Methods of sputum processing for cell counts, immunocytochemistry and in situ hybridisation.

Since the first attempts to use standardised methods for sampling induced airways sputum, two methods for processing the expectorate have evolved. The first involves selecting all viscid or denser portions from the expectorated sample with the aid of an inverted microscope. This method has been extensively evaluated and reported in detail. The second approach involves processing the entire expectorate, comprising sputum plus variable amounts of saliva. Recent modifications to this method include collecting saliva and sputum separately in order to reduce salivary contamination. Both methods have advantages and disadvantages. The advantages of using selected sputum are: squamous cell contamination is v5%, making cell counting easier and quicker to perform, the total cell count (TCC) can be expressed per gram of lower airway secretions, and concentrations of chemicals in the fluid phase are unaffected by the confounding influence of saliva, and can be accurately corrected for dilution. The disadvantage is that selection takes a few minutes longer to perform and requires an inverted microscope. The advantage of using the entire expectorate is that the technique is quicker to perform, but there are some disadvantages that require consideration. The expectorate contains a variable mixture of sputum plus saliva which maydilute the sputum and confound its analysis. The reproducibility of cell counts has been reported to be lower if squamous cell contamination represents w20% of all recovered cells. There is conflicting data as to whether or not differential cell counts (DCCs) differ between the two methods. One study reported a higher percentage of eosinophils in sputum processed by the selection method compared to the entire expectorate but this has not been confirmed in other studies. Although, both the selected sputum and the entire expectorate methods have the same ability to distinguish asthmatics or bronchitics from healthy subjects, they are not interchangeable, and, once a technique has been adopted for a given study, it should always be applied

Broad band X-ray spectral properties of Gamma-ray bursts with BeppoSAX

In about one year, five gamma-ray bursts were simultaneously observed with the Wide Field Cameras and Gamma Ray Burst Monitor aboard the BeppoSAX satellite. From some of them X-ray afterglow emission has been clearly detected with the same satellite. In order to understand how GRB emission is related to the X-ray afterglow, we are performing a systematic study of the spectral properties of these events. We report here preliminary results of this study.Comment: 5 pages, 3 figures. To appear in the Proceedings of the 4th Huntsville Gamma-ray Burst Symposiu

Comparing the effects of two inhaled glucocorticoids on allergen-induced bronchoconstriction and markers of systemic effects, a randomised cross-over double-blind study

<p>Abstract</p> <p>Background</p> <p>Inhaled glucocorticoids are efficient in protecting against asthma exacerbations, but methods to compare their efficacy vs systemic effects have only been attempted in larger multi-centre studies. The aim of the current study was therefore to directly compare the effects of two separate inhaled glucocorticoids, mometasone and budesonide, to compare the effects on the early and late asthmatic responses to inhaled allergen in patients with mild allergic asthma, and sputum eosinophils, and to relate the clinical positive effects to any systemic effects observed.</p> <p>Methods</p> <p>Twelve patients with documented early and late asthmatic responses (EAR and LAR) to inhaled allergen at a screening visit were randomized in a double-blind fashion to treatment with mometasone (200 μg × 2 or 400 μg × 2), budesonide (400 μg × 2) or placebo in a double-blind crossover fashion for a period of seven days. Challenge with the total allergen dose causing both an EAR and LAR was given on the last day of treatment taken in the morning. Lung function was assessed using FEV1, and systemic glucocorticoid activity was quantified using 24 h urinary cortisol.</p> <p>Results</p> <p>Mometasone and budesonide attenuate both EAR and LAR to allergen to a similar degree. No significant dose-related effects on the lung function parameters were observed. Both treatments reduced the relative amount of sputum eosinophils (%) after allergen. At the dose of 800 μg daily, mometasone reduced 24 h urinary cortisol by approximately 35%. Both drugs were well tolerated.</p> <p>Conclusions</p> <p>Mometasone and budesonide are equieffective in reducing early and late asthmatic responses induced by inhaled allergen challenge. Mometasone 800 μg given for seven days partially affects the HPA axis.</p