Decreased H2O2 in exhaled breath condensate during pregnancy--feasible effect of 17beta-estradiol.

Since pregnancy is known to favor systemic generation of reactive oxygen species, this study was designed to assess the levels of exhaled hydrogen peroxide (eH2O2), serum progesterone (PG), 17beta-estradiol (E2) and systemic oxidative parameters in 20 pregnant women between 15th and 28th gestation week and 23 healthy, eumenorrheic women. Exhaled breath condensate H2O2 was assessed fluorometrically with homovanillic acid. Exhaled H2O2 levels were lowered in pregnancy (median Me 0.13 microM) compared with follicular (Me 0.29 microM) or luteal phase (Me 0.26 microM; p<0.05 vs. both). The follicular H2O2 tended to exceed luteal phase. Whole blood chemiluminescence was increased approximately ten fold in pregnancy. E2 markedly decreased chemiluminescence of isolated polymorphonuclear leukocytes. In vitro ferric reducing ability of plasma and 2,2-diphenyl-1-picryl-hydrazyl scavenging assay were not affected by E2 or PG. Decreased exhaled H2O2 during pregnancy, despite of the increased oxidative capacity of peripheral phagocytes, might be ascribed to the magnitude of increased 17beta-estradiol levels.

Elevated exhalation of hydrogen peroxide and circulating IL-18 in patients with pulmonary tuberculosis.

Mycobacteria are the strong stimulators of respiratory burst, resulting in production of reactive oxygen species and nitrogen intermediates. The aim of our study was to assess the concentration of hydrogen peroxide (H(2)O(2)) in expired breath condensate (EBC) and the serum level of interleukin-18 (IL-18) in patients with active pulmonary tuberculosis (TB) before introduction of chemotherapy and after 2 months of treatment. Sixteen patients, current cigarette smokers, with advanced pulmonary TB were enrolled into the study. As a control served two groups: I group--16 asymptomatic cigarette smokers, II group--17 healthy never smoked subjects. The level of H(2)O(2) in EBC was significantly higher in patients with TB (1.3+/-0.7 microM) as compared to cigarette are healthy nonsmoker subjects (0.4+/-0.1 and 0.2+/-0.1 microM, respectively, P<0.05). Two months of treatment significantly decreased the level of H(2)O(2) exhalation in TB patients (0.5+/-0.3 microM) to the value that was not different from that in asymptomatic smokers but was still higher than in never smoked subjects. Serum concentration of IL-18 in TB patients was higher than that found in both control groups either before and after antituberculous treatment (P<0.05). Exhaled H(2)O(2) did not correlate with circulating IL-18 in TB patients before or after treatment. These results demonstrated the occurrence of oxidative stress in the airways of TB patients completely attenuating after 2 months of successful antituberculous treatment.

Increased exhaled H2O2 and impaired lung function in patients undergoing bioincompatible hemodialysis.

BACKGROUND: Chronic renal failure (CRF) and hemodialysis (HD) accumulate an inflammatory milieu, contributing to increased systemic and airway oxidative stress that may lead to lung damage. OBJECTIVES: This study was designed to assess exhaled hydrogen peroxide (H2O2), lung function and whole blood chemiluminescence in HD and CRF patients and healthy controls. METHODS: The study included 59 patients (Polyamide STM or Hemophan membranes--19, cuprophane--16, hemodiafiltration--14, continuous ambulatory peritoneal dialysis--10), 16 CRF and 16 healthy controls. The assessment of lung function included FVC (forced vital capacity), FEV1 (forced expiratory volume in the first second) and DLCOc (single breath CO diffusing capacity). Exhaled H2O2 was determined fluorometrically and resting and n-formyl-methionyl-leucyl-phenylalanine (fMLP) luminol-dependent whole blood chemiluminescence (LBCL) were measured simultaneously. RESULTS: Only cuprophane HD patients presented decreased lung function (FVC 63.8+/-17.4%, FEV1 55.9+/-20.3 and DLCOc 72.1+/- 9.3 % of predicted; p<0.05 vs. controls). These patients exhaled the highest H2O2 levels in comparison to CRF (p<0.01): median 0.36 microM (range R: 0.09-0.56 microM) and controls (p<0.05): 0.17 microM (0.2-17.8 microM). These levels were not decreased during the HD session: preHD 1.25 microM (0.2-16.5 microM) and postHD 1.3 microM (0.2-17.8 microM). As a marker of systemic oxidative stress, fMLP-induced LBCL (total light emission) was increased in these patients (1570.6 aUxs/10(4) phagocytes; R: 274.2-8598.9) and in the CRF group (2389.4 aUxs /10(4) phagocytes; R: 491.5- 6184; p<0.05 vs. controls). Other patient groups did not express elevated LBCL and revealed decreased exhaled H2O2 after a session. CONCLUSIONS: An increased oxidative burden in the lungs may contribute to functional lung impairment in patients dialyzed with a cellulose membrane. Biocompatible dialysis with other modalities might reduce airway-borne oxidative stress and is not related with lung damage.

Decreased ratio of circulatory vascular endothelial growth factor to endostatin in patients with systemic sclerosis--association with pulmonary involvement.

OBJECTIVE: Vascular endothelial growth factor (VEGF) and endostatin appear to be involved in development of systemic sclerosis (SSc). We undertook this study to determine ratios of serum concentrations of VEGF to endostatin in SSc patients, healthy controls, assessments between cytokines, and lung-diffusing capacity (DLCO) as lung injury measurements related to interstitial lung disease (ILD). MATERIALS AND METHODS: Serum VEGF and endostatin levels were measured with ELISA in 28 SSc patients (16 with lcSSc) and 20-matched healthy volunteers, evaluating correlation and balance. DLCO was corrected for hemoglobin, alveolar volume, and determined with a single breath technique. RESULTS: SSc serum concentrations (median; range) of endostatin were higher than controls (107.2; 13.6-261.2 vs. 77.8; 18.0-110.4 ng/ml, p < 0.05); VEGF levels did not differ (151.2; 4.5-836.4 vs. 286.4; 23.7-708.5 pg/ml, p < 0.05). Ratios of VEGF to endostatin were 2.6 and 3.6 times lower (p < 0.05) in SSc and dcSSc in comparison to healthy subjects. There were significant negative correlations between VEGF, endostatin in SSc (r = -0.51), and controls (r = -0.57). SSc with ILD (n = 20) had similar concentrations of VEGF, endostatin, and ratios of VEGF to endostatin compared to SSc alone. No correlations were seen between DLCO, VEGF, endostatin and their ratios in the whole SSc group. Negative correlations were noted between DLCO and VEGF (r = -0.82), with DLCO and the ratio of VEGF to endostatin (-0.62) in lcSSc with ILD (n = 10). CONCLUSION: Decreased ratios of VEGF to endostatin may reflect imbalances between serum angiogenic, and anti-angiogenic activity in SSc, explaining impaired neoangiogenesis.

Elevated whole blood chemiluminescence in patients with systemic sclerosis.

OBJECTIVE: Systemic sclerosis (SSc) is accompanied by oxidative stress that in turn may accelerate endothelium degeneration and thus disease progression. We tested whether phagocytes from SSc patients release more reactive oxygen species (ROS) and whether this release correlates with some clinical parameters. METHODS: ROS production by blood phagocytes was measured with the luminol enhanced whole blood chemiluminescence (CL). Resting and N-formyl-methionyl-leucyl-phenylalanine -induced CL (fMLP-induced CL) was measured in 30 patients with SSc and 30 healthy controls matched as to age, sex, and level of cigarette smoking. RESULTS: Resting CL and fMLP-induced CL calculated per 10(4) phagocytes present in the assayed blood sample were higher in patients with systemic sclerosis than in healthy controls (median; range, 0.88; 0.47-1.39 vs. 0.73; 0.13-1.07 aU/10(4)p and 621; 293-3522 vs. 411; 289-810 aUxs/10(4)p, p<0.02). Patients treated with cyclophosphamide and/or prednisone for 11; 3-168 months did not differ in respect to CL from those that never received the medications. Similarly, no significant differences were found between patients with limited and diffuse SSc. Resting CL correlated (p<0.05) with clinically manifested interstitial lung disease (r=0.59), single breath carbon monoxide diffusing capacity (r= -0.56) and serum autoantibodies titre (r= 0.43). CONCLUSIONS: Blood phagocytes from patients with systemic sclerosis, especially from those with interstitial lung disease, generate elevated amounts of ROS as assessed with CL. This confirms the presence of systemic oxidative stress in SSc patients.

Influence of cerebral ventricles perfusion with hexapeptide derivatives of substance P on evoked tongue jerks in rats.

The effects of perfusion through cerebral ventricles with two hexapeptides, C-terminal derivatives of substance P, on evoked tongue jerks (ETJ), were studied on male rats. During perfusion, stimulation of the infraorbital nerve caused retractive movements (ETJ) of the stretched tongue, the amplitudes of which were recorded. The mean amplitudes of ETJ recorded during each 10 min period of perfusion with McIlwain-Rodnight's solution and solution containing hexapeptides were compared. The biologically most active C-terminal hexapeptide derivative of substance P (pyro-Glu6)SP6-11, perfused through cerebral ventricles in a concentration of 5 nmol/ml increased the ETJ. This effect was dose-dependent. A 10 times higher concentration in the perfusion fluid of the less biologically active hexapeptide SP6-11 was not effective. The possibility that substance P, and its active fragments present in the fluid in cerebral ventricles, could modulate the reflex centers in the medulla is discussed.

Use of a common food frequency questionnaire (FFQ) to assess dietary patterns and their relation to allergy and asthma in Europe: pilot study of the GA2LEN FFQ.

BACKGROUND/OBJECTIVES: Comparable international data on food and nutrient intake is often hindered by the lack of a common instrument to assess food intake. The objective of this study was within the Global Allergy and Asthma European Network of Excellence (GA(2)LEN), we developed and piloted a food frequency questionnaire (FFQ) to assess its validity in Europe. SUBJECTS/METHODS: Five countries participating in GA(2)LEN took part in the pilot study. A total of 200 adults aged 31-75 years were invited to complete a FFQ in two occasions and to give a blood sample. The intra-class correlation coefficient (ICC) was used to assess repeatability of the FFQ. Plasma phospholipid fatty acids (FAs) were analysed by gas chromatography. Pearson correlation was used to analyse the correlation between estimated dietary FA intake and plasma phospholipid FA levels. RESULTS: A total of 177 participants (89%) had complete data on FFQ(1) and plasma phospholipid FAs. In all, 152 participants (76%) completed both FFQs. ICCs between macronutrients ranged from 0.70 (saturated FAs) to 0.78 (proteins) and between 0.70 (retinol) and 0.81 (vitamin D) for micronutrients. Dietary n-3 FAs showed a good correlation with total plasma phospholipid n-3 FAs and with docosahexaenoic acid in the whole sample (0.40) and in individual countries. Poor correlations were observed for other FAs. CONCLUSIONS: The GA(2)LEN FFQ is an appropriate tool to estimate dietary intake for a range of nutrients across Europe regardless of cultural and linguistic differences. The FFQ seems to be useful to estimate the intake of n-3 FAs but not other FAs.

Inhibition of evoked tongue jerks during the rat's cerebral ventricles perfusion with gamma-aminobutiric acid and beta-phenyl-gamma-aminobutiric acid.

The supraorbital branch of the trigeminal nerve of rats in chloralose anaesthesia was stimulated with electrical pulses. The stimulation of the afferent fibres of the trigeminal nerve evoked the retractive jerks of the outstretched tongue. These evoked tongue jerks (ETJ) were recorded on the tape of the line recorder through an isotonic transducer. The caniulae were inserted into the lateral cerebral ventricles by means of streotaxic apparatus and perfusion was performed with inflow of GABA or beta-phenyl-GABA-containing fluid into the right lateral ventricle and outflow through the left lateral ventricle. Perfusion with solution of 0.01 mol/l GABA reduced the amplitude of ETJ 25.9% while perfusion with hypertonic NaCl solution increased it by 19.5%, and perfusion with solution of 0.2 mol/l beta-phenyl-GABA reduced that amplitude by 39.9%.

Elevated exhalation of hydrogen peroxide in patients with systemic sclerosis.

BACKGROUND: Systemic sclerosis is accompanied by an influx of activated phagocytes into distal airways. These cells release H2O2, which may evaporate from the airways surface and be detected in expired breath condensate. We tested whether patients with systemic sclerosis exhale more H2O2 than healthy subjects and whether breath condensate H2O2 levels correlate with some clinical parameters. MATERIAL AND METHODS: H2O2 was measured fluorimetrically in the expired breath condensate of 27 patients (22 women, five men, mean age 49 +/- 13.1 years) with systemic sclerosis and 27 age- and sex- matched healthy controls. RESULTS: Exhaled H2O2 levels were 3.5-fold higher (0.88 +/- 0.62 microM vs. 0.25 +/- 0.17 microM, P < 0.001) in the patients with systemic sclerosis than in the controls. Treatment with cyclophosphamide and/or prednisone (29 +/- 50 months, range 3-168 months) did not significantly decrease H2O2 exhalation (0.78 +/- 0.50 microM, n= 10 vs. 0.94 +/- 0.67 microM, n= 17, P > 0.05). No significant difference was found between patients with limited and diffuse scleroderma (1.03 +/- 0.69 microM, n= 17 vs. 0.63 +/- 0.41 microM, n= 10, P > 0.05). H2O2 levels correlated with disease duration (r = 0.38, P < 0.05) and time from the first Raynaud's episode (r = 0.44, P < 0.05). CONCLUSIONS: Patients with systemic sclerosis exhale more H2O2 than healthy controls, suggesting involvement of reactive oxygen species in disease processes. Lack of significant intergroups differences in H2O2 levels may have resulted from the small number of patients analyzed.