BackgroundRecent studies have suggested that excessive formation of neutrophil extracellular traps (NETs) plays a critical role in the pathogenesis of sepsis. Although elevation of the plasma level of cell-free DNA (cf-DNA) has been reported in sepsis patients, there has been little direct measurement of circulating free NETs such as myeloperoxidase-conjugated DNA (MPO-DNA). The objectives of this study were to detect NETs in the bloodstream of patients with septic shock, and to assess the correlations of circulating NET levels with organ dysfunction, disease severity, and mortality.MethodsFifty-five patients with septic shock admitted to the intensive care units (ICUs) of 35 Japanese hospitals were studied. Septic shock was diagnosed according to the 1997 definition of the American College of Chest Physicians/Society of Critical Care Medicine. To detect circulating NETs, plasma levels of MPO-DNA and cf-DNA were measured by sandwich enzyme-linked immunosorbent assay and by fluorometric assay on days 1, 3, and 7 after the onset of septic shock. Physiological and mortality data were collected from the clinical database.ResultsOn days 1, 3, and 7, the patients showed a marked increase in plasma MPO-DNA levels compared with healthy volunteers, whereas the plasma cf-DNA level was only increased significantly on day 1 and then decreased rapidly. A high MPO-DNA level on days 3 and 7 were associated with 28-day mortality.On days 3 and 7, the MPO-DNA levels were inversely correlated with both the mean arterial pressure and the PaO2/FIO2 ratio, whereas the cf-DNA level was not correlated with either parameter. There was a positive correlation between the plasma MPO-DNA level and the sepsis-related organ failure assessment score on days 3 and 7. Neither cf-DNA nor MPO-DNA levels were correlated with the disseminated intravascular coagulation (DIC) score or the platelet count.ConclusionThe increase in circulating MPO-DNA in patients with septic shock indicates acceleration of NET formation in the early stages of sepsis. High MPO-DNA levels are associated with the severity of organ dysfunction and 28-day mortality due to septic shock, but not with the DIC score. These results suggest that excessive NET formation contributes to the pathogenesis of septic shock.Electronic supplementary materialThe online version of this article (10.1186/s13054-018-2109-7) contains supplementary material, which is available to authorized users.

Background The purpose of this study is to investigate the time course of syndecan-1 (Syn-1) plasma levels, the correlation between Syn-1 and organ damage development, and the associations of Syn-1 level with cumulative fluid balance and ventilator-free days (VFD) in patients with septic shock. Methods We collected blood samples from 38 patients with septic shock upon their admission to ICU and for the first 7 days of their stay. Syn-1 plasma level, acute respiratory distress syndrome (ARDS), other organ damage, VFD, and cumulative fluid balance were assessed daily. Results Over the course of 7 days, Syn-1 plasma levels increased significantly more in patients with ARDS than in those without ARDS. Patients with high levels of Syn-1 in the 72 h after ICU admission had significantly higher cumulative fluid balance, lower PaO2/FiO2, and fewer VFD than patients with low levels of Syn-1. Syn-1 levels did not correlate with sequential organ failure assessment score or with APACHE II score. Conclusions In our cohort of patients with septic shock, higher circulating level of Syn-1 of cardinal glycocalyx component is associated with more ARDS, cumulative positive fluid balance, and fewer VFD. Measurement of Syn-1 levels in patients with septic shock might be useful for predicting patients at high risk of ARDS.

Components of neutrophil extracellular traps (NETs) are released into the circulation by neutrophils and contribute to microcirculatory disturbance in sepsis. Removing NET components (DNA, histones, and proteases) from the circulation could be a new strategy for counteracting NET-dependent tissue damage. We evaluated the effect of hemoperfusion with a polymyxin B (PMX) cartridge, which was originally developed for treating gram-negative infection, on circulating NET components in patients with septic shock, as well as the effect on phorbol myristate acetate (PMA)-stimulated neutrophils obtained from healthy volunteers. Ex vivo closed loop hemoperfusion was performed through PMX filters in a laboratory circuit. Whole blood from healthy volunteers (incubated with or without PMA) or from septic shock patients was perfused through the circuit. For in vivo experiment blood samples were collected before and immediately after hemoperfusion with PMX to measure the plasma levels of cell-free NETs. The level of cell-free NETs was assessed by measuring myeloperoxidase-associated DNA (MPO-DNA), neutrophil elastase-associated DNA (NE-DNA), and cell-free DNA (cf-DNA). Plasma levels of MPO-DNA, NE-DNA, and cf-DNA were significantly increased after 2 h of PMA stimulation. When the circuit was perfused with blood from septic shock patients or PMA-stimulated neutrophils from healthy volunteers, circulating levels of MPO-DNA, NE-DNA, and cf-DNA were significantly reduced after 1 and 2 h of perfusion with a PMX filter compared with perfusion without a PMX filter. In 10 patients with sepsis, direct hemoperfusion through filters with immobilized PMX significantly reduced plasma levels of MPO-DNA and NE-DNA. These ex vivo and in vivo findings demonstrated that hemoperfusion with PMX removes circulating NET components. Selective removal of circulating NET components from the blood could be effective for prevention/treatment of NET-related inappropriate inflammation and thrombogenesis in patients with sepsis.

BackgroundVenous thromboembolism (VTE) and the subsequent development of pulmonary embolism (PE) is a major cause of post-operative mortality in total knee arthroplasty (TKA). We evaluated whether the addition of an ultrasound-guided femoral nerve block with general anesthesia affected the incidence in the development of VTE following TKA.MethodsThis was a retrospective non-randomized comparative study with patients assigned to groups based on the surgery date (pre-femoral nerve block versus post-femoral nerve block periods). All anesthesia and medical records of the patients who had undergone computer-navigated TKA in our facility between January 2009 and March 2010 were retrospectively reviewed.ResultsForty patients were identified; 15 patients underwent computer-navigated TKA under general anesthesia alone (Group G) and 25 patients underwent surgery under general anesthesia combined with ultrasound-guided femoral nerve block (Group F). The incidence of development of VTE post-operatively was significantly lower in Group F (P = 0.037). Logistic regression analysis identified the use of a femoral nerve block as the most significant variable correlating with the incidence of post-operative development of VTE, and the odds ratio for VTE development in Group G was 3.12 (95% CI, 0.57-20.56).ConclusionsWe suggest the possibility that the addition of a femoral nerve block on general anesthesia may reduce the incidence of the development of VTE following TKA.

TiN film was coated onto SK 5 substrate by RF ion plating under different bias voltages, and X-ray diffraction analysis and ESCA measurement were performed on this film to determine the X-ray intensities of the (111) and (200) planes and the composition of the film. To evaluate the orientation of the TiN film using this X-ray intensity and composition data, the relation ship between the composition and the X-ray intensity ratio of the (111) plane to the (200) plane for randomly oriented TiN (TiC) was investigated.It was found that the composition of the TiN (TiC ) affects the X-ray intensity ratio, and that the orientation of the film changed with the bias voltage.The composition also affects the packing density of each plane and, therefore, is considered to be one of the parameters which influences the orientation of the film. In addition, the orientations of previously reported TiN and TiC films coated by the PVD method were re-examined.