Rabea K L Bouhafs

Karolinska Institutet, Solna, Stockholm, Sweden

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Publications (10)25.78 Total impact

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    ABSTRACT: We hypothesized that aggregates of ultrafine carbon and washed diesel particles impair the ability of alveolar macrophages (AM) to kill bacteria and enhance the AM lipid peroxidation (LPO) of lung surfactant. Rat AM were exposed, 5h, to particles 20 microg/ml. The AM, containing carbon or washed diesel particles, were incubated 2h, with Streptococcus pneumoniae, an American Type Culture Collection (ATCC) strain or clinical isolates. Surviving bacteria were quantified. Surfactant was incubated, 5h, with carbon or washed diesel loaded AM and LPO was measured. The particle load was approximately 1 microg/10(6) AM, representing accepted exposure to ambient particles in Europe. Metal concentrations were 10 to 100 fold higher in washed diesel--than in carbon particles. There was a dose dependent increase in bacterial survival with carbon-loaded macrophages, but not with washed diesel-loaded AM. Clinical isolates had a higher survival rate with carbon-loaded macrophages than the ATCC strain. Surfactant LPO was increased with washed diesel-loaded macrophages (95%) and with carbon-loaded macrophages (55%) compared to controls. High LPO caused by washed diesel-loaded AM reflects their increased oxidative metabolism, probably caused by particle metals. The additional oxygen metabolites maintained bactericidal activity of AM, while corresponding activity was decreased in carbon-loaded AM. Altered functions of AM may explain health problems related to air pollution.
    Environmental Research 07/2007; 104(2):250-7. · 3.24 Impact Factor
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    ABSTRACT: In preterm infants with respiratory distress syndrome, surfactant administration followed by immediate extubation to spontaneous breathing with nasal continuous positive airway pressure reduces the need for mechanical ventilation. With this treatment approach, repeated doses of surfactant are rarely indicated. We used a rabbit model to test the hypothesis that exogenous surfactant therapy followed by spontaneous breathing results in a more sustained initial treatment response compared with treatment followed by mechanical ventilation. Preterm rabbits (gestational age 28.5 d) were treated with pharyngeal deposition of 200 mg/kg radiolabeled surfactant (14C-Curosurf) and randomized to 4 h of spontaneous breathing or mechanical ventilation or to a control group, killed immediately after surfactant administration. With pharyngeal deposition, 46 +/- 10% (mean +/- SEM) of the administered surfactant reached the lungs. The dynamic lung-thorax compliance was higher in spontaneously breathing compared with mechanically ventilated animals (median, 9.9 and 0.75 ml x cm H2O(-1) x kg(-1), respectively; p < 0.05). The relative distribution of 14C-Curosurf in bronchoalveolar lavage fluid and homogenized lung tissue showed a higher degree of tissue association in the spontaneously breathing animals [53 +/- 4 versus 26 +/- 3% (mean +/- SEM)] than in mechanically ventilated animals (p < 0.01), the latter figure being very similar to that of the control group (25 +/- 5%). There was a higher degree of lipid peroxidation and fewer microbubbles in bronchoalveolar lavage fluid from mechanically ventilated animals. We conclude that the initial lung tissue association of exogenous surfactant is impaired by mechanical ventilation. This is associated with a reduction of dynamic compliance and evidence of increased surfactant inactivation.
    Pediatric Research 05/2005; 57(5 Pt 1):624-30. · 2.67 Impact Factor
  • R K L Bouhafs, C Jarstrand, B Robertson
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    ABSTRACT: Group B streptococcal (GBS) pneumonia, with neutrophilic granulocytes immigrating into the lungs, may occur in neonates. The incidence is particularly high among preterm infants, who often are treated with exogenous surfactant. We have previously demonstrated in vitro that neutrophils stimulated by GBS cause lipid peroxidation (LPO) and functional impairment of lung surfactant. The present study aimed at evaluating LPO of exogenous lung surfactant (Curosurf) and the protective effect of the natural antioxidant, vitamin E in immature ventilated newborn rabbits with experimental neonatal GBS pneumonia. There was a prominent proliferation of GBS in the lungs of animals treated with surfactant and ventilated for 5 h. GBS-infected rabbits had a higher LPO of lung lavage fluid than non-infected ones. The LPO could be diminished using vitamin E, which, however, did not affect bacterial proliferation. During the 5-h incubation period, mean lung-thorax compliance values were significantly lower in GBS-infected than in noninfected animals. We speculate that addition of vitamin E to exogenous surfactant preparations may improve their resistance to LPO and make them more suitable for treatment of neonates with pneumonia.
    Beiträge zur Klinik der Tuberkulose 02/2004; 182(2):61-72. · 2.06 Impact Factor
  • Rabea K L Bouhafs, Anders Samuelson, Connie Jarstrand
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    ABSTRACT: We used Pseudomonas aeruginosa, Burkholderia cepacia and Stenotrophomonas maltophilia, live or heat-killed, isolated from the airways of children with Cystic Fibrosis, to stimulate human neutrophils (PMN) and rat alveolar macrophages (AM) to produce reactive oxygen metabolites in the presence or absence of Curosurf, a natural porcine lung surfactant. We determined: (1) the amount of lipid peroxidation (LPO) as assessed by the amounts of malondialdehyde (MDA) and 4-hydroxyalkenals (4-HNE) using the LPO 586 test kit; (2) the production by AM of superoxide with the nitroblue tetrazolium test and (3) of nitric oxide (NO) with the Griess reaction. Stimulation of PMN or AM increases LPO of Curosurf and cell wall lipids. In both types of phagocytes, B. cepacia induced the highest LPO levels followed by P. aeruginosa and S. maltophilia. PMN, stimulated by live bacteria, induced higher LPO than those stimulated by heat-killed bacteria. B. cepacia stimulated AM to produce more superoxide and NO than did P. aeruginosa and S. maltophilia. The high phagocyte-stimulating ability of B. cepacia and its higher surfactant LPO than those of the other bacteria used in this in vitro study may play a role in vivo in the serious clinical condition known as the "Cepacia syndrome".
    Free Radical Research 10/2003; 37(9):909-17. · 3.28 Impact Factor
  • Rabea K L Bouhafs, Connie Jarstrand
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    ABSTRACT: Production of oxygen radicals by stimulated phagocytes followed by surfactant lipid peroxidation (LPO) and loss of surfactant function have all been implicated in the pathogenesis of acute lung injury. We studied the interactions between natural lung surfactant (Curosurf) and neutrophils in vitro, and compared various antioxidants; (superoxide dismutase (SOD), vitamin E, vitamin C, ebselen and melatonin), or combinations of them in duplicate and triplicate regarding their ability to decrease superoxide production and the peroxidation level of surfactant caused by activated phagocytes. The superoxide production of neutrophils activated by Candida albicans was measured with the nitroblue tetrazolium (NBT) test. The subsequent LPO was estimated as the content of malondialdehyde (MDA) and 4-hydroxyalkenals (4-HNE). We found that lung surfactant decreased the superoxide production by activated neutrophils (29.7%) and that Curosurf was peroxidized with elevated MDA/4-HNE values. With supplements of antioxidants (except vitamin C), superoxide radical production and the surfactant LPO level fell in a dose-dependent manner. The protective effect of the antioxidants differed in each test. SOD had a slight effect in both tests. The findings with vitamin E, melatonin and ebselen were similar. The best combination was that of a natural and a synthetic antioxidant (melatonin-ebselen) with a 60% decrease in comparison to the corresponding control. These findings suggest that antioxidants, particularly in combination, prevent LPO of lung surfactant.
    Free Radical Research 08/2002; 36(7):727-34. · 3.28 Impact Factor
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    ABSTRACT: The possible contribution of bacteria and polymorphonuclear neutrophils (PMN) to the disease process of periodontitis was evaluated. Fusobacterium nucleatum has been associated with chronic adult periodontitis. Intracellular production and extracellular release of reactive oxygen species (ROS) by PMN stimulated by fusobacteria were evaluated. To estimate the potential extracellular damage that might be caused by the ROS, the lipid peroxidation (LPO) of an exogenous phospholipid, Intralipid, was assayed. The ROS production of PMN was studied by the nitroblue tetrazolium and chemiluminescence tests. The levels of malonaldehyde (MDA) and 4-hydroxyalkenals were used to indicate LPO. Fusobacterium nucleatum strains stimulated neutrophils to produce a large amount of ROS, independently of plasma complement factors. The two strains tested induced considerable intracellular, but no extracellular chemiluminescence responses during the first hour, indicating that ROS were released into phagosomes. However an incubation period of 4 h, in the presence of the extracellular lipid resulted in a high degree of LPO, presumably caused by ROS release from the Fusobacterium-stimulated PMN. ROS production and lipid peroxidation could be counteracted by vitamin E. In periodontitis local bacteria might stimulate PMN to release ROS, which cause inflammation and destruction.
    Oral Diseases 02/2001; 7(1):41-6. · 2.38 Impact Factor
  • Rabea K.L. Bouhafs, Connie Jarstrand
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    ABSTRACT: The major etiologic agent in neonatal pneumonia and meningitis is group B streptococci (GBS). Nitric oxide (NO) production by alveolar macrophages (AM) in response to Gram-positive bacteria such as GBS and the effect of surfactant on this production have received little attention. We studied production of NO by GBS-stimulated AM using the Griess reaction, the effect of lung surfactant on this NO production, and the possible lipid peroxidation (LPO) of surfactant caused by NO. The LPO test was used to measure surfactant peroxidation. Heat-killed and live GBS were found to stimulate NO production by rat alveolar macrophages, and the presence of interferon gamma (IFN-gamma) increased this stimulation in a synergistic manner. Curosurf(R), the natural surfactant used in our study, significantly reduced NO production in various sets of experiments. Lipid peroxidation of surfactant was noted when NO was produced by stimulated AM, a phenomenon that could be suppressed by NG-monomethyl L-arginine (L-NMMA), the inhibitor of NO synthase. In the lung of GBS-infected neonates, nitric oxide produced by AM might contribute to the destruction of surfactant caused by inflammatory cells. Pediatr Pulmonol. 2000; 30:106- 113.
    Pediatric Pulmonology 09/2000; 30(2):106-13. · 2.38 Impact Factor
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    ABSTRACT: In newborn infants, group B streptococci (GBS) often cause pneumonia, with polymorphonuclear leukocytes (PMN) migrating into the lungs. Because surfactant therapy may be needed in such patients, we evaluated the interaction between GBS or GBS-stimulated PMN and a surfactant preparation (Curosurf) in vitro. The superoxide production of GBS strains or GBS-activated PMN was measured, using the nitroblue tetrazolium (NBT) test and the subsequent lipid peroxidation (LPO) as the content of malondialdehyde (MDA) and 4-hydroxyalkenals (4-HNE). The growth of GBS in surfactant was determined and related to the LPO. Finally, the effect of LPO on surfactant activity, caused by GBS-stimulated PMN, was assessed by measuring dynamic surface tension in a pulsating bubble surfactometer. Curosurf diminished the NBT reduction by both live GBS and GBS-stimulated PMN. Surfactant was peroxidized by reactive oxygen species (ROS) from both GBS and GBS-stimulated PMN in a time-dependent manner. Vitamin E significantly reduced the peroxidation level of surfactant in both cases. Surfactant peroxidation was associated with a reduction in the number of live bacteria. The biophysical activity of Curosurf was impaired by GBS-stimulated PMN, as reflected by increased minimum surface tension during cyclic compression. These findings indicate that Curosurf undergoes LPO by ROS produced by GBS and/or PMN. We speculate that exogenous surfactant preparations should be supplemented with vitamin E or another antioxidant, when given to infants with GBS pneumonia.
    Beiträge zur Klinik der Tuberkulose 02/2000; 178(5):317-29. · 2.06 Impact Factor
  • Rabea K.L. Bouhafs, Connie Jarstrand
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    ABSTRACT: Surfactant therapy is given routinely to premature newborns with respiratory failure. However, alterations in surfactants have been shown to be a significant factor in some forms of respiratory failure in newborns in animal models of lung injury. To investigate whether antioxidant supplementation might help to protect exogenous surfactant from damage by oxygen free radicals, we examined the influence of vitamin E in combination with surfactant on superoxide production as estimated by the nitroblue tetrazolium reduction test, and measured surfactant peroxidation with a new colorimetric method with or without addition of superoxide dismutase (SOD) or vitamin E. Our results showed that surfactant interacts with free radicals; surfactant reduced superoxide production by neutrophils and was peroxidized when incubated with resting and with stimulated cells. Vitamin E supplementation decreased superoxide radical production and in a dose-dependent manner decreased surfactant peroxidation. The decrease in lipid peroxidation by SOD was not significant. These findings suggest that phagocytes induce lipid peroxidation of lung surfactant, a reaction that might be prevented by antioxidants.
    Pediatric Pulmonology 06/1999; 27(5):322-7. · 2.38 Impact Factor
  • R. K. L. Bouhafs, C Jarstrand
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    ABSTRACT: The epithelium of the lung is lined with extracellular pulmonary surfactant. This is the surface that invading bacteria first come into contact with when they enter the alveoli. As bacteria become established and interact with this layer, various characteristics of surfactant may become altered. We studied free radical production by three bacterial species, group B streptococci, Escherichia coli, and Pseudomonas aeruginosa, as well as the effect of two concentrations of lung surfactant (Curosurf at 0.04 and 0.4 mg/ml) on this production estimated by the nitro blue tetrazolium reduction test. We also measured the lipid peroxidation of surfactant at various incubation times (0-20 h), using a LPO-586 test kit. In addition, the effect of vitamin E as an antioxidant in a concentration of 0.5 microM was determined by the lipid peroxidation test. We found that the nitro blue tetrazolium reduction by the three bacterial species and lipid peroxidation of lung surfactant increased with time. Vitamin E reduced the lipid peroxidation of this surfactant. By measuring bacterial growth at various incubation times we showed that lung surfactant was bactericidal to group B streptococcal and E. coli strains and that P. aeruginosa strains were resistant to surfactant. We conclude that bacteria, probably by their production of reactive oxygen species, cause lipid peroxidation of lung surfactant.
    Beiträge zur Klinik der Tuberkulose 02/1999; 177(2):101-10. · 2.06 Impact Factor