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PubMed1996

Measurement of inflammatory indices in induced sputum: effects of selection of sputum to minimize salivary contamination.

Sputum examination is being used increasingly as a noninvasive method to assess airway inflammation. Expectorated sputum has variable contamination with saliva. Methods of processing have included the selection of portions of the sample considered to be representative of pulmonary origin versus use of the whole specimen, which is confounded by varying volumes of saliva. We compared cell profiles and eosinophilic cationic protein (ECP) concentration in sputum selected from the expectorate and in the usually discarded residual portion to determine to what degree salivary contamination is minimized and if the results are representative of lower respiratory secretions. Sputum was induced with hypertonic saline in six healthy and nine asthmatic subjects. All portions considered to be of pure lower respiratory tract origin were selected from the residual. The selected and residual portions were treated with dithiothreitol, total cell counts and cell viability were obtained, cytospins were made for differential cell counts and supernatant was collected for ECP assay. Selected portions of the specimens, in comparison with the residual portion showed: little squamous cell contamination (median 1.2 vs 70%; p < 0.001); higher total cell counts.mL-1 (5.1 vs 0.5 x 10(6) cells.mL-1; p < 0.001); higher number of viable nonsquamous cells per sample (1.9 vs 0.6 x 10(6) cells; p < 0.001); higher slide quality score (7 vs 4; p < 0.001); and higher levels of ECP (768 vs 136 micrograms.L-1; p < 0.001). There were no differences in the differential cell counts of eosinophils (1.3 vs 3.8%), neutrophils (44 vs 32%), and lymphocytes (0.6 vs 0.6%). While the proportion of macrophages was lower (36 vs 54%; p < 0.05), the absolute number (41 vs 19 x 10(4) cells; p < 0.05) was higher in the selected portion. In summary, selection of all portions of induced sputum from the expectorate minimized the confounding influence of saliva. Loss of nonsquamous cells in the residual portion was variable but usually less than one third of those in the selected portion. With one exception, this loss had little influence on the differential counts of inflammatory cells. Similar observations apply to eosinophilic cationic protein levels. We conclude that, in healthy subjects and treated asthmatics, inflammatory markers in the selected portion of the expectorate can be used to represent those in the lower respiratory tract in general.

Adult↗

PubMed1996

Effects of native and oxidation-resistant secretory leukoprotease inhibitor on cystic fibrosis sputum: inhibition of neutrophil elastase activity and of sputum-induced secretion from porcine tracheal submucosal glands.

Because the antiprotease defense in cystic fibrosis (CF) airways is overwhelmed by neutrophil elastase (NE), substitution of antiproteases such as secretory leukoprotease inhibitor (SLPI) seems to be a reasonable therapeutic approach. Knowing, however, that native antiproteases may be liable to rapid inactivation by the locally abundant oxidants, we comparatively investigated the interactions of CF sputum with recombinant native SLPI (rSLPI) and its partially oxidation-resistant variant (rSLPI-242), respectively, to estimate their therapeutic potentials. NE activity in supernatants from diluted CF sputum samples was dose-dependently inhibited by both rSLPI and rSLPI-242, with comparable potency. Addition of the oxidant N-chlorosuccinimide resulted in significant superiority of rSLPI-242 over rSLPI. When fresh neutrophil-rich CF sputum was incubated with rSLPI and rSLPI-242, respectively, rSLPI-242 inhibited sputum NE activity significantly more potently than did rSLPI; addition of the antioxidant superoxide dismutase significantly improved the effect of rSLPI. Furthermore, secretion of radiolabeled macromolecules from porcine tracheal glands induced by purified NE or by CF sputum was inhibited dose-dependently by rSLPI and even better by rSLPI-242. We conclude that both rSLPI and rSLPI-242 effectively inhibit NE activity and NE-induced gland hypersecretion in vitro. In vivo effects of CF remain to be analyzed; an advantage of the partially oxidation-resistant rSLPI-242 can be expected.

Adolescent↗

PubMed2001

Relationship between sputum inflammatory markers and osmotic airway hyperresponsiveness during induction of sputum in asthmatic patients.

Hypertonic saline aerosols are being used increasingly for bronchial provocation testing and induction of sputum. The aims of this study were to assess the response to challenge with 3% hypertonic saline administered via a ultrasonic nebulizer in patients with asthma, and to evaluate relationship between % fall of FEV1 during induction of sputum (osmotic airway hyperresponsiveness; osmotic AHR) and biochemical markers of induced sputum. We investigated changes in FEV1 in response to inhaling ultrasonically nebulized 3% saline in 25 patients with asthma and 10 control subjects. FEV1 was measured before, during, and after induction of sputum. We used fluoroimmunoassay to detect eosinophil cationic protein (ECP), immunohistochemical staining to detect EG2+ (secretory form of ECP) eosinophils, and a sandwich ELISA to detect interleukin (IL)-5. Protein concentration was determined by using bicinchoninic acid protein assay reagent. Asthmatics, compared with controls, had significantly higher osmotic AHR. Moderate to severe asthmatics had significantly higher osmotic AHR compared to mild asthmatics. Osmotic AHR was significantly correlated with the proportion of eosinophils, the levels of ECP, EG2+ eosinophils, IL-5, and proteins. These data suggest that osmotic AHR is closely related to the clinical status and biochemical markers of sputum supernatant in asthmatic patients.

Adult↗

PubMed2000

[Study on pathogenicity of sputum from cavity of sputum negative patients with pulmonary tuberculosis after short course chemotherapy].

OBJECTIVE: To find out whether or not the sputa of pulmonary tuberculosis(PTB) patients with cavitation but sputum negative have pathogenicity after short course chemotherapy. METHODS: Guinea pigs were inoculated with sputa of PTB patients who were with cavitation but sputum negative after having finished short course chemotherapy. Then their body weight, enlargement of local lymph nodes and other ordinary symptom were observed. Six weeks later, pathological changes of TB in the internal organs were examined by dissecting these guinea pigs. Culture and drug resistance test of tubercle bacillus were also conducted. All of which were with negative and positive controls. RESULTS: Of the 63 cases included, 3(5%) patients' sputa resulted in tuberculous nodulation varying in amount in lung, liver and spleen of these guinea pigs, and the culture for tubercle bacillus of these sputa was positive too. CONCLUSIONS: 5% of sputum collected from PTB patients with cavitation but sputum negative still show pathogenicity after short course chemotherapy. For the cases with drug-resistant PTB and slow sputum negative conversion, the treatment should be prolonged and tubercle bacilli in their sputa should be monitored.

Animals↗

PubMed2004

Diagnostic value of microscopic examination of Gram-stained sputum and sputum cultures in patients with bacteremic pneumococcal pneumonia.

Clinicians continue to question the usefulness of microscopic examination of Gram-stained sputum specimens ("Gram staining") and sputum culture for diagnosis of pneumonia. We analyzed the sensitivity of these techniques in 105 patients with pneumococcal pneumonia proven by blood culture. Gram staining revealed gram-positive cocci in pairs and chains, and culture yielded pneumococci in only 31% and 44% of all cases, respectively. However, sputum specimens were never submitted for examination in 31 cases; in 16 others, the specimen was inadequate and a culture was not done. Excluding these cases, the sensitivities of Gram staining and culture were 57% and 79%, respectively. If patients receiving antibiotics for >24 h had been excluded, Gram staining would have suggested pneumococci in 63%, and culture results would have been positive in 86%. Sensitivity increased in inverse proportion to the duration of antibiotic therapy (P<.05). Microscopic examination of sputum samples before antibiotics were administered and performance of culture within 24 h of receipt of such treatment yielded the correct diagnosis in >80% of cases of pneumococcal pneumonia.

Anti-Bacterial Agents↗

PubMed1975

Recovery of Cryptococcus neoformans from sputum using new technics for the isolation of fungi from sputum.

Three sputum-digesting agents, N-acetyl-l-cytseine, dithiothreitol, and pancreatin-trypsin, were shown to be equally effective in allowing for the isolation of Cryptococcus neoformans from sputum samples in quantitative comparisons. By quantitative plating on bird-seed medium it was also shown the centrifugation after digestion concentrated C. neoformans into a platable sediment and, further, that the organisms, when present in concentrations as low as 10 yeasts per ml. of sputum, could be isolated with much higher frequencies than when no digestion-centrifugation procedure was used.

Acetylcysteine↗

PubMed2001

Can semi-automated image cytometry on induced sputum become a screening tool for lung cancer? Evaluation of quantitative semi-automated sputum cytometry on radon- and uranium-exposed workers.

The correlations between semi-automated sputum cytometry (ASC), conventional cytology and the final diagnosis was investigated in industrially-exposed workers. Slides of sputum samples from 201 former uranium miners with silicosis, 100 patients with asbestosis, 103 workers resected for lung cancer, and 200 controls (50% smokers), were stained using the Papanicolaou (Pap) method and the Feulgen reaction with thionin. Cytometry was performed using the Cyto-Savant automated system. Atypical nuclei were found in 72 of 404 patient samples, 327 samples were normal and five were inadequate for ASC analysis. Thirteen tumours (Pap IV, Pap V) and 11 cases of severe dysplasia were identified by cytology. Lung cancer was confirmed in 20 patients. Compared to the final diagnosis of lung cancer, the sensitivity of ASC was 75% (15 out of 20) and specificity 89.8% (520 out of 579). The results represent a diagnostic efficiency of 89.3%. The combination of ASC with cytology increased sensitivity to 80% (16 out of 20) without significant loss of specificity (89.7% or 523 out of 581). In this investigation of a limited number of patients with occupational radon or asbestos exposure, semi-automated sputum cytometry appears to be sensitive and reliable for the detection of malignant changes in the tracheobronchial mucosa. Together with conventional cytology, it would be reasonable to test the validity of the combined methods in a large-scale feasibility study of early lung cancer detection.

Adult↗

PubMedDate not supplied

Factors significant in the diagnostic accuracy of lung cytology in bronchial washing and sputum samples. II. Sputum samples.

Some factors influencing the detection of malignant cells in sputum samples were evaluated in 449 consecutive cases of primary lung carcinoma seen between 1959 and 1974. Diagnostic accuracy increased during the years under study; the reasons are discussed. The overall accuracy was 82.8%. Detection of malignant cells was 85% for small-cell carcinoma, squamous-cell carcinoma and large-cell carcinoma, 75% for adenocarcinoma, bronchioloalveolar carcinoma and adenosquamous carcinoma and 64% for the uncommon tumors. Accuracy was 87% for central tumors and 42% for peripheral lesions. Tumors less than 2 cm in diameter yielded only 39% accuracy as compared to 90% for larger tumors. The specificity of diagnosis of cell type in those specimens with malignant cells was 95% for small-cell carcinoma and squamous-cell carcinoma, more than 80% for adenocarcinoma and large-cell carcinoma, 65% for bronchioloalveolar-cell carcinoma and adenosquamous carcinoma and less than 30% for the uncommon tumors. Diagnostic accuracy was optimal in those cases with three or more sputum samples: 83% for those with three samples and 90% for those with five or more samples per case. The use of both sputum and bronchial specimens was complementary and increased the accuracy further. Reasons for unsatisfactory specimens included no deep cough, limited cellular material, excessive blood or leukocytes and drying artifacts; the first two were the most common causes.

Adenocarcinoma↗

PubMedDate not supplied

Sputum cells from lung tumor patients carry a cell surface marker not found in normal sputum.

Cells were collected from sites of known lung tumours and corresponding control areas of these lungs. Fluorescent staining demonstrated that the tumour cells and epithelial cells (cytologically these cells appeared normal) both possessed a receptor for these fluorescent probes. Fluorescent labelling of sputum cells from these tumour patients also resulted in fluorescent labelling of these "cyto logically normal" epithelial cells. No such fluorescent epithelial cells were observed in sputum samples collected from control subjects or in cells collected from the control areas of the tumour patients' lungs. We conclude that a cell surface protein receptor is expressed in lung tumour-associated epithelial cells but is absent from control sputum epithelial cells.

Biomarkers, Tumor↗

PubMed1988

Measurement of Pseudomonas aeruginosa phenazine pigments in sputum and assessment of their contribution to sputum sol toxicity for respiratory epithelium.

The phenazine pigments pyocyanin and 1-hydroxyphenazine were resolved by high-pressure liquid chromatography from the sputum sol phase from 9 of 13 patients with cystic fibrosis or bronchiectasis colonized by Pseudomonas aeruginosa. The concentrations measured were each sufficient to inhibit ciliary beating in vitro and contributed a significant proportion of sol phase toxicity for respiratory epithelium.

Cilia↗

PubMed2005

Lot quality assurance sampling of sputum acid-fast bacillus smears for assessing sputum smear microscopy centers.

Assessment of 12 microscopy centers in a tuberculosis unit by blinded checking of eight sputum smears selected by using a lot quality assurance sampling (LQAS) method and by unblinded checking of all positive and five negative slides, among the slides examined in a month in a microscopy centre, revealed that the LQAS method can be implemented in the field to monitor the performance of acid-fast bacillus microscopy centers in national tuberculosis control programs.

Bacteriological Techniques↗

PubMed1985

Diagnosis of sputum smear- and sputum culture-negative pulmonary tuberculosis.

A group of patients with new or enlarging apical lung lesions thought to be tuberculous was randomly selected for observation without treatment. All were tuberculin-positive and all had sputum negative for Mycobacterium tuberculosis on direct examination and on culture. Of the 152 patients, 88 (58%) developed bacteriologically confirmed pulmonary tuberculosis during a follow-up period of up to 65 months.

Follow-Up Studies↗

PubMed1960