Activation of circulating polymorphonuclear neutrophils during exercise-induced muscle damage
; Croisier, Jean-Louis ; Chapelle, Jean-Paul et al
in Pflugers Arch – Eur J Physiol (2003, November), 447Detailed reference viewed: 7 (3 ULg)
Oxygen consumption and electron spin resonance studies of free radical production by alveolar cells exposed to anoxia: inhibiting effects of the antibiotic ceftazidime.
; ; et al
in Redox Report : Communications in Free Radical Research (2002), 7(2), 85-94
By EPR spectroscopy, we investigated free radical production by cultured human alveolar cells subjected to anoxia/re-oxygenation (A/R), and tested the effects of ceftazidime, an antibiotic previously ... [more ▼]
By EPR spectroscopy, we investigated free radical production by cultured human alveolar cells subjected to anoxia/re-oxygenation (A/R), and tested the effects of ceftazidime, an antibiotic previously demonstrated to possess antioxidant properties. Two A/R models were performed on type II pneumocytes (A549 cell line), either on cells attached to culture dishes (monolayer A/R model; 3.5 h of anoxia, 30 min of re-oxygenation) or after cell detachment (suspension A/R model; 1 h of anoxia, 10 min of re-oxygenation). Ceftazidime and selective inhibitors (SOD, Tiron, L-NMMA) were added before anoxia. Free radical production was assessed by the EPR spin trapping technique. Oxygen consumption was monitored, in parallel with EPR studies, in the suspension A/R model. The production of free radical species was demonstrated by the generation of PBN-radical adducts: (a(N) = 15.2 G) in the monolayer A/R model and a six-line EPR spectrum (a(N) = 15.7 G and a(H) = 2.7 G) in the suspension A/R model. A kinetic study performed by oximetry, in parallel with EPR spectroscopy, demonstrated marked alterations of the cell respiratory function and that the free radical production started during anoxia and increased during re-oxygenation. In the suspension A/R model, the amplitude of EPR spectra were decreased upon the addition of 200 U/ml SOD (37% inhibition), 0.1 mM Tiron (67% inhibition) and 1 mM L-NMMA (43% inhibition). Addition of 1 mM ceftazidime decreased the amplitude of EPR spectra (37% inhibition) in both A/R models. Complementary in vitro EPR studies demonstrated that CAZ scavenged the hydroxyl radical (produced by the Fenton reaction). The protective effect of ceftazidime in the cell model could thus be linked to its ability to scavenge superoxide anions, nitrogen-derived species and hydroxyl radicals. [less ▲]Detailed reference viewed: 12 (1 ULg)
Marqueurs humoraux d'activation de l'endothélium vascualire et des polymorphonucléaires neutrophiles circulants lors de l'exercice intense
; ; Nys, Monique et al
Poster (2001, May)Detailed reference viewed: 10 (2 ULg)
In vitro study of the antioxydant properties of the nimesulide and 4OH nimesulide : effect on HRP- and luminol-dependent chemiluminescence produced by human chondrocytes
Henrotin, Yves ; ; Mouithys-Mickalad, Ange et al
in Annals of the Rheumatic Diseases (2000), 59(S1), 265Detailed reference viewed: 6 (2 ULg)
In vitro study of the antioxydant properties of nonsteroidal anti-inflammatory drugs by chemiluminescence and electron spin resonance (ESR)
Henrotin, Yves ; Mouithys-Mickalad, Ange ; et al
in Annals of the Rheumatic Diseases (2000), 59(S1), 187Detailed reference viewed: 7 (1 ULg)
In vitro study of the antioxidant properties of non steroidal anti-inflammatory drugs by chemiluminescence and electron spin resonance (ESR).
Mouithys-Mickalad, Ange ; ; et al
in Free Radical Research (2000), 33(5), 607-21
OBJECTIVES: To determine the antioxidant activities of nonsteroidal anti-inflammatory drugs (NSAIDS), we examined by chemiluminescence (CL) and electron spin resonance (ESR) their scavenging properties ... [more ▼]
OBJECTIVES: To determine the antioxidant activities of nonsteroidal anti-inflammatory drugs (NSAIDS), we examined by chemiluminescence (CL) and electron spin resonance (ESR) their scavenging properties towards lipid peroxides, hypochlorous acid and peroxynitrite. METHODS: The antioxidant properties of nimesulide (NIM), 4-hydroxynimesulide (4-HONIM), aceclofenac (ACLO), 4-hydroxyaceclofenac (4-HOA-CLO), diclofenac (DICLO) and indomethacin (INDO) were tested on four different reactive oxygen species (ROS) generating systems: (I) phorbol-myristate acetate (PMA)-activated neutrophils, (II) Fe2+/ascorbate-induced lipid peroxidation, (III) HOCl-induced light emission, (IV) the kinetics of ONOO- decomposition followed by spectrophotometry. ROS production was monitored by luminol-enhanced CL or by ESR using two different spin traps. RESULTS: At 10 microM, ACLO, NIM, 4-HONIM, 4-HOA-CLO, and DICLO decreased luminol-enhanced CL generated by PMA-activated neutrophils. Inversely, INDO increased the luminol enhanced CL. Interestingly, hydroxylated metabolites were more potent antioxidants than the parent drugs. Furthermore, all drugs tested, excepted ACLO, lowered lipid peroxidation induced by Fe2+/ascorbate system. ACLO and DICLO, even at the highest concentration tested (100 microM), did not significantly lower HOCl induced CL, whereas the other drugs were potent scavengers. Finally, all the NSAIDS accelerated decomposition of ONOO-, suggesting a potential capacity of the molecules to scavenge peroxynitrite. CONCLUSION: The NSAIDs possess variable degrees of antioxidant activities, linked to their ability to react with HOCl, lipid peroxides or ONOO-. These antioxidant activities could offer interesting targeted side-effects in the treatment of joint inflammatory diseases. [less ▲]Detailed reference viewed: 12 (4 ULg)
Oxygen therapy in intensive care patients: a vital poison?
Deby, Ginette ; ; Lamy, Maurice
in Vincent, Jean-Louis (Ed.) Yearbook of intensive care and emergency medicine (1999)Detailed reference viewed: 25 (0 ULg)
Oxidant-Scavenging Activities of Beta-Lactam Agents
; ; et al
in European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology (1998), 17(1), 43-6
The relative antioxidant effect of ampicillin, ceftazidime, ceftriaxone, and cefuroxime on oxygen-reactive species was examined in vitro using stimulated human polymorphonuclear neutrophils. There was no ... [more ▼]
The relative antioxidant effect of ampicillin, ceftazidime, ceftriaxone, and cefuroxime on oxygen-reactive species was examined in vitro using stimulated human polymorphonuclear neutrophils. There was no evidence that any of the beta-lactam agents tested had an effect on superoxide or H2O2 generation. In contrast, all of the beta-lactam agents prevented hypochlorous acid (HOCI) chlorination of 1,1-dimethyl-4-chloro-3,5-cyclo-hexanedione in a cell-free system at concentrations of < 10 microg/ml. Furthermore, all antibiotics provided dose-dependent protection against HOCI cytotoxicity to 16HBE140 bronchial epithelial cells. Taken together, these data indicate a possible therapeutic role for beta-lactam agents in protecting host tissues from HOCI-induced oxidative damage. [less ▲]Detailed reference viewed: 8 (0 ULg)
Purification of myeloperoxidase from equine polymorphonuclear leucocytes.
Mathy, Marianne ; ; Grulke, Sigrid et al
in Canadian Journal of Veterinary Research = Revue Canadienne de Recherche Vétérinaire (1998), 62(2), 127-32
Increases of plasma concentrations of neutrophil myeloperoxidase (MPO) can be used as markers of polymorphonuclear leucocytes (PMN) activation in pathological situations (sepsis, acute lung injury, acute ... [more ▼]
Increases of plasma concentrations of neutrophil myeloperoxidase (MPO) can be used as markers of polymorphonuclear leucocytes (PMN) activation in pathological situations (sepsis, acute lung injury, acute inflammation). To develop an assay for measurement of plasma MPO in horses during the above-mentioned infectious and inflammatory conditions, MPO was purified from equine PMN isolated from blood anticoagulated with citrate. PMN were extracted in a saline milieu (0.2 M Na acetate, 1 M NaCl, pH 4.7) to eliminate most of cellular proteins. Pellets were then extracted in the same buffer containing cationic detergent (1% cetyltrimethyl ammonium bromide). The supernatant was further purified by ion exchange chromatography (Hiload S Sepharose HP column 0.5 x 26 cm, equilibrated with 25 mM Na acetate, 0.2 M NaCl, pH 4.7) with a NaCl gradient (until 1 M). Most of the peroxidase activity of MPO (spectrophotometrically measured by the oxidation of orthodianisidine by hydrogen peroxide) was eluted at 0.65 M NaCl. MPO was further purified by gel filtration chromatography (Sephacryl S 200 column 2.6 x 42 cm with 25 mM Na acetate, 0.2 M NaCl, pH 4.7). MPO (specific activity: 74.3 U/mg) was obtained with a yield of 30% from the detergent extraction supernatant. Electrophoresis (non-reducing conditions) showed 3 bands identified, by comparison with human MPO, (i) the mature tetrameric enzyme (150 kDa) with 2 light and 2 heavy subunits, (ii) the precursor form (88 kDa) and (iii) a form of the heavy subunit without the prosthetic heme group (40 kDa). The mature enzyme and its precursor were glycosylated and possessed peroxidase activity. Equine MPO showed strong similarities with human and bovine MPO, with an absorption peak at 430 nm (Soret peak) characteristic of ferrimyeloperoxidase. Enzymatic activity was pH dependent (optimal value at pH 5.5). [less ▲]Detailed reference viewed: 37 (1 ULg)
Equine neutrophil myeloperoxidase in plasma: design of a radio-immunoassay and first results in septic pathologies.
Deby, Ginette ; Grulke, Sigrid ; et al
in Veterinary Immunology and Immunopathology (1998), 66(3-4), 257-71
The strangulated intestinal pathologies of horses are accompanied by a local activation of the neutrophils, that can be revealed by measuring the tissular enzymatic activity of the granulocytic enzyme ... [more ▼]
The strangulated intestinal pathologies of horses are accompanied by a local activation of the neutrophils, that can be revealed by measuring the tissular enzymatic activity of the granulocytic enzyme myeloperoxidase (MPO). To estimate the possible spreading of this neutrophil activation to the systemic circulation, we designed a radioimmunoassay (RIA) for equine neutrophil myeloperoxidase (MPO) (EC 126.96.36.199) using a specific rabbit antiserum. MPO was labeled with 1 mCi 125I by a technique of self-labeling in the presence of 10(-4) M hydrogen peroxide. The RIA was performed by incubation of 100 microl diluted antiserum, 100 microl labeled MPO (+/-30,000 cpm) and 100 microl of the reference molecule (unlabeled MPO) solution or the unknown sample, at room temperature for 18 h. The antibody-antigen complexes were isolated by double antibody precipitation. The sensitivity of the RIA was 2 ng/ml. The RIA showed good precision and accuracy with intra- and inter-assay coefficients of variation 6% and 8%, respectively, for MPO concentrations ranging from 2 ng/ml to 60 ng/ml. The best sampling technique for MPO measurement in plasma was to collect blood into EDTA, which allowed us to get a plasmatic value stable with time. The mean MPO value in normal horses was 69.5 +/- 19.4 ng/ml in EDTA anticoagulated plasma (n = 48). The stress of transport and anaesthesia did not modify the mean plasmatic value of MPO. No significant increase of plasma MPO was observed in 17 horses submitted to surgery for pathologies without systemic impact. But, in 25 horses with obstructive intestinal pathologies, persistent abnormal MPO concentrations were measured (until 740 ng/ml). [less ▲]Detailed reference viewed: 38 (1 ULg)
The antibiotic ceftazidime is a singlet oxygen quencher as demonstrated by ultra-weak chemiluminescence and by inhibition of AAP consumption.
Deby, Ginette ; ; Mouithys-Mickalad, Ange et al
in Biochimica et Biophysica Acta (1998), 1379(1), 61-8
We demonstrated that the cephalosporin antibiotic ceftazidime (CAZ) deactivated singlet oxygen (1O2). We then studied the mechanisms of the CAZ effects on the ultra weak chemiluminescence (uwCL ... [more ▼]
We demonstrated that the cephalosporin antibiotic ceftazidime (CAZ) deactivated singlet oxygen (1O2). We then studied the mechanisms of the CAZ effects on the ultra weak chemiluminescence (uwCL) associated with the energy decay of 1O2 generated by the Mallet reaction (H2O2 + HOCl --> HCl + H2O + 1O2), and on the anthracene-9,10-dipropionic acid (AAP) consumption by 1O2 generated by irradiation of Rose Bengal (RB). The uwCL generated by the Mallet reaction was amplified (6.2 times) by CAZ. The use of red and blue filters, which absorb radiation below 610 nm and between 470 and 700 nm respectively, demonstrated that CAZ increased the uwCL by a radiation emission at wavelengths shorter than the 633 and 704 nm wavelength emissions of 1O2. CAZ was excited by scavenging the energy excess of 1O2, which so returned to its fundamental state, while CAZ deactivated with light emission between 430-480 nm. CAZ also inhibited in a dose-dependent manner the consumption of AAP by 1O2 generated by the irradiation of RB. The protection of AAP by 5 x 10(-3) M CAZ was equivalent to that of 10(-3) M histidine and 3 X 10(-6) M sodium azide. This process of 1O2 deactivation will be useful in diseases characterized by an excessive PMN activation with a release of activated oxygen species. [less ▲]Detailed reference viewed: 57 (8 ULg)
Propofol reacts with peroxynitrite to form phenoxyl radicals. Demonstration by ESR
Mouithys-Mickalad, Ange ; ; et al
Conference (1998)Detailed reference viewed: 4 (2 ULg)
Effects of sphingosine and sphingosine analogues on the free radical production by stimulated neutrophils: ESR and chemiluminescence studies.
Mouithys-Mickalad, Ange ; Deby, Ginette ; Hoebeke, Maryse et al
in Mediators of Inflammation (1997), 6(5-6), 327-33
Sphingolipids inhibit the activation of the neutrophil (PMN) NADPH oxidase by protein kinase C pathway. By electron spin resonance spectroscopy (ESR) and chemiluminescence (CL), we studied the effects of ... [more ▼]
Sphingolipids inhibit the activation of the neutrophil (PMN) NADPH oxidase by protein kinase C pathway. By electron spin resonance spectroscopy (ESR) and chemiluminescence (CL), we studied the effects of sphingosine (SPN) and ceramide analogues on phorbol 12-myristate 13-acetate (PMA, 5 x 10(-7)M) stimulated PMN (6 x 10(6) cells). By ESR with spin trapping (100 mM DMPO: 5,5-dimethyl-1-pyrroline-Noxide), we showed that SPN (5 to 8 x 10(-6)M), C(2)-ceramide (N-acetyl SPN) and C(6)-ceramide (N-hexanoyl SPN) at the final concentration of 2 x 10(-5) and 2 x 10(-4)M inhibit the production of free radicals by stimulated PMN. The ESR spectrum of stimulated PMN was that of DMPO-superoxide anion spin adduct. Inhibition by 5 x 10(-6)M SPN was equivalent to that of 30 U/ml SOD. SPN (5 to 8 x 10(-6)M) has no effect on in vitro systems generating superoxide anion (xanthine 50 mM/xanthine oxidase 110 mU/ml) or hydroxyl radical (Fenton reaction: 88 mM H(2)O(2), 0.01 mM Fe(2+) and 0.01 mM EDTA). SPN and N-acetyl SPN also inhibited the CL of PMA stimulated PMN in a dose dependent manner (from 2 x 10(-6) to 10(-5)M), but N-hexanoyl SPN was less active (from 2 x 10(-5) to 2 x 10(-4)M). These effects were compared with those of known PMN inhibitors, superoxide dismutase, catalase and azide. SPN was a better inhibitor compared with these agents. The complete inhibition by SPN of ESR signal and CL of stimulated PMN confirms that this compound or one of its metabolites act at the level of NADPH-oxidase, the key enzyme responsible for production of oxygen-derived free radicals. [less ▲]Detailed reference viewed: 38 (8 ULg)
Effects of training on myocellular enzyme leakage and delayed onset muscle soreness following maximal isokinetic eccentric exercise
Croisier, Jean-Louis ; ; et al
in Mediators of Inflammation (1997), 6Detailed reference viewed: 7 (0 ULg)
Experimental model for the study by chemiluminescence of the activation of isolated equine leucocytes.
; Deby, Ginette ; et al
in Research in Veterinary Science (1996), 61(1), 59-64
The activation of human polymorphonuclear leucocytes (the respiratory burst) can be studied by measuring their chemiluminescent response. This technique was adapted to equine leucocytes to investigate the ... [more ▼]
The activation of human polymorphonuclear leucocytes (the respiratory burst) can be studied by measuring their chemiluminescent response. This technique was adapted to equine leucocytes to investigate the effects of cell number, activator concentration, enhancers of chemiluminescence, pH, temperature and inhibitors. Leucocytes were isolated from citrated blood from healthy horses and chemiluminescence was measured with a Bio-Orbit luminometer sensitive to 900 nm light. The optimal cell density for the maximal chemiluminescent response ranged from 10(6) to 10(7) leucocytes 600 microliters-1. Chemiluminescence increased as a function of temperature, and the concentrations of luminol, lucigenin and phorbol myristate acetate (PMA), and was pH related (optimal pH value = 8.0 for lucigenin and 8.5 for luminol). The inhibition of chemiluminescence by 5 x 10(-5) M azide was 88 per cent for luminol and 37 per cent for lucigenin. Superoxide dismutase (100 IU) totally inhibited the chemiluminescence response. Approximately 30 per cent variability in chemiluminescence was observed under the same assay conditions, depending on the origin of the leucocytes. Based on these results, the conditions selected for the measurement of equine leucocyte chemiluminescence were: 10(6) to 10(7) leucocytes 600 microliters-1, 1 x 10(-6)M PMA, 1 mM luminol or 0.4 mM lucigenin, physiological pH (7.4) and physiological temperature (37.8 degrees C). These conditions were similar to those used for measuring the chemiluminescent response of human leucocytes. [less ▲]Detailed reference viewed: 30 (1 ULg)
Bactericidal activity against Pseudomonas aeruginosa is acquired by cultured human monocyte-derived macrophages after uptake of myeloperoxidase.
; Deby, Ginette ; Melin, Pierrette et al
in Experientia (1996), 52(2), 167-74
Myeloperoxidase (MPO) is an enzyme located within polymorphonuclear neutrophils capable of producing cytotoxic oxidant species that are particularly active against bacteria with polysaccharide capsules ... [more ▼]
Myeloperoxidase (MPO) is an enzyme located within polymorphonuclear neutrophils capable of producing cytotoxic oxidant species that are particularly active against bacteria with polysaccharide capsules. Pseudomonas aeruginosa (10(6) bacteria per 1ml) are killed within 1 h in vitro by a MPO/H2O2/C1- system (48mU=132ng of MPO). The question arose as to whether human macrophages would acquire cytotoxic activity when loaded with this enzyme. Monocytes were therefore isolated from human blood and cultured for up to ten days to induce maturation to macrophages. These cells lost endogenous MPO within five days while H2O2 production in response to stimulation by phorbol myristate acetate (10(-6)M) decreased to 23% within ten days. On the other hand, their capacity to take up exogenous MPO increased fourfold from day three to day ten. Human macrophages cultured from eight days (when both H2O2 production and MPO uptake were sufficient) were therefore used to study the effects of MPO uptake on cytocidal activity against Pseudomonas aeruginosa. After a 1 h MPO loading period, macrophages (5X10(5) cells per ml) were incubated in the presence of bacteria (0.5 to 2X10(6) bacteria per ml) for 2 h at 37 degrees C. At a bacteria/macrophage ratio of 1, only 34.8+/-7.0% of bacteria survived (compared to killing by non-loaded macrophages), while 74.4+/-9.3% survived at a ratio of 4. From these results, we conclude that loading macrophages with exogenous MPO could enhance their microbicidal activity, suggesting a potentially useful therapeutic application. [less ▲]Detailed reference viewed: 148 (0 ULg)
Piroxicam fails to reduce myocellular enzyme leakage and delayed onset muscle soreness induced by isokinetic eccentric exercise
Croisier, Jean-Louis ; ; et al
in Pflügers Archiv : European Journal of Physiology (1996), 431Detailed reference viewed: 3 (0 ULg)
Piroxicam fails to reduce myocellular enzyme leakage and delayed onset muscle soreness induced by isokinetic eccentric exercise.
Croisier, Jean-Louis ; ; et al
in Mediators of Inflammation (1996), 5(3), 230-4
To test the hypothesis that delayed onset muscular soreness (DOMS) following intense eccentric muscle contraction could be due to increased production of prostaglandin E(2) (PGE(2)), ten healthy male ... [more ▼]
To test the hypothesis that delayed onset muscular soreness (DOMS) following intense eccentric muscle contraction could be due to increased production of prostaglandin E(2) (PGE(2)), ten healthy male subjects were studied. Using a double-blind randomized crossover design, each subject performed two isokinetic tests separated by a period of at least 6 weeks: once with placebo, and once with piroxicam (Feldene((R))). They were given one capsule containing either placebo or piroxicam (20 mg) per day for 6 days with initial doses given starting 3 days prior to isokinetic testing. Exercise consisted of eight stages of five maximal contractions of the knee extensor and flexor muscle groups of both legs separated by 1 min rest phases, on a Kin Trex device at 60( degrees )/s angular velocity. The subjective presence and intensity of DOMS were evaluated using a visual analogue scale immediately after, and 24 and 48 h after each test. The mean plasma concentration of PGE(2) measured at rest and after exercise was significantly lower in the group treated with piroxicam (p < 0.05). However, statistical analysis (two-way ANOVA test) revealed that exercise did not cause any significant change of mean plasma PGE(2) over time in either of the two groups. Eccentric work was followed by severe muscle pain in extensor and flexor muscle groups. Maximal soreness was noted 48 h postexercise. Serum creatine kinase activity and the serum concentration of myoglobin increased significantly, and reached peak values 48 h after exercise in both experimental conditions (p < 0.001). By paired t-test, it appeared that there were no significant differences in the serum levels of these two markers of muscle damage between the two groups at any time point. We conclude that: (1) oral administration of piroxicam fails to reduce muscle damage and DOMS caused by strenuous eccentric exercise; and (2) the hypothetical role of increased PGE(2) production in eccentric exercise-induced muscle damage, DOMS, and reduced isokinetic performance is not substantiated by the present results. [less ▲]Detailed reference viewed: 52 (7 ULg)
Antioxidant Defense and Free Radical Production in a Rabbit Model of Kidney Ischemia-Reperfusion
Franssen, Colette ; Defraigne, Jean-Olivier ; Detry, Olivier et al
in Transplantation Proceedings (1995), 27(5), 2880-3Detailed reference viewed: 34 (0 ULg)