RESUMEN
The prevalence of asthma has increased worldwide. The reasons for this rise remain unclear. Oxidative stress plays an important role in the pathogenesis of asthma. Glutathione (GSH) is the major representative of the class of nonprotein thiols and plays a pivotal role in a variety of enzymatic and nonenzymatic reactions that protect tissues against oxidative stress. In antioxidative reactions, GSH is converted into its oxidized form, glutathione disulfide (GSSG) that in its turn is enzymatically reduced into GSH to maintain a physiological redox balance. We used a guinea pig model of asthma to assess whether the early asthmatic reaction is associated with decreased lung levels of glutathione, and whether decreased glutathione is implicated in the increased airway smooth muscle reactivity that is associated with exposure of the lungs to allergen. Lung glutathione levels were decreased immediately after the onset of the early asthmatic reaction in vivo and associated with the release of 8-iso-PGF(2alpha), an indicator for oxidative stress. Glutathione ethylester, a glutathione precursor, blunted the airway obstruction during an early asthmatic reaction in a perfusion model and glutathione depletion rendered the airways hyperreactive. Glutathione ethyl ester in the buffer prevented this hyperreactivity. These results indicate that glutathione can modulate the early asthmatic reaction as well as the airway hyperresponsiveness.
Asunto(s)
Asma/metabolismo , Asma/prevención & control , Hiperreactividad Bronquial/metabolismo , Hiperreactividad Bronquial/prevención & control , Glutatión/fisiología , Animales , Asma/fisiopatología , Hiperreactividad Bronquial/fisiopatología , Modelos Animales de Enfermedad , Glutatión/antagonistas & inhibidores , Cobayas , Histamina/farmacología , Masculino , Ovalbúmina/administración & dosificación , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/fisiología , Factores de TiempoRESUMEN
Our understanding of asthma severity was advanced by the identification of biomarkers which account for differences in lung function impairment. We tried to examine the effects of corticosteroid treatment on known correlates of asthma severity including peripheral eosinophil counts, total IgE, IL-5, and eotaxin Levels in plasma. We compared these biomarkers among groups of stable asthmatics categorized by the dose of corticosteroid (N: steroid-free, n = 25; L: low-dose inhaled, n = 27; MH: medium or high-dose inhaled, n= 19; O: inhaled plus oral, n= 8). Next we compared these markers and peak expiratory flow rate (PEFR) in unstable asthmatics before and after treatment with steroids (n = 22). Eotaxin levels in the O group were higher than those in the N and MH groups (P < 0.05). Logistic regression analysis demonstrated that plasma eotaxin level was correlated with the severity of asthma defined by treatment intensity (P = 0.01) and % FEV1 (P = 0.04) while the other markers were not. Eotaxin levels did not change after steroid treatment in unstable patients, whereas eosinophil counts decreased in parallel with PEFR. Among biomarkers of asthma severity studied, plasma eotaxin level was not significantly affected by corticosteroid treatment, and was associated with the severity even in the presence of steroid therapy.
Asunto(s)
Corticoesteroides/administración & dosificación , Asma/tratamiento farmacológico , Quimiocinas CC/sangre , Administración por Inhalación , Administración Oral , Anciano , Asma/sangre , Asma/fisiopatología , Biomarcadores/sangre , Quimiocina CCL11 , Estudios Transversales , Femenino , Volumen Espiratorio Forzado/fisiología , Humanos , Masculino , Persona de Mediana Edad , Ápice del Flujo Espiratorio/fisiología , Análisis de RegresiónRESUMEN
BACKGROUND: Eotaxin, a CC chemokine expressed in the asthmatic lung, has been associated with impaired lung function. The role of its variant form is unknown. OBJECTIVE: The purpose of this study was to detect the population frequency and effects of a known single-nucleotide polymorphism in the eotaxin gene in which a threonine residue (THR(23)) is substituted for the wild-type alanine (ALA(23)) at the 23rd amino acid at the terminus of the peptide leader sequence. METHODS: We measured eotaxin protein secretion in 293 cells transfected with expression vectors and in PBMCs obtained from individuals bearing the alternative forms of the gene. A case-control study of plasma eotaxin levels and eosinophil counts, a comparison of baseline lung function by genotype in a population of 806 subjects with asthma, and a comparison of the allele frequency with a nonasthmatic population were performed. RESULTS: Human 293 cells and PBMCs with THR(23) variant eotaxin secreted significantly less eotaxin protein than did ALA(23)-bearing cells. In the case-control study, THR(23)-THR(23) individuals had lower plasma levels of eotaxin (310 [240-350] vs 420 [270-700] pg/mL; P < .05) and eosinophil counts (120 [5-220] vs 190 [110-470] cells/microL; P < .05) than ALA(23)-ALA(23) subjects; heterozygous subjects had intermediate levels. Higher levels of lung function were associated with THR(23) eotaxin (percent of predicted FEV(1), 65% +/- 3.5% [THR(23)-THR(23)] vs 58% +/- 0.9% [THR(23)-ALA(23)] and 56% +/- 0.5% [ALA(23)-ALA(23)]; P < .05). CONCLUSION: The THR(23) variant is associated with both decreased eosinophil counts and higher levels of lung function in subjects with asthma.
Asunto(s)
Asma/genética , Quimiocinas CC/genética , Adulto , Estudios de Casos y Controles , Células Cultivadas , Quimiocina CCL11 , Clonación Molecular , Femenino , Genotipo , Humanos , Masculino , Mutación , Fenotipo , Polimorfismo Conformacional Retorcido-SimpleAsunto(s)
Quimiocinas/fisiología , Lipopolisacáridos/farmacología , Receptores de Quimiocina/metabolismo , Animales , Quimiocina CCL11 , Quimiocinas/biosíntesis , Quimiocinas/clasificación , Quimiocinas/genética , Quimiocinas CC/fisiología , Factores Quimiotácticos/farmacología , Factores Quimiotácticos/fisiología , Quimiotaxis/efectos de los fármacos , Quimiotaxis/fisiología , Eosinófilos/fisiología , Humanos , Hipersensibilidad Inmediata/metabolismo , Lactante , Inflamación/metabolismo , Ligandos , Ratones , Ratones Noqueados , Eosinofilia Pulmonar/metabolismo , Receptores CCR3 , Especificidad por SustratoRESUMEN
To investigate the mechanisms of eosinophil recruitment in allergic airway inflammation, we examined the effects of interleukin (IL)-4, a Th2-type cytokine, on eotaxin and monocyte chemoattractant protein-4 (MCP-4) expression in human peripheral blood mononuclear cells (PBMCs; n = 10), in human lower airway mononuclear cells (n = 5), in the human lung epithelial cell lines A549 and BEAS-2B, and in human cultured airway epithelial cells. IL-4 inhibited eotaxin and MCP-4 mRNA expression induced by IL-1 beta and tumor necrosis factor-alpha in PBMCs but did not significantly inhibit expression in epithelial cells. Eotaxin and MCP-4 mRNA expression was not significantly induced by proinflammatory cytokines in lower airway mononuclear cells. IL-1 beta-induced eotaxin and MCP-4 protein production was also inhibited by IL-4 in PBMCs, whereas IL-4 enhanced eotaxin protein production in A549 cells. In contrast, dexamethasone inhibited eotaxin and MCP-4 expression in both PBMCs and epithelial cells. The divergent effects of IL-4 on eotaxin and MCP-4 expression between PBMCs and epithelial cells may create chemokine concentration gradients between the subepithelial layer and the capillary spaces that may promote the recruitment of eosinophils to the airway in Th2-type responses.
Asunto(s)
Quimiocinas CC , Citocinas/metabolismo , Leucocitos Mononucleares/metabolismo , Pulmón/metabolismo , Proteínas Quimioatrayentes de Monocitos/metabolismo , Mucosa Respiratoria/metabolismo , Northern Blotting , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Línea Celular , Células Cultivadas , Quimiocina CCL11 , Factores Quimiotácticos Eosinófilos/genética , Factores Quimiotácticos Eosinófilos/metabolismo , Citocinas/genética , Relación Dosis-Respuesta a Droga , Ensayo de Inmunoadsorción Enzimática , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Humanos , Interleucina-1/farmacología , Interleucina-4/farmacología , Interleucina-8/genética , Interleucina-8/metabolismo , Interleucinas/metabolismo , Linfocitos/metabolismo , Proteínas Quimioatrayentes de Monocitos/genética , Estabilidad del ARN , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factores de Tiempo , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
Prior studies have suggested that angiotensin I-converting enzyme (ACE) genotype correlates with superior physical performance in highly selected populations. This study assessed whether such an association exists in a heterogeneous population. Using polymerase chain reaction techniques, we determined the ACE genotypes (insertion/insertion, deletion/insertion, or deletion/deletion) of 62 male and 85 female US Army recruits. Before and after 8 wk of basic training, we determined peak oxygen uptake and performance on the Army Physical Fitness Test (APFT), which includes standardized measures of muscular endurance (sit-ups, push-ups) and a 2-mile run. Subjects of different ACE genotypes had similar peak oxygen uptakes and APFT scores, both before and after training. Subjects with genotype II had higher APFT scores than others, but the differences were not statistically significant. Furthermore, no ACE genotype group had a performance advantage in analyses that adjusted for baseline fitness. We conclude that ACE genotype does not have a strong effect on aerobic power or muscular endurance in healthy, young American adults drawn from an ethnically and geographically diverse population.
Asunto(s)
Ejercicio Físico/fisiología , Peptidil-Dipeptidasa A/genética , Adulto , Estudios de Cohortes , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Masculino , Personal Militar , Consumo de Oxígeno/genética , Resistencia Física/genéticaRESUMEN
Chronic eosinophilic pneumonia (CEP) is characterized by chronic or recurrent pulmonary infiltrates with eosinophils, but the precise mechanism of eosinophil accumulation has not been fully elucidated. Eotaxin is one of the CC chemokines that selectively recruits eosinophils and contributes to the pathogenesis of allergic airway diseases including asthma, but its roles in pathogenesis of CEP have not been fully elucidated. The authors measured concentrations of eotaxin and other CC chemokines, monocyte chemoattractant protein-1 (MCP-1), regulated on activation, normal T-cell expressed and secreted, macrophage inflammatory protein-1alpha, and the eosinophil activating Th2 cytokine interleukin (IL)-5 in bronchoalveolar lavage (BAL) fluid from CEP patients (n=11), and compared these concentrations with those from control subjects (n = 6). The eotaxin (904 +/- 203 versus 29 +/- 7 pg x mL(-1), p = 0.0001), MCP-1 (194 +/- 57 versus 15 +/- 2 pg x mL(-1), p < 0.05), and IL-5 (7.8 +/- 2.0 versus 2.7 +/- 0.6 pg x mL(-1), p < 0.05) levels were significantly higher for cases with CEP in comparison to those serving as controls. Proportions of eosinophil and lymphocyte counts were greater in BAL fluid from CEP patients. Eotaxin and IL-5 levels correlated with the proportion of eosinophils in BAL fluid from CEP patients. MCP-1 correlated with the relative lymphocyte numbers. In short, eotaxin, interleukin-5, and monocyte chemoattractant protein-1 levels were higher in the BAL fluid of CEP patients and these levels may contribute to eosinophil and lymphocyte recruitment and activation in the airways as found with this disorder.
Asunto(s)
Quimiocina CCL2/metabolismo , Quimiocinas CC/metabolismo , Eosinofilia Pulmonar/inmunología , Adulto , Anciano , Líquido del Lavado Bronquioalveolar/inmunología , Quimiocina CCL11 , Eosinófilos/inmunología , Femenino , Humanos , Interleucina-5/metabolismo , Recuento de Leucocitos , Activación de Linfocitos/inmunología , Recuento de Linfocitos , Masculino , Persona de Mediana EdadRESUMEN
STUDY OBJECTIVES: To measure the prevalence of exercise-induced bronchospasm (EIB) and to determine its effect on the physical performance response to training in otherwise healthy young adults. DESIGN: Observational, retrospective study. SETTING: Fort Jackson, SC, May to July 1998. PARTICIPANTS: One hundred thirty-seven ethnically diverse US Army recruits undergoing an 8-week Army basic training course. MEASUREMENTS AND RESULTS: Subjects underwent exercise challenge testing at the end of basic training to evaluate for EIB (defined as a decrease in FEV(1) of > or = 15%, 1 or 10 min after running to peak oxygen uptake on a treadmill). Those subjects who were unable to run to peak oxygen uptake, or who were unable to perform two baseline FEV(1) maneuvers the results of which were within 5% of each other, were excluded from analysis. We measured peak oxygen uptake on a treadmill and the scores achieved on the components of the US Army physical fitness test (APFT). Of 137 subjects, 121 (58 men and 63 women) met our inclusion criteria. Eight subjects (7%) had EIB. Subjects who experienced EIB and unaffected control subjects both showed statistically significant gains in performance on the APFT events during basic training. At the end of basic training, peak oxygen uptake levels and APFT event scores were not significantly different between subjects with EIB and unaffected control subjects. CONCLUSIONS: Seven percent of the US Army recruits who were tested had EIB, but this did not hinder their physical performance gains during basic training. EIB per se should not be an absolute reason to exclude individuals from employment in jobs with heavy physical demands.
Asunto(s)
Asma Inducida por Ejercicio/epidemiología , Asma Inducida por Ejercicio/fisiopatología , Ejercicio Físico , Personal Militar , Adulto , Femenino , Humanos , Masculino , Consumo de Oxígeno , Educación y Entrenamiento Físico , Aptitud Física , Prevalencia , Estados Unidos/epidemiologíaRESUMEN
Expression of pulmonary eotaxin protein and mRNA was determined in six subjects with atopic asthma and five nonatopic normal subjects. Levels of eotaxin expression and eosinophil mobilization were compared before and after segmental allergen challenge in subjects with atopic asthma. In the absence of allergen challenge, we found significantly higher levels of eotaxin in the bronchoalveolar lavage (BAL) fluid of subjects with asthma than in that of normal subjects (25 +/- 3 versus 15 +/- 2 pg/ml, p < 0.05). BAL eotaxin levels increased after segmental allergen challenge in all six subjects with atopic asthma tested, with a mean increase from 22 +/- 4 to 53 +/- 10 pg/ml (p = 0.013). Segmental allergen challenge was associated with a significant increase in the percentage of BAL macrophages and eosinophils that were immunopositive for eotaxin. Eotaxin mRNA was detectable by northern analysis in BAL cells exclusively from allergen-challenged segments. Allergen- induced increases in eotaxin levels were strongly associated with increases in BAL eosinophil recovery (r(2) = 0.88, p = 0.0036). Segmental allergen challenge also increased eotaxin expression in airway epithelial and endothelial cells obtained by endobronchial biopsy. These findings demonstrate, for the first time, that the airways of subjects with allergic asthma respond to allergen by increasing eotaxin expression. The tissue loci of eotaxin expression, the levels of eotaxin recovered in BAL fluid, and the association of eotaxin levels with eosinophil mobilization suggest either that eotaxin plays a mechanistic role in allergen-induced airway eosinophilia or that it serves as a biomarker for the causal mechanisms.
Asunto(s)
Alérgenos/administración & dosificación , Asma/inmunología , Pruebas de Provocación Bronquial , Quimiocinas CC , Citocinas/metabolismo , Hipersensibilidad Inmediata/inmunología , Adulto , Alérgenos/inmunología , Asma/metabolismo , Asma/patología , Bronquios/metabolismo , Bronquios/patología , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Broncoscopía , Recuento de Células , Quimiocina CCL11 , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunohistoquímica , Masculino , ARN/análisis , Pruebas CutáneasRESUMEN
Peribronchial smooth muscle constriction causes airway stretch, an important mechanical force in developing lung. Little is known about factors influencing these spontaneously active muscle elements. We measured contractile activity of neurokinin (NK) receptors on fetal intrapulmonary smooth muscle by tracheal perfusion assay (n = 11). Injecting either capsaicin or the NK(2) receptor agonist [NLE(10)]NKA resulted in significant (P < 0.05) bronchoconstriction. A specific NK(2) receptor antagonist inhibited constriction caused by endogenous tachykinins released by capsaicin. We then examined NK(2) receptor (n = 44) and NKA (n = 23) ontogeny in human lung. NKA immunostaining was identified in peribronchial nerves in samples with gestational age >12 wk. NK(2) receptor protein was identified in peribronchial and perivascular smooth muscle. These results indicate that endogenous tachykinins released by the developing lung act via NK(2) receptors to cause smooth muscle constriction. We speculate that tachykinins could modulate lung development.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica/fisiología , Pulmón/metabolismo , Neuroquinina A/metabolismo , Receptores de Neuroquinina-2/metabolismo , Adulto , Compuestos de Bario/farmacología , Broncoconstricción/efectos de los fármacos , Broncoconstricción/fisiología , Broncoconstrictores/farmacología , Capsaicina/farmacología , Cloruros/farmacología , Feto , Humanos , Inmunohistoquímica , Técnicas In Vitro , Pulmón/citología , Pulmón/efectos de los fármacos , Pulmón/embriología , Cloruro de Metacolina/farmacología , Músculo Liso/embriología , Músculo Liso/inervación , Músculo Liso/metabolismo , Neuroquinina A/análogos & derivados , Neuroquinina A/farmacología , Péptidos/farmacología , Perfusión , ARN Mensajero/metabolismo , Receptores de Neuroquinina-1/agonistas , Receptores de Neuroquinina-1/metabolismo , Receptores de Neuroquinina-2/agonistas , Receptores de Neuroquinina-2/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tráquea/irrigación sanguínea , Tráquea/efectos de los fármacos , Tráquea/embriología , Tráquea/metabolismoRESUMEN
Nitric oxide (NO) is a molecular gas that can be recovered in higher levels from the exhaled gas of subjects with asthma than from subjects without asthma. However, the precise mechanisms responsible of promoting increased fraction of expired nitric oxide (FE(NO)) in asthma are unknown. As leukotriene antagonism has been shown to reduce FE(NO) in patients with asthma, we hypothesized that leukotrienes mediate the increased FE(NO) encountered in this condition. Furthermore, because leukotriene antagonism stabilizes serum eosinophil markers during reductions in inhaled corticosteroid doses, and FE(NO) has been shown to correlate with sputum eosinophils in asthma, we reasoned that the effect of leukotrienes on FE(NO) might be mediated by eosinophils recruited to the airway by leukotrienes. To test this hypothesis, we performed methacholine and leukotriene (LT) E(4) bronchoprovocation challenges in 16 subjects with atopic asthma and measured FE(NO) and sputum differential counts before and after bronchoprovocation. We then compared FE(NO) in the seven subjects who developed increased sputum eosinophils following LTE(4) inhalation with values measured after methacholine inhalation in these seven subjects. Following LTE(4) inhalation, eosinophils rose from 4.01 +/- 0.89% pre-LTE(4) to 8.33 +/- 1.52% post-LTE(4). The mean change in sputum eosinophils from baseline after LTE(4) inhalation was larger than that after methacholine inhalation (+4.31 +/- 1.25% versus -1.14 +/- 0.93%). After LTE(4) inhalation, FE(NO) levels did not differ from prechallenge baseline or from levels following methacholine inhalation (ANOVA p > 0.05). These data indicate that neither LTE(4) nor recruitment of eosinophils into the airway by LTE(4) is a sufficient stimulus to acutely increase FE(NO) in subjects with asthma.
Asunto(s)
Asma/metabolismo , Pruebas Respiratorias , Leucotrieno E4/administración & dosificación , Cloruro de Metacolina/administración & dosificación , Óxido Nítrico/metabolismo , Administración por Inhalación , Adulto , Resistencia de las Vías Respiratorias , Asma/patología , Asma/fisiopatología , Pruebas de Provocación Bronquial , Recuento de Células , Eosinófilos , Femenino , Humanos , Leucotrieno E4/farmacología , Masculino , Cloruro de Metacolina/farmacología , Persona de Mediana Edad , Esputo/citologíaRESUMEN
Eotaxin is an asthma-related C-C chemokine that is produced in response to interleukin-1beta (IL-1beta). We detected an increase in newly transcribed eotaxin mRNA in IL-1beta-stimulated airway epithelial cells. Transient transfection assays using promoter-reporter constructs identified a region as essential for IL-1beta-induced increases in eotaxin transcription. Using site-directed mutagenesis, we found that a nuclear factor-kappaB (NF-kappaB) site located 46 bp upstream from the transcriptional start site was both necessary and sufficient for IL-1beta induction of reporter construct activity. Electrophoretic mobility shift assay demonstrated that IL-1beta-stimulated airway epithelial cells produced p50 and p65 protein that bound this site in a sequence-specific manner. The functional importance of the NF-kappaB site was demonstrated by coexpression experiments in which increasing doses of p65 expression vector were directly associated with reporter activity exclusively in constructs with an intact NF-kappaB site (r(2) = 0.97, P = 0.002). Moreover, IL-1beta-induced increases in eotaxin mRNA expression are inhibited by inhibitors of NF-kappaB. Our findings implicate NF-kappaB and its binding sequence in IL-1beta-induced transcriptional activation of the eotaxin gene.
Asunto(s)
Quimiocinas CC , Citocinas/genética , Interleucina-1/farmacología , FN-kappa B/genética , Mucosa Respiratoria/fisiología , Activación Transcripcional/efectos de los fármacos , Adenocarcinoma Bronquioloalveolar , Quimiocina CCL11 , Células Epiteliales/efectos de los fármacos , Células Epiteliales/fisiología , Regulación de la Expresión Génica/efectos de los fármacos , Genes Reporteros , Prueba de Complementación Genética , Humanos , Neoplasias Pulmonares , Mutagénesis Sitio-Dirigida/fisiología , FN-kappa B/metabolismo , Regiones Promotoras Genéticas/fisiología , ARN Mensajero/metabolismo , Mucosa Respiratoria/citología , Células Tumorales CultivadasRESUMEN
BACKGROUND: Unstable atherosclerotic lesions typically have an abundant inflammatory cell infiltrate, including activated T cells, macrophages, and mast cells, which may decrease plaque stability. The pathophysiology of inflammatory cell recruitment and activation in the human atheroma is incompletely described. METHODS AND RESULTS: We hypothesized that differential gene expression with DNA microarray technology would identify new genes that may participate in vascular inflammation. RNA isolated from cultured human aortic smooth muscle cells treated with tumor necrosis factor-alpha (TNF-alpha) was examined with a DNA microarray with 8600 genes. This experiment and subsequent Northern analyses demonstrated marked increases in steady-state eotaxin mRNA (>20 fold), a chemokine initially described as a chemotactic factor for eosinophils. Because eosinophils are rarely present in human atherosclerosis, we then studied tissue samples from 7 normal and 14 atherosclerotic arteries. Immunohistochemical analysis demonstrated overexpression of eotaxin protein and its receptor, CCR3, in the human atheroma, with negligible expression in normal vessels. Eotaxin was predominantly located in smooth muscle cells. The CCR3 receptor was localized primarily to macrophage-rich regions as defined by immunopositivity for CD 68; a minority of mast cells also demonstrated immunopositivity for the CCR3 receptor. CONCLUSIONS: Eotaxin and its receptor, CCR3, are overexpressed in human atherosclerosis, suggesting that eotaxin participates in vascular inflammation. These data demonstrate how genomic differential expression technology can identify novel genes that may participate in the stability of atherosclerotic lesions.
Asunto(s)
Arteriosclerosis/metabolismo , Quimiocinas CC , Citocinas/biosíntesis , Músculo Liso Vascular/metabolismo , Receptores de Quimiocina/biosíntesis , Vasculitis/metabolismo , Actinas/metabolismo , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Aorta Torácica/metabolismo , Aorta Torácica/patología , Arteriosclerosis/genética , Arteriosclerosis/patología , Northern Blotting , Arterias Carótidas/metabolismo , Arterias Carótidas/patología , Células Cultivadas , Quimiocina CCL11 , Citocinas/genética , Expresión Génica , Humanos , Inmunohistoquímica , Mastocitos/metabolismo , Mastocitos/patología , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/patología , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Mensajero/metabolismo , Receptores CCR3 , Receptores de Quimiocina/genética , Factor de Necrosis Tumoral alfa/farmacología , Vasculitis/genética , Vasculitis/patologíaRESUMEN
PURPOSE: We sought to determine the effects of a communication process that was designed to encourage the use of advanced supportive technology when it is of benefit, but to limit its burdens when it is ineffective. We compared usual care with a proactive, multidisciplinary method of communicating that prospectively identified for patients and families the criteria that would determine whether a care plan was effective at meeting the goals of the patient. This process allowed caregivers to be informed of patient preferences about continued advanced supportive technology when its continuation would result in a compromised functional outcome or death. MATERIALS AND METHODS: We performed a before-and-after study in 530 adult medical patients who were consecutively admitted to a university tertiary care hospital for intensive care. Multidisciplinary meetings were held within 72 hours of critical care admission. Patients, families, and the critical care team discussed the care plan and the patients' goals and expectations for the outcome of critical care. Clinical "milestones" indicative of recovery were identified with time frames for their occurrence. Follow-up meetings were held to discuss palliative care options when continued advanced supportive technology was not achieving the patient's goals. We measured length of stay, mortality, and provider team and family consensus in 134 patients before the intensive communication intervention and in 396 patients after the intervention. RESULTS: Intensive communication significantly reduced the median length of stay from 4 days (interquartile range, 2 to 11 days) to 3 days (2 to 6 days, P = 0.01 by survival analysis). This reduction remained significant after adjustment for acute physiology and chronic health evaluation (APACHE) 3 score [risk ratio (RR) = 0.81; 95% confidence interval (CI), 0.66 to 0.99; P = 0.04). Subgroup analysis revealed that this reduction occurred in our target group, patients with acuity scores in the highest quartile who died (RR = 0.60; 95% CI, 0.38 to 0.92; P = 0.02). The intervention, which allowed dying patients earlier access to palliative care, was not associated with increased mortality. CONCLUSIONS: Intensive communication was associated with a reduction in critical care use by patients who died. Our multidisciplinary process targeted advanced supportive technology to patients who survived and allowed the earlier withdrawal of advanced supportive technology when it was ineffective.
Asunto(s)
Comunicación , Cuidados Críticos/métodos , Cuidados Críticos/normas , Ciencia del Laboratorio Clínico , Planificación de Atención al Paciente , Pautas de la Práctica en Medicina/normas , APACHE , Anciano , Boston/epidemiología , Manejo de Caso , Femenino , Investigación sobre Servicios de Salud , Mortalidad Hospitalaria , Humanos , Unidades de Cuidados Intensivos/estadística & datos numéricos , Tiempo de Internación , Modelos Logísticos , Masculino , Massachusetts/epidemiología , Persona de Mediana Edad , Oportunidad Relativa , Cuidados Paliativos , Grupo de Atención al Paciente , Pautas de la Práctica en Medicina/estadística & datos numéricos , Modelos de Riesgos Proporcionales , Estudios Prospectivos , Índice de Severidad de la Enfermedad , Análisis de SupervivenciaRESUMEN
We performed an association study of plasma eotaxin levels, eosinophil counts, total IgE levels, asthma diagnosis, and lung function in an ethnically diverse and geographically dispersed population. We studied 515 asthmatic and 519 normal subjects, none of whom was taking inhaled or oral corticosteroids. Logistic regression analysis demonstrated a direct relationship between asthma diagnosis and eotaxin levels (p < 0.0001). The odds of an asthma diagnosis increased with eotaxin quartile, with the highest quartile having an odds ratio of 5.4 (95% CI 3.2 to 9.2, p < 0.001) compared with the lowest eotaxin quartile. Eotaxin levels were inversely related to lung function (p < 0.001), with the mean percent predicted FEV(1) in the highest eotaxin quartile being 13.5 percentage points (SEM 2.1, p < 0.001) less than that in the lowest quartile. Plasma eotaxin levels were associated with asthma and inversely related to lung function independent of age, race, sex, or smoking status. When combined with eosinophil counts and IgE levels, eotaxin levels contributed to the odds of an asthma diagnosis and of impaired lung function. Our results are the first to associate eotaxin levels with asthma diagnosis and compromised lung function in a large geographically and ethnically diverse population.
Asunto(s)
Asma/diagnóstico , Quimiocinas CC , Factores Quimiotácticos Eosinófilos/sangre , Citocinas/sangre , Mecánica Respiratoria , Adulto , Asma/sangre , Asma/inmunología , Asma/fisiopatología , Biomarcadores/sangre , Quimiocina CCL11 , Intervalos de Confianza , Eosinófilos , Femenino , Volumen Espiratorio Forzado , Humanos , Inmunoglobulina E/sangre , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Análisis de RegresiónRESUMEN
BACKGROUND: The eosinophil chemotactic and activating effects of eotaxin and the known association of eosinophils with asthma suggest that eotaxin expression is increased during asthma exacerbations. OBJECTIVE: We sought to determine whether plasma eotaxin levels are elevated in patients presenting for emergency treatment of acute asthma and to correlate eotaxin levels with disease activity and responses to treatment. METHODS: A case-control study of plasma eotaxin levels was performed in the 46 patients who presented for emergency asthma treatment and 133 age-, sex-, and ethnicity-matched subjects with stable asthma. RESULTS: Plasma eotaxin levels were significantly higher in 46 patients with acute asthma symptoms and airflow obstruction (520 pg/mL [250, 1100 pg/mL]; geometric mean [-1 SD, +1 SD]) than in 133 subjects with stable asthma (350 pg/mL [190, 620 pg/mL]; P =.0008). Among the patients with emergency asthma flares, those who responded to asthma treatment with an increase in peak expiratory flow rate by an amount equal to at least 20% of their predicted normal value had lower eotaxin levels than those who did not (410 pg/mL [210, 800 pg/mL] and 660 pg/mL [300, 1480 pg/mL], respectively; P =.04). CONCLUSION: These findings imply that eotaxin either is mechanistically involved in acute asthma or serves as a biomarker for activity of the CCR3 receptor ligand system, which is functionally linked to asthma.
Asunto(s)
Asma/sangre , Quimiocinas CC , Factores Quimiotácticos Eosinófilos/sangre , Citocinas/sangre , Enfermedad Aguda , Adulto , Asma/epidemiología , Asma/terapia , Boston , Estudios de Casos y Controles , Quimiocina CCL11 , Medicina de Emergencia , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pruebas de Función RespiratoriaRESUMEN
Recent family-based studies have revealed evidence for linkage of human chromosome 5q31 to the diagnosis of asthma, elevated serum IgE levels, and bronchial hyperresponsiveness. Among the candidate genes in this region is the gene encoding for human interleukin-4 (IL-4). We reasoned that this gene could also serve as a candidate gene with respect to asthma severity as indicated by the FEV(1) measured when bronchodilator treatment was withheld. To test this hypothesis, we examined a large population of patients with asthma (ascertained without respect to genetic characteristics), for associations between a genetic variant in the IL-4 promoter region (C-589T) and asthma severity, as indicated by FEV(1). We used amplification by the polymerase chain reaction followed by BsmF1 restriction digestion to assign genotypes at the IL-4 promoter C-589T locus. We compared genotypes at this locus in 772 Caucasian and African American patients with asthma of varying severity, and we used multiple regression analysis to relate genotypic findings to FEV(1). Among white individuals, the homozygous presence of the C-589T IL-4 promoter genotype (TT) was associated with a FEV(1) below 50% of predicted (p = 0.013; OR, 1.44; 95% CI: 1.09 to 1.90). Subjects with the TT genotype had mean FEV(1) (% predicted) values 4.5% lower than those of subjects with the wild-type (CC) genotype at this locus. FEV(1) values of white patients with a CC or CT genotype were broadly distributed, whereas the TT genotype was associated with a narrow distribution of low FEV(1) values. The frequency of the T allele was significantly greater (p = 1 x 10(-)(23)) among African American asthmatics (0.544) than among white asthmatics (0.183). These data provide the first evidence associating FEV(1) in patients with asthma and genetic determinants at any locus. Our data are consistent with the idea that the FEV(1) in asthma is the result of multiple factors; one of these factors is the genotype at the IL-4 C-589T locus. This locus is associated with a small but significant decrement in pulmonary function among white asthmatic subjects.
Asunto(s)
Asma/genética , Volumen Espiratorio Forzado , Genes/genética , Interleucina-4/genética , Adulto , Alelos , Asma/sangre , Asma/fisiopatología , Estudios de Cohortes , Interpretación Estadística de Datos , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Inmunoglobulina E/sangre , Masculino , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas/genéticaRESUMEN
Endogenous nitric oxide (NO) is an important modulator of airway function, but its role in the regulation of airway microvascular leak (AMVL) remains unclear. Thus we assessed the effects of NO synthase (NOS) inhibition on expired NO (ENO) levels and on AMVL measured by the Evans blue dye technique in guinea pigs. In control unsensitized animals, systemic NG-nitro-L-arginine methyl ester (L-NAME) reduced ENO by 70 +/- 8% (P < 0.01) and reduced AMVL by 92 +/- 1 and 44 +/- 17% (P < 0.05 for both) in the extrapulmonary and intrapulmonary airways, respectively. In animals sensitized and challenged with intratracheal antigen, markedly increased levels of AMVL and ENO were similarly attenuated by L-NAME. In contrast, aminoguanidine, a relatively selective type II NOS inhibitor, reduced ENO in both antigen-sensitized and control unsensitized animals by 39 +/- 3% (P < 0.01) but had no effect on AMVL. These data indicate that endogenous pulmonary NO contributes to both basal and antigen-stimulated levels of AMVL in guinea pigs and that this NO-dependent activity does not appear to be derived from type II NOS.
Asunto(s)
Presión Sanguínea/efectos de los fármacos , Permeabilidad Capilar/fisiología , Microcirculación/fisiología , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico/fisiología , Circulación Pulmonar/fisiología , Animales , Permeabilidad Capilar/efectos de los fármacos , Guanidinas/administración & dosificación , Guanidinas/farmacología , Cobayas , Homeostasis , Infusiones Intravenosas , Pulmón/enzimología , Masculino , Óxido Nítrico/análisis , Óxido Nítrico Sintasa/antagonistas & inhibidores , Mecánica Respiratoria/fisiologíaRESUMEN
Regulation of eotaxin expression was investigated in U-937 cells, a human monocyte-like cell line. Eotaxin mRNA was induced by tumor necrosis factor-alpha (TNF-alpha; 0.1-100 ng/ml) and phorbol 12-myristate 13-acetate (PMA; 0.01-1 microM). PMA-induced eotaxin mRNA expression was of greater magnitude and was maximal at a later time point than TNF-alpha-induced expression (16 h vs. 2 h after stimulation), which was consistent with eotaxin protein expression detected by immunocytochemistry. Dexamethasone (0.01-10 microM) decreased eotaxin mRNA expression in both TNF-alpha- and PMA-stimulated U-937 cells. PMA-induced eotaxin mRNA expression was inhibited by cycloheximide (10 microg/ml), whereas TNF-alpha-induced expression was not. The protein kinase C (PKC) inhibitor staurosporine (10-50 nM) inhibited PMA-induced eotaxin mRNA expression, whereas TNF-alpha-induced expression was enhanced by this reagent. These results suggest that eotaxin expression can be induced by more than one mechanism: the PMA-triggered pathway is mediated by PKC activation and requires new protein synthesis, whereas the TNF-alpha-triggered pathway is independent of PKC and protein synthesis. TNF-alpha- and PMA-induced pathways are both associated with nuclear factor-kappaB, because its binding activity was enhanced in the presence of these stimuli, and both pathways were limited by its inhibitor, diethyldithiocarbamate.
Asunto(s)
Quimiocinas CC , Citocinas/genética , Células Epiteliales/inmunología , Regulación de la Expresión Génica , Monocitos/inmunología , Factor de Necrosis Tumoral alfa/farmacología , Adenocarcinoma , Línea Celular , Células Cultivadas , Quimiocina CCL11 , Factores Quimiotácticos Eosinófilos/genética , Citocinas/biosíntesis , Células Epiteliales/citología , Células Epiteliales/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Interferón gamma/farmacología , Interleucina-1/farmacología , Interleucina-4/farmacología , Cinética , Lipopolisacáridos/farmacología , Neoplasias Pulmonares , Monocitos/citología , Monocitos/efectos de los fármacos , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Acetato de Tetradecanoilforbol/farmacología , Factores de Tiempo , Transcripción Genética/efectos de los fármacos , Células Tumorales Cultivadas , Factor de Necrosis Tumoral alfa/fisiologíaRESUMEN
Eotaxin is the major eosinophil chemoattractant found in bronchoalveolar lavage (BAL) fluid from sensitized guinea pigs after antigen challenge. In this study we have performed immunostaining for eotaxin in airways obtained from challenged animals and examined purified guinea pig lung cells (epithelial cells > 98% purity, mast cells > 90% purity) for eotaxin mRNA and protein. In the airways of antigen (ovalbumin) challenged animals, significant amounts of epithelial cell eotaxin immunostaining were observed. Northern analysis of total RNA obtained from unchallenged, freshly isolated airway epithelial cells contained high levels of eotaxin mRNA. Semi-pure and high purity lung mast cell preparations (challenged or unchallenged) did not express eotaxin mRNA. Western analysis of supernatant fluids obtained from incubated airway epithelial cells demonstrated detectable amounts of eotaxin protein, with the majority of the protein being cell-associated. Thus, airway epithelial cells are identified as a major cellular source of eotaxin in the guinea pig pulmonary system.