Repeatability of peak nasal inspiratory flow measurements and utility for assessing the severity of rhinitis.

BACKGROUND: The measurement of peak nasal inspiratory flow (PNIF) provides a simple, cheap, fast and readily available tool for determining the extent of nasal airway patency. However, there are questions regarding its repeatability when used to assess the degree of nasal obstruction in large populations. Therefore, this study aimed to evaluate the repeatability of PNIF measurements and to assess their association with the signs and symptoms of rhinitis. METHODS: The PNIF, rhinitis symptoms, judged by Meltzer questionnaire and rhinitis signs, as determined by anterior rhinoscopy, were assessed in 283 adults representative of the general population. One training and two test PNIF measurements were recorded during the same session. RESULTS: The PNIF was highly reproducible (ICC = 0.92; 95% limits of agreement: +/-36 l/min). The PNIF was strongly correlated with rhinitis signs, measured by anterior rhinoscopy (rs= -0.38, P < 0.0001) but was not correlated with rhinitis symptoms, measured by questionnaire (rs= -0.11, P = 0.057). Differences in PNIF for subjects categorized as asymptomatic, mild or moderate/severe on the basis of rhinitis signs, were highly significant (P < 0.0001), but less significant on the basis of rhinitis symptoms (P = 0.04). A PNIF cut-off of 115 l/min had moderately high specificity (72%) and sensitivity (65%) and a high negative predictive value (90%) for moderate/severe signs of rhinitis. CONCLUSION: In a large general population-based sample of young adults, PNIF was highly reproducible and closely related to the signs of rhinitis, as determined by clinical examination. The PNIF provides information that is qualitatively different to that provided by symptom scores and may be useful to measure the extent of nasal obstruction.

Eosinophilia, interleukin-5, and tumour necrosis factor-alpha in asthmatic children.

BACKGROUND: There are few paediatric studies of the interrelationships between inflammatory markers and asthma severity. We therefore assessed the relationships between eosinophil-associated markers, cytokines, and asthma severity in asthmatic children aged 8-12 years. METHODS: Forty-five children were tested twice, 2 weeks apart. Asthma severity was measured in terms of symptoms, lung function, medication needs, and histamine responsiveness. Peripheral inflammatory markers measured included eosinophil numbers, serum ECP, IL-5, and TNF-alpha and mononuclear cell IL-5, and TNF-alpha production. RESULTS: Histamine responsiveness was correlated with circulating eosinophils (r = 0.56, P = 0.0001) and serum ECP (r = 0.54, P = 0.003). Eosinophilia was increased in children with severe as opposed to mild airway hyperresponsiveness (P = 0.02) and those who lost days at school as opposed to those who did not (P = 0.01). There were no other associations between markers of asthma severity and inflammation. Children taking inhaled corticosteroids had lower serum IL-5 levels than those on beta-agonists +/- cromolyn (mean and 95% CI: 20.5 [11.7-35.7] pg/ml vs 64.3 [26.6-155.4] pg/ml; P = 0.04). Cellular IL-5 production correlated with serum TNF-alpha (r = 0.63, P = 0.0062) and IL-5 (r = -0.59, P = 0.005). CONCLUSION: Serum levels of TNF-alpha and IL-5 were not related to peripheral eosinophilia and asthma severity in these children but were related to their own cellular production ex vivo. This study confirms that eosinophilia is the index of inflammation that is most closely related to the clinical severity of childhood asthma.

Demonstration of a Na(+)/H(+) exchanger NHE1 in fresh bovine corneal endothelial cell basolateral plasma membrane.

Apical and basolateral plasma membranes of fresh bovine corneal endothelial cells were isolated using positively charged polyacrylamide beads. Marker enzyme assays demonstrated that the isolated apical and basolateral plasma membrane domains could be isolated and separated with relative purity. Western blotting with a polyclonal anti-NHE1 antibody detected a protein of 70 kDa in the basolateral plasma membrane isolate. NHE1 immunoreactivity was not detected in the apical membrane sample. This suggests that the Na(+)/H(+) exchanger, NHE1, is strictly localised to the basolateral membrane of fresh bovine corneal endothelial cells.

Effect of regular inhaled salbutamol on airway responsiveness and airway inflammation in rhinitic non-asthmatic subjects.

BACKGROUND: Regular, inhaled beta 2 agonists may increase airway responsiveness in asthmatic subjects. The mechanism is not known but may be via an increase in airway inflammation. A study was undertaken to examine the effect of regular inhaled salbutamol on airway responsiveness to methacholine and hypertonic saline, on the maximal response plateau to methacholine, and on inflammatory cells in induced sputum in rhinitic non-asthmatic subjects. METHODS: Thirty subjects with a baseline maximal response plateau of > 15% fall in forced expiratory volume in one second (FEV1) entered a randomised, placebo controlled, parallel trial consisting of two weeks run in, four weeks of treatment, and two weeks washout. Methacholine challenges were performed at the beginning of the run in period, before treatment, after treatment, and after washout. Hypertonic saline challenges were performed before and after treatment and induced sputum samples were collected for differential cell counting. RESULTS: There was no change in airway responsiveness, maximal response plateau to methacholine, or in induced sputum eosinophils or mast cells. The maximum fall in FEV1 after hypertonic saline increased in the salbutamol group (median change 6.0%, interquartile range (IQR) 11.0) but did not change in the placebo group (median change 1.3%, IQR 5.5). CONCLUSIONS: Regular inhaled salbutamol for four weeks increases airway responsiveness to hypertonic saline but does not alter airway responsiveness to methacholine or cells in induced sputum in non-asthmatic individuals with rhinitis. The relevance of these findings to asthmatic subjects has not been established.

Monoclonal antibodies specific for guinea pig eosinophil major basic protein: their use in an ELISA, immunocytochemistry and flow cytometry.

Eosinophil major basic protein (MBP), purified from guinea pig eosinophil granules was used to raise five monoclonal antibodies (MoAbs). Their reactivity with MBP was confirmed by immunoblotting and indirect ELISA. Two of the MoAbs were used to develop a sensitive and specific antigen capture (sandwich) ELISA for guinea pig eosinophil MBP which gives an accurate and reproducible standard curve over the range of 10-10,000 ng/ml. The specificity of the ELISA for MBP was confirmed and its suitability for testing biological samples ascertained by measurement of MBP in bronchoalveolar lavage fluid (BALF) and plasma from guinea pigs sensitized and challenged with ovalbumin. The ELISA was also capable of detecting MBP in culture supernatants from purified eosinophil preparations challenged with calcium ionophore in vitro. One of the monoclonals could be used to strongly and specifically stain guinea pig eosinophils in immunocytochemistry, whilst all five could be used to visualize eosinophils in suspension in BALF or peritoneal lavage fluid by flow cytometry. There was no staining of other guinea pig leucocyte types, nor crossreactivity with human eosinophils by immunocytochemistry or flow cytometry.

Density profile of bronchoalveolar lavage eosinophils in the guinea pig model of allergen-induced late-phase allergic responses.

Inhalation of aerosolized ovalbumin by guinea pigs both during sensitization and upon challenge induces a pulmonary eosinophilia as assessed by cells recovered in bronchoalveolar lavage fluid (BALF). In comparison with BALF eosinophil numbers in naive animals of 0.82 +/- 0.2 x 10(6) cells, those in sensitized animals before challenge and 17 and 72 h after challenge were 1.48 +/- 0.2 x 10(6), 2.60 +/- 0.6 x 10(6), and 4.2 +/- 0.7 x 10(6) cells, respectively. BALF eosinophils from all these groups were notable for their heterogeneity with respect to density, size, and appearance under the electron microscope. In comparison with peritoneal eosinophils, which had a single mean density peak of 1.088 +/- 0.001 g/ml, BALF cells comprised hypodense (less than 1.080 g/ml), normodense (1.080 to 1.096 g/ml), and hyperdense (greater than 1.096 g/ml) eosinophils. The percentage of hypodense eosinophils rose from 25% in naive animals to 63% in sensitized animals (P less than 0.001) and fell after challenge. In contrast, challenge induced the appearance of hyperdense eosinophils, which rose from 6% in sensitized animals to 42% 72 h after challenge (P less than 0.001). Blood eosinophils in naive animals showed a similar profile to those in the lung, but after sensitization and challenge no gross changes in the proportion of either hypodense or hyperdense eosinophils were observed. Flow cytometric analysis of BALF eosinophils indicated that hypodense eosinophils, with a mean diameter of 15.8 microns, were larger than both normodense and hyperdense eosinophils, which had mean diameters of 14.3 and 11.6 microns, respectively. Although the numbers and size of granules were not reduced in hypodense BAL eosinophils, electron microscopy morphology indicated a reduced granular content.(ABSTRACT TRUNCATED AT 250 WORDS)

Studies of cellular mechanisms for the generation of superoxide by guinea-pig eosinophils and its dissociation from granule peroxidase release.

Guniea-pig peritoneal eosinophils generated superoxide anions in response to opsonized zymosan, platelet activating factor, sodium fluoride, digitonin, phorbol ester and calcium ionophore, but were refractory to fMLP. These agonists did not stimulate release of eosinophil peroxidase. The phospholipase inhibitor, mepacrine, and the protein kinase inhibitor, trifluoperazine, were effective inhibitors of superoxide production. Activators of protein kinase C, such as exogenously added phorbol ester and endogenously derived diacylglycerol, stimulate superoxide production, which is therefore proposed to be via pathways dependent on phospholipase and protein kinase activity.