• Cardiac conduction abnormalities
• Cx43 gap junction
• IFN-γ
• IL-18
• Neutrophil extracellular traps

• Animals
• Doxorubicin/adverse effects
• Connexin 43/metabolism
• Connexin 43/genetics
• Cardiotoxicity/prevention & control
• Cardiotoxicity/immunology
• Interleukin-18/metabolism
• Interferon-gamma/metabolism
• Extracellular Traps/metabolism
• Extracellular Traps/immunology
• Mice
• Mice, Inbred C57BL
• Mice, Knockout
• Male
• Arrhythmias, Cardiac/chemically induced
• Antibiotics, Antineoplastic

• Humans
• Bacteremia/blood
• Bacteremia/diagnosis
• Bacteremia/mortality
• Biomarkers/blood
• Cell-Free Nucleic Acids/blood
• Prognosis
• ROC Curve
• Sepsis/blood
• Sepsis/diagnosis
• Sepsis/mortality

• Humans
• Arthritis, Rheumatoid/blood
• Arthritis, Rheumatoid/diagnosis
• DNA, Mitochondrial/blood
• Female
• Male
• Middle Aged
• Biomarkers/blood
• Aged
• Prospective Studies
• Adult
• Inflammation/blood
• Inflammation/diagnosis

• University of Basel

• acute kidney injury
• biomarkers
• cell-free DNA
• neutrophil gelatinase-associated lipocalin
• sepsis

• NETs
• histones
• horses
• immunofluorescence
• infection
• peritoneal fluid
• synovial fluid

• Animals
• Horses
• Extracellular Traps
• Ascitic Fluid
• Synovitis/veterinary
• Inflammation/veterinary
• Peritonitis/veterinary
• Microscopy, Fluorescence/veterinary
• Neutrophils
• Synovial Fluid
• Horse Diseases/diagnosis

• Australian Government Research Training Program (AGRTP) Scholarship
• Doctor of Veterinary Studies Scholarship

• biomarker
• crystal arthritis
• gout
• hot joint
• infectious arthritis
• point-of-care testing
• septic arthritis
• synovial fluid

• Humans
• Sensitivity and Specificity
• Arthritis, Infectious/diagnosis
• Biomarkers
• Predictive Value of Tests
• Leukocyte Count
• Synovial Fluid/chemistry

• Wellcome Trust

• Chemotherapy-induced peripheral neuropathy
• Microcirculatory disturbance
• Neutrophil extracellular traps
• SHp-DNase1

• peptidylarginine deiminase
• neutrophil extracellular trap
• citrullinated histone h3
• trauma
• inflammation
• traumatic brain injury
• burn
• hemorrhage

• Protein-Arginine Deiminases/genetics
• Protein-Arginine Deiminases/metabolism
• Extracellular Traps/metabolism
• Citrullination
• Neutrophils/metabolism
• Histones/metabolism

• F32 HL162378 NHLBI NIH HHS
• R01 HL155116 NHLBI NIH HHS

• biomarkers
• cell-free DNA
• diagnosis
• periprosthetic joint infection
• synovial fluid

• cell‐free circulating DNA
• lateral lumbar interbody fusion
• low back pain
• lumbar canal stenosis
• numeric rating scale

• Tokai University

• autoimmune diseases
• inflammation
• lupus erythematosus
• systemic

• Neutrophil extracellular trap (NET)
• bronchoalveolar lavage
• lung injury
• one-lung ventilation

• Cytokines
• Endotoxin
• Innate immunity
• Intensive care
• Low tidal volume ventilation

Over a decade ago, the formation of neutrophil extracellular traps (NETs) was described as a novel mechanism employed by neutrophils to tackle infections. Currently applied methods for NETs release quantification are often limited by the use of unspecific dyes and technical difficulties. Therefore, we aimed to develop a fully automatic image processing method for the detection and quantification of NETs based on live imaging with the use of DNA-staining dyes. For this purpose, we adopted a recently proposed Convolutional Neural Network (CNN) model called Mask R-CNN. The adopted model detected objects with quality comparable to manual counting—Over 90% of detected cells were classified in the same manner as in manual labelling. Furthermore, the inhibitory effect of GW 311616A (neutrophil elastase inhibitor) on NETs release, observed microscopically, was confirmed with the use of the CNN model but not by extracellular DNA release measurement. We have demonstrated that a modern CNN model outperforms a widely used quantification method based on the measurement of DNA release and can be a valuable tool to quantitate the formation process of NETs.

• automatic image analysis
• chronic granulomatous disease
• convolutional neural networks (CNN), mask R-CNN
• neutrophil extracellular traps (NETs) quantification
• neutrophils
• nitric oxide
• peroxynitrite
• reactive nitrogen species

According to the World Health Organization (WHO), trauma is responsible for 10% of deaths and 16% of disabilities worldwide. This is considerably higher than those for malaria, tuberculosis, and HIV/AIDS combined. While the human suffering and death caused by injury is well-recognized, injury has a significant medical care cost. Better prediction of the state of trauma patients in the days immediately after trauma may reduce costs. Traumatic injuries to multiple organs can cause dysfunction in all systems of the body especially the immune system placing patients at high risk of infections and inflammatory complications which are often fatal. Neutrophils are the most abundant leukocyte in the human circulation and are crucial for the prevention of microbial disease. Significant changes in neutrophil functions such as enhanced chemotaxis, Neutrophil extracellular trap (NET)-induced cell death (NETosis), and phagocytosis occur early after injury followed by prolonged functional defects such as phagocytosis, killing mechanisms, and receptor expression. Analysis of these changes may improve the prediction of the patient's condition over time. We provide a comprehensive and up-to-date review of the literature investigating the effect of trauma on neutrophil phenotype with an underlying goal of using this knowledge to examine the predictive potential of neutrophil alterations on secondary complications in patients with traumatic injuries. We conclude that alterations in neutrophil surface markers and functions may be potential biomarkers that predict the outcome of trauma patients.

• injury
• neutrophil subtype
• neutrophils
• survival
• trauma

Cell-free DNA (cf-DNA) concentration in human plasma is often increased after burn and trauma injuries. Two major sources of cf-DNA are the parenchymal cells damaged by the injury and various circulating cells indirectly altered by the response to injury. The cf-DNA originating from neutrophils, also known as circulating neutrophil extracellular traps (cNETs), is of notable interest because cNETs have been associated with pathological processes in other conditions, including cancer, autoimmunity, etc. Both intact chromatin and oligonucleotides, which are the by-product of cf-DNA degradation, are assumed to contribute to the cf-DNA in patients. However, traditional assays for cf-DNA quantification do not distinguish between cNETs and cf-DNA of other origins and do not differentiate between intact chromatin and oligonucleotides. Here we measure the amount of intact cNETs in the circulation, using a microfluidic device that mechanically traps chromatin fibers directly from blood and an immunofluorescence protocol that detects neutrophil-specific proteins associated with chromatin. In a rat model of burn injury, we determined that the chromatin fibers in the circulation after injury originate exclusively from neutrophils and are cNETs. We found that the concentration of cNETs surges the first day after injury and then decreases slowly over several days. In a secondary sepsis model, which involved a burn injury followed by cecal-ligation-puncture, we measured additional increases in cNETs in the days after sepsis was induced. These results validate a microfluidic assay for the quantification of cNETs and will facilitate fruther studies probing the contribution of cNETs to complications after burns and sepsis.

• Autophagy
• Bioinformatics
• Cytokine
• Neutrophil
• Rheumatoid arthritis

Liver dysfunction (LD) and liver failure are associated with poor outcome in critically ill patients. In patients with severe sepsis or septic shock, LD occurred in nearly 19% of patients. An early diagnosis of LD at time of initial damage of the liver can lead to a better prognosis of these patients because an early start of therapy is possible. We performed a second prospective study with septic patients to test a new cell-based cytotoxicity device (biosensor) to evaluate clinical relevance for early diagnosis of LD and prognostic capacity. In the clinical study, 99 intensive care unit patients were included in two groups. From the patients of the septic group (n = 51, SG), and the control (non-septic) group [n = 49, control group (CG)] were drawn 20 ml blood at inclusion, after 3, and 7 days for testing with the biosensor. Patients’ data were recorded for hospital survival, organ function, and demographic data, illness severity [acute physiology and chronic health evaluation (APACHE) II-, sepsis-related organ failure assessment (SOFA) scores], cytokines, circulating-free deoxyribonucleic acid/neutrophil-derived extracellular traps (cf-DNA/NETs), microbiological results, and pre-morbidity. For the developed cytotoxicity test, the human liver cell line HepG2/C3A was used. Patients’ plasma was incubated in a microtiter plate assay with the test cells and after 6 days incubation the viability (trypan blue staining, XTT-test) and functionality (synthesis of albumin, cytochrome 1A2 activity) was analyzed. An impairment of viability and functionality of test cells was only seen in the SG compared with the CG. The plasma of non-survivors in the SG led to a more pronounced impairment of test cells than the plasma of survivors at inclusion. In addition, the levels of cf-DNA/NETs were significantly higher in the SG at inclusion, after 3, and after 7 days compared with the CG. The SG showed an in-hospital mortality of 24% and the values of bilirubin, APACHE II-, and SOFA scores were markedly higher at inclusion than in the CG. Hepatotoxicity of septic plasma was already detected with the liver cell-based biosensor at inclusion and also in the course of disease. The biosensor may be a tool for early diagnosis of LD in septic patients and may have prognostic relevance.

• biosensing techniques
• cytotoxicity
• hepatocytes
• inflammation
• liver failure

• Staphylococcus aureus
• chemoattractant
• infection
• neutrophil
• septic arthritis
• tissue damage

• Anti-Bacterial Agents/therapeutic use
• Arthritis, Infectious/immunology
• Arthritis, Infectious/microbiology
• Arthritis, Infectious/surgery
• Arthritis, Infectious/therapy
• Arthrocentesis/methods
• Arthroscopy/methods
• Cell Movement/immunology
• Chemokines/immunology
• Chemokines/metabolism
• Humans
• Inflammation
• Joints/drug effects
• Joints/immunology
• Joints/microbiology
• Joints/surgery
• Leukotrienes/immunology
• Leukotrienes/metabolism
• Neutrophil Infiltration/drug effects
• Neutrophils/drug effects
• Neutrophils/immunology
• Neutrophils/pathology
• Staphylococcal Infections/immunology
• Staphylococcal Infections/microbiology
• Staphylococcal Infections/surgery
• Staphylococcal Infections/therapy
• Staphylococcus aureus
• Suction/methods

• CXCR2
• Chemokine
• Neutrophil
• Septic arthritis
• Staphylococcus aureus

• Dermatomyositis
• Interstitial lung disease
• Melanoma differentiation-associated gene 5
• Neutrophil extracellular traps

• DNA
• Imaging.
• Inflammation
• Ischemia
• Nanoprobe
• RNA

• Animals
• Anti-Inflammatory Agents/administration & dosage
• Benzothiazoles/administration & dosage
• Cytoprotection
• Dextrans/administration & dosage
• Mice
• Myocardial Reperfusion Injury/diagnostic imaging
• Myocardial Reperfusion Injury/drug therapy
• Myocardial Reperfusion Injury/pathology
• Nanostructures/administration & dosage
• Nucleic Acids/metabolism
• Optical Imaging
• Quinolines/administration & dosage
• Theranostic Nanomedicine/methods
• Treatment Outcome

• R01 HL093038 NHLBI NIH HHS
• R01 HL112831 NHLBI NIH HHS
• R00 HL121152 NHLBI NIH HHS
• R01 GM117233 NIGMS NIH HHS
• K99 HL121152 NHLBI NIH HHS
• R01 EB011996 NIBIB NIH HHS

Sepsis is one of the most common causes of death in severely injured patients worldwide. The early detection of sepsis still has to be solved in clinical practice. The delayed diagnosis often contributes to inappropriate antimicrobial treatment and subsequent high mortality. Sepsis biomarkers are produced during the host response to infection. Traditional biomarkers are polypeptides and/or proteins derived from this response. Omics-based biomarkers are screening out from all kinds of molecules of host response while high-throughout omics technologies are emerging. This review describes traditional and potential omics-based sepsis biomarkers from currently available literatures. The combination of these biomarkers would refine the identification of sepsis for further clinical and experimental sepsis studies.

Background. This study aimed to determine the perioperative change in serum double-strand DNA (dsDNA) as a marker potentially reflecting neutrophil extracellular trap concentration in samples from patients undergoing cardiac surgery and to analyze a relationship between serum dsDNA concentrations and perioperative renal dysfunction. Methods. Serum dsDNA concentrations in samples that were collected during a previously conducted, prospective, multicenter, observational study were measured. Eighty patients undergoing elective cardiac surgery were studied. Serum samples were collected at baseline, immediately after surgery, and the day after surgery (POD-1). Results. Serum dsDNA concentration was significantly increased from baseline (median, 398 ng/mL [interquartile range, 372–475 ng/mL]) to immediately after surgery (median, 540 ng/mL [437–682 ng/mL], p < 0.001), and they were reduced by POD-1 (median, 323 ng/mL [256–436 ng/mL]). The difference in serum creatinine concentration between baseline and POD-1 was correlated with dsDNA concentration on POD-1 (r_s = 0.61, p < 0.001). Conclusions. In patients undergoing cardiac surgery, serum dsDNA concentration is elevated postoperatively. Prolonged elevation in dsDNA concentration is correlated with perioperative renal dysfunction. Further large-scale studies are needed to determine the relationship between serum concentration of circulating dsDNA and perioperative renal dysfunction.

Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a group of life-threatening disorders, and frequently affects the kidneys. This study investigated whether the circulating neutrophil extracellular traps (NETs) levels were associated with disease activity of AAV. We collected serum samples from 34 patients with AAV in active stage and 62 patients with AAV in remission. Cell free DNA in serum was quantified using the Quant-iT PicoGreen assay. NETs associated MPO-DNA complexes, citrullinated-histone H3-DNA (cit-H3-DNA) complexes and the concentration of deoxyribonuclease I (DNase I) were quantified using ELISA. The activity of DNase I was quantified using radial enzyme-diffusion method. Associations between circulating levels of NETs with clinico-pathological parameters were analyzed. Serum levels of NETs in active AAV patients were significantly higher than those in healthy controls, and the level of cell free DNA correlated with C-reactive protein (CRP). However, no correlation was found between MPO-DNA complexes or cit-H3-DNA complexes level and CRP. Also there was no significant correlation between NETs level and initial serum creatinine, estimated glomerular filtration rate (eGFR), crescents formation or Birmingham Vasculitis Activity Score (BVAS). Furthermore, there was no significant difference of serum levels of cell free DNA or MPO-DNA complexes between active stage and remission of AAV. In conclusion, circulating levels of NETs cannot be used as a biomarker to assess disease activity in AAV patients.

• Adult
• Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis/blood
• Biomarkers/blood
• DNA/blood
• Extracellular Traps/metabolism
• Female
• Humans
• Male
• Peroxidase/blood

Recently, it has been reported that circulating free DNA (cf-DNA) in the blood is increased in various infectious diseases, including sepsis. Moreover, a relationship between cf-DNA and neutrophil extracellular traps (NETs) has been suggested. However, it is still unclear what the source and physiological role of cf-DNA in sepsis are. In this study, we examined the source of cf-DNA by detecting citrullinated histone H3, a characteristic feature of NET formation, in cecal ligation and puncture- (CLP-)operated mice. In addition, neutrophil depletion using anti-Ly6G antibodies was performed to assess the association between neutrophils and cf-DNA. Increased cf-DNA levels were observed only in CLP mice and not in the control groups; the qPCR findings revealed that the cf-DNA was mainly host-derived, even in bacteremic conditions. Citrullinated histone H3 was not increased in the neutrophils upon CLP, and the depletion of neutrophils showed limited effects on decreasing the amount of cf-DNA. Taken together, these results suggested that elevated cf-DNA levels during early-phase sepsis may represent a candidate biomarker for the severity of sepsis and that, contrary to previous findings, cf-DNA is not derived from neutrophils or NETs.

• Adaptive Immunity/immunology
• Apoptosis/immunology
• Bacteria/immunology
• Extracellular Traps/immunology
• Granulomatous Disease, Chronic/immunology
• Humans
• Hydrolases/immunology
• Immunity, Innate/immunology
• Neutrophils/immunology
• Neutrophils/microbiology
• Periodontitis/immunology
• Periodontitis/microbiology
• Porphyromonas gingivalis/immunology
• Protein-Arginine Deiminases

• Aged
• Bacterial Infections/immunology
• Citrulline/metabolism
• Critical Illness
• Extracellular Traps
• Female
• Histones/blood
• Histones/metabolism
• Humans
• Male
• Middle Aged
• Neutrophils/immunology
• Prospective Studies
• Protein Processing, Post-Translational
• Respiratory Tract Infections/immunology
• Trachea/microbiology

• inflammatory marker
• interleukin-6
• lactate
• periprosthetic joint infection
• synovial fluid
• synovial fluid white blood cell count

• Burn
• Gut permeability
• Mucosal barrier
• NETs
• SIRS
• Sepsis
• Statin

• S06 GM008043 NIGMS NIH HHS
• SC3 GM099632 NIGMS NIH HHS

• Gouty arthritis
• Inflammatory marker
• Interleukin-6
• Lactic acid
• Septic arthritis
• Synovial fluid

• Animals
• Cattle
• DNA/metabolism
• Macrophages/immunology
• Macrophages/microbiology
• Microbial Viability
• Microscopy, Confocal
• Microscopy, Electron, Scanning
• Neutrophils/immunology
• Neutrophils/microbiology
• Pasteurellaceae/immunology

Although the formation of neutrophil (PMN) extracellular traps (NETs) has been detected during infection and sepsis, their role in vivo is still unclear. This study was performed in order to evaluate the influence of NETs depletion by administration of recombinant human (rh)DNase on bacterial spreading, PMN tissue infiltration and inflammatory response in a mouse model of polymicrobial sepsis.

In a prospective controlled double-armed animal trial, polymicrobial sepsis was induced by cecal ligation and puncture (CLP). After CLP, mice were treated with rhDNase or phosphate buffered saline, respectively. Survival, colony forming unit (CFU) counts in the peritoneal cavity, lung, liver and blood were determined. PMN and platelet counts, IL-6 and circulating free (cf)-DNA/NETs levels were monitored. PMN infiltration, as well as organ damage, was analyzed histologically in the lungs and liver. Capability and capacity of PMN to form NETs were determined over time.

cf-DNA/NETs were found to be significantly increased 6, 24, and 48 hours after CLP when compared to the levels determined in sham and naïve mice. Peak levels after 24 hours were correlated to enhanced capacity of bone marrow-derived PMN to form NETs after ex vivo stimulation with phorbol-12-myristate-13-acetate at the same time. rhDNase treatment of mice resulted in a significant reduction of cf-DNA/NETs levels 24 hours after CLP (P < 0.001). Although overall survival was not affected by rhDNase treatment, median survival after 24 hours was significantly lower when compared with the CLP group (P < 0.01). In mice receiving rhDNase treatment, CFU counts in the lung (P < 0.001) and peritoneal cavity (P < 0.05), as well as serum IL-6 levels (P < 0.001), were found to be already increased six hours after CLP. Additionally, enhanced PMN infiltration and tissue damage in the lungs and liver were found after 24 hours. In contrast, CFU counts in mice without rhDNase treatment increased later but more strongly 24 hours after CLP (P < 0.001). Similarly, serum IL-6 levels peaked after 24 hours (P < 0.01).

This study shows, for the first time, that depletion of NETs by rhDNase administration impedes the early immune response and aggravates the pathology that follows polymicrobial sepsis in vivo.

Introduction. Neutrophil extracellular traps (NET) consist of a DNA scaffold that can be destroyed by Deoxyribonuclease (DNase). Thus DNases are potential prerequisites for natural counter regulation of NETs formation. In the present study, we determined the relationship of NETs and DNase after major trauma. Methods. Thirty-nine major trauma patients, 14 with and 25 without sepsis development were enrolled in this prospective study. Levels of cell-free (cf)-DNA/NETs and DNase were quantified daily from admission until day 9 after admission. Results. Levels of cf-DNA/NETs in patients who developed sepsis were significantly increased after trauma. In the early septic phase, DNase values in septic patients were significantly increased compared to patients without sepsis (P < 0.05). cf-DNA/NETs values correlated to values of DNase in all trauma patients and patients with uneventful recovery (P < 0.01) but not in septic patients. Recombinant DNase efficiently degraded NETs released by stimulated neutrophils in a concentration-dependent manner in vitro. Conclusions. DNase degrades NETs in a concentration-dependent manner and therefore could have a potential regulatory effect on NET formation in neutrophils. This may inhibit the antibacterial effects of NETs or protect the tissue from autodestruction in inadequate NETs release in septic patients.

• Antigen Presentation/immunology
• Chemotaxis, Leukocyte/physiology
• Cytokines/biosynthesis
• Humans
• Inflammation/immunology
• Inflammation/metabolism
• Leukocyte Elastase/metabolism
• Myeloblastin/metabolism
• NADPH Oxidases/metabolism
• Neutropenia/immunology
• Neutrophil Infiltration/immunology
• Neutrophils/physiology

• R01 CA022556 NCI NIH HHS
• R37 CA022556 NCI NIH HHS
• R37 CA022556-34 NCI NIH HHS
• CA022556 NCI NIH HHS

• Circulating free DNA
• Inflammation
• Neutrophil extracellular traps (NETs)
• Neutrophils
• Outcome