Airway inflammation in the elite athlete and type of sport.

BACKGROUND: The prevalence of asthma and bronchial hyper-responsiveness is greater in elite athletes than in the general population, and its association with mild airway inflammation has recently been reported. OBJECTIVE: To study the relationship between the type of sport practised at the highest levels of competition (on land or in water) and sputum induction cell counts in a group of healthy people and people with asthma. MATERIAL AND METHODS: In total, 50 athletes were enrolled. Medical history, results of methacholine challenge tests and sputum induced by hypertonic saline were analysed RESULTS: Full results were available for 43 athletes, who were classified by asthma diagnosis and type of sport (land or water sports). Nineteen were healthy (10 land and 9 water athletes) and 24 had asthma (13 land and 11 water athletes). Although the eosinophil counts of healthy people and people with asthma were significantly different (mean difference 3.1%, 95% CI 0.4 to 6.2, p = 0.008), analysis of variance showed no effect on eosinophil count for either diagnosis of asthma or type of sport. However, an effect was found for neutrophil counts (analysis of variance: F = 2.87, p = 0.04). There was also a significant correlation between neutrophil counts and both duration of training and bronchial hyper-responsiveness among athletes exposed to water (Spearman's rank correlations, 0.36 and 0.47, p = 0.04 and 0.04, respectively). CONCLUSIONS: Elite athletes who practice water sports have mild neutrophilic inflammation, whether or not asthma is present, related to the degree of bronchial hyper-reactivity and the duration of training in pool water.

Anti-inflammatory effects of high-dose inhaled fluticasone versus oral prednisone in asthma exacerbations.

The objective of the present study was to investigate the kinetics of high doses of inhaled steroid fluticasone in comparison with oral steroid prednisone on plasma protein leakage and bronchial eosinophilia in adults with moderate asthma exacerbations. The study design was a randomised, double-blind, placebo-controlled prospective trial. In total, 45 patients treated at the emergency department for moderate asthma exacerbations were recruited and 39 were assigned to receive fluticasone and placebo of prednisone (19 patients), or prednisone and placebo of fluticasone (20 patients). Medication was administered to all patients via a metered-dose inhaler and spacer (16 puffs; 4,000 microg.day(-1) or placebo) plus one pill (prednisone 30 mg.day(-1) or placebo). Spirometry and induced sputum for differential cell counts, albumin and alpha(2)-macroglobulin levels and blood eosinophils, interleukin-5 and granulocyte-macrophage colony-stimulating factor levels were obtained before treatment and at 2, 6 and 24 h after treatment. Symptoms clearly improved after 24 h in both groups. No differences were seen between groups in peak expiratory flow or forced expiratory flow in one second, which improved progressively but then decayed slightly after 24 h. Eosinophil counts in sputum also improved over time in both groups. The effect was faster with fluticasone than with prednisone, but was partially lost at 24 h. However, plasma proteins in sputum and eosinophil count in blood both decreased until 24 h, with no significant differences between groups. There was no correlation between eosinophil counts and plasmatic protein levels. In conclusion, both treatments improved symptoms, airway obstruction and inflammation, and plasma protein leakage at 24 h. Prednisone reduced blood eosinophil counts, while fluticasone reduced airway eosinophil counts, suggesting that the anti-inflammatory performance of fluticasone is exerted locally.

[Markers of airway remodeling in induced sputum from healthy smokers]. / Marcadores de remodelado bronquial en el esputo inducido de fumadores sanos.

OBJECTIVE: Airway remodeling in chronic obstructive pulmonary disease (COPD) has been linked to the equilibrium between matrix metalloproteinase (MMP) 9 and its inhibitor, tissue inhibitor of metalloproteinase (TIMP) 1. However, that equilibrium has not been analyzed in healthy smokers. The aim of this study was to assess the equilibrium between MMP-9 and TIMP-1 in induced sputum from healthy smokers, healthy nonsmokers (controls), and patients with COPD. PATIENTS AND METHODS: Samples of induced sputum were obtained from 35 individuals: 12 healthy smokers, 12 controls, and 11 patients with COPD. In each sample, a differential cell count was performed and enzyme-linked immunosorbent assays were used to analyze the concentrations of MMP-9 (total and active fraction) and TIMP-1. RESULTS: Compared with controls, healthy smokers were found to have a higher mean (SD) concentration of total MMP-9 (273 [277] ng/mL vs 128 [146] ng/mL) and a higher ratio of total MMP-9 to TIMP-1 (0.16 [0.14] vs 0.08 [0.06]). However, the ratio of active MMP-9 to TIMP-1 was similar in the 2 groups. Samples from patients with COPD had the highest concentrations of total MMP-9 (477 [262] ng/mL) and active MMP-9 (178 [126] ng/mL) and the lowest concentrations of TIMP-1 (1.044 [1.036] microg/mL). When all groups were considered together, there was an inverse relationship between the MMP-9/TIMP-1 ratio and the forced expiratory volume in the first second (FEV1). The relationship between the active MMP-9/TIMP-1 ratio and FEV1 was even stronger, and the relation of both ratios with FEV1 became stronger still when smoking was considered. CONCLUSIONS: Healthy smokers had a higher concentration of total MMP-9 and that concentration was correlated with their exposure to tobacco smoke. Maintenance of the active MMP-9/TIMP-1 ratio in healthy smokers may explain the absence of progressive airway obstruction. Measurement of active MMP-9 concentration could be useful for assessment of airway remodeling.

Marcadores de remodelado bronquial en el esputo inducido de fumadores sanos / Markers of airway remodeling in induced sputum from healthy smokers

Objetivo: El remodelado bronquial en la enfermedad pulmonar obstructiva crónica (EPOC) se ha relacionado con el equilibrio entre la metaloproitenasa (MMP) 9 y su inhibidor, el inhibidor tisular de MMP tipo 1 (TIMP-1). Dicho equilibrio no se ha analizado en fumadores sanos. Nuestro objetivo ha sido estudiar dicho equilibrio en el esputo inducido de fumadores sanos respecto a sanos no fumadores (controles) y pacientes con EPOC. Pacientes y métodos: Se obtuvieron 35 muestras de esputo inducido, de las que 12 provenían de fumadores sanos, otras 12 de controles y 11 de pacientes con EPOC. Se estudiaron la celularidad de las muestras y la concentración de MMP-9 (total y fracción activa) y TIMP-1 mediante enzimoinmunoanálisis. Resultados: Los fumadores sanos mostraron mayor concentración media (± desviación estándar) de MMP-9 total (273 ± 277 ng/ml) y una ratio mayor (0,16 ± 0,14) que los controles (128 ± 146 ng/ml y 0,08 ± 0,06, respectivamente). Sin embargo, la ratio MMP-9 activa/TIMP-1 fue equiparable en ambos grupos. Los pacientes con EPOC mostraron los valores más altos de MMP-9 total (477 ± 262 ng/ml) y activa (178 ± 126 ng/ml) y los más bajos de TIMP-1 (1.044 ± 1.036 ng/ml). Globalmente, la ratio mostró una relación inversa con el volumen espiratorio forzado en el primer segundo. Dicha relación fue aún superior con la MMP-9 activa y con el grado de tabaquismo. Conclusiones: Los fumadores sanos presentaron una mayor concentración de MMP-9 total en relación con el grado de exposición tabáquica. Una ratio MMP-9 activa/TIMP-1 conservada en fumadores sanos podría explicar la ausencia de obstrucción progresiva de la vía aérea. La medida de la MMP-9 activa puede ser útil en la determinación del remodelado bronquial

Objective: Airway remodeling in chronic obstructive pulmonary disease (COPD) has been linked to the equilibrium between matrix metalloproteinase (MMP) 9 and its inhibitor, tissue inhibitor of metalloproteinase (TIMP) 1. However, that equilibrium has not been analyzed in healthy smokers. The aim of this study was to assess the equilibrium between MMP-9 and TIMP-1 in induced sputum from healthy smokers, healthy nonsmokers (controls), and patients with COPD. Patients and methods: Samples of induced sputum were obtained from 35 individuals: 12 healthy smokers, 12 controls, and 11 patients with COPD. In each sample, a differential cell count was performed and enzyme-linked immunosorbent assays were used to analyze the concentrations of MMP-9 (total and active fraction) and TIMP-1. Results: Compared with controls, healthy smokers were found to have a higher mean (SD) concentration of total MMP-9 (273 [277] ng/mL vs 128 [146] ng/mL) and a higher ratio of total MMP-9 to TIMP-1 (0.16 [0.14] vs 0.08 [0.06]). However, the ratio of active MMP-9 to TIMP-1 was similar in the 2 groups. Samples from patients with COPD had the highest concentrations of total MMP-9 (477 [262] ng/mL) and active MMP-9 (178 [126] ng/mL) and the lowest concentrations of TIMP-1 (1.044 [1.036] µg/mL). When all groups were considered together, there was an inverse relationship between the MMP-9/TIMP-1 ratio and the forced expiratory volume in the first second (FEV1). The relationship between the active MMP-9/TIMP-1 ratio and FEV1 was even stronger, and the relation of both ratios with FEV1 became stronger still when smoking was considered. Conclusions: Healthy smokers had a higher concentration of total MMP-9 and that concentration was correlated with their exposure to tobacco smoke. Maintenance of the active MMP-9/TIMP-1 ratio in healthy smokers may explain the absence of progressive airway obstruction. Measurement of active MMP-9 concentration could be useful for assessment of airway remodeling

[Expansion of primary bronchial epithelial cell cultures]. / Método para amplificar cultivos primarios de células epiteliales bronquiales.

OBJECTIVE: Cell cultures provide a good model for studying lung diseases but they are difficult to reproduce and the number of cells obtained is limited. The aim of this study was to develop a way to increase the production of human bronchial epithelial cells (BEC) in primary cultures. MATERIAL AND METHODS: A total of 12 samples (9 from surgical specimens and 3 from endoscopic biopsies) were processed on plates coated with type I collagen with growth medium supplemented for BEC. When cell proliferation started, the explants were removed for successive subculturing. The remaining cells were left to proliferate and were trypsinized after 50% confluence. We recorded the number of cells obtained, cell viability, and the percentage positive for cytokeratin 7. RESULTS: The total number of cells obtained by this method was 3-fold the number of human BEC obtained with simple primary cultures. The maximum number of subcultures was 5, mean (SD) cell viability was 91.9% (11.7%), and the percentage of cells positive for cytokeratin 7 was 30.71% (10.68%). CONCLUSIONS: The described method for expanding primary BEC cultures increases cell production.

Método para amplificar cultivos primarios de células epiteliales bronquiales / Expansion of primary epithelial cell cultures

Objetivo: Los cultivos celulares son un buen modelo para el estudio de las enfermedades pulmonares, pero son difíciles de reproducir y producen un número limitado de células. El objetivo de este estudio ha sido desarrollar un método que incrementase la producción de células epiteliales bronquiales (CEB) humanas en cultivos primarios. Material y métodos: Se procesó un total de 12 muestras (9 procedentes de muestras quirúrgicas y 3 de biopsias endoscópicas) en placas recubiertas de colágeno tipo I con medio suplementado para CEB. Al iniciarse la proliferación celular a su alrededor, los explantes se extrajeron y subcultivaron sucesivamente. Las células restantes se dejaron proliferar y se tripsinizaron tras alcanzar más del 50% de confluencia. Se valoraron el número de células obtenidas, la viabilidad y la citoqueratina 7. Resultados: El número total de células obtenidas con este método superó en una media de 3 veces el número de CEB humanas obtenidas en cultivos primarios simples. El número máximo de subcultivos fue de 5, la viabilidad media (± desviación estándar) fue de 91,9 ± 11,7% y el porcentaje de células positivas para la citoqueratina 7 del 30,71 ± 10,68%. Conclusiones: El método descrito para amplificar cultivos primarios de CEB permite incrementar la producción de células obtenidas

Objective: Cell cultures provide a good model for studying lung diseases but they are difficult to reproduce and the number of cells obtained is limited. The aim of this study was to develop a way to increase the production of human bronchial epithelial cells (BEC) in primary cultures. Material and methods: A total of 12 samples (9 from surgical specimens and 3 from endoscopic biopsies) were processed on plates coated with type I collagen with growth medium supplemented for BEC. When cell proliferation started, the explants were removed for successive subculturing. The remaining cells were left to proliferate and were trypsinized after 50% confluence. We recorded the number of cells obtained, cell viability, and the percentage positive for cytokeratin 7. Results: The total number of cells obtained by this method was 3-fold the number of human BEC obtained with simple primary cultures. The maximum number of subcultures was 5, mean (SD) cell viability was 91.9% (11.7%), and the percentage of cells positive for cytokeratin 7 was 30.71% (10.68%). Conclusions: The described method for expanding primary BEC cultures increases cell production

[Bronchial exudate of serum proteins during asthma attack]. / Exudado bronquial de proteínas séricas en las crisis de asma.

OBJECTIVE: Although altered vascular permeability and edema of the bronchial mucosa are associated with asthma attack, their influence on its severity remains unknown. We address this issue by comparing relative indices for the concentration of albumin (RIAlb) and alpha2-macroglobulin (RIalpha2M) in induced sputum and peripheral blood from patients with exacerbated asthma, patients with stable asthma, and control subjects. PATIENTS AND METHODS: Forty-six volunteers participated in the study: 14 with exacerbated asthma (forced expiratory volume in the first second [FEV1] 74.3% [SD, 20.8%] of reference), 23 with stable asthma (FEV1 93.6% [7.5%]), and 9 controls (FEV1 101.1% [9.9%]). The concentrations of albumin and alpha2-macroglobulin were quantified by immunoturbidimetry and immunonephelometry, respectively. The relative index was then calculated by dividing the concentration in sputum supernatant by the concentration in peripheral blood. RESULTS: The mean RIAlb was 1.2 (1.1) in the control group, 2.9 (3.1) in the stable asthma group, and 6.0 (6.7) in the exacerbated asthma group. The RIalpha2M values were 11.7 (10.9), 11.9 (14.7), and 3.2 (3.8) for the control group and stable and exacerbated asthma groups, respectively. The increases in the RIAlb values between all groups, and the decrease in the RIalpha2M value between the exacerbated asthma and control groups were statistically significant (P<.05). The percentage of neutrophils, but not of eosinophils, in sputum was correlated with the RIAlb (r=0.39; P=.008) but not the RIalpha2M (r=-0.035; P=.82). FEV1 displayed an inverse relationship with the RIAlb (r=-0.43; P=.009) but not with the RIalpha2M (r=-0.206; P=.24). No correlation was found between oxyhemoglobin saturation and either the RIAlb (r=-0.33; P=.19) or the RIalpha2M (r=-0.12; P=.84). CONCLUSIONS: Vascular permeability is altered during asthma exacerbations and appears to be correlated with the presence of neutrophils and the degree of bronchial obstruction.

Exudado bronquial de proteínas séricas en las crisis de asma / Bronchial exudate of serum proteins during asthma attack

Objetivo: La alteración de la permeabilidad vascular y el edema de la mucosa bronquial se asocian con las crisis de asma. Hay pocos datos publicados y además no se conoce su relación con la gravedad de éstas. Así se propuso comparar los índices relativos de albúmina (IRAlb) y de macroglobulina α2 (IRMα2), entre esputo inducido y sangre periférica, de asmáticos agudizados (AA), asmáticos estables (AE) y controles. Pacientes y métodos: Se estudió a 46 voluntarios: 14 del grupo AA (volumen espiratorio forzado en el primer segundo [FEV1]: 74,3 ± 20,8), 23 del AE (FEV1: 93,6 ± 7,5) y 9 controles (FEV1: 101,1 ± 9,9). Se cuantificó la concentración de albúmina (turbidimetría inmunoquímica) y de macroglobulina α2 (nefelometría inmunoquímica) en el sobrenadante del esputo y en sangre venosa periférica y se calcularon los índices relativos. Resultados: La media ± desviación estándar del IRAlb fue de 1,2 ± 1,1 en el grupo control, de 2,9 ± 3,1 en AE y de 6,0 ± 6,7 en AA. El IRMα2 fue 11,7 ± 10,9, 11,9 ± 14,7 y 3,2 ± 3,8, respectivamente. El incremento del IRAlb entre los grupos AA, AE y control, y el descenso del IRMα2 entre AA y control fueron estadísticamente significativos (p < 0,05). Se relacionó el porcentaje de neutrófilos, y no el de eosinófilos, con el IRAlb (r = 0,39; p = 0,008), pero no con el IRMα2 (r = -­0,035; p = 0,82). El FEV1 se relacionó inversamente con el IRAlb (r = ­-0,43; p = 0,009) y no con el IRMα2 (r = -­0,206; p = 0,24), y tampoco se relacionó la saturación de oxihemoglobina con el IRAlb (r = ­0,33; p = 0,19) o el IRMα2 (r = ­-0,12; p = 0,84). Conclusiones: La permeabilidad vascular está alterada en las agudizaciones de asma y parece relacionarse con la presencia de neutrófilos y el grado de obstrucción bronquial

Objective: Although altered vascular permeability and edema of the bronchial mucosa are associated with asthma attack, their influence on its severity remains unknown. We address this issue by comparing relative indices for the concentration of albumin (RIAlb) and α2-macroglobulin (RIα2M) in induced sputum and peripheral blood from patients with exacerbated asthma, patients with stable asthma, and control subjects. Patients and methods: Forty-six volunteers participated in the study: 14 with exacerbated asthma (forced expiratory volume in the first second [FEV1] 74.3% [SD, 20.8%] of reference), 23 with stable asthma (FEV1 93.6% [7.5%]), and 9 controls (FEV1 101.1% [9.9%]). The concentrations of albumin and α2-macroglobulin were quantified by immunoturbidimetry and immunonephelometry, respectively. The relative index was then calculated by dividing the concentration in sputum supernatant by the concentration in peripheral blood. Results: The mean RIAlb was 1.2 (1.1) in the control group, 2.9 (3.1) in the stable asthma group, and 6.0 (6.7) in the exacerbated asthma group. The RIα2M values were 11.7 (10.9), 11.9 (14.7), and 3.2 (3.8) for the control group and stable and exacerbated asthma groups, respectively. The increases in the RIAlb values between all groups, and the decrease in the RIα2M value between the exacerbated asthma and control groups were statistically significant (P<.05). The percentage of neutrophils, but not of eosinophils, in sputum was correlated with the RIAlb (r=0.39; P=.008) but not the RIα2M (r=-­0.035; P=.82). FEV1 displayed an inverse relationship with the RIAlb (r=­-0.43; P=.009) but not with the RIα2M (r=­-0.206; P=.24). No correlation was found between oxyhemoglobin saturation and either the RIAlb (r=­-0.33; P=.19) or the RIα2M (r=­-0.12; P=.84). Conclusions: Vascular permeability is altered during asthma exacerbations and appears to be correlated with the presence of neutrophils and the degree of bronchial obstruction

[Total IgE in the sputum and serum of patients with asthma]. / IgE total en el esputo y suero de pacientes con asma.

BACKGROUND: The correlation between total IgE in induced sputum (IS) and serum is not well defined. The aim of this study was to investigate the relationship between total IgE in IS and total IgE in serum and airway inflammation. METHODS: Twenty-one patients with stable asthma and thirteen healthy controls were studied. Clinical and spirometric data were collected and a skin prick test to the 13 most common aeroallergens in our area was performed in all subjects. Total IgE in IS and serum was determined by the UNICAP immunoanalysis system (Pharmacia Uppsala, Sweden) while albumin concentration in IS and serum was determined using the Cobas Integra turbidimetric method (Roche Diagnostics, Basel, Switzerland). RESULTS: The percentage of eosinophils in EI was 8.7 (11.8) in asthmatic subjects and was 0.5 (1) in healthy controls. Total IgE (KU/L) was 43.2 (23) in asthmatics vs 25.6 (3) in healthy controls in IS, and was 329 (413) in asthmatics vs 57 (78) in controls in serum. Total IgE in IS was significantly correlated with total IgE in serum; r = 0.71 (p = 0.048), but not with the albumin relative index. No correlation was found between IgE and the number of eosinophils in IS. CONCLUSIONS: Total IgE can be measured in IS. Total IgE in IS is mildly correlated with total IgE measured in serum. The lack of correlation between total IgE and albumin in IS suggests that IgE in IS could be locally produced, at least in part.

IgE total en el esputo y suero de pacientes con asma / Total IgE in the sputum and serum of patients with asthma

Antecedentes: La IgE total en esputo inducido (EI) y su relación con la IgE total en el suero (S) no están bien definidas. El objetivo de este trabajo fue estudiar la relación entre la concentración de IgE total en EI con la concentración de IgE total en S y la inflamación de las vías aéreas. Métodos: Se estudiaron 21 pacientes con asma estable y 13 controles sanos. De cada paciente se recogieron los datos clínicos y espirométricos, y a todos se les hicieron pruebas cutáneas (prick test) con 13 aeroalergenos comunes en nuestra área. Se determinó la concentración de IgE total en EI y en S por inmuno-ensayo con el sistema UNICAP (Pharmacia-Uppsala, Suecia) y la concentración de albúmina por el método de turbidimetría Cobas Integra (Roche Diagnostics, Basel, Suiza). Resultados: El porcentaje de eosinófilos (%) en EI de asmáticos fue de 8,7 (11,8) y 0,5 (1) en sanos. La concentración de IgE (KU/L) en EI fue 43,2 (23) en asmáticos vs 25,6 (3) en sanos, y en S fue 329 (413) en asmáticos vs 57 (78) en sanos. La correlación entre IgE total en EI e IgE total en S fue de r = 0,71 (p = 0,048) pero no se relacionó con el índice relativo de albúmina. No hubo relación entre IgE en EI y número de eosinófilos en EI. Conclusiones: La IgE total puede medirse en esputo inducido y su concentración se relaciona con la IgE determinada en sangre. La ausencia de relación con la albúmina sugiere que la IgE en esputo no proviene exclusivamente de la extravasación, sino que podría intervenir una cierta producción local

Background: The correlation between total IgE in induced sputum (IS) and serum is not well defined. The aim of this study was to investigate the relationship between total IgE in IS and total IgE in serum and airway inflammation. Methods: Twenty-one patients with stable asthma and thirteen healthy controls were studied. Clinical and spirometric data were collected and a skin prick test to the 13 most common aeroallergens in our area was performed in all subjects. Total IgE in IS and serum was determined by the UNICAP immunoanalysis system (Pharmacia Uppsala, Sweden) while albumin concentration in IS and serum was determined using the Cobas Integra turbidimetric method (Roche Diagnostics, Basel, Switzerland). Results: The percentage of eosinophils in EI was 8.7 (11.8) in asthmatic subjects and was 0.5(1) in healthy controls. Total IgE (KU/L) was 43.2 (23) in asthmatics vs 25.6 (3) in healthy controls in IS, and was 329 (413) in asthmatics vs 57 (78) in controls in serum. Total IgE in IS was significantly correlated with total IgE in serum; r = 0.71 (p = 0.048), but not with the albumin relative index. No correlation was found between IgE and the number of eosinophils in IS. Conclusions: Total IgE can be measured in IS. Total IgE in IS is mildly correlated with total IgE measured in serum. The lack of correlation between total IgE and albumin in IS suggests that IgE in IS could be locally produced, at least in part