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86 results for “Neutrophil extracellular traps”
Fig. 3 in Bottlenose dolphins (Tursiops truncatus) do also cast neutrophil extracellular traps against the apicomplexan parasite Neospora caninum
Fig. 3. Dose, kinetic and functional inhibition assays of N. caninum tachyzoites-triggered NET formation in dolphins. PMN were incubated with tachyzoites, zymosan (1 mg/ ml, positive control) or plain medium (negative control) at different ratios (a; PMN: tachyzoites = 1:1, 1:2, 1:3) and time periods (b; 30, 60 and 90 min). To prove the DNA nature of NETs, the samples were treated with DNase I (a; 15 min). Moreover, cetacean PMN cells were pre-treated with NOX-inhibitor (b; DPI, 10 MM) for 30 min prior to N. caninum stimulation (1:3 ratio; 90 min). After incubation, all samples were analyzed for extracellular DNA by quantifying Pico Green ®-derived fluorescence intensities. Each condition was performed in duplicates. Geometric means of three PMN donors. Differences were regarded as significant at a level of p <0.05 (*) and p <0.01 (**).
Fig. 2 in Bottlenose dolphins (Tursiops truncatus) do also cast neutrophil extracellular traps against the apicomplexan parasite Neospora caninum
Fig. 2. Neospora caninum tachyzoite-triggered dolphin NET structures (SEM) and co-localization of extracellular DNA with histones (H1, H2A/H2B, H3 and H4), NE, MPO and PTX. (a‾d) Scanning electron microscopy (SEM) analyses revealed NETs being formed by dolphin PMN after co-culture with N. caninum tachyzoites. (a) Mesh of DNA-structures (white arrow) derived from dolphin PMN attached to N. caninum-tachyzoites (black arrows). (b) Intact cetacean-PMN (black stars) derived a fine filaroid structure (white arrow) being attached to tachyzoites (black arrows). (c) Conglomerates of several tachyzoites (black arrow) being entrapped in a rather chunky meshwork of cetacean-PMN-released thicker extracellular filaments (white arrow) (d) Dolphin PMN activated (black star) entrapping diverse N. caninum-tachyzoites (black arrows). (e‾l) Co-cultures of dolphin PMN and N. caninum tachyzoites were fixed, permeabilized, stained for analysis of co-localization (i-l; merge, white arrows) of extracellular DNA (e-h; red; Sytox Orange ®) and classical NETs components (all green, white arrows) such as histones (i), NE (j), MPO (k) and pentraxin (l). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 1 in Bottlenose dolphins (Tursiops truncatus) do also cast neutrophil extracellular traps against the apicomplexan parasite Neospora caninum
Fig. 1. Minimally-invasive blood extraction method for cetaceans. (a) Puncture of the ventral superficial fluke plexus with a fine needle attached to infusion system and one syringe to create a vacuum for blood extraction. (b) Professional trainers performed physical restraint of one dolphin using whistle to give a positive reinforcement during sampling.
Trapalyzer: A computer program for quantitative analyses in fluorescent live-imaging studies of Neutrophil Extracellular Trap formation.
<p>This data set contains a set of fluorescent microscopy images of a co-culture of neutrophil cells and E. coli bacteria used to study the Neutrophil Extracellular Trap (NET) formation stimulated by bacteria. </p> <p>NETs and live cells were visualized with a double fluorescent staining of DNA using Hoechst 33342 and SYTOX Green. </p> <p><strong>Reagents.</strong></p> <p>Roswell Park Memorial Institute (RPMI) 1640 medium, HEPES, SYTOX<sup>TM</sup> Green, and Hoechst 33342 were purchased from Thermo Fisher Scientific (Waltham, USA). LB broth was purchased from Sigma Aldrich (St Louis, MO, USA).</p> <p><strong>Preparation of blood neutrophils.</strong></p> <p>Neutrophils were obtained from peripheral blood of one healthy blood donor. Blood sample was purchased at Local Blood Donation Centre and according to local regulations, the blood donor enabled blood donation center to sell their blood samples for scientific purposes and the consent of bioethical committee was not required. Blood was collected into a citrate tube and processed within 2 hours from collection. Neutrophils were isolated using density gradient centrifugation followed by polyvinyl alcohol sedimentation, exactly as described in [1]. Isolated neutrophils were suspended in RPMI 1640 medium with 10 mM HEPES (RH). </p> <p><strong>Preparation of bacteria.</strong></p> <p><em>Escherichia coli</em> (American Type Culture Collection(ATCC) 25922 strain) were grown overnight in LB broth with shaking. In the morning, an aliquot of bacterial culture was taken, diluted 100 x in a fresh LB medium and grown for subsequent 2-3 hours. Subsequently, bacterial cultures were washed and resuspended in RH medium.</p> <p><strong>Co-culture of neutrophils with bacteria</strong><br> Neutrophils were seeded into the wells of 48-well plates at the density of 2 ⨉ 10<sup>4</sup> cells/well and allowed to settle for 30 minutes at 37°C, 5% CO2. Subsequently, <em>E. coli</em> was added into the appropriate wells at the multiplicity of infection of 4 or 1 (<em>E.coli</em>: neutrophil). Neutrophils incubated without bacteria were used as a control group. A technical duplicate for each condition was prepared. <br> For each intended timepoint (t=0, 60, 90, 120, 180 minutes), a separate 48 well plate was prepared. The plates were centrifuged for 5 minutes at 250 g to allow the contact of bacteria with neutrophils. The plates were incubated at 37°C, 5\% CO2 for a specified time and then the samples were stained with SYTOX<sup>TM</sup> Green (100 nM) and Hoechst 33342 (1.25 μM) for 10 minutes. Four images of each well were taken with Leica DMi8 fluorescent microscope equipped with a 10× magnification objective (Leica, Wetzlar, Germany). Overall, 120 images have been obtained.</p> <p> </p> <p><strong>2019_04_24--ecoli_neu_tiff_channel_merged.zip:</strong> Images in .tif format, each containing 5 channels: channel 1 for SYTOX Green fluorescent stain (green fluorescence), channel 2 for Hoechst 33342 fluorescent stain (blue fluorescence), and three channels for transmission light encoded in RGB values. </p> <p> </p> <p><strong>2019_04_24--ecoli_neu_tiff_raw_exported.zip:</strong> Images split by different light sources: transmission light (_ch00.tif), SYTOX Green fluorescence (_ch01.tif), Hoechst 33342 fluorescence (_ch02.tif).</p> <p> </p> <p>[1] Bystrzycka W, Moskalik A, Sieczkowska S, Manda-Handzlik A, Demkow U, Ciepiela O. The effect of clindamycin and amoxicillin on neutrophil extracellular trap (NET) release. <em>Cent Eur J Immunol</em>. 2016;41(1):1-5. doi:10.5114/ceji.2016.58811</p>
Randomized Study Evaluating the Effect of Danirixin on Neutrophil Extracellular Traps (NETs) in Chronic Obstructive Pulmonary Disease (COPD)
ClinicalTrials.gov study NCT03250689. IPD Sharing: YES. Countries: 1. Publications: 1.
Neutrophil extracellular traps are present in the airways of ENaC-overexpressing mice with cystic fibrosis-like lung disease
<p>Background: Neutrophils are key components of the exacerbated inflammation and tissue damage in cystic fibrosis (CF) airways. Neutrophil extracellular traps (NETs) trap and kill extracellular pathogens. While NETs are abundant in the airways of CF patients and have been hypothesized to contribute to lung damage in CF, the in vivo role of NETs remains controversial, partially due to lack of appropriate animal models. The goal of this study was to detect NETs and to further characterize neutrophilmediated inflammation in the airways of mice overexpressing the epithelial sodium channel (βENaC-Tg mice on C57BL/6 background) in their lung with CF-like airway disease, in the absence of any apparent bacterial infections.</p> <p>Methods: Histology scoring of lung tissues, flow cytometry, multiplex ELISA, immunohistochemistry and immunofluorescence were used to characterize NETs and the airway environment in uninfected, βENaC-Tg mice at 6 and 8 weeks of age, the most chronic time points so far studied in this model.</p> <p>Results: Excessive neutrophilic infiltration characterized the lungs of uninfected, βENaC-Tg mice at 6 and 8 weeks of age. The bronchoalveolar lavage fluid (BALF) of βENaC-Tg mice contains increased levels of CF-associated cytokines and chemokines: KC, MIP-1α/β, MCP-1, G-CSF, IL-5, and IL-6. The BALF of βENaC-Tg mice contain MPO-DNA complexes, indicative of the presence of NETs. Immunofluorescence and flow cytometry of BALF neutrophils and lung tissues demonstrated increased histone citrullination, a NET-specific marker, in βENaC-Tg mice.</p> <p>Conclusions: NETs are detected in the airways of βENaC-Tg mice, in the absence of bacterial infections. These data demonstrate the usefulness of the βENaC-Tg mouse to serve as a model for studying the role of NETs in chronic CF airway inflammation.</p> <p>Keywords Cystic fibrosis; neutrophil extracellular traps; NET; ENaC, neutrophil.</p>
CCR. Low-dose ionizing-radiation elicits the extrusion of neutrophil extracellular traps
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Melanoma-derived soluble mediators modulate neutrophil plasticity and the release of Neutrophil Extracellular Traps
<p>Background: polymorphonuclear neutrophils (PMNs) are main effector cells in the inflammatory responses. The significance of PMN infiltration in tumor microenvironment remains to be clarified. Metastatic melanoma is the most lethal type of skin cancer with an increasing incidence over the last decades. Few studies investigated the role of PMNs in human melanoma. The aim of this study was to investigate the role of PMNs and their mediators in human melanoma; (2) Methods: highly purified human PMNs from healthy donors were stimulated, in vitro, with conditioned media derived from the melanoma cell lines SKMEL28 and A375 (melanoma-CM) as well as from primary melanocytes as control. PMN functions (chemotaxis, survival, activation, cell tracking, morphology and NETs release) were evaluated; (3) Results: we found that the A375 cell line produced soluble factors able to promote PMN chemotaxis, survival, activation, and to modify PMN morphological changes and kinetic properties. Furthermore, both melanoma cell lines CM induced activation and the release of neutrophil extracellular traps (NETs) from PMNs. By contrast, primary melanocytes CM did not modify any PMN biological behavior. In addition, serum levels of myeloperoxidase (MPO), matrix metalloprotease-9 (MMP-9), CXCL8/IL-8, granulocyte and monocyte colony stimulating factor (GM-CSF) and NETs components were significantly increased in advanced melanoma patients compared to healthy controls; (4) Conclusions: melanoma cell lines produce soluble factors able to ‘educate’ PMNs towards an activated functional state. Metastatic melanoma patients display increased circulating levels of neutrophil-related mediators and NETs, suggesting that a neutrophil-related signature exists in metastatic melanoma patients. Further investigations are needed to better understand the role of these “tumor-educated neutrophils” in modifying melanoma cell behavior.</p>
Covid-19: Possible Role of Neutrophil Extracellular Traps
ClinicalTrials.gov study NCT04412382. IPD Sharing: YES. Countries: 1. Publications: 1.
Neutrophil Extracellular Traps in Different Forms of Systemic Sclerosis
ClinicalTrials.gov study NCT06462768. IPD Sharing: Not stated. Countries: 1. Publications: 1.
Transfusion-related Inflammatory Cytokine and Neutrophil Extracellular Trap Quantification in Neonates
ClinicalTrials.gov study NCT01735552. IPD Sharing: Not stated. Countries: 1. Publications: 9.
Neutrophil Extracellular Trap Formation in Patients Undergoing Bone Marrow Transplant
ClinicalTrials.gov study NCT01735565. IPD Sharing: Not stated. Countries: 1. Publications: 15.
Neutrophil Extracellular Traps and Thrombolysis in the Acute Stroke
ClinicalTrials.gov study NCT02476188. IPD Sharing: Not stated. Countries: 1. Publications: 3.
Pleural Fluid Neutrophil Extracellular Traps Exacerbate Disease Severity and Risk of One-year Mortality in Pleural Infection (TORPIDS-3)
ClinicalTrials.gov study NCT07194915. IPD Sharing: NO. Countries: 1. Publications: 1.
Impact of Neutrophil Extracellular Traps on Tissue Plasminogen Activator Induced Thrombolysis in Acute Ischemic Stroke Patients
ClinicalTrials.gov study NCT02907736. IPD Sharing: Not stated. Countries: 1. Publications: 1.
Neutrophil extracellular traps are present in the airways of ENaC-overexpressing mice with cystic fibrosis-like lung disease
Open the record for dataset details and reuse information.
Neutrophil Extracellular Trap Formation in Newborn Infants at Risk for Inflammatory Syndromes
ClinicalTrials.gov study NCT01106209. IPD Sharing: Not stated. Countries: 1. Publications: 0.
Cell Cycle Proteins Control Production of Neutrophil Extracellular Traps (NETs)
GEO Series GSE103755. Homo sapiens. 8 samples. Type: Expression profiling by array; Non-coding RNA profiling by array.
Effect of Neutrophil Extracellular Traps and Psoriasis-Associated TRAF3IP2 Genotype on Induction of Human Th17 Cells
GEO Series GSE121315. Homo sapiens. 92 samples. Type: Expression profiling by high throughput sequencing.
mRNA, ncRNA-seq and miRNA-seq of AGS cellline and AGS cellline with neutrophil extracellular traps
GEO Series GSE188741. Homo sapiens. 12 samples. Type: Expression profiling by high throughput sequencing; Non-coding RNA profiling by high throughput sequencing.
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