Introduction Increased blood pressure (BP), especially sudden, unexpected and severe rises in women with preeclampsia considerably increases the risk of peripartum complications. A specific quantitative criterion for diagnosing labile blood pressure does not exist and minimal evidence is available to guide therapeutic decisions. Objective To assess the effect of antihypertensives commonly used to manage hypertension in pregnant women on blood pressure lability at rest in an experimental model of preeclampsia (EPE). Methods EPE was established in Papio hamadryas ( n =6) by surgically ligating the non-dominant uterine branch artery to reduce perfusion by 30–40%. EPE was confirmed by assessing proteinuria (urine protein:creatinine ratio mmol/L), BP changes (intra-arterial radiotelemetry) and soluble fms-like tyrosine kinase receptor-1 (sFLT-1) (ELISA). BP readings were collected every 20s before and after antihypertensive treatment. Equipotent doses of labetalol, methyldopa and hydralazine were given. Lability was defined by SD1 and SD2 values as measures of short-term variability (STV) and long-term variability (LTV) respectively. These values were generated using Poincare plots (Spike2 v8.06). Prism GraphPad6 v6.05 was used for parametric testing, data expressed as mean±SEM and significance set at p Results All animals developed EPE, demonstrated by systolic BP rise of 7.1mmHg ( p p p p p p p =0.03). Diastolic STV was not significantly affected. The three drugs did not significantly affect systolic or diastolic LTV. Conclusions Systolic STV was affected by drug choice but not diastolic STV or LTV in EPE.

Increased blood pressure (BP), especially sudden, unexpected and severe rises in women with preeclampsia considerably increases the risk of peripartum complications. These risks are even more acute in the context of general anaesthesia. To assess the effects of antihypertensive drugs on short and long term blood pressure variability when used concurrently with propofol and ketamine anaesthesia. Blood Pressure was measured during anaesthesia in six, pregnant Papio hamadryas (baboons) with experimental preeclampsia (EPE). Animals were given antihypertensives commonly used to manage preeclampsia (labetalol, methyldopa and hydralazine) at equipotent doses equivalent to mild starting dose rates commonly used in women with preeclampsia. When anaesthetised with ketamine (most commonly used anaesthetic agent in EPE), systolic BP increased significantly by 4.9 mmHg when animals were on labetalol (n = 3, p < 0.05), decreased by 0.5 mmHg when animals were on methyldopa (n = 3, NS) and decreased by 4.9mmHg when animals were on hydralazine (n = 2, NS) as compared to systolic BP under ketamine anaesthesia prior to receiving any medication. With propofol anaesthesia (commonly used when anaesthesia required in women with preeclampsia), systolic BP decreased by 1.4 mmHg when animals were on labetalol (n = 3, NS), decreased by 7.2 mmHg when animals were on methyldopa (n = 2, NS) and increased by 2.8 mmHg when animals were on hydralazine (n = 3, NS) as compared to systolic BP during propofol anaesthesia prior to receiving any medication. These results show that in the two anaesthetic agents studied so far the greatest reduction in BP was achieved with hydralazine during ketamine anaesthesia and methyldopa during propofol anaesthesia. It is likely that antihypertensive treatment affects blood pressure during anaesthesia, and that the interaction between antihypertensive and anaesthetic agent is of clinical importance in managing sudden, unexpected and severe rises in BP in women with preeclampsia.

1. Increases in soluble fms-like tyrosine kinase 1 (sFlt-1) and soluble endoglin (sEng) contribute to the pathogenesis of pre-eclampsia. Soluble Flt-1 binds to circulating free vascular endothelial growth factor and placenta growth factor and this is associated with endothelial dysfunction. Soluble endoglin, a transforming growth factor (TGF)-beta coreceptor, was reported to synergize with sFlt-1 to amplify endothelial dysfunction by inhibiting TGF-beta1-mediated vasorelaxation. 2. The aim of the present study was to examine whether the antihypertensive drugs clonidine (0.08-1.3 microg/mL), diazoxide (25-300 microg/mL), frusemide (60-1000 microg/mL) and hydralazine (6.3-100 microg/mL) have any effect on placental production of sFlt-1 and sEng in placentas from normal and pre-eclamptic pregnancies. 3. Explants were taken from non-laboured term placentas of normal pregnancy (n = 5) and women with pre-eclampsia (n = 5). Villous explants were cultured with increasing doses of antihypertensive drugs. Placental sFlt-1 and sEng production was examined using ELISA. 4. Baseline sFlt-1 production was higher in placentas from women with pre-eclampsia than from normal pregnancy (4.5 +/- 1.4 vs 3.2 +/- 0.6 ng/mg of total protein, respectively; P < 0.001), as was sEng production (9.0 +/- 2.3 vs 4.1 +/- 0.6 ng/mg of total protein, respectively; P < 0.001). With the exception of frusemide, none of the antihypertensive drugs tested had any effect on sFlt-1 and sEng production from placental explants of normal pregnancy and women with pre-eclampsia. Increasing frusemide concentrations were correlated with increased sEng production in normal pregnancy (P < 0.005). 5. In conclusion, placental sFlt-1 and sEng production was higher in pre-eclampsia and antihypertensive drugs had no effect on placental production of sFlt-1 and sEng in vitro.

Background: Rhinovirus infection is responsible for considerable morbidity and mortality as the major cause of exacerbations of asthma, and is also known to induce exacerbations of cystic fibrosis and chronic obstructive pulmonary disease. Exacerbations of these diseases are also frequently associated with bacterial and atypical bacterial infection. Alveolar macrophages are the major immune cells in the airways and are important in defence against bacterial infections. Methods: The authors investigated whether rhinovirus modifies cytokine release, the pattern recognition receptor expression and phagocytosis by human alveolar macrophages in response to bacterial products. Results: Viable rhinovirus was detected in macrophages up to 3 days after exposure and viral RNA expression persisted for 10 days. Infectious but not UV inactivated rhinovirus increased tumour necrosis factor α (TNFα) and interleukin (IL)8 release by macrophages. In contrast, infectious rhinovirus impaired lipopolysaccharide and lipoteichoic acid induced TNFα and IL8 secretion by macrophages. Rhinovirus induced impairment of macrophage antibacterial immune responses did not involve IL10, prostaglandin E 2 or downregulation of Toll-like receptor 2. Furthermore, the macrophage phagocytic response to labelled bacterial particles, but not to latex beads, was impaired. Conclusion: The authors have identified impairment of cytokine responses to bacterial lipopolysaccharide and lipoteichoic acid by alveolar macrophages in response to infectious rhinovirus. Virus induced impairment of antibacterial host defence has important implications in the pathogenesis of exacerbations of respiratory diseases.

Background: Neutrophils, in addition to eosinophils, are prominent in the airways of patients with severe asthma who are usually on long-term oral and inhaled corticosteroid treatment. We determined whether inhaled or oral corticosteroid therapy can induce airway neutrophilia.Methods: We performed two separate placebo-controtted studies in which patients with mild asthma were treated with either prednisolone (30mg per day for 7 days; n = 9) or placebo tablets (n = 8), or with either inhaled budesonide (800 mug twice daily for 4 weeks; n = 6) or inhaled placebo (n = 6). Fiberoptic bronchoscopy was performed before treatment and at day 7 of oral treatment, and at day 28 of inhaled therapy. Bronchial sections were immunostained with an antibody to major basic protein for eosinophils, and with an antibody to neutrophil elastase for neutrophils. Induced sputum was obtained in the prednisolone study.Results: Neutrophils in airway submucosa increased after prednisolone from median 76 to 140/mm(2) (P = 0.05); this change was higher than that after placebo (P = 0.04). Eosinophils decreased from 24 to 9/mm(2) (P = 0.03), but this was not significantly different from placebo. Eosinophils and neutrophits, and Levels of IL-8 and myeloperoxidase in induced sputum did not change after prednisolone. There was no change in neutrophil. counts after budesonide, but the reduction in eosinophils was greater than placebo (P = 0.05). Budesonide improved bronchial responsiveness, but prednisolone did not.Conclusion: Corticosteroid therapy by the oral but not inhaled route can induce neutrophil recruitment into the airways of patients with mild asthma. This could explain the increase in airway neutrophits observed in severe asthmatics treated with oral corticosteroids. (C) 2004 Elsevier Ltd. Alt rights reserved.

Background: Increased levels of bacterial lipopolysaccharide (LPS) have been demonstrated in chronic heart failure (CHF). LPS can induce cellular desensitization, with specific down-regulation of LPS-mediated cellular tumor necrosis factor (TNF-alpha) production which does not affect other cytokine parameters. It is not known if LPS desensitization occurs in CHF.Methods and results: Mononuclear cells from 24 CHF patients (mean age 70 +/- 2 years, age range 58 to 78 years, NYHA class 3.0 +/- 0.2) and I I healthy controls (mean age 53 +/- 3 years, age range 39 to 75 years) were separated frorn venous blood and Cultured for 24 It with LPS (E, coli, 0-10 ng/mL). Culture supernatants were tested for TNF-alpha and interleukin receptor antagonist (IL- IRA). Patients were subgrouped into mild (n=1 0), moderate (n=5), and severe (n=9) CHF. Independently of age, rnononuclear cells from patients with severe heart failure produced less TNF-alpha than controls (p < 0.05) and patients with mild (p < 0.001) or moderate CHF (p < 0.05). IL- I RA release was higher for CHF patients as a group, compared with controls (p < 0.05). There was no significant difference in IL-1RA release between CHF patient subgroups.Conclusions: Mononuclear cells from patients with severe heart failure produce significantly less TNF-a than healthy controls or patients with mild to moderate disease. Production of IL-1RA is not affected. This resembles a picture indicative of LPS desensitization occurring in patients with severe CHF. (c) 2004 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.

Inflammatory cytokine production by alveolar macrophages (AMs) is regulated by transcriptional activation and may be increased by cigarette smoking. The smoking-induced regulation of interleukin (IL)-8 by extracellular signal-regulated kinase (ERK)-1 and -2, p38 mitogen-activated protein kinase (MAPK) and the transcription factor nuclear factor-kappaB (NF-kappaB) in lipopolysaccharide-stimulated AMs was assessed in nine smokers compared with nine healthy nonsmokers. IL-8 production was dependent on phosphorylation of ERK-1 and -2 and p38 MAPK, as examined by PD 098059 (10 microM), an inhibitor of the upstream activator of MAPK kinase (MKK)-1, and SB 203580 (10 microM), an inhibitor of p38 MAPK. IL-8 release and the inhibitory effect of PD 098059 were increased in AMs from smokers. Moreover, ERK-2 messenger ribonucleic acid expression, as examined by reverse transcriptase polymerase chain reaction and phosphorylation of ERK-2 using Western blots, were increased in AMs from smokers, indicating a smoking-induced modulatory role of ERK-1 and -2. Lipopolysaccharide-induced IL-8 production was dependent on activation of NF-kappaB, as examined by SN 50 (100 microM), an inhibitor of NF-kappaB translocation, and the specific NF-kappaB inhibitor kinase-2 inhibitor, AS 602868 (10 microM), with no differences in AMs from smokers and nonsmokers. SN 50 but not PD 098059 and SB 203580 blocked NF-kappaB deoxyribonucleic acid-binding, and this occurred to the same extent in AMs from smokers and nonsmokers, as examined by electromobility shift assay. It is concluded that cigarette smoking enhances mitogen-activated protein kinase activation more than nuclear factor-kappaB activation to increase lipopolysaccharide-induced interleukin-8 production in alveolar macrophages.

We investigated the effect of chronic cigarette smoking on the expression of haem-oxygenase (HO)-1 and HO-2. Normal subjects and asymptomatic young current smokers with normal lung function tests underwent bronchoalveolar lavage for recovery of macrophages. Reverse transcription/polymerase chain reaction (RT-PCR) analysis showed no significant difference in HO-1 and HO-2 mRNA expression between the two groups. On the other hand, Western blot analysis showed a significant (P<0.05) reduction of HO-2 protein, but not of HO-1, in alveolar macrophages from smokers compared to normal. There was no significant differences by immunocytochemistry for HO-1 and HO-2 expression between the groups. We concluded that HO-2 expression is reduced in alveolar macrophages of smokers, possibly due to the oxidative stress of cigarette smoke. This may in turn lead to reduced protection against further oxidative insults.

BACKGROUND:Interleukin (IL)-10 is a pleiotropic cytokine with a broad spectrum of immunosuppressive and anti-inflammatory effects. IL-10 secretion from alveolar macrophages is defective in patients with asthma and lower concentrations of IL-10 are found in bronchoalveolar lavage (BAL) from asthmatic patients than in normal control subjects. Reduced IL-10 may result in exaggerated and more prolonged inflammatory responses in asthmatic airways. IL-10 acting through the IL-10 receptor (IL-10R) stimulates the transcription factors STAT1 and STAT3.METHODS:We investigated IL-10 and IL-10R expression in normal and asthmatic bronchial epithelium and BAL macrophages using reverse transcription-polymerase chain reaction, immunohistochemistry and Western blotting. The functional effect of IL-10 was examined using granulocyte-macrophage-colony stimulating factor, enzyme-linked immunosorbent assay and Western blotting for phosphorylated STAT1 and STAT3.RESULTS:IL-10 was not expressed in epithelial cells; furthermore these cells did not express the IL-10R and had no functional response to exogenous IL-10. Bronchial epithelial cells expressed variable levels of phosphorylated STAT1 and STAT3 with no change in expression between normal subjects and asthmatics. IL-10 protein and IL-10R expression was detected in alveolar macrophages from all subjects.CONCLUSION:Our study suggests that the bronchial epithelium is not a source of IL-10 and cannot respond to exogenous IL-10 because of a lack of IL-10R expression.

Inflammation has long been recognized as a major pathologic feature of the airways and lung parenchyma in chronic obstructive pulmonary disease (COPD)1van den Berge M. Polosa R. Kerstjens H.A.M. Postma D.S. The role of endogenous and exogenous AMP in asthma and chronic obstructive pulmonary disease.J Allergy Clin Immunol. 2004; 114: 737-746Abstract Full Text Full Text PDF PubMed Scopus (39) Google Scholar, 2Sutherland E.R. Outpatient treatment of chronic obstructive pulmonary disease: comparisons with asthma.J Allergy Clin Immunol. 2004; 114: 715-724Abstract Full Text Full Text PDF PubMed Scopus (15) Google Scholar (Fig 1, Fig 2, Fig 3). Increased numbers of T cells have been demonstrated by many groups in the small and large airways in all forms of COPD. CD8+ cells, the most predominant T-cell subtype, usually invade the submucosa but can also be demonstrated within the epithelium, mucous glands, and the smooth muscle layer (Fig 4). The other types of cells that have been reported to be increased in COPD are macrophages; these are mainly visualized in small airways and lung parenchyma (Fig 5). Different cytokines and chemokines were reported to be highly expressed in COPD, including IL-8, TNF-α, and MCPs. Recent studies have shown that extensive airway remodeling is a feature of COPD pathology (Fig 6, Fig 7, Fig 8, Fig 9, Fig 10). Increased smooth muscle mass, mucous gland hyperplasia, and subepithelial fibrosis are also features of large and small airways pathology in COPD.3Hirst S.J. Martin J.G. Bonacci J.V. Chan V. Fixman E.D. Hamid Q.A. et al.Proliferative aspects of airway smooth muscle.J Allergy Clin Immunol. 2004; 114: S2-S17Abstract Full Text Full Text PDF PubMed Scopus (193) Google Scholar Lymphoid aggregation containing follicles has recently been described in COPD4Hogg J.C. Chu F. Utokaparch S. Woods R. Elliott W.M. Buzatu L. et al.The nature of small-airway obstruction in chronic obstructive pulmonary disease.N Engl J Med. 2004; 350: 2645-2653Crossref PubMed Scopus (2793) Google Scholar; this feature is not restricted to small airways and can be demonstrated in large and intermediate-sized airways (Fig 11).Fig 2Small airway in a smoker with centrilobular emphysema. Thickened wall and abundant muscle contribute to the markedly narrowed and deformed lumen.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 3Wall of an airway less than 2 mm. in diameter in a smoker with centrilobular emphysema. The epithelium shows goblet cells, not normally found in airways of this caliber; the wall is thickened with infiltration of T cells and increase in smooth muscle mass.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 4Immunostaining of section from a biopsy specimen of a patient with COPD using antibodies to CD8. Note the large number of CD8+ cells in the upper mucosa. (ABC, original magnification ×200.)View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 5Section from a proximal airway in COPD showing CD68 immunoreactivity.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 6Small airway in panlobular emphysema (PLE). The abnormalities in the small airways are less prominent in PLE than in centrilobular emphysema, and the wall of the airways shows significant less remodeling.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 7Biopsy specimen from a patient with COPD shows mucous gland hyperplasia. (Hematoxylin and eosin, original magnification ×200.)View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 8Masson-Trichrome stain of an airway wall in a smoker with centrilobular emphysema showing a thickening of basement membrane.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 9Section from the proximal airway of a patient with COPD showing an extensive subepithelial fibrosis. (Hematoxylin and eosin, original magnification ×200.)View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 10Section from the proximal airway of a patient with COPD shows an impressive increase in smooth muscle mass that extends close to the epithelium.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Fig 11Section from a patient with COPD shows a lymphoid aggregation containing a follicle in a proximal airway. (Hematoxylin and eosin, original magnification ×200.)View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Neutrophils and mononuclear cells are implicated in the pathogenesis of several inflammatory conditions including chronic obstructive pulmonary disease (COPD). Neutrophil-derived serine proteases, such as cathepsin G (CG) and neutrophil elastase (NE), may interact with mononuclear cells via protease-activated receptors (PARs) which are seven-transmembrane G protein-coupled receptors activated by proteolytic cleavage of the extracellular N-terminus, and which, on activation, induce the release of several mediators and cytokines. We determined whether CG and NE could affect PAR-1 expression and function in mononuclear cells. Human blood mononuclear cells were isolated from 20 healthy donors. Surface and intracellular receptor expression and calcium mobilisation (using the calcium chelator, FLUO3-AM) were studied by fluorescence-assisted cell sorting (FACS analysis). Positive controls, i.e. thrombin (0.1-100 mU/ml) and the PAR-1-activating peptide SFLLRN (100 microM) induced a rapid and transient intemalisation of PAR-1 in monocytes and lymphocytes. CG but not NE had a similar effect. By contrast, in monocytes intracellular calcium mobilisation was induced by thrombin and SFLLRN but not by CG and NE. Thus, CG can induce intracellular PAR-1 sequestration without activation of the receptor, and may act as an antagonist and prevent subsequent activation of PAR-1 in mononuclear cells. These findings may be of relevance to the pathogenesis of COPD.

Granulocyte macrophage-colony-stimulating factor (GM-CSF), released from alveolar macrophages (AM), is an important regulator of eosinophil, T cell, and macrophage function and survival. We determined the mechanisms of GM-CSF regulation in AM from normal volunteers activated by lipopolysaccharide (LPS) by examining the role of nuclear factor-kappaB (NF-kappaB), and of p38 mitogen-activated protein (MAP) kinase and MAP kinase kinase (MKK-1). PD 098059 (10 microM), an inhibitor of upstream activator of MKK-1, inhibited GM-CSF expression, but the expression of GM-CSF was not inhibited by SB 203580 (10 microM), an inhibitor of p38-MAP kinase. Phosphorylation of extracellular signal-regulated kinase-1 (ERK-1), ERK-2, and p38 MAP kinase by LPS were demonstrated on Western blot analysis. LPS increased NF-kappaB:DNA binding as examined by electrophoretic mobility shift assay, but this was not suppressed by PD 098059 or by SB 203580. LPS induced an increase in NF-kappaB activation as examined by p50 translocation assay without suppression by PD 098059 or by SB 203580. SN50 (100 microM), an inhibitor of NF-kappaB translocation and the specific IKK-2-Inhibitor (AS602868; 10 microM), also prevented GM-CSF expression and release induced by LPS, indicating that GM-CSF release is NF-kappaB-dependent. PD 098059, but not SB 203580, inhibited LPS-induced histone acetyltransferase (HAT) activity, indicating chromatin modification. Furthermore, AS602868 and SN 50 suppressed LPS-induced HAT activity. TSA (10 ng/ml), an inhibitor of histone deacetylase (HDAC), reversed the inhibitory effect of PD 098059, SB 203580, SN 50 and AS602868 on GM-CSF release. GM-CSF expression and release in AM is controlled by NF-kappaB activation, and this is modulated by phosphorylation of MKK-1 and p38 MAP kinase acting on histone acetylation.

BACKGROUNDProtease-activated receptors 1 and 2 (PAR-1 and PAR-2) are 7-transmembrane G protein-coupled receptors activated by serine proteases in many cell types, including monocytes-macrophages, leading to the production of pro-inflammatory mediators, cytokines, and growth factors.OBJECTIVEWe determined the influence of chronic smoking and asthma on the expression of PAR-1 and PAR-2 receptors on alveolar macrophages (AMs).METHODSWe used RT-PCR and immunocytochemistry with confocal microscopy to determine mRNA and protein expression of PAR-1 and PAR-2 in AMs obtained from healthy smokers, asthmatic patients, and healthy subjects. In addition, we examined the effect of IL-1beta and LPS.RESULTSPAR1 mRNA was decreased, whereas PAR2 mRNA was increased in 24-hour cultured AMs from smokers when compared with values in AMs from healthy subjects. Paradoxically, there was a higher degree of PAR-1 protein staining in AMs from smokers, whereas PAR-2 staining was similar in smokers and healthy subjects. PAR-1 and PAR-2 mRNA and protein expression were similar in asthmatic patients and control subjects. IL-1beta and LPS had no effect on PAR1 and PAR2 gene expression by AMs.CONCLUSIONSThere is a dissociation between gene and protein expression of PAR-1 and PAR-2. PAR-1 protein overexpression in AMs from smokers might be important in the pathophysiology of chronic airways disease.