Reliability of a rapid hematology stain for sputum cytology.

OBJECTIVE: To determine the reliability of a rapid hematology stain for the cytological analysis of induced sputum samples. METHODS: This was a cross-sectional study comparing the standard technique (May-Grünwald-Giemsa stain) with a rapid hematology stain (Diff-Quik). Of the 50 subjects included in the study, 21 had asthma, 19 had COPD, and 10 were healthy (controls). From the induced sputum samples collected, we prepared four slides: two were stained with May-Grünwald-Giemsa, and two were stained with Diff-Quik. The slides were read independently by two trained researchers blinded to the identification of the slides. The reliability for cell counting using the two techniques was evaluated by determining the intraclass correlation coefficients (ICCs) for intraobserver and interobserver agreement. Agreement in the identification of neutrophilic and eosinophilic sputum between the observers and between the stains was evaluated with kappa statistics. RESULTS: In our comparison of the two staining techniques, the ICCs indicated almost perfect interobserver agreement for neutrophil, eosinophil, and macrophage counts (ICC: 0.98-1.00), as well as substantial agreement for lymphocyte counts (ICC: 0.76-0.83). Intraobserver agreement was almost perfect for neutrophil, eosinophil, and macrophage counts (ICC: 0.96-0.99), whereas it was moderate to substantial for lymphocyte counts (ICC = 0.65 and 0.75 for the two observers, respectively). Interobserver agreement for the identification of eosinophilic and neutrophilic sputum using the two techniques ranged from substantial to almost perfect (kappa range: 0.91-1.00). CONCLUSIONS: The use of Diff-Quik can be considered a reliable alternative for the processing of sputum samples.

Reliability of a rapid hematology stain for sputum cytology / Confiabilidade da coloração hematológica rápida para citologia de escarro

Objective: To determine the reliability of a rapid hematology stain for the cytological analysis of induced sputum samples. Methods: This was a cross-sectional study comparing the standard technique (May-Grünwald-Giemsa stain) with a rapid hematology stain (Diff-Quik). Of the 50 subjects included in the study, 21 had asthma, 19 had COPD, and 10 were healthy (controls). From the induced sputum samples collected, we prepared four slides: two were stained with May-Grünwald-Giemsa, and two were stained with Diff-Quik. The slides were read independently by two trained researchers blinded to the identification of the slides. The reliability for cell counting using the two techniques was evaluated by determining the intraclass correlation coefficients (ICCs) for intraobserver and interobserver agreement. Agreement in the identification of neutrophilic and eosinophilic sputum between the observers and between the stains was evaluated with kappa statistics. Results: In our comparison of the two staining techniques, the ICCs indicated almost perfect interobserver agreement for neutrophil, eosinophil, and macrophage counts (ICC: 0.98-1.00), as well as substantial agreement for lymphocyte counts (ICC: 0.76-0.83). Intraobserver agreement was almost perfect for neutrophil, eosinophil, and macrophage counts (ICC: 0.96-0.99), whereas it was moderate to substantial for lymphocyte counts (ICC = 0.65 and 0.75 for the two observers, respectively). Interobserver agreement for the identification of eosinophilic and neutrophilic sputum using the two techniques ranged from substantial to almost perfect (kappa range: 0.91-1.00). Conclusions: The use of Diff-Quik can be considered a reliable alternative for the processing of sputum samples. .

Objetivo: Determinar a confiabilidade da coloração hematológica rápida para a análise do escarro induzido. Métodos: Estudo transversal comparando a técnica padrão (coloração May-Grünwald-Giemsa) com a coloração hematológica rápida (panótico rápido). Participaram do estudo 50 indivíduos (21 asmáticos, 19 portadores de DPOC e 10 controles). Após a coleta do escarro induzido, foram preparadas 4 lâminas, sendo 2 coradas por May-Grünwald-Giemsa e 2 por panótico rápido. As lâminas foram lidas de forma independente por dois pesquisadores capacitados para o exame de escarro induzido e cegados para a identificação das lâminas. A confiabilidade para as contagens celulares dos dois métodos foi avaliada pela determinação dos coeficientes de correlação intraclasse (CCI) para as concordâncias intraobservador e interobservador. As concordâncias na identificação de escarro neutrofílico e eosinofílico entre observadores e entre as duas colorações foram calculadas por estatística kappa. Resultados: Nas duas colorações, os CCI apontaram concordância interobservador quase perfeita para as contagens de neutrófilos, eosinófilos e macrófagos (variação do CCI: 0,98-1,00) e substancial para as contagens de linfócitos (variação do CCI: 0,76-0,83). Na análise intraobservador, a concordância foi quase perfeita para as contagens de neutrófilos, eosinófilos e macrófagos (variação do CCI: 0,96-0,99) e de moderada a substancial para as contagens de linfócitos (CCI = 0,65 e 0,75 para observadores 1 e 2, respectivamente). A concordância interobservador na identificação de escarro eosinofílico e neutrofílico para os dois métodos de ...

ABCC transporter inhibition reduces zymosan-induced peritonitis.

Inflammatory mediators are released from injured tissues being responsible for the first steps of inflammatory processes. Multidrug efflux transporters, members of the ATP-binding cassette (ABC) family, are ubiquitously expressed. ABCC molecules transport several endogenous substances, including leukotriene C4 (LTC4) and PGE2, which are involved in zymosan-induced inflammation. The present study investigated the role played by ABCC transporters on zymosan-induced peritonitis in mice. Most of the resident peritoneal cells were macrophages, based on their morphology and membrane-activated complex 3 expression. RT-PCR demonstrated that these cells expressed ABCC, and ABCC activity was analyzed in vivo via the s.c. injection of ABCC inhibitors [probenecid (PROB) 200 mg/kg or MK571 20 mg/kg], followed by an i.v. injection of carboxyfluorescein diacetate (CFDA), an ABCC fluorescent substrate. Both inhibitors increased CFDA accumulation, suggesting ABCC impairment. Moreover, ABCC reversors decreased zymosan-induced plasma exudation by 86.6 +/- 7.4 and 97.6 +/- 2.3%, a feature related to a diminished secretion of LTC(4) (65.1+/-11 and 47.8+/-9.9%) and PGE(2) (under basal levels). Cell migration was inhibited similarly. Furthermore, PROB and MK571 inhibited IL-1ss by 83.4 +/- 13 and 71.2 +/- 13.4% and TNF-alpha content by 47 +/- 4.5 and 28.9 +/- 0.8%, respectively. NO metabolites and reactive oxygen species production were also reduced. The present results suggest that ABCC molecules have a relevant role in the acute inflammatory response produced by zymosan in mice.

Airway inflammation in steroid-naïve asthmatics: characteristics of induced sputum

BACKGROUND: Airway inflammation, acknowledged as an important feature of asthma, can be assessed by the examination of induced sputum. OBJECTIVE: To determine the pattern of inflammatory cells in induced sputum from stable steroid-naïve asthmatics, in Florianópolis, Santa Catarina. METHOD: The induced sputum from 34 asthmatics using exclusively inhaled bronchodilators on demand was examined. The patients' clinical characteristics were obtained at visit 1, and sputum was induced at visit 2. Differential cell count was performed on Giemsa-stained cytospins. Sputum was considered to be eosinophilic if there were 3 percent eosinophils, and neutrophilic if there were 65 percent neutrophils. RESULTS: Results are expressed by median and interquartile range. The total cell count was 3.4 (3.7) x 10(6) cells/ml, and cell viability was 80.0 (16.4) percent. The proportion of neutrophils was 14.4 (22.1) percent, of eosinophils 6.4 (17.2) percent, of macrophages 60.3 (37.5) percent, and of lymphocytes 1.1 (1.2) percent. Eosinophilic sputum was observed in 24 subjects (70.6 percent); none of them had neutrophilic sputum. There were no significant differences between the eosinophilic and non-eosinophilic groups concerning the measured clinical outcomes, total cell count and proportions of cells in the sputum, except for the proportion of eosinophils (14.4 [19.3] vs 0.4 [1.1], p < 0.001). CONCLUSIONS: In our environment, steroid-naïve asthmatics present a higher proportion of sputum eosinophils, as compared to the established reference values. The clinical and physiological parameters analyzed were unable to predict the presence of eosinophilic inflammation of the airways.