IL-10 does not affect oxidative burst and expression of selected surface antigen on human blood phagocytes in

Summary Cytokines play a major role in the control of inflammatory responses, participate in the regulation of blood phagocyte activities and as such are used for immunomodulatory therapy. In the present study, the influence of IL-10 on human blood phagocyte activity in the presence/absence of IL-6, IL-8 and TNF-α was tested in vitro. Our research analyzed the effects of cytokines on the production of reactive oxygen species measured by chemiluminescence and flow cytometry, and on the expression of surface molecules (CD11b, CD15, CD62L, CD31) measured by flow cytometry. IL-10 had no inhibitory effect on reactive oxygen species production and the expression of any examined adhesion molecule by resting or stimulated blood phagocytes within 3 h of incubation. Conversely, TNF-α, IL-6, and IL-8 increased reactive oxygen species production and the expression of CD11b and CD15 on both neutrophils and monocytes and decreased the expression of CD62L. These priming effects of the tested pro-inflammatory cytokines were not affected by IL-10. The obtained results suggest that IL-10 does not directly control blood phagocyte activation. These results also provide better information about the contribution of IL-6, IL-8 and TNF-α to the regulation of blood phagocyte-mediated inflammatory processes

Influence of polysulfone and hemophan hemodialysis membranes on phagocytes.

Abstract. The aim of the study was to compare the effect of hemophane and polysulfone membranes on the phagocyte-derived production of reactive oxygen species (ROS) as well as on neutrophil CD11b and CD62L expression in patients undergoing regular hemodialysis. The effects of hemodialysis membranes were also studied in in vitro conditions after coincubating them with differentiated HL-60 cells. ROS production was measured using chemiluminometric and flow cytometric methods. Expression of CD11b, CD62L and mitochondrial membrane potential were detected by monoclonal antibodies and by the JC-1 fluorescent probe, respectively. Depressed ROS production was observed in patients already before dialysis. Further decrease in ROS production and an increase in CD11b expression were observed especially in patients after hemophan hemodialysis. Decreased ROS production and increased CD11b expression were observed also after incubation of HL-60 cells with hemophan membranes. Mitochondrial membrane potential dropped only after incubating cells with hemophan membranes proving its more serious adverse effects in comparison with the polysulfone membrane. In conclusion, deleterious effects of hemodialysis on the metabolic activity of phagocytes were proved. Combining chemiluminescent and flow cytometric methods for the detection of ROS production and determining mitochondrial membrane potential can be useful tools for the analysis of material biocompatibility. Key words: Hemodialysis -Hemophan -Polysulfone -Reactive oxygen species -CD11b Abbreviations: ROS, reactive oxygen species; DCFH-DA, 2',7'-dichlorodihydrofluorescein diacetate; HE, hydroethidine; PMA, phorbol myristate acetate; FMLP, N-formyl-L-methionyl-L-leucyl-L-phenylalanine; OZP, zymosan opsonized in human serum; MRF, median of relative fluorescence

Effect of an Acute Iron Overdose on PMN Cells Chemiluminescence and Indices of Inner Environment in a Swine Model

The objective of the study was to evaluate the effect of an acute iron dextran administration on polymorphonuclear cells chemiluminescence and indices of inner environment in piglets. Piglets in the experimental group (n = 16) were given 2000 mg (20 ml) Fe3+ in the form of iron dextran i.m. Piglets in the control group (n = 13) were given the same amount of physiological saline. Blood was taken before administration, and 1, 4 and 24 hours after. Iron application resulted in a ten times higher concentration of iron in blood plasma in the experimental group. No differences in spontaneous, PMA- and OZP-stimulated chemiluminescence were recorded between the two groups. The increased activities of CK, AST and LDH are indicative of a muscle injury at the injection site. No changes indicating liver or kidney injury were reported. Our study provides evidence of a good safety of iron dextran after parenteral administration

New Role for L-Arginine in Regulation of Inducible Nitric-Oxide-Synthase-Derived Superoxide Anion Production in Raw 264.7 Macrophages

Dietary supplementation with L-arginine was shown to improve immune responses in various inflammatory models. However, the molecular mechanisms underlying L-arginine effects on immune cells remain unrecognized. Herein, we tested the hypothesis that a limitation of L-arginine could lead to the uncoupled state of murine macrophage inducible nitric oxide synthase and, therefore, increase inducible nitric-oxide-synthase-derived superoxide anion formation. Importantly, we demonstrated that L-arginine dose- and time dependently potentiated superoxide anion production in bacterial endotoxin-stimulated macrophages, although it did not influence NADPH oxidase expression and activity. Detailed analysis of macrophage activation showed the time dependence between LPS-induced iNOS expression and increased O2∙− formation. Moreover, downregulation of macrophage iNOS expression, as well as the inhibition of iNOS activity by NOS inhibitors, unveiled an important role of this enzyme in controlling O2∙− and peroxynitrite formation during macrophage stimulation. In conclusion, our data demonstrated that simultaneous induction of NADPH oxidase, together with the iNOS enzyme, can result in the uncoupled state of iNOS resulting in the production of functionally important levels of O2∙− soon after macrophage activation with LPS. Moreover, we demonstrated, for the first time that increased concentrations of L-arginine further potentiate iNOS-dependent O2∙− formation in inflammatory macrophages

Chemiluminescence response induced by mesenteric ischaemia/reperfusion: effect of antioxidative compounds ex vivo

Ischaemia and reperfusion (I/R) play an important role in human pathophysiology as they occur in many clinical conditions and are associated with high morbidity and mortality. Interruption of blood supply rapidly damages metabolically active tissues. Restoration of blood flow after a period of ischaemia may further worsen cell injury due to an increased formation of free radicals. The aim of our work was to assess macroscopically the extent of intestinal pathological changes caused by mesenteric I/R, and to study free radical production by luminol enhanced chemiluminescence (CL) of ileal samples. In further experiments, the antioxidative activity of the drugs tested was evaluated spectrophotometrically by the use of the DPPH radical. We studied the potential protective ex vivo effect of the plant origin compound arbutin as well as of the pyridoindole stobadine and its derivative SMe1EC2. I/R induced pronounced haemorrhagic intestinal injury accompanied by increase of myeloperoxidase (MPO) and N-acetyl-β-D-glucosaminidase (NAGA) activity. Compared to sham operated (control) rats, there was only a slight increase of CL response after I/R, probably in association with neutrophil increase, indicated by enhanced MPO activity. All compounds significantly reduced the peak values of CL responses of the ileal samples ex vivo, thus reducing the I/R induced increase of free radical production. The antioxidants studied showed a similar inhibitory effect on the CL response influenced by mesenteric I/R. If proved in vivo, these compounds would represent potentially useful therapeutic antioxidants

NA61/SHINE measurements of anisotropic flow relative to the spectator plane in Pb+Pb collisions at 30A GeV/c

We present an analysis of the anisotropic flow harmonics in Pb+Pb collisions at beam momenta of 30A GeV/c collected by the NA61/SHINE experiment in the year 2016. Directed and elliptic flow coefficients are measured relative to the spectator plane estimated with the Projectile Spectators Detector (PSD). The flow coefficients are reported as a function of transverse momentum in different classes of collision centrality. The results are compared with a new analysis of the NA49 data for Pb+Pb collisions at 40A GeV using forward calorimeters (VCal and RCal) for event plane estimation

Search for the critical point by the NA61/SHINE experiment

NA61/SHINE is a fixed target experiment operating at CERN SPS. Its main goals are to search for the critical point of stronglyinteractingmatterandtostudytheonsetofdeconfinement. Forthesegoalsascanofthetwodimensionalphase diagram (T-μB) is being performed at the SPS by measurements of hadron production in proton-proton, proton-nucleus and nucleus-nucleus interactions as a function of collision energy. In this paper the status of the search for the critical point of strongly interacting matter by the NA61/SHINE Collaboration is presented including recent results on proton intermittency, strongly intensive fluctuation observables of multiplicity and transverse momentum fluctuations. These measurements are expected to be sensitive to the correlation length and, therefore, have the ability to reveal the existence of the critical point via possible non-monotonic behavior. The new NA61/SHINE results are compared to the model predictions