Inflammatory markers in exhaled breath condensate from patients with asthma

Background and objective:   Evaluation of airway inflammation is important for the diagnosis and treatment of asthma. Exhaled breath condensate (EBC) is a minimally invasive method for assessing inflammation and may be useful for monitoring airway inflammation in asthma. The aims of this study were to establish an EBC collection method, to assess biomarkers reflecting asthmatic airway inflammation, and to determine the relationship of these biomarkers with asthma severity and lung function.
Methods:   Fifty-eight non-smoking healthy subjects, seven asymptomatic smokers, nine subjects with common cold and 55 asthmatics with disease severity ranging from mild intermittent to severe persistent were studied. The efficacy of a pipette method was compared with that of a commercial collecting device. pH, CRP, albumin, hydrogen peroxide (H₂O₂) and nitrite/nitrate levels were measured in EBC.
Results:   Except for the quantity of EBC collected and albumin levels, there were no differences between the commercial method and the pipette method in levels of biomarkers measured. Levels of CRP, H₂O₂ and nitrite/nitrate were significantly higher in the asthma group than that in the control group. In terms of asthma severity, pH and levels of CRP, H₂O₂ and nitrate were significantly higher in the mild persistent group than that in the other groups. In addition, H₂O₂ levels in EBC correlated significantly with the level of nitrite/nitrate. FEV₁ and PEF showed significant negative correlations with H₂O₂ and nitrite/nitrate levels.
Conclusion:   Measurement of EBC biomarkers is a non-invasive and useful way to evaluate airway inflammation in patients with asthma.     

Inflammatory markers in exhaled breath condensate following lung resection for bronchial carcinoma

Background and objective: Pulmonary resection may cause inflammatory changes with subsequent injury to the remaining lung and deterioration in respiratory function. This study investigated the pattern of serum inflammatory markers and exhaled breath condensate (EBC) in patients undergoing major lung resection due to bronchial carcinoma compared with minimally invasive thoracic surgery. Methods: The pro-inflammatory markers IL-1-β, IL-6, IL-8, tumor necrosis factor-α (TNF-α) and soluble intercellular adhesion molecule-1 (sICAM-1) were measured preoperatively (day −1) and on three postoperative days (day 1, 3, 7) in serum and EBC in patients after lobectomy or pneumonectomy due to bronchial carcinoma (test group) and in patients undergoing thoracoscopy with minimal wedge resection (control group). Results: All mediators were detectable in serum and all but IL-8 were detectable in EBC. No patient suffered postoperative respiratory failure. In the test group, serum IL-6 was significantly higher postoperatively compared with day −1 (P < 0.001). For EBC (test group), the postoperative values of IL-1-β were significantly higher compared with day −1 (P = 0.005). In EBC (test group), day −1 TNF-α and sICAM-1 were significantly higher compared with controls (P < 0.029 and P = 0.032, respectively). There was no correlation between the levels of mediators and the extent of resection. Conclusions: Pro-inflammatory markers are detected in EBC following pulmonary surgery. Mediators are detectable in both serum and EBC in patients with bronchial carcinoma undergoing pulmonary resection, but the levels are higher in EBC.     

Bronchoalveolar lavage, sputum and exhaled clinically relevant inflammatory markers: values in healthy adults.

Bronchoalveolar lavage (BAL), induced sputum and exhaled breath markers (exhaled nitric oxide and exhaled breath condensate) can each provide biological insights into the pathogenesis of respiratory disorders. Some of their biomarkers are also employed in the clinical management of patients with various respiratory diseases. In the clinical context, however, defining normal values and cut-off points is crucial. The aim of the present review is to investigate to what extent the issue of defining normal values in healthy adults has been pursued for the biomarkers with clinical value. The current authors reviewed data from literature that specifically addressed the issue of normal values from healthy adults for the four methodologies. Most studies have been performed for BAL (n = 9), sputum (n = 3) and nitric oxide (n = 3). There are no published studies for breath condensate, none of whose markers yet has clinical value. In healthy adult nonsmokers the cut-off points (mean+2sd) for biomarkers with clinical value were as follows. BAL: 16.7% lymphocytes, 2.3% neutrophils and 1.9% eosinophils; sputum: 7.7 x 10(6).mL(-1) total cell count and 2.2% eosinophils; nitric oxide: 20.2 ppb. The methodologies differ concerning the quantity and characteristics of available reference data. Studies focusing on obtaining reference values from healthy individuals are still required, more evidently for the new, noninvasive methodologies.     

Inflammatory and microbiologic markers in induced sputum after intravenous antibiotics in cystic fibrosis

Induced sputum has been used to study airway inflammation. We sought to determine whether markers of infection and inflammation in induced sputum were a useful and safe outcome measure in cystic fibrosis. We hypothesized that bacterial density and inflammatory content of induced sputum would decrease after antibiotic therapy. Induced sputum was assayed for bacterial density, cell count, and differential and inflammatory markers before and after treatment with intravenous antibiotics. Fifty-five of the 72 subjects enrolled (mean age +/- SD 18.2 +/- 7.9 years) completed the study. FEV1 increased by an average 0.3 +/- 0.3 L (10.4 +/- 8.7% predicted FEV1), p<0.0001; density of Pseudomonas aeruginosa and Staphylococcus aureus decreased by 2.4 +/- 3.1 log10 cfu/g (p<0.0005) and 4.0 +/- 2.3 log10 cfu/ml (p<0.0001), respectively; neutrophil count decreased by 0.4 +/- 0.6 log10 cells/ml (p<0.0001), interleukin-8 concentration by 0.5 +/- 1.3 log10 pg/ml (p<0.05), and neutrophil elastase by 0.4 +/- 0.7 log10 microg/ml (p<0.005). Seven of 127 (6%) sputum induction procedures showed a decrease in FEV1 of 20% or more. We conclude that markers in induced sputum may be useful, noninvasive outcome measures to assess response to therapies in cystic fibrosis studies.     

支气管哮喘患者呼出气冷凝液和诱导痰中炎性指标检测的临床意义

目的 通过测定支气管哮喘(以下简称哮喘)患者呼出气冷凝液(EBC)和诱导痰中炎性指标的浓度,分析其与临床指标的关系,探讨炎性指标用于病情和疗效评估的价值.方法 选四川大学华西医院呼吸内科门诊接受吸入糖皮质激素联合长效β2受体激动剂治疗的中重度慢性持续期哮喘患者,记录治疗前和治疗1个月后哮喘症状积分,测定第1秒钟用力呼气容积(FEV1)占预计值百分比,采集诱导痰和EBC标本,测定标本中过氧化氧(H2O2)、硝酸盐/哑硝酸盐(NO3-/N2-)和半胱氨酰白三烯E4(LTE4)浓度.结果 共有25例中重度哮喘患者按研究方案完成治疗和随访.哮喘患者经联合治疗1个月后临床症状积分和FEV1占预计值百分比明显改善(P<0.01);EBC和诱导痰中H2O2、NO3-/NO2-和LTE4浓度均降低,但仍高于健康对照者;H2O2和NO3-/NO2-的下降比LTE4明显;哮喘患者EBC中的H2O2和NO3-/NO2-;浓度与FEV1呈负相关(P<0.01),与症状积分呈正相关(P<0.01),LTE4与症状积分和FEV1均无相关性.诱导痰中H2O2浓度与FEV1占预计值百分比呈负相关(P<0.01),与症状积分呈正相关(P<0.01);NO3-/NO2-浓度与FEV1占预计值百分比呈负相关(P<0.01),与症状积分无相关性;LTE4浓度与症状积分和FEV1占预计值百分比均无相关性.治疗后FEV1占预计值百分比增高程度与EBC和诱导痰中H2O3、NO3-/NO2-浓度降低水平呈正相关(P<0.01).诱导痰和EBC对应的炎性指标之间有相关性(P值均小于0.01).EBC和诱导痰中H2O2浓度与NO3-/NO2-浓度呈正相关(P<0.01),与LIE4之间无相关性.结论 中重度哮喘患者联合治疗后在临床症状和肺功能改善的同时,气道炎症显著减轻.EBC的安全性和可重复性优于诱导痰,H2O2和NO3-/NO2-的敏感性优于LTE4.     

诱导痰液分析在咳嗽变异性哮喘诊断中的应用

目的　探讨咳嗽变异性哮喘 (CVA)患者是否存在气道炎症。方法　采用 3%～ 5 %的高渗盐水超声雾化吸入诱导排痰 ,对 35例临床诊断为CVA的患者痰液进行肥大细胞 (MC)百分率、嗜酸性粒细胞 (EC)百分率、嗜酸性粒细胞阳离子蛋白 (ECP)及中性粒细胞 (N)进行检测 ,并同步测定 1s用力呼气容积 (FEV1)及 1s用力呼气占预计值百分比 (FEV1占预计值 % )。结果　 35例临床诊断为CVA的患者 ,其痰液中MC、EC百分率、ECP浓度、FEV1及FEV1占预计值 %与对照组比较 ,差异均具有显著性 (P <0 .0 5 ) ,与典型支气管哮喘比较差异无显著性 (P >0 .0 5 )。经抗炎治疗后咳嗽症状很快缓解 ,复测痰液中MC、EC百分率、ECP浓度、FEV1及FEV1占预计值 %与治疗前比较 ,差异有显著性。结论　痰液分析从实验室角度阐明CVA存在气道炎症 ,从而为其诊断及治疗提供了一项可靠的评价指标     

Inflammatory markers and stroke

Basic and animal research implicate inflammatory mechanisms in the pathogenesis and progression of atherosclerosis, plaque rupture, thrombosis, and stroke. Inflammatory biomarkers, particularly high-sensitivity C-reactive protein and lipoproteinassociated phospholipase A2, have been identified as potential predictors of stroke risk and prognosis. Infections may also precipitate stroke. Medications, especially hydroxymethylglutaryl coenzyme A reductase inhibitors (statins), reduce inflammatory marker levels independently of lipid effects, and the ability of statins to reduce coronary events and stroke correlates with their effect on inflammatory biomarkers. Vaccination against influenza may also reduce stroke risk. Determining whether reduction of biomarkers reduces risk of recurrent stroke, however, requires further study before inflammatory markers become a routine part of the evaluation of stroke patients.     

Validity and safety of sputum induction by inhaled uridine 5'-triphosphate

Inhalation of hypertonic saline during sputum induction causes bronchoconstriction. We studied the validity and safety of sputum induction by uridine 5'-triphosphate (UTP). Sputum was induced by a 5-min inhalation of hypertonic saline (3%) on Day 1 and UTP (5 mg/ml in 0.9% saline) on Days 8 and 15 in 16 healthy subjects and 16 patients with mild-to-moderate asthma. Inhaled UTP produced twofold greater amounts of sputum than did hypertonic saline. There were significant differences in oxygen desaturation and bronchoconstriction during the procedure between the two methods: the maximal fall in Sa(O(2)), the AUC of the Sa(O(2))-time response, and the fall in PEF were less in the subjects who received UTP than in those who received hypertonic saline. Sputum total cell and differential cell counts, with a high proportion of eosinophils in asthmatics, were similar between specimens obtained by hypertonic saline and UTP. When we compared two consecutive measurements on the UTP-induced sputum samples, the reproducibility calculated by the intraclass correlation coefficient was high for the proportion of eosinophils, neutrophils, and macrophages. Therefore, inhalation of UTP aerosols may provide an effective, relatively noninvasive, valid, and reproducible method of sputum induction for the assessment of airway inflammation in asthma. Keywords: uridine triphosphate; induced sputum; airway inflammation; bronchoconstriction; asthma     

Influenza vaccination: changes in exhaled nitric oxide levels and sputum cytology

OBJECTIVE: Influenza vaccination is routinely recommended for patients with chronic lung disease, but has been reported to cause a small increase in airway reactivity. The use of simple, non-invasive methods of assessing changes in airway inflammation could potentially allow improved understanding of the cellular mechanisms underlying such changes. METHODOLOGY: We studied a group of 44 hospital workers before and after routine influenza vaccination, using spirometry, and exhaled nitric oxide (eNO) as a marker of pulmonary inflammation (Group A). In addition, methacholine challenge and sputum induction were also performed in a subgroup (subgroup B, n = 7) at baseline and at 7 days after vaccination. RESULTS: Spirometry did not change in either group. In Group A there was a small but significant rise in mean peak eNO from 47.32 +/- 4.5 (mean NO p.p.b. +/- SEM) to 53.2 +/- 4.9 between days 0 and 7 (P < 0.05). A non-significant rise in eNO was seen in subgroup B. In subgroup B, when the differential cell counts in induced sputum were compared between baseline and sampling 1 week later, there was a significant rise in the percentage of lymphocytes, from 3.9 (1.8-9.8: median %total (range)) to 11.9 (6.0-18.5, P < 0.02) and a non-significant increase in shed respiratory epithelial cells from 3.1 (0.6-5.8) to 13.0 (1-30.3, P=0.06). There was a corresponding significant fall in the percentage of macrophages. Methacholine challenge in subgroup B showed no change in reactivity in these normal subjects. CONCLUSION: Influenza vaccination causes a small increase in exhaled NO, and is accompanied by increased sputum lymphocytosis and respiratory epithelial shedding. An influx of inflammatory cells may help to explain the induction of increased airway reactivity that has been described in other studies.     

Sputum induction leads to a decrease of exhaled nitric oxide unrelated to airflow.

Measurement of exhaled nitric oxide (eNO) and analysis of induced sputum are both established noninvasive methods for studying airway inflammation in asthma. Both methods are often used sequentially within short time frames. The aim of the present study was to evaluate the influence of sputum induction on eNO in adults and to follow the kinetics of airway eNO production after induction in relation to forced expiratory volume in one second (FEV1). eNO and FEV1 were measured in 41 adult patients (aged 29 (range 23-50) yrs, 56% male) with asymptomatic atopy or mild asthma (mean FEV1 97.2+/-3% predicted) prior to and immediately after sputum induction with hypertonic saline (4%). Sputum induction with isotonic saline was also performed in 13 subjects (control group). Repeatability of eNO decrease after sputum induction was also studied in 27 patients on separate occasions and kinetics of eNO production after sputum induction were followed over 24 h in another 10 patients. Sputum induction with hypertonic, but not isotonic, saline led to a marked decrease in eNO (log) immediately after the procedure (pre: 3.85+/-0.13 parts per billion (ppb); post: 3.24+/-0.14 ppb). This decrease was shown to be highly reproducible and not related to a fall in FEV1 following sputum induction. While FEV1 returned to baseline within 1 h, decreased eNO levels were observed over 4 h and returned to baseline after 24 h. Hypertonic saline sputum induction leads to a prolonged reduction in exhaled nitric oxide in adult atopics that is reproducible within subjects and not related to a reduction in airflow following sputum induction. This methodological interference should be taken into account when sputum induction and exhaled nitric oxide measurements are performed in the same subject.     

Inflammatory markers in metabolic syndrome

The Metabolic Syndrome (MS) is associated with a systemic inflammatory response that plays an important pathogenetic role in atherothrombotic disease. Highly sensitive C-reactive protein (hsCRP) and fibrinogen are acute phase reactants and indicate underlying inflammatory state. We studied inflammatory markers in 50 Indian subjects with MS diagnosed by IDF criteria and 24 healthy age and sex matched controls. Clinical evaluation included anthropometry, body fat analysis by bio-impedance, biochemical, hsCRP, fibrinogen and insulin measurement. Subjects with MS had higher hsCRP (4.07?±?1.72 vs. 2.09?±?0.98?mg/L, P?=?0.0006); and fibrinogen levels (336?±?77 vs. 193?±?43?mg/dl, P <0.001) than controls. hsCRP and fibrinogen levels increased with number of metabolic abnormalities. Both inflammatory markers were positively associated with body mass index, body fat mass, percent body fat, HOMA-IR and all parameters of MS, except HDL with which only fibrinogen was negatively associated. Waist hip ratio was an independent predictor of hsCRP and fibrinogen in multiple regression analysis. hsCRP level of 2.6?mg/L predicted the MS with sensitivity, specificity and accuracy of 71?%, 78?% and 75?% respectively. Subjects with MS have increased inflammatory markers compared to healthy controls.     

Nasal response to inhaled histamine measured by acoustic rhinometry in infants

Aerosolized histamine, delivered via a face mask, is commonly used to evaluate bronchial reponsiveness in infants. To investigate nasal response to inhaled histamine we have measured nasal passage geometry in 32 infants by the use of acoustic reflections. Satisfactory data were obtained from only 17 infants (12 males, 5 females, 6.6 ± 4.4 months), because of awakening prior to completing the study in the remaining 15 infants. Acoustic rhinometry provided nasal cavity volume at 4 cm from the entrance of the nostril (VO4), the minimum cross-sectional area (A_min), and the distance from the nostril to A_min (D_min). Nasal geometry and lung function (maximum expiratory functional residual capacity [V_maxFRC] were measured before and immediately after a histamine challenge test using rapid thoratic compression. The histamine aerosols decreased both VO4 and A_min significantly by a mean of 17% and 13%, respectively (P < 0.001). There was a small, but significant increase (mean = 0.19 cm) of D_min in the right side only, indicating a posterior dislocation of the narrowest site with swelling of the mucous membrane. In general, we found a dose-response relationship in grouped data, with a greater fall in VO4 with increasing dose of histamine, but there was no correlation between percent fall in VO4 and V_maxFRC. This pilot study suggests that histamine aerosol affects nasal cavity geometry and that of acoustic rhinometry in infants and children warrants further investigation. Pediatr Pulmonol. 1994; 17:312–319. © 1994 Wiley-Liss, Inc.     

Inflammatory markers in cystic fibrosis

Plasma neutrophil elastase-alpha 1 antiproteinase complex, lactoferrin and C-reactive protein (CRP) were determined over a 15-month period in 26 patients with cystic fibrosis, of whom 21 were chronically infected with Pseudomonas aeruginosa. Median concentrations of both neutrophil products and CRP were greater in patients who were clinically stable than in healthy subjects without cystic fibrosis. CRP concentrations increased further at the onset of symptomatic exacerbations. Thirty-five courses of intravenous antibiotics and 22 courses of oral ciprofloxacin were reviewed and revealed similar improvements in clinical scores and lung function tests for both forms of treatment. Intravenous antibiotics reduced the plasma concentrations of both neutrophil products and CRP, while oral ciprofloxacin only significantly reduced the concentration of neutrophil elastase-alpha 1 antiproteinase complex. Plasma concentrations of inflammatory markers were significantly greater in exacerbations associated with fever and leukocytosis. Statistical modelling demonstrated negative within-patient relationships between lung function and both CRP and lactoferrin, and positive relationships between the three inflammatory markers. Neutrophil granule products and CRP reflect the pulmonary inflammatory state in cystic fibrosis and may be of value in monitoring treatment.