Background: Induced sputum is widely used in assessing airway inflammation. However, its utility as a diagnostic tool in the diagnosis of lung disease in immunosuppressed patients merits further investigation. Objectives: To determinate the diagnostic yield of sputum induction in the diagnosis of lung diseases in HIV-positive patients. Method: Subjects were selected from among HIV-positive patients older than 14 years who were evaluated at a reference hospital between January 2001 and September 2002. Those with respiratory symptoms for 7 days or longer with normal or abnormal chest X-rays, as well as those without respiratory symptoms but with abnormal chest X-rays, were included. All subjects were submitted to clinical examination, radiologic evaluation, sputum induction and laboratory testing. Subsequently, flexible fiberoptic bronchoscopy, bronchoalveolar lavage and transbronchial lung biopsy were performed. Samples were processed for Gram and Ziehl-Neelsen staining, quantitative culture for pyogenic bacteria, direct staining for fungi, culture for mycobacteria and fungi, silver stain for Pneumocystis jiroveci, as well as for total and differential cellularity determination. Results: A total of 54 patients were included. Upon testing negative for any etiologic agent, 7 patients were excluded, resulting in a total of 54 patients studied. A total of 60 infectious agents were isolated. Among the etiologic agents isolated, 46.7% were P. jiroveci, 33.5 were pyogenic bacteria and 16.7% were Mycobacterium tuberculosis. Sputum induction presented 57.5% sensitivity, 42.9% specificity, 87.1% predictive positive value, 13% predictive negative value and 55.6% overall accuracy. Conclusions: In this population, sputum induction proved to be a technique that is safe and easily performed, with a good diagnostic yield.

Keywords: HIV, Acquired Immunodeficiency Syndrome, Bronchoscopy, Sputum, Lung Disease/diagnosi.

Objective: To determine the diagnostic sensitivity of bacteriological analyses in induced sputum (IS) for the diagnosis of pulmonary tuberculosis (TB) and to identify the clinical characteristics associated with the confirmed diagnosis, as well as to determine the diagnostic yield of bronchoscopy carried out when IS tests negative for AFB in smear microscopy. Methods: A retrospective, cross-sectional study of patients suspected of having active pulmonary TB and referred to our clinic for sputum induction. We consecutively reviewed the laboratory data of all patients submitted to sputum induction between June of 2003 and January of 2006, as well as their electronic medical records. In addition, the results of the bacteriological analysis of bronchoscopic specimens collected from the patients whose AFB tests were negative in IS were reviewed. Results: Of the 417 patients included in the study, 83 (19.9%) presented IS samples that tested positive for TB (smear microscopy or culture). In the logistic regression analysis, radiological findings of cavitation (OR = 3.8; 95% CI: 1.9-7.6) and of miliary infiltrate (OR = 3.7; 95% CI: 1.6-8.6) showed the strongest association with the diagnosis of pulmonary TB. In 134 patients, bronchoscopy was carried out after negative AFB results in IS and added 25 (64.1%) confirmed diagnoses of pulmonary TB. Conclusions: In our clinical practice, the frequency of confirmed diagnosis of pulmonary TB using IS (19.9%) was lower than that previously reported in controlled trials. Cavitation and miliary infiltrate increase the diagnostic probability of pulmonary TB using IS. The use of bronchoscopy when IS tests negative for AFB significantly increases sensitivity in the diagnosis of pulmonary TB.

Keywords: Tuberculosis, pulmonary; Diagnosis; Sputum.

Abstract Objective: To evaluate whether respiratory therapy techniques influence the number of cells within and quantity of induced sputum in patients with asthma or chronic obstructive pulmonary disease (COPD). Methods: Randomized clinical trial, in which patients with asthma or COPD under intervention (n = 16 and 10, respectively) were compared with control groups (n = 16 and 10). Patients in the asthma/intervention (A/I) and COPD/intervention (C/I) groups were submitted to oscillating positive expiratory pressure maneuvers for 5 min, followed by 10 forced expiratory technique sequences. These patients were also submitted to an induced sputum protocol with inhaled hypertonic saline (3%, 4% or 5%; A/I group) or inhaled isotonic saline (C/I group). The asthma/control (A/C) and COPD/control (C/C) groups were submitted only to the standard induced sputum protocol. Results: The final mean weight of the sputum samples was significantly greater in the A/I group than in the A/C group (2,767.25 ± 998.08 mg vs. 1,689.17 ± 1,189.96 mg; p = 0.03). The mean/median total cell counts (×106/mL) were higher in the A/I and C/I groups than in the A/C and C/C groups (4.06/0.95 and 0.63/0.39, p = 0.05, vs. 5.08/1.77 and 0.64/0.40, p = 0.02). There were no statistically significant differences among the groups in terms of cell viability. Conclusions: The use of respiratory therapy techniques can increase sputum sample weight in asthma patients, as well as increasing total cell counts in patients with asthma or COPD.

Keywords: Asthma; Pulmonary disease, obstructive chronic; Sputum; Physical therapy modalities.

Objective: To evaluate the diagnostic accuracy of bronchoscopy in patients with clinical or radiological suspicion of tuberculosis who were unable to produce sputum or with negative sputum smear microscopy results. Methods: A prospective cross-sectional study involving 286 patients under clinical or radiological suspicion of having pulmonary tuberculosis and submitted to bronchoscopy-BAL and transbronchial biopsy (TBB). The BAL specimens were submitted to direct testing and culture for AFB and fungi, whereas the TBB specimens were submitted to histopathological examination. Results: Of the 286 patients studied, 225 (79%) were diagnosed on the basis of bronchoscopic findings, as follows: pulmonary tuberculosis, in 127 (44%); nonspecific chronic inflammation, in 51 (18%); pneumocystis, fungal infections, or nocardiosis, in 20 (7%); bronchiolitis obliterans organizing pneumonia, alveolites, or pneumoconiosis, in 14 (5%); lung or metastatic neoplasms, in 7 (2%); and nontuberculous mycobacterium infections, in 6 (2%). For the diagnosis of tuberculosis, BAL showed a sensitivity and a specificity of 60% and 100%, respectively. Adding the TBB findings significantly increased this sensitivity (to 84%), as did adding the post-bronchoscopy sputum smear microscopy results (total sensitivity, 94%). Minor post-procedure complications occurred in 5.6% of the cases. Conclusions: Bronchoscopy is a reliable method for the diagnosis of pulmonary tuberculosis, with low complication rates. The combination of TBB and BAL increases the sensitivity of the method and facilitates the differential diagnosis with other diseases.

Keywords: Bronchoscopy; Tuberculosis, pulmonary; Sputum; Bronchoalveolar lavage; Biopsy.

Objective: To establish reference values for cellularity in induced sputum samples collected from healthy adults. Methods: Induced sputum samples were obtained from 88 healthy adult never-smokers (39 males). The mean age was 36 years (range, 18-68 years). The participants had been residing in the city of Florianópolis, Brazil (a medium-sized non-industrial city) for at least two years. After the samples had been processed, we obtained total and differential cell counts. Results: The mean total cell count was 4.8 ± 4.2 × 106 cells/g. There was a predominance of macrophages (mean, 77.5 ± 14.7%) and neutrophils (mean, 23.4 ± 14.3%). Eosinophils were virtually absent (mean, 0.1 ± 0.3%). Lymphocytes and bronchial epithelial cells were scarce. Neither age nor atopy had any effect on the total or differential cell counts. Conclusions: In the induced sputum of this healthy adult population, macrophages and neutrophils predominated. However, the proportion of neutrophils was lower than that reported in previous studies, which suggests that reference values might vary depending on geographic location.

Keywords: Sputum; Reference values; Brazil.

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 3% eosinophils, and neutrophilic if there were ³ 65% neutrophils. Results: Results are expressed by median and interquartile range. The total cell count was 3.4 (3.7) x 106 cells/ml, and cell viability was 80.0 (16.4) %. The proportion of neutrophils was 14.4 (22.1) %, of eosinophils 6.4 (17.2) %, of macrophages 60.3 (37.5) %, and of lymphocytes 1.1 (1.2) %. Eosinophilic sputum was observed in 24 subjects (70.6%); 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.

Keywords: Asthma. Inflammation. Sputum. Eosinophils. Neutrophils.

Objective: To evaluate the usefulness of determining the inflammatory component of airway diseases (inflammometry) by induced sputum cell counts, as well as its influence on treatment decisions in a tertiary facility for the treatment of respiratory diseases. Methods: We analyzed 151 sputum samples from 132 consecutive patients referred for clinical sputum induction by five pulmonologists between July of 2006 and February of 2007. A structured questionnaire related to the reasons for requesting the test and to the therapeutic decision making based on test results was completed by each attending physician upon receiving the test results. Induced sputum was obtained and processed according to a technique previously described. Results: The principal motives for ordering the test were inhaled corticosteroid dose titration in patients with moderate-to-severe asthma (in 54.3%), investigation of chronic cough (in 30.5%), and monitoring airway inflammation in patients with bronchiectasis (in 7.3%) or chronic obstructive pulmonary disease (in 6%). Of the 82 patients with asthma, 47 (57%) presented eosinophilic bronchitis (>3% eosinophils). Nonasthmatic eosinophilic bronchitis was diagnosed in 9 (19%) of the 46 patients with chronic cough. Neutrophilic bronchitis (>65% neutrophils) was found in 13 patients, of which 5 had asthma, 2 had chronic cough, and 6 had chronic obstructive pulmonary disease/bronchiectasis. Based on the induced sputum results, the corticosteroid dose was modified in 48 asthma patients (64.7%). Conclusions: The systematic application of inflammometry using induced sputum cell counts can be beneficial for patients with airway diseases, particularly those with asthma or chronic cough.

Keywords: Sputum; Asthma; Bronchitis.

Objective: To determine cell profiles, as well as to identify CD4+ and CD8+ lymphocyte subgroups, in induced sputum (IS) and peripheral venous blood (PVB) of patients with chronic obstructive pulmonary disease (COPD). Methods: Total cell counts and counts of individual cell types, including CD4+ and CD8+ T lymphocytes, were determined in the IS and PVB of 85 subjects (38 with COPD without exacerbation, 29 smokers without obstruction and 18 nonsmokers). Mann-Whitney and Spearman non-parametric tests were used in the statistical analysis, and values of p < 0.05 were considered statistically significant. Results: Comparing the IS of subjects with COPD to that of nonsmokers, neutrophil, eosinophil and CD8+ T lymphocyte counts were higher (respectively p = 0.005, p < 0.05 and p < 0.05), whereas the percentage of macrophages was lower (p = 0.003). There were weak linear correlations (r2 < 0.1) between each cell type in IS and forced expiratory volume in one second (FEV1), forced vital capacity (FVC) and FEV1/FVC ratio. Eosinophil and CD8+ T lymphocyte counts were also higher in PVB (p = 0.04 and p = 0.02). Conclusions: In patients with stable COPD, CD8+ T lymphocyte counts were higher in PVB, whereas total leukocyte counts were similar to those of the other two groups analyzed, suggesting systemic inflammatory involvement. The CD8+ T lymphocyte count in blood can be a useful marker of systemic inflammation and can help identify smokers who already present a COPD inflammatory pattern.

Keywords: Sputum; T-lymphocytes; Neutrophils; Macrophages; Eosinophils; Pulmonary disease, chronic obstructive.

Objective: To propose a methodology for characterizing proficiency in sputum smear microscopy for acid-fast bacilli (AFB) in the diagnosis of tuberculosis and to determine the number of microscopies necessary to establish this proficiency, as well as the quality of the transcription of results, the causes of the discrepancies in the readings (rater or microscope used), and the criterion for classification of microscopy results that poses the most difficulty in characterizing proficiency. Methods: Four hundred sputum smear microscopies for the diagnosis of tuberculosis were analyzed through double-blind readings by six professionals who usually read/supervise microscopies performed in public health care facilities. The sample was stratified to obtain, at least, a reliability of 90% in the double-blind readings, an α error of 5%, and a precision of 3%. The results were analyzed using observed reliability and the kappa index. Results: Thirteen errors (0.27%) were found in the transcription of results. Reliability increased when the three distinct categories of positive results (AFB+, AFB++, and AFB+++) were grouped or when inconclusive results were excluded from the analysis. The quantification of the bacterial load was the classification criterion that posed the most difficulty in establishing proficiency. Using higher quality microscopes increased reliability. Reliability values stabilized only from the reading of 75 microscopies onward. Conclusions: Double-blind sputum smear microscopy readings using a panel containing 75 slides (36 negative, 4 inconclusive, and 35 positive) proved to be appropriate for characterizing proficiency in sputum smear microscopy for the diagnosis of tuberculosis when such proficiency is intended to reproduce laboratory routine.

Keywords: Tuberculosis; Microscopy; Reproducibility of results.

A comparative study to evaluate contamination in cultures of morning sputum samples, comparing those collected at home under currently recommended conditions and those collected under supervision after patient orientation and education. The home and supervised collection groups produced 43 and 76 sputum samples, respectively. The contamination rate was nearly 3-times higher among samples collected at home than among those collected under supervision (37% vs. 13%, p < 0.05; OR = 0.25). The simple educational and hygiene measures described can decrease the contamination rate among sputum samples collected for diagnostic culture.

Keywords: Tuberculosis/diagnosis; Sputum/microbiology; Culture techniques.

Objective: To evaluate the role of video-assisted thoracoscopy in the diagnosis of diffuse parenchymal lung diseases. Methods: The medical charts of patients suspected of having diffuse parenchymal lung disease were retrospectively reviewed, as were the results of the anatomopathological examination of lung biopsy specimens collected through video-assisted thoracoscopy. Results: Of the 48 patients included in the study, 25 (52.08%) were female and 23 (47.92%) were male. The mean age was 58.77 years (range, 20-76 years). A total of 54 biopsy fragments were submitted to anatomopathological examination: 24 (44.44%) from the lingula; 10 (18.52%) from the left lower lobe; 7 (12.96%) from the right middle lobe; 6 (11.11%) from the right lower lobe; 5 (9.26%) from the left upper lobe; and 2 (3.71%) from the right upper lobe. The mean duration of thoracic drainage was 2.2 days. Adverse events included conversion to thoracotomy, in 2 patients (4.17%), and residual pneumothorax, in 1 (2.08%). The definitive diagnosis was made in 46 patients (95.83%), and idiopathic interstitial pneumonia was the predominant diagnosis (in 54.18%). The most common diagnoses were usual interstitial pneumonia (in 29.27%), nonspecific interstitial pneumonia (in 16.67%) and hypersensitivity pneumonia (in 12.50%). Conclusions: Lung biopsy through video-assisted thoracoscopy is a safe, effective and viable procedure for the diagnosis of diffuse parenchymal lung diseases.

Keywords: Lung diseases, interstitial; Thoracoscopy; Diagnosis.

Background: Tuberculosis continues to be a global health problem. Objective: To evaluate an automated system designed to diagnose tuberculosis, comparing it to sputum microscopy and culture in Löwenstein-Jensen medium. Method: A comparative study using 844 sputum samples, collected between September and December of 1999 at a reference center for tuberculosis in São Paulo, Brazil, to draw distinctions between the results obtained through the use of the automated system and those obtained through sputum microscopy and culture in Löwenstein-Jensen medium. Results: Of the 844 samples evaluated, 27.1% tested positive for acid-fast bacilli, and 72.9% tested negative. In Löwenstein-Jensen culture, 34.7% were positive and 63% were negative, compared with 37.1% positivity and 56.9% negativity using the automated system. Sensitivity was 98.1% for the automated system and 91.9% for Löwenstein-Jensen culture. Specificity and positive predictive value were 100% for both methods. Negative predictive value was 98.9% for the automated system and 95.5% for Löwenstein-Jensen culture. The degree of accuracy was 99.3% for the automated system and 97% for Löwenstein-Jensen culture, and the Kappa was 0.99 for the automated system and 0.94 for Löwenstein-Jensen culture. The difference between the mean time to detection of mycobacteria using the automated system (10.5 days) and that found using Löwenstein-Jensen culture (34.7 days) was statistically significant. Conclusion: The difference between the culture yield obtained using the automated system and that achieved with Löwenstein-Jensen culture was statistically significant. Mean time to detection of mycobacteria was significantly shorter with the automated system. The higher yield provided by this new system justifies its use in a reference center for tuberculosis in São Paulo

Keywords: Mycobacterium tuberculosis. Diagnostic. Automated systems.

Drug-induced lung disease (DILD) encompasses a broad, highly heterogeneous group of conditions that may occur as a result of exposure to numerous agents, such as antineoplastic drugs, conventional or biological disease-modifying antirheumatic drugs, antiarrhythmics, and antibiotics. Between 3% and 5% of prevalent cases of interstitial lung diseases are reported as DILDs. The pathogenesis of lung injury in DILD is variable, multifactorial, and often unknown. Acute presentation is the most common, can occur from days to months after the start of treatment, and ranges from asymptomatic to acute respiratory failure. The CT patterns are varied and include ground-glass opacities, organizing pneumonia, and diffuse alveolar damage. Notably, there are no clinical manifestations or CT patterns specific to DILD, which makes the diagnosis quite challenging and necessitates a high index of suspicion, as well as the exclusion of alternative causes such as infection, cardiac-related pulmonary edema, exacerbation of a preexisting ILD, and neoplastic lung involvement. Discontinuation of the offending medication constitutes the cornerstone of treatment, and corticosteroid treatment is usually necessary after the onset of clinical manifestations. The prognosis varies widely, with high mortality rates in severe cases. A history of medications related to pulmonary toxicity in patients with new-onset respiratory symptoms should prompt consideration of DILD as a potential underlying cause.

Keywords: Lung diseases, interstitial/chemically induced; Lung diseases, interstitial/diagnostic imaging; Immunotherapy/adverse effects.

Objective: To study the clinical, radiological, and histopathological patterns of transbronchial biopsy (TBB) used in order to confirm the diagnosis in patients with clinical suspicion of interstitial lung disease (ILD) treated at a tertiary-care university hospital. Methods: We reviewed the medical records, radiology reports, and reports of transbronchial biopsies from all patients with suspected ILD who underwent TBB between January of 1999 and December of 2006 at the Hospital das Clínicas de Botucatu, located in the city of Botucatu, Brazil. Results: The study included 56 patients. Of those, 11 (19.6%) had a definitive diagnosis of idiopathic pulmonary fibrosis (IPF), the rate of which was significantly higher in the patients in which ILD was a possible diagnosis in comparison with those in which ILD was the prime suspect (p = 0.011), demonstrating the contribution of TBB to the diagnostic confirmation of these diseases. The histopathological examination of the biopsies revealed that 27.3% of the patients with IPF showed a pattern of organizing pneumonia, which suggests greater disease severity. The most common histological pattern was the indeterminate pattern, reflecting the peripheral characteristic of IPF. However, the fibrosis pattern showed high specificity and high negative predictive value. For CT scan patterns suggestive of IPF, the ROC curve showed that the best relationship between sensitivity and specificity occurred when five radiological alterations were present. Honeycombing was found to be strongly suggestive of IPF (p = 0.01). Conclusions: For ILDs, chest CT should always be performed, and TBB should be used in specific situations, according to the suspicion and distribution of lesions.

Keywords: Lung diseases, interstitial; Diagnosis, differential; Bronchoscopy.

The role of immunohistochemistry is to recognize antigens and, consequently, to identify and classify specific cells within a cell population whose morphology is heterogenous or apparently homogenous. The visualization of the antigen-antibody complex is made possible through the addition of either a fluorochrome conjugate or an enzyme to the antibody, which is then viewed under microscopy. Immunohistochemistry can be used in the routine diagnosis of lung cancer, in order to identify biological markers (diagnostic and prognostic). The essential immunohistochemistry panels will be discussed in this review.

Keywords: Immunohistochemistry; Tumor markers, biological; Lung neoplasms.