces chemokine production by signaling through toll-like receptor 2...
2018: Frontiers in Immunology
Jingjing Tang, Jeremy M Frey, Carole L Wilson, Angela Moncada-Pazos, Clémence Levet, Matthew Freeman, Michael E Rosenfeld, E Richard Stanley, Elaine W Raines, Karin E Bornfeldt
Macrophages are prominent cells in acute and chronic inflammatory diseases. Recent studies highlight a role for macrophage proliferation post monocyte recruitment in inflammatory conditions. Using an acute peritonitis model, we identify a significant defect in macrophage proliferation in mice lacking leukocyte transmembrane protease ADAM17. The defect is associated with decreased levels of macrophage colony-stimulating factor-1 (CSF-1) in the peritoneum, and is rescued by intraperitoneal injection of CSF-1...
June 11, 2018: Molecular and Cellular Biology
L L K Leung, J Morser
Two basic carboxypeptidases, carboxypeptidase B2 (CPB2) and carboxypeptidase N (CPN) are present in plasma. CPN is constitutively active, whereas CPB2 circulates as a precursor, procarboxypeptidase B2 (proCPB2), that needs to be activated by the thrombin-thrombomodulin complex or plasmin bound to glycosaminoglycans. The substrate specificities of CPB2 and CPN are similar; they both remove C-terminal basic amino acids from bioactive peptides and proteins, thereby inactivating them. The complement cascade is a cascade of proteases and cofactors activated by pathogens or dead cells, divided into two phases, with the second phase only being triggered if sufficient C3b is present...
June 8, 2018: Journal of Thrombosis and Haemostasis: JTH
Mona H C Biermann, Sebastian Boeltz, Elmar Pieterse, Jasmin Knopf, Jürgen Rech, Rostyslav Bilyy, Johan van der Vlag, Angela Tincani, Jörg H W Distler, Gerhard Krönke, Georg Andreas Schett, Martin Herrmann, Luis E Muñoz
Deficient clearance of apoptotic cells reportedly contributes to the etiopathogenesis of the autoimmune disease systemic lupus erythematosus (SLE). Based on this knowledge, we developed a highly specific and sensitive test for the detection of SLE autoantibodies (AAb) utilizing secondary NEcrotic cell (SNEC)-derived material as a substrate. The goal of the present study was to validate the use of SNEC as an appropriate antigen for the diagnosis of SLE in large cohort of patients. We confirmed the presence of apoptotically modified autoantigens on SNEC (dsDNA, high mobility group box 1 protein, apoptosis-associated chromatin modifications, e...
2018: Frontiers in Immunology
Päivi Sirniö, Anne Tuomisto, Taina Tervahartiala, Timo Sorsa, Kai Klintrup, Toni Karhu, Karl-Heinz Herzig, Jyrki Mäkelä, Tuomo J Karttunen, Tuula Salo, Markus J Mäkinen, Juha P Väyrynen
BACKGROUND: Matrix metalloproteinase-8 (MMP-8) is a protease mainly expressed by neutrophils that cleaves numerous substrates, including collagens and cytokines. We have previously shown that serum MMP-8 levels increase in colorectal cancer (CRC) and correlate with distant metastasis. However, short follow-up in our prospective cohort did not enable survival analyses at the time of the first publication. METHODS: Preoperative serum MMP-8 levels were measured by immunofluorometric assay in 271 CRC patients and related to clinicopathological parameters, markers of systemic inflammation (modified Glasgow Prognostic Score, mGPS; serum levels of C-reactive protein (CRP), albumin and 13 cytokines), the density of six types of tumour-infiltrating immune cells and survival...
May 29, 2018: British Journal of Cancer
Dimitry Y Sorokin, Alexander Y Merkel, Ben Abbas, Kira S Makarova, W Irene C Rijpstra, M Koenen, Jaap S Sinninghe Damsté, Erwin A Galinski, Eugene V Koonin, Mark C M van Loosdrecht
Methanogenic enrichments from hypersaline lakes at moderate thermophilic conditions have resulted in the cultivation of an unknown deep lineage of euryarchaeota related to the class Halobacteria. Eleven soda lake isolates and three salt lake enrichment cultures were methyl-reducing methanogens that utilize C1 methylated compounds as electron acceptors and H2 or formate as electron donors, but they were unable to grow on either substrates alone or to form methane from acetate. They are extreme halophiles, growing optimally at 4 M total Na+ and the first representatives of methanogens employing the 'salt-in' osmoprotective mechanism...
May 21, 2018: International Journal of Systematic and Evolutionary Microbiology
Nathalie Burg, Steven Swendeman, Stefan Worgall, Timothy Hla, Jane E Salmon
OBJECTIVE: Immune complex (IC) deposition activates neutrophils (PMN), increases vascular permeability and leads to organ damage in SLE and RA. The bioactive lipid sphingosine-1-phosphate (S1P), acting via S1P receptor 1 (S1P1 ), is a key regulator of endothelial cell (EC) barrier function. We hypothesized that augmenting EC integrity via S1P1 signaling would attenuate inflammatory injury mediated by ICs. METHODS: In vitro barrier function was assessed in human umbilical vein endothelial cells (HUVECs) by Electric Cell-substrate Impedance Sensing (ECIS)...
May 21, 2018: Arthritis & Rheumatology
Youssef Z Wadghiri, Dung Minh Hoang, Anita Leporati, Matthew J Gounis, Aurora Rodríguez-Rodríguez, Mary L Mazzanti, John P Weaver, Ajay K Wakhloo, Peter Caravan, Alexei A Bogdanov
Progress in clinical development of magnetic resonance imaging (MRI) substrate-sensors of enzymatic activity has been slow partly due to the lack of human efficacy data. We report here a strategy that may serve as a shortcut from bench to bedside. We tested ultra high-resolution 7T MRI (µMRI) of human surgical histology sections in a 3-year IRB approved, HIPAA compliant study of surgically clipped brain aneurysms. µMRI was used for assessing the efficacy of MRI substrate-sensors that detect myeloperoxidase activity in inflammation...
May 16, 2018: Scientific Reports
C M Scholte, D C Nelson, M Garcia, S B Linden, T H Elsasser, S Kahl, Y Qu, K M Moyes
Mastitis is the leading cause of antimicrobial use on dairy farms. The potential for antimicrobial resistance has led to the examination of alternative strategies for controlling mastitis. One such alternative is PlyC, a potent peptidoglycan hydrolase derived from the streptococcal C1 bacteriophage that causes targeted lysis of the cell wall of Streptococcus uberis. At a concentration of 1.0 μg/mL, recombinant PlyC can induce lytic activity, suggesting that a low dose may successfully eliminate infection. We evaluated the dose effect of PlyC (1-50 µg/mL) on cytotoxicity and oxidative response on bovine blood polymorphonuclear leukocytes (PMN) obtained from 12 healthy, mid-lactation primiparous dairy cows...
July 2018: Journal of Dairy Science
Michael Thorpe, Zhirong Fu, Gurdeep Chahal, Srinivas Akula, Jukka Kervinen, Lawrence de Garavilla, Lars Hellman
Human neutrophils express at least four active serine proteases, cathepsin G, N-elastase, proteinase 3 and neutrophil serine protease 4 (NSP4). They have all been extensively studied due to their importance in neutrophil biology and immunity. However, their extended cleavage specificities have never been determined in detail. Here we present a detailed cleavage specificity analysis of human cathepsin G (hCG). The specificity was determined by phage display analysis and the importance of individual amino acids in and around the cleavage site was then validated using novel recombinant substrates...
2018: PloS One
Mamatha Kunder, V Lakshmaiah, A V Moideen Kutty
Dengue fever (DF) is characterized by systemic inflammatory response including neutrophil activation leading to uncontrolled elastase activity. This study was aimed to measure the activity of plasma neutrophil elastase (NE), its endogenous inhibitors α1 -antitrypsin (α1 -AT) and α2 -macroglobulin (α2 -MG) and elastase in complex with α1 -AT (NE-α1 -AT complex) in DF. 50 dengue patients [39 DF and 11 dengue hemorrhagic fever (DHF)] and 52 healthy subjects were included in the study. NE was measured using N -succinyl-tri-alanine- p -nitroanilide as substrate...
April 2018: Indian Journal of Clinical Biochemistry: IJCB
Jeong Yoon Kim, Yan Wang, Zia Uddin, Yeong Hun Song, Zuo Peng Li, Janar Jenis, Ki Hun Park
Flemingia philippinensis has been used throughout history to cure rheumatism associated with neutrophil elastase (NE). In this study, we isolated sixteen NE inhibitory flavonoids (1-16), including the most potent and abundant prenyl isoflavones (1-9), from the F. philippinensis plant. These prenyl isoflavones (2, 3, 5, 7, and 9) competitively inhibited NE, with IC50 values of 1.3-12.0 μM. In addition, they were reversible, simple, slow-binding inhibitors according to their respective parameters. Representative compound 3 had an IC50  = 1...
August 2018: Bioorganic Chemistry
Paulo C Maffía, Diego Guerrieri, Ximena Villalonga, Fiorella Caro, Sonia Gómez, Nancy Tateosian, Betiana P Bogado, Mercedes L Sánchez, Nella Ambrosi, Eduardo Chuluyan
Secretory Leukocyte Proteinase Inhibitor (SLPI) is an antiinflammatory peptide that blocks the activity of serine proteases, primarily the neutrophil elastase. In an attempt to direct the activity of SLPI on inflamed sites, a chimera consisting of the transglutaminase II substrate domain of trappin 2 (cementoin), and the mature SLPI protein was constructed. Cell attachment and biological activity were compared between SLPI and this chimera. By using whole cell ELISA, fluorescence microscopy and flow cytometry assays we observed that the cementoin-SLPI fusion protein (FP) but not SLPI attached to a human lung epithelial cell line and monocytes...
March 28, 2018: Scientific Reports
Sarah L King, Christoffer K Goth, Ulrich Eckhard, Hiren J Joshi, Amalie D Haue, Sergey Y Vakhrushev, Katrine T Schjoldager, Christopher M Overall, Hans H Wandall
Proteolytic processing is an irreversible post-translational modification functioning as a ubiquitous regulator of cellular activity. Protease activity is tightly regulated via control of gene expression, enzyme and substrate compartmentalization, zymogen activation, enzyme inactivation, and substrate availability. Emerging evidence suggests that proteolysis can also be regulated by substrate glycosylation and that glycosylation of individual sites on a substrate can decrease or, in rare cases, increase its sensitivity to proteolysis...
May 18, 2018: Journal of Biological Chemistry
Svetlana N Pleskova, Ekaterina N Gorshkova, Ruslan N Kriukov
In the process of performing their protective functions, neutrophils can form neutrophil extracellular traps (NETs), consisting of DNA in combination with enzymes and histones. The aim of the study was to determine the dynamics of the formation of NETs under the influence of opsonized Staphylococcus aureus and to determine the morphological features of their development in real time by atomic force microscopy. It was found that the maximum formation of NETs was observed after 3 hours of co-incubation of neutrophils and opsonized S...
July 2018: Journal of Molecular Recognition: JMR
Catherine M Dickinson, Brian W LeBlanc, Muhammad M Edhi, Daithi S Heffernan, Mohd Hafeez Faridi, Vineet Gupta, William G Cioffi, Xian O'Brien, Jonathan S Reichner
Background: Multi-organ failure occurs during critical illness and is mediated in part by destructive neutrophil-to-endothelial interactions. The β2 integrin receptor, CR3 (complement receptor 3; Mac-1; CD11b/CD18), which binds endothelial intercellular adhesion molecule-1 (ICAM-1), plays a key role in promoting the adhesion of activated neutrophils to inflamed endothelia which, when prolonged and excessive, can cause vascular damage. Leukadherin-1 (LA-1) is a small molecule allosteric activator of CR3 and has been shown to promote adhesion of blood neutrophils to inflamed endothelium and restrict tissue infiltration...
2018: Journal of Intensive Care
Emmet A Francis, Volkmar Heinrich
Global bursts in free intracellular calcium (Ca2+ ) are among the most conspicuous signaling events in immune cells. To test the common view that Ca2+ bursts mediate rearrangement of the actin cytoskeleton in response to the activation of G protein-coupled receptors, we combined single-cell manipulation with fluorescence imaging and monitored the Ca2+ concentration in individual human neutrophils during complement-mediated chemotaxis. By decoupling purely chemotactic pseudopod formation from cell-substrate adhesion, we showed that physiological concentrations of anaphylatoxins, such as C5a, induced nonadherent human neutrophils to form chemotactic pseudopods but did not elicit Ca2+ bursts...
March 13, 2018: Science Signaling
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