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neutrophil substrate

Christina N Como, Andrew N Bubak, Anna M Blackmon, Dallas Jones, Niklaus H Mueller, Richard Davidson, Maria A Nagel
Purpose: While VZV DNA and antigen have been detected in acute and chronic VZV keratitis, it is unclear whether productive infection of corneal cells is ongoing or whether residual, noninfectious VZV antigens elicit inflammation. Herein, we examined VZV-infected primary human corneal epithelial cells (HCECs) and keratocytes (HKs) to elucidate the pathogenesis of VZV keratitis. Methods: HCECs and HKs were mock- or VZV infected. Seven days later, cells were examined for morphology, proinflammatory cytokine and matrix metalloproteinase (MMP) release, ability to recruit peripheral blood mononuclear cells (PBMCs) and neutrophils, and MMP substrate cleavage...
February 1, 2019: Investigative Ophthalmology & Visual Science
Qiang Gu, Jin-Chao Hou, Xiang-Ming Fang
Background/aim: Sphingosine 1-phosphate receptor 2 (S1PR2), a member of the seven-transmembrane receptor family, can be activated by its natural ligand sphingosine 1-phosphate (S1P) to initiate signal transduction and is involved in a wide range of biological effects such as immune cell migration and vascular permeability. Its relationship with neuropathic pain (NP) has not been reported. In this study, the effects of S1PR2 on the development of NP were studied. Materials and methods: We generated a model of NP by partial sciatic nerve ligation (pSNL)...
February 11, 2019: Turkish Journal of Medical Sciences
Shuzheng Song, Yingyan Yu
The citrullination modification (Cit) of proteins has received increasing attention in recent years. This kind of protein modification was first discovered in autoimmune diseases such as rheumatoid arthritis. The citrullination modification process is catalyzed by the peptidyl arginine deiminases (PADIs) family. A well-known citrullination of histone involves the key mechanism of neutrophil extracellular traps (NETs) of inflammation in the peripheral blood. Further studies revealed that citrullination modification of proteins also involves in carcinogenesis in human being...
2019: Frontiers in Oncology
Charmaine N Nganje, Scott A Haynes, Christine M Qabar, Rachel C Lent, Elsa N Bou Ghanem, Mara G Shainheit
Streptococcus pneumoniae (Spn) is an asymptomatic colonizer of the human nasopharynx but can also cause disease in the inner ear, meninges, lung and blood. Although various mechanisms contribute to the effective clearance of Spn, opsonophagocytosis by neutrophils is perhaps most critical. Upon phagocytosis, Spn is exposed to various degradative molecules, including a family of neutrophil serine proteases (NSPs) that are stored within intracellular granules. Despite the critical importance of NSPs in killing Spn, the bacterial proteins that are degraded by NSPs leading to Spn death are still unknown...
2019: PloS One
Chien-Neng Wang, Yu-Chao Lin, Bo-Chun Chang, Ching-Hsien Chen, Reen Wu, Chen-Chen Lee
BACKGROUND: Myristoylated alanine-rich C kinase substrate (MARCKS), a protein kinase C (PKC) substrate, facilitates mucus production and neutrophil migration. However, the effects of therapeutic procedures targeting the phosphorylation site of MARCKS on steroid-resistant asthma and the mechanisms underlying such effects have not yet been investigated. We designed a peptide that targets the MARCKS phosphorylation site (MPS peptide) and assessed its therapeutic potential against steroid-resistant asthma...
February 1, 2019: British Journal of Pharmacology
Deborah Rudin, Angelo Lanzilotto, Fabio Bachmann, Catherine E Housecroft, Edwin C Constable, Jürgen Drewe, Manuel Haschke, Stephan Krähenbühl
Metamizole is an analgesic and antipyretic, but can cause neutropenia and agranulocytosis. We investigated the toxicity of the metabolites N-methyl-4-aminoantipyrine (MAA), 4-aminoantipyrine (AA), N-formyl-4-aminoantipyrine (FAA) and N-acetyl-4-aminoantipyrine (AAA) on neutrophil granulocytes and on HL60 cells (granulocyte precursor cell line). MAA, FAA, AA, and AAA (up to 100 µM) alone were not toxic for HL60 cells or granulocytes. In the presence of the myeloperoxidase substrate H2 O2 , MAA reduced cytotoxicity for HL60 cells at low (<50 µM), but increased cytotoxicity at 100 µM H2 O2 ...
January 14, 2019: Biochemical Pharmacology
Shi-Yu Liu, Hao Xiong, Rong-Rong Li, Wen-Chao Yang, Guang-Fu Yang
Neutrophil elastase (NE), a typical hematopoietic serine protease, has significant roles in inflammatory and immune responses, and thus highly associated with various diseases such as acute lung injury (ALI) and lung cancer. Rapid and accurate measurement of NE activity in biological systems is particularly important for understanding the role of NE in inflammatory diseases, as well as the clinic diagnosis. However, the specific detection and non-invasive imaging of NE in vivo remains a challenge. To address this issue, a small-molecule substrate-based near-infrared fluorogenic probe (NEP) for NE was constructed via incorporating pentafluoroethyl as the recognition group with a hemicyanine dye-based fluorophore...
January 10, 2019: Analytical Chemistry
Sheela Nagarkoti, Samreen Sadaf, Deepika Awasthi, Tulika Chandra, Kumaravelu Jagavelu, Sachin Kumar, Madhu Dikshit
Recent report from this lab has shown role of Rac2 in the translocation of iNOS to the phagosomal compartment of PMNs following phagocytosis of beads. Present study was undertaken to further assess the status and role of tetrahydrobiopterin (BH4 ), a redox sensitive cofactor, and L-arginine, the substrate of NOS, in sustained nitric oxide (.NO) production in killing of phagocytosed microbes ( E.coli ) by human PMNs. Time dependent study revealed consistent. NO and reactive oxygen species (ROS) production in the PMNs following phagocytosis of beads...
January 8, 2019: Free Radical Research
Farzaneh Atashrazm, Deborah Hammond, Gayathri Perera, Marc F Bolliger, Elie Matar, Glenda M Halliday, Birgitt Schüle, Simon J G Lewis, R Jeremy Nichols, Nicolas Dzamko
BACKGROUND: Leucine-rich repeat kinase 2 is a potential therapeutic target for the treatment of Parkinson's disease, and clinical trials of leucine-rich repeat kinase 2 inhibitors are in development. The objective of this study was to evaluate phosphorylation of a new leucine-rich repeat kinase 2 substrate, Rab10, for potential use as a target engagement biomarker and/or patient enrichment biomarker for leucine-rich repeat kinase 2 inhibitor clinical trials. METHODS: Peripheral blood mononuclear cells and neutrophils were isolated from Parkinson's disease patients and matched controls, and treated ex vivo with a leucine-rich repeat kinase 2 inhibitor...
December 30, 2018: Movement Disorders: Official Journal of the Movement Disorder Society
J J B C van Beers, J Vanderlocht, C Roozendaal, J Damoiseaux
The eventual presence of anti-neutrophil cytoplasmic antibodies (ANCA) can initially be screened with indirect immunofluorescence (IIF). The majority of laboratories that facilitate ANCA testing use commercial kits. Although in-house assays are not encouraged in routine clinical laboratories, knowledge on the methodological aspects of the assay remains of importance. These aspects include choice of substrate, choice of fixative, staining procedure, and interpretation procedure. In this paper details on the methodology are provided and discussed in the context of the clinical application...
2019: Methods in Molecular Biology
Michaela Finsterbusch, M Ursula Norman, Pam Hall, A Richard Kitching, Michael J Hickey
Platelet-leukocyte interactions promote acute glomerulonephritis. However, neither the nature of the interactions between platelets and immune cells nor the capacity of platelets to promote leukocyte activation has been characterized in this condition. We used confocal intravital microscopy to define the interactions of platelets with neutrophils, monocytes, and endothelial cells in glomerular capillaries in mice. In the absence of inflammation, platelets underwent rapid on/off interactions with immune cells...
February 2019: Kidney International
Consuelo Pasten, Cristóbal Alvarado, Jocelyn Rocco, Luis Contreras, Paula Aracena, Jéssica Liberona, Cristian Suazo, Luis Fernando Michea, Carlos E Irarrázabal
Upon renal ischemia-reperfusion (IR) injury, recruitment of neutrophils during the inflammatory process promotes local generation of oxygen and nitrogen reactive species which, in turn, are likely to exacerbate tissue damage. The mechanism by which iNOS is involved in IR has not been completely elucidated. In this work, the selective iNOS inhibitor L-N6 -(1-iminoethyl)lysine (L-NIL) and the NOS substrate L-Arginine (L-Arg), were employed to understand the role of NOS activity on the expression of particular target genes and the oxidative stress elicited after a 30 min of bilateral renal ischemia, followed by 48h reperfusion in BALB/c mice...
December 5, 2018: American Journal of Physiology. Renal Physiology
Jonathan W Handing, Stephanie A Ragland, Urmila V Bharathan, Alison K Criss
The mucosal inflammatory response to Neisseria gonorrhoeae (Gc) is characterized by recruitment of neutrophils to the site of infection. Gc survives exposure to neutrophils by limiting the ability of neutrophils to make antimicrobial products and by expressing factors that defend against these products. The multiple transferable resistance (Mtr) system is a tripartite efflux pump, comprised of the inner membrane MtrD, the periplasmic attachment protein MtrC, and the outer membrane channel MtrE. Gc MtrCDE exports a diverse array of substrates, including certain detergents, dyes, antibiotics, and host-derived antimicrobial peptides...
2018: Frontiers in Microbiology
Colin G Buss, Jaideep S Dudani, Reid T K Akana, Heather E Fleming, Sangeeta N Bhatia
BACKGROUND: Respiratory tract infections represent a significant public health risk, and timely and accurate detection of bacterial infections facilitates rapid therapeutic intervention. Furthermore, monitoring the progression of infections after intervention enables 'course correction' in cases where initial treatments are ineffective, avoiding unnecessary drug dosing that can contribute to antibiotic resistance. However, current diagnostic and monitoring techniques rely on non-specific or slow readouts, such as radiographic imaging and sputum cultures, which fail to specifically identify bacterial infections and take several days to identify optimal antibiotic treatments...
November 26, 2018: EBioMedicine
Yu Su, Shuting Xu, Jinan Zhang, Xiang-Jian Chen, Li-Ping Jiang, Tingting Zheng, Jun-Jie Zhu
Cardiorenal syndrome (CRS) has posed tremendous challenges in patient management, and the detection of serum biomarkers may provide opportunities for early diagnosis and effective treatment. Herein, we introduce a novel surface-enhanced Raman scattering (SERS)-based sandwich immunoassay platform to simultaneously detect cardiac troponin I (cTnI), N-terminal prohormone of brain natriuretic peptide (NT-ProBNP) and neutrophil gelatinase-associated lipocalin (NGAL) for the early diagnosis of CRS by using Raman reporter molecules-labeled Ag-Au nanostars (Ag-Au NSs) as nanotags and three-dimensional ordered macroporous (3DOM) Au-Ag-Au plasmonic array as substrate...
November 30, 2018: Analytical Chemistry
Camille V Goemans, François Beaufay, Isabelle S Arts, Rym Agrebi, Didier Vertommen, Jean-François Collet
Hypochlorous acid (bleach), an oxidizing compound produced by neutrophils, turns the Escherichia coli chaperedoxin CnoX into a powerful holdase protecting its substrates from bleach-induced aggregation. CnoX is well conserved in bacteria, even in non-infectious species unlikely to encounter this oxidant, muddying the role of CnoX in these organisms. Here, we found that CnoX in the non-pathogenic aquatic bacterium Caulobacter crescentus functions as a holdase that efficiently protects 50 proteins from heat-induced aggregation...
November 27, 2018: MBio
Zhirong Fu, Michael Thorpe, Srinivas Akula, Gurdeep Chahal, Lars T Hellman
Serine proteases are major granule constituents of several of the human hematopoietic cell lineages. Four proteolytically active such proteases have been identified in human neutrophils: cathepsin G (hCG), N-elastase (hNE), proteinase 3 (hPR-3), and neutrophil serine protease 4 (hNSP-4). Here we present the extended cleavage specificity of two of the most potent and most abundant of these enzymes, hNE and hPR-3. Their extended specificities were determined by phage display and by the analysis of a panel of chromogenic and recombinant substrates...
2018: Frontiers in Immunology
André Raposo Moreira Dias, Arianna Pina, Amelia Dean, Hans-Georg Lerchen, Michele Caruso, Fabio Gasparri, Ivan Fraietta, Sonia Troiani, Daniela Arosio, Laura Belvisi, Luca Pignataro, Alberto Dal Corso, Cesare Gennari
We took advantage of one of the hallmarks of cancers, i.e. the presence of tumor infiltrating cells of the immune system and leukocyte-secreted enzymes, to promote the activation of an anticancer drug at the tumor site. The peptidomimetic integrin ligand cyclo(DKP-RGD) was found to accumulate on the surface of αvβ3 integrin-expressing human renal cell carcinoma 786-O cells. The ligand was conjugated to the anticancer drug paclitaxel through a Asn-Pro-Val (NPV) tripeptide linker, which is a substrate of neutrophil-secreted elastase...
November 19, 2018: Chemistry: a European Journal
Benedetta Terziroli Beretta-Piccoli, Giorgina Mieli-Vergani, Diego Vergani
Autoimmune serology is key to the diagnosis and management of autoimmune liver diseases. Its correct use in clinical practice requires a basic knowledge of the laboratory techniques used for autoantibody detection. Indirect immunofluorescence (IIF) on triple rodent tissue is still the gold standard screening procedure for liver-relevant autoantibodies, while HEp2 cells and human ethanol-fixed neutrophils are used as substrates to characterize nuclear reactivities and to detect anti-neutrophil cytoplasm antibody, respectively...
December 2018: Journal of Autoimmunity
Svetlana I Galkina, Natalia V Fedorova, Alexander L Ksenofontov, Vladimir I Stadnichuk, Ludmila A Baratova, Galina F Sud'Ina
Neutrophils release branched-chain (valine, isoleucine, leucine), aromatic (tyrosine, phenylalanine) and positively charged free amino acids (arginine, ornithine, lysine, hydroxylysine, histidine) when adhere and spread onto fibronectin. In the presence of agents that impair cell spreading or adhesion (cytochalasin D, fMLP, nonadhesive substrate), neutrophils release the same amino acids, except for a sharp decrease in hydroxylysine and an increase in phenylalanine, indicating their special connection with cell adhesion...
October 29, 2018: Cell Adhesion & Migration
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