Hydrogen peroxide (H2O2) production in exhaled air was measured in ventilated preterm newborns at 5, 24 and 48 hours after delivery, using originally designed method of exhaled breath condensate (EBC) collection. H2O2 production in expired gas was 812±34 pmol/20 min during the first measurement and then declined to 389±21 at 24 hours and 259±26 pmol/20 min at 48 hours.
Postnatal heart development is characterized by critical periods of heart remodeling. In order to characterize the changes in the lipophilic fraction induced by free radicals, fatty acids and t heir oxidized products, lipofuscin -like pigments (LFP), were investigated. Fatty acids were analyz ed by gas chromatography and LFP were studied by fluorescence techniques. A fluorophore characterized by spectral methods was further resolved by HPLC. Major changes in the composition of fatty acids occurred immediately after birth and then during maturation. Fluorescence of LFP changed markedly on postnatal days 1, 4, 8, and 14, and differed from the adult animals. LFP comprise several fluorophores that were present since fetal state till adulthood. No new major fluorophores were formed during development, just the abundances of individual fluorophores have been modulated which produced changes in the shape of the spectral arrays. HPLC resolved the fluorophore with excitation maximum at 360 nm and emission maximum at 410 nm. New chromatographically distinct species appeared immediately on postnatal day 1, and then on days 30 and 60. Consumption of polyunsaturated fatty acids immediately after birth and subseque nt formation of LFP suggests that oxidative stress is involved in normal heart development., J. Wilhelm, J. Ivica, Z. Veselská, J. Uhlík, L. Vajner., and Obsahuje bibliografii
Phagocytosis is associated with respiratory burst producing reactive oxygen and nitrogen species. Several studies imply that erythrocytes can inhibit the respiratory burst during erythrophagocytosis. In this work we studied the mechanisms of this effect using control and in vitro peroxidized erythrocyte membranes. We demonstrated that autofluorescence of peroxidation products can be used for visualization of phagocytozed membranes by fluorescence microscopy. We also found that respiratory burst induced by a phorbol ester was inhibited by control membranes (5 mg/ml) to 63 % (P0.001), and to 40 % by peroxidized membranes (P0.001). We proved that this effect is not caused by the direct interaction of membranes with free radicals or by the interference with luminol chemiluminescence used for the detection of respiratory burst. There are indications of the inhibitory effects of iron ions and free radical products. Macrophages containing ingested erythrocyte membranes do not contain protein-bound nitrotyrosine. These observations imply a specific mechanism of erythrocyte phagocytosis.
Increased generation of reactive oxygen species results in the formation of fluorescent end-products of lipid peroxidation – lipofuscin-like pigments (LFP). LFP increased up to six-fold from the fetal value in the rat heart immediately after birth. In the experimental design of this study the fetuses were sampled 1 day before birth, and then the samples were collected on postnatal days 1, 4, 7, 10, 15, 30, and 60. Males and females were compared on day 30 and 60 when the difference between right and left ventricle was studied as well. Four LFP fluorophores were analyzed: F355/440, F310/470, F350/450, F315/450 (excitation/emission, nm). All fluorophores decreased on day 4 relative to day 1, subsequent transient increases ended in a significant decrease on day 60. However, the LFP levels on day 60 are still about threefold higher than those in fetuses. Differences between male and female hearts were observed on day 30. The corresponding male ventricles contained by one third higher concentration of LFP than the female counterparts. The increase in LFP concentration in male ventricles on day 30 was only transient, no difference between corresponding male and female ventricles was found on day 60. The most distinguished feature in the male heart was a sharp LFP decrease in the right ventricle on day 60., J. Wilhelm, I. Ošťádalová., and Obsahuje seznam literatury
We have studied in vitro alveolar macrophages (AMs) obtained by tracheobronchial lavage from rats exposed to subacute (3 hours and 3 days) and chronic (3 weeks) hypoxia (Fi02 = 0.1) and from rats recovering from chronic hypoxia. Hydrogen peroxide production by AMs was measured by luminol- depcndent chemiluminescence after AMs adhered to the walls of the measuring cuvette, after stimulation with phorbol-myristate-acetate (PMA), and when N-formyl-methionyl-leucyl-phenylanine (FMLP) was added subsequently to the cells which had been previously stimulated by adherence or PMA. H2O2 production after cell adherence and adherence combined with FMLP stimulation did not differ between the groups. The increase of H2O2 production after adding PMA, and FMLP in addition to PMA was significantly higher in AMs from rats exposed to hypoxia for 3 days than in the controls. Other experimental groups did not differ from their controls. It is concluded that 3 days’ hypoxia primes AMs for enhanced production of H2O2 upon stimulation. The mechanism is probably at the level of synthesis of proteins involved in H2O2 production, or the shift to a more reactive phenotype of alveolar macrophages subpopulations.
Production of hydrogen peroxide by rat lung alveolar macrophages represents one of the key events in the inflammatory process. For the interpretation of the in vitro measurements it is important to control all possible interfering influences. The present work documents that the type of anaesthesia might critically influence the observed results. H2O2 production was measured in isolated rat alveolar macrophages by luminol chemiluminescence catalyzed by horseradish peroxidase. Three different mechanisms of H2O2 production were observed after stimulation of cells with a chemotactic peptide (FMLP), phorbol ester (PMA), and during cell adherence. All these activities were influenced independently by the treatment with barbiturates, which both stimulated or inhibited the H2O2 production, depending on the barbiturate concentration. As the effective barbiturate concentrations were found to be within the range used for the anaesthesia of experimental animals, the presented results imply that barbiturates are not suitable for experiments in which the production of reactive oxygen species by phagocytes is measured, and that other anaesthetics should be tested.
Hydrogen peroxide production was measured in non-elicited rat peritoneal macrophages using luminol-dependent chemiluminescence (LDCL). Isolated cells were activated by a chemotactic peptide (FMLP) or by a phorbol ester (PMA) or by the combination of both. A hundred-fold higher LDCL intensity was achieved with PMA relative to FMLP. However, when FMLP was added subsequently to PMA it produced approximately the same response as did PMA. These measurements were carried out with cells isolated from controls and from animals exposed to normobaric hypoxia (10 % O2) for 3 hours, 3 days, or 21 days. Hypoxia had a dual effect. Acutely (within 3 hours) it attenuated the production of hydrogen peroxide triggered by PMA, whilst during longer exposure (3 or 21 days) it increased the response induced by FMLP. Hypoxia can thus modulate the capacity of respiratory burst in peritoneal macrophages.
Hydrogen peroxide injected into the inflow cannula of isolated ventilated rat lungs produced a dose-dependent vasoconstriction in the range 0.25-10 mM, with maximum response between 2 - 5 mM. The effects of H2O2 can be influenced by ionophores or specific inhibitors of ionic channels or pumps. A key role is played by sodium ions which govern the subsequent inflow or outflow of calcium, an ion mediating the vasoconstriction. A physiological role for H2O2 generated by NADPH oxidase is postulated.
The effect of three-day fasting on cardiac ischemic tolerance was investigated in adult male Wistar rats. Anesthetized open-chest animals (pentobarbitone 60 mg/kg, i.p.) were subjected to 20-min left anterior descending coronary artery occlusion and 3-h reperfusion for infarct size determination. Ventricular arrhythmias were monitored during ischemia and at the beginning (3 min) of reperfusion. Myocardial concentrations of beta-hydroxybutyrate and acetoacetate were measured to assess mitochondrial redox state. Short-term fasting limited the infarct size (48.5±3.3 % of the area at risk) compared to controls (74.3±2.2 %) and reduced the total number of premature ventricular complexes (12.5±5.8) compared to controls (194.9±21.9) as well as the duration of ventricular tachycardia (0.6±0.4 s vs. 18.8±2.5 s) occurring at early reperfusion. Additionally, fasting increased the concentration of beta-hydroxybutyrate and betahydroxybutyrate/acetoacetate ratio (87.8±27.0) compared to controls (7.9±1.7), reflecting altered mitochondrial redox state. It is concluded that three-day fasting effectively protected rat hearts against major endpoints of acute I/R injury. Further studies are needed to find out whether these beneficial effects can be linked to altered mitochondrial redox state resulting from increased ketogenesis., M. Šnorek ... [et al.]., and Obsahuje seznam literatury