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PubMed2006

Methylation profile in tumor and sputum samples of lung cancer patients detected by spiral computed tomography: a nested case-control study.

We evaluated the aberrant promoter methylation profile of a panel of 3 genes in DNA from tumor and sputum samples, in view of a complementary approach to spiral computed tomography (CT) for early diagnosis of lung cancer. The aberrant promoter methylation of RARbeta2, p16(INK4A) and RASSF1A genes was evaluated by methylation-specific PCR in tumor samples of 29 CT-detected lung cancer patients, of which 18 had tumor-sputum pairs available for the analysis, and in the sputum samples from 112 cancer-free heavy smokers enrolled in a spiral CT trial. In tumor samples from 29 spiral CT-detected patients, promoter hypermethylation was identified in 19/29 (65.5%) cases for RARbeta2, 12/29 (41.4%) for p16(INK4A) and 15/29 (51.7%) for RASSF1A. Twenty-three of twenty-nine (79.3%) samples of the tumors exhibited methylation in at least 1 gene. In the sputum samples of 18 patients, methylation was detected in 8/18 (44.4%) for RARbeta2 and 1/18 (5%) for both RASSF1A and p16(INK4A). At least 1 gene was methylated in 9/18 (50%) sputum samples. Promoter hypermethylation in sputum from 112 cancer-free smokers was observed in 58/112 (51.7%) for RARbeta2 and 20/112 (17.8%) for p16, whereas methylation of the RASSF1A gene was found in only 1/112 (0.9%) sputum sample. Our study indicates that a high frequency of hypermethylation for RARbeta2, p16(INK4A) and RASSF1A promoters is present in spiral CT-detected tumors, whereas promoter hypermethylation of this panel of genes in uninduced sputum has a limited diagnostic value in early lung cancer detection.

Aged↗
PubMed2002

Relationship between sputum inflammatory markers, lung function, and lung pathology on high-resolution computed tomography in children with cystic fibrosis.

High-resolution computed tomography (HRCT) is a sensitive technique for early visualisation and location of cystic fibrosis (CF) bronchopathology, and has been shown to detect acute reversible and chronic changes. It would be expected to correlate with markers of the underlying pathological processes, such as sputum cytokines and cytology, as well as with pulmonary function tests (PFTs). Our aim was to study the relationship between PFTs, sputum cytology, and sputum cytokine interleukin-8 (IL-8) and HRCT in CF patients. Prospective standardized collection of sputum samples was performed at the time of routine annual high-resolution CT scans. Forced expired volume in 1 sec (FEV(1)) and forced vital capacity (FVC) were recorded. Sputum processing was selective, with dispersal by the three-enzyme technique. IL-8 measurements were by kit assay. HRCT scans were scored by a pediatric radiologist, blinded to clinical condition, using a modified Bhalla score.Forty-three CT scans were performed on 34 children with CF between March 1998 and April 2000. Mean age was 12.3 years (range, 6-21 years), FEV(1) (% predicted) was 67% (range, 23-120%), and mean modified Bhalla score was 11.2 (range, 0-22). Sputum IL-8 concentration (mean, 86; range, 4-150 ng/mL) and total cell count (mean, 31.9 x 10(6)/mL; range, 21.8-42.0 x 10(6)/mL) were high. FEV(1) and FVC correlated with modified Bhalla score (r = -0.66, P < 0.0001 for both), and most individual components of the score, especially mosaic perfusion (r = -0.64, r = -0.61 respectively, P < 0.0001) and extent of bronchiectasis (r = -0.61, P < 0.0001 for both). The combination of these two predicted 58% of the variability in FEV(1) on analysis of variance (P < 0.0001). Sputum total cell count correlated weakly with modified Bhalla score (r = 0.38, P < 0.05) and with FEV(1) and FVC (r = -0.36, P < 0.05; and r = -0.46, P < 0.01). Differential cell counts, cell viability, and IL-8 did not correlate with modified Bhalla scores, or with reversible components such as mucus plugging, centrilobular nodules, or peribronchial thickening. In conclusion, pathological changes on HRCT correlated with lung function but not with sputum markers of inflammation.

Adolescent↗
PubMed2004

Detection of DNA aneuploidy in exfoliated airway epithelia cells of sputum specimens by the automated image cytometry and its clinical value in the identification of lung cancer.

To evaluate the value of detection of DNA aneuploidy in exfoliated airway epithelia cells of sputum specimens by the automated image cytometry for the identification of lung cancer, 100 patients were divided into patient group (50 patients with lung cancer) and control group (30 patients with tuberculosis and 20 healthy people). Sputum was obtained for the quantitative analysis of DNA content of exfoliated airway epithelial cells with the automated image cytometry, together with the examinations of brush cytology and conventional sputum cytology. Our results showed that DNA aneuploidy (DI>2.5 or 5c) was found in 20 out of 50 sputum samples of lung cancer, 1 out of 30 sputum samples from tuberculosis patients, and none of 20 sputum samples from healthy people. The positive rates of conventional sputum cytology and brush cytology were 16% and 32%, which was lower than that of DNA aneuploidy detection by the automated image cytometry (P<0.01, P>0.05). Our study showed-that automated image cytometry, which uses DNA aneuploidy as a marker for tumor, can detect the malignant cells in sputum samples of lung cancer and it is a sensitive and specific method serving as a complement for the diagnosis of lung cancer.

Adult↗
PubMed2005

Assessing abnormal gene promoter methylation in paraffin-embedded sputum from patients with NSCLC.

Aberrant methylation of CpG islands is an important pathway for regulation of gene expression. Recent data suggest that epigenetic abnormalities may occur very early in lung carcinogenesis. We studied the promoters of the four genes, HOX A9, p16(INK4a) (p16), MAGE A1 and MAGE B2 by methylation-specific PCR in matched normal tissue, tumour, and cytological negative sputum samples obtained from 22 patients with non-small cell lung cancer (NSCLC). We further report methylation abnormalities in sputum samples from 56 smokers with differential cytology readouts (negative, inflammatory changes, suspicious, and cancer). Our method was successfully performed on formalin-fixed and paraffin-embedded (FFPE) samples, and was fit to study only few cells obtained by a convenient non-invasive sputum collection and handling. The promoters of MAGE A1 and MAGE B2 had abnormal methylation patterns in, respectively, 50% and 41% of the cytologically negative sputum samples from NSCLC patients, whereas methylation abnormalities of p16 was observed in 27% of negative sputum samples. Interestingly, 95.5% (21 of 22) of the cytologically negative sputum samples from NSCLC patients had abnormal methylation in at least one of the four genes indicating a high sensitivity of this marker system. Moreover, a higher frequency of methylation abnormalities was observed in sputum samples from smokers with high cytological grade compared to low cytological grade. We conclude that the identification of abnormal gene methylation of a limited number of markers in FFPE sputum samples is feasible and may be investigated as a potential system for population-based screening of early stages of lung cancer.

Aged↗
PubMed2004

Measurement of eotaxin (CCL11) in induced sputum supernatants: validation and detection in asthma.

BACKGROUND: Induced sputum is widely used in asthma research; however, for many mediators, the detection methods have not been validated. OBJECTIVE: We sought to optimize the method of detection of eotaxin, an important chemokine acting through the CCR3 receptor on eosinophils, basophils, and T(H)2 cells. METHODS: Induced sputum from normal and asthmatic subjects was processed with dithioerythritol (DTE) or PBS; recovery of eotaxin was assessed by means of ELISA before and after spiking with recombinant eotaxin. Furthermore, the effects of removing DTE by means of ultrafiltration or the addition of protease inhibitors and high-speed centrifugation on endogenous levels and spiking recovery of eotaxin were assessed. RESULTS: Endogenous eotaxin was undetectable in DTE-processed samples, with a mean of only 30% (SD, 13%) spike recovery. DTE had no effect on the immunoassay capture antibody but dramatically reduced the detection of recombinant eotaxin. Removal of DTE from sputum before immunoassay did not improve detection, although it restored the recovery of a subsequent eotaxin spike. In contrast, PBS-processed sputum resulted in an eotaxin spike recovery of 101% (SD, 20%). Addition of protease inhibitors or high-speed centrifugation had no effect on eotaxin detection. By using this optimized protocol, eotaxin levels in PBS-processed sputum samples were found to be significantly increased in asthmatic sputum (P<.05). CONCLUSION: Measurement of eotaxin by means of immunoassay is adversely affected by DTE, possibly through irreversible denaturation of epitopes, which makes eotaxin undetectable by using the immunoassay antibody. Sputum samples should be processed into PBS for assessment of eotaxin, which is present at increased levels in asthmatic sputum.

Adult↗
PubMed2004

K-ras and p16(INK4A)alterations in sputum of NSCLC patients and in heavy asymptomatic chronic smokers.

NSCLC rates among the most frequent and lethal neoplasm world-wide and a significant decrease in morbidity and mortality relies only upon effective early diagnostic strategies. We investigated K-ras mutations and p16(INK4A) hypermethylation in tumor tissue and sputum of 50 patients with NSCLC and correlated them with sputum cytology and with tumor staging, grading and location, to ascertain, in sputum, their potential diagnostic impact. The same genetic/epigenetic abnormalities and cytological features were also evaluated in sputum from 100 chronic heavy smokers. Genetic analysis identified molecular abnormalities in 64% tumors (14/50 K-ras mutations and 24/50 p16(INK4A) hypermethylation) and in 48% sputum (11/50 K-ras mutations and 16/50 p16(INK4A) hypermethylation). In tumors K-ras mutations and p16(INK4A) hypermethylation were mostly mutually exclusive, being found in the same patients in 3 cases only. Genetic abnormalities in sputum were detected only in molecular abnormal tumors. Molecular changes in sputum had rates of detection similar to cytology (42%) but the cyto-molecular combination increased the diagnostic yield up to 60%. Interestingly, the rate of detection of genetic changes in sputum of tumors at early stage (T1) was not significantly different from that of tumors at more advanced stage (T2-T4). In fact K-ras point mutations were frequently recognised in tumors at early stage while p16(INK4A) inactivation prevailed in tumors at advanced stage ( P=0.0063). As expected, diagnostic cytological findings were more frequently found in tumors at advanced stage (P=0.004). No correlation was found between tumor grading and location (central versus peripheral) and molecular changes. p16(INK4A) hypermethylation, but not K-ras mutations, was documented in sporadic cases of asymptomatic heavy smokers (4%) where it was uncoupled from cytological abnormalities. In conclusion the cyto-molecular diagnostic strategy adopted in this study was able to detect the majority of tumors but in order to be proposed as effective and early diagnostic tool, this molecular panel needs to be tested in prospective studies with adequate follow-up.

Aged↗
PubMed2005

Sputum substance P and neurokinin A are reduced during exacerbations of chronic obstructive pulmonary disease.

Involvement of tachykinins in airway inflammation has been demonstrated in animal models, but evidence in humans is sparse. The aim of this study was to quantify the levels of substance P and neurokinin A in induced sputum of patients with chronic obstructive pulmonary disease (COPD) and to compare them with the levels in smokers with normal lung function and healthy nonsmokers. Content of tackykinins was measured in 12 sputum samples collected during stable condition and nine sputum samples collected during exacerbations from 13 COPD patients, in eight sputum samples from smokers with normal lung function and in nine from healthy nonsmokers. Patients with COPD exacerbations had a lower sputum content of substance P compared with the other 3 groups (p<0.05). No differences were found between patients with stable COPD, smokers with normal lung function, and nonsmokers. Sputum levels of neurokinin A were trending in the same direction of substance P, but the significant difference was reached for the paired sputum samples collected from the same COPD patients (n=8) during exacerbation and in stable condition. COPD exacerbations are associated with a reduced sputum content of substance P and neurokinin A. These tackykinins might be involved in COPD exacerbations.

Aged↗
PubMed2005

Inhaled fluticasone propionate is effective as well as oral prednisone in reducing sputum eosinophilia during exacerbations of asthma which do not require hospitalization.

The aim of this study was to evaluate whether fluticasone propionate (FP) is effective as well as prednisone (P) in reducing sputum eosinophilia and in improving airway obstruction due to asthma exacerbations not requiring hospitalization. We measured, in a parallel-group, double-blind double-dummy, randomized study, sputum and blood inflammatory cell counts and soluble mediators in 37 asthmatic subjects during a spontaneous exacerbation of asthma (Visit 1) and after a 2 week (Visit 2) treatment with inhaled FP (1000microg bid) (Group A, n=18) or a reducing course of oral P (Group B, n=19). Asthma exacerbation was accompanied by sputum eosinophilia (eosinophils >2%) in almost all patients (95%). FP improved FEV(1) (from 53.9%+/-16.8 at Visit 1 to 76.4%+/-21.2 at Visit 2, p=0.0001) and reduced the percentage of sputum eosinophils (from 38%[0-78] to 3%[1-31, p=0.0008) as well as oral P (FEV(1): from 51.5%+/-14.4 to 83.6%+/-21.1, p=0.0001; sputum eosinophils: from 52%[1-96] to 11%[0-64], p=0.0003). At Visit 2, sputum eosinophils were significantly lower in Group A than in Group B. P but not FP induced significant decrease in blood and sputum ECP. Oxygen saturation, PEF variability, symptom score and use of rescue medication similarly improved in both groups. We conclude that FP is effective at least as well as P in reducing sputum eosinophilia and in improving airway obstruction due to asthma exacerbation. However, the cost/effectiveness ratio of this option should be further evaluated.

Acute Disease↗
PubMed2001

[Usefulness of induced sputum cytology in the study of central masses in elderly patients].

OBJECTIVE: To analyze the diagnostic yield of cytology obtained by means of sputum induction in elderly patients with central masses. METHODS: A total of 23 patients older than 75 years with centrally located pulmonary masses identified by computerized tomography were studied. All patients collected three spontaneously produced sputum specimens and underwent three sputum-induction sessions. Induction was considered satisfactory when the patient expectorated within the following 15 minutes. All sputum specimens were processed by May-Grünwald-Giemsa and Papanicolau staining techniques. The definite diagnostic was considered that obtained by means of bronchial biopsy or chest percutaneous aspirative puncture (PAP). RESULTS: With sputum induction, 19 patients (83%) collected satisfactory specimens and in 14 cases cytology was positive for malignancy (sensitivity 74%) and negative in five. No false-positive cases were recorded and in no patient did complications occur in association with sputum induction. The analysis of agreement between induced sputum and definite diagnosis showed a kappa index of 0.66. CONCLUSIONS: Among elderly patients with central pulmonary masses, the cytology of induced sputum should be considered as the first diagnostic procedure because of its harmlessness, remarkable sensitivity and high histologic correlation.

Aged↗
PubMed1988

Sputum changes associated with therapy for endobronchial exacerbation in cystic fibrosis.

We sought to define objective indicators of the resolution of Pseudomonas aeruginosa endobronchial infection in patients with cystic fibrosis. We prospectively studied 75 patients admitted for treatment of a pulmonary exacerbation and quantitated sputum bacterial density, DNA content, and the concentration of albumin and total protein in sputum, and compared these values with clinical evaluation. Eleven of the 75 patients had systemic signs, fever, and leukocytosis, which we arbitrarily defined as due to endobronchial infection. At the end of hospitalization, these 11 patients were afebrile, had peripheral leukocyte counts in the normal range, and were judged improved. Sputum P. aeruginosa density, DNA content, and total protein content on admission were similar in the two illness groups. Hospitalization and parenteral antibiotic administration for an average of 14.6 days were associated with improved pulmonary function in all 75 subjects (P values for forced vital capacity, forced expiratory volume at 1 second, and peak expiratory flow rate were all less than 0.001). With improvement, there was a decrease in sputum P. aeruginosa density (mean of both groups decreased from 10(7.80) CFU/g on admission to 10(5.96) CFU/g; P less than 0.001), and a decreased DNA concentration (overall mean 4.73 +/- 4.75 on admission to 2.76 +/- 2.49 mg/g; P less than 0.002). The decrease in sputum total protein concentration for both groups was not significant (overall mean 60.5 +/- 48.4 to 43.9 +/- 38.2 mg/g; P = 0.06). Sputum albumin concentrations did not change in either group. We conclude that in cystic fibrosis subjects with a pulmonary exacerbation, bacterial density, sputum DNA and protein content decrease with hospitalization and parenteral antibiotic therapy. At the end of treatment, these indices of sputum infection and inflammation correlate with improved pulmonary function and clinical improvement. These changes are independent of the presence or absence of fever on admission.

Adolescent↗
PubMed1999

Inhaled IL-5 increases concentrations of soluble intracellular adhesion molecule-1 in sputum from atopic asthmatic subjects.

BACKGROUND: Both IL-5 and intracellular adhesion molecule-1 (ICAM-1) have been shown to play important roles in the production of allergic inflammation, including atopic asthma. OBJECTIVE: The aim of this study was to investigate the effects of recombinant human IL-5 on changes of soluble (s) ICAM-1 concentrations in induced sputum from allergic asthmatic subjects. METHODS: In a randomized, double-blind, placebo-controlled study design, 8 nonsmoking patients with allergic asthma and 6 nonallergic normal subjects were administered recombinant human IL-5 by nebulization, and the concentrations of sICAM-1 in induced sputum from each subject were determined before and at 2, 24, 48, and 72 hours after inhalation. RESULTS: sICAM-1 levels in sputum within the control group did not appear to change from baseline at any time throughout the study. In allergic asthmatic subjects vehicle challenge was not able to cause any changes in sputum sICAM-1 concentrations. However, there were increases in sputum sICAM-1 concentrations after IL-5 inhalation, which increased with time to significantly greater levels than those at baseline, reaching a maximum at 48 hours and lasting no less than 72 hours. The concentrations of sICAM-1 in sputum after IL-5 challenge exceeded levels that could be accounted for by passive transudation from the circulation on the basis of the magnitude of increases in sputum albumin concentration. CONCLUSION: Our results indicated that IL-5 inhalation was capable of inducing an elevation of sputum sICAM-1 concentration by stimulating its local release in allergic asthmatic, but not in nonallergic normal, subjects.

Administration, Inhalation↗
PubMed2002

Asthma exacerbations and sputum eosinophil counts: a randomised controlled trial.

BACKGROUND: Treatment decisions in asthma are based on assessments of symptoms and simple measures of lung function, which do not relate closely to underlying eosinophilic airway inflammation. We aimed to assess whether a management strategy that minimises eosinophilic inflammation reduces asthma exacerbations compared with a standard management strategy. METHODS: We recruited 74 patients with moderate to severe asthma from hospital clinics and randomly allocated them to management either by standard British Thoracic Society asthma guidelines (BTS management group) or by normalisation of the induced sputum eosinophil count and reduction of symptoms (sputum management group). We assessed patients nine times over 12 months. The results were used to manage those in the sputum management group, but were not disclosed in the BTS group. The primary outcomes were the number of severe exacerbations and control of eosinophilic inflammation, measured by induced sputum eosinophil count. Analyses were by intention to treat. FINDINGS: The sputum eosinophil count was 63% (95% CI 24-100) lower over 12 months in the sputum management group than in the BTS management group (p=0.002). Patients in the sputum management group had significantly fewer severe asthma exacerbations than did patients in the BTS management group (35 vs 109; p=0.01) and significantly fewer patients were admitted to hospital with asthma (one vs six, p=0.047). The average daily dose of inhaled or oral corticosteroids did not differ between the two groups. INTERPRETATION: A treatment strategy directed at normalisation of the induced sputum eosinophil count reduces asthma exacerbations and admissions without the need for additional anti-inflammatory treatment.

Adult↗
PubMed1997

Comparison of induced versus expectorated sputum for diagnosis of pulmonary tuberculosis by acid-fast smear.

OBJECTIVE: To evaluate the sensitivity of induced versus expectorated sputum for the diagnosis of pulmonary tuberculosis by acid-fast smear. METHODS: We performed a retrospective review of data on patients with cultures from respiratory secretions that were positive for Mycobacterium tuberculosis. We analyzed data on the 114 patients with positive cultures during a 4-year period in an urban tertiary care academic medical center. RESULTS: Sputum samples had been obtained for 103 of the 114 patients with cultures positive for tuberculosis. Forty-four of the 114 patients were HIV seropositive. Overall 33 of 79 patients (42%) had positive acid-fast smears of expectorated sputum, and 6 of 24 (25%) had positive smears of induced sputum (p = 0.21). When data were analyzed for subgroups by HIV serostatus, there was no significant difference in the results. Estimated cost of sputum induction for 1 year was approximately $45,000. CONCLUSIONS: We found sputum induction to be costly, and induced sputum offered no advantage over routine expectorated sputum for the diagnosis of pulmonary tuberculosis.

Coloring Agents↗
PubMed1998

Leukocyte counts in hypertonic saline-induced sputum in subjects with occupational asthma.

We measured markers of eosinophilic inflammation in the blood and in the sputum induced by hypertonic saline (HS) inhalation of 24 subjects with occupational asthma who were still exposed to high molecular weight compounds (HMWCs, n = 8) or to low molecular weight compounds (LMWCs, n = 16); all subjects were symptomatic and showed bronchial hyperresponsiveness to methacholine at the time of study. Sputum cell counts were also measured in 14 normal subjects and in 24 subjects with non-occupational asthma with asthma severity similar to that of occupational asthmatics. Both occupational and non-occupational asthmatic subjects showed higher neutrophil percentages in HS-induced sputum than normal subjects, asthmatics with LMWC-induced asthma showing the highest values. Eosinophil percentages in HS-induced sputum were higher in non-occupational asthmatics and in asthmatics with HMWC-induced asthma than in normal subjects and in subjects with occupational asthma due to LMWCs. No difference in bronchial responsiveness, peak expiratory flow variability and serum eosinophil cationic protein (ECP) levels were observed among the different asthma groups. Although sputum eosinophil percentages significantly correlated with blood eosinophil percentages, sputum allowed the detection of a higher number of subjects with eosinophilic inflammation than blood. Serum ECP levels were normal in most asthmatic subjects. A significant correlation between sputum eosinophil percentages and bronchial hyperresponsiveness to HS was observed. Despite a similar degree of functional abnormalities, subjects with asthma due to LMWCs and still exposed to the occupational sensitizer showed a lower degree of eosinophilic inflammation and a higher degree of neutrophilic inflammation in the airways than subjects with occupational asthma due to HMWCs or non-occupational asthmatics. Furthermore, sputum eosinophil counts detect, better than blood indices, the degree of airway inflammation in both occupational and non-occupational asthma.

Adult↗
PubMed1999

The safety and success rate of sputum induction using a low output ultrasonic nebuliser.

Induced sputum differential cell counts have been advocated as a method of non-invasively assessing airway inflammation in asthma and other airway diseases. Since sputum induction usually involves delivering hypertonic saline via a high output ultrasonic nebulizer there have been concerns about its safety in asthma. There are relatively little data on the effects of sputum induction in large numbers of patients. We have examined the success rate and effect of sputum induction on forced expiratory volume in 1 sec (FEV1) in 100 inductions performed on 79 patients using a low output nebulizer. Thirty-seven patients had asthma, 29 had miscellaneous conditions (mainly chronic cough) and 13 were subjects without respiratory symptoms. Sputum was induced 10 min after 200 micrograms of inhaled salbutamol by sequential 5-min inhalations of 3, 4 and 5% saline delivered via a Fisoneb ultrasonic nebulizer and FEV1 was measured after each inhalation. Sputum induction resulted in a sample suitable for analysis in 92% of asthmatics, 90% of those with miscellaneous conditions and 100% of normal subjects. The mean (SEM) maximum per cent fall in FEV1 was 5.4% (0.1), 4.3%, (1.0) and 2.6% (1.1) in subjects with asthma, miscellaneous conditions and in asymptomatic subjects respectively. Only 13 inductions resulted in a > 10% fall in FEV1, and only three of these resulted in a > 20% fall. The maximum per cent fall in FEV1 did not correlate with baseline FEV1 % predicted (r = -0.17), the log sputum eosinophil count (r = -0.12), or the methacholine PC20 (r = -0.14). We conclude that sputum induction using a relatively low output ultrasonic nebulizer with premedication with salbutamol is successful and safe in the majority of patients with asthma and other airway conditions.

Adolescent↗
PubMed1996

Sputum smear conversion during directly observed treatment for tuberculosis.

SETTING: Treatment program for tuberculosis in a refugee camp in Thailand. OBJECTIVES: To determine the cumulative frequency of conversion of sputum smears examined by direct microscopy by month of treatment and to identify factors predicting failure to convert. METHODS: Analysis of conversion based on three sputum smear examinations (performed monthly) in a cohort of patients with sputum smear-positive tuberculosis treated with a directly observed daily regimen containing rifampicin throughout. Nested case-control study of patients failing to convert definitively within four months compared to controls who did convert. RESULTS: Sputum conversion after the 2-month intensive phase was 75.0%, with a range from 61.7% to 90.9% in patients with initially strongly- and weakly-positive smears, respectively. The strongest predictor identified for no definitive conversion within four months of treatment was a positive sputum smear result at the end of the 2-month intensive phase (adjusted relative odds 4.2, 95% confidence interval 1.5-11.4). Of those patients who did not convert, positive smears were an isolated phenomenon in 15, repeatedly in four who definitely converted with a prolongation of treatment, and persistently positive in two requiring a re-treatment regimen. CONCLUSIONS: Definitive sputum smear conversion is judged to be slower if a strict program of sputum smear examination is undertaken than under routine program conditions, but positive results late in the course are commonly an isolated phenomenon and possibly of little significance. Sputum smear results at two months strongly predict bacteriologic results beyond three months of treatment, and thus identify cases who might benefit from a prolongation of the intensive phase.

Case-Control Studies↗
PubMed2004

Sputum tests in the diagnosis and monitoring of asthma.

OBJECTIVE: To review the techniques of sputum analysis with relevance to the diagnosis and monitoring of asthma. DATA SOURCES: MEDLINE databases were searched to identify all publications involving sputum studies related to the diagnosis and treatment of asthma from 1990 to October 2003. We also used internal reference files related to sputum and searched bibliographies of relevant articles. The review was limited to human data in English-language publications. STUDY SELECTION: Studies were selected by the expert opinions of the authors for quality and relevance to the evaluation of asthmatic inflammation by induced sputum. RESULTS: During the past 10 years, there have been an increasing number of publications concerning the diagnosis and treatment of asthma using sputum analysis. Analysis of induced sputum provides similar data to secretions obtained through bronchial wash, bronchoalveolar lavage, and, to some extent, bronchial biopsy. The techniques of cellular counting and immunochemical analysis are described along with potential problems and pitfalls of these methods. Clinical application of sputum analysis is discussed as it pertains to the diagnosis and monitoring of asthma and asthma-related conditions. CONCLUSIONS: Analysis of induced sputum is increasingly being considered as a noninvasive means of evaluating airway inflammation and may provide useful information with regard to the diagnosis and monitoring of asthma in select individuals.

Asthma↗
PubMed2000

Reproducibility of measurements of exhaled NO, and cell count and cytokine concentrations in induced sputum.

Sputum induction is a noninvasive, well-tolerated method for studying airway inflammation. When induction with hypertonic saline is repeated at short time-intervals (<24 h), the cell profile of sputum has not been reproducible. To determine the proper interval between sampling cell profiles and cytokine contents of sputum samples that had been induced 48 h apart, were compared. In addition, the inducible nitric oxide synthase (iNOS) expression of sputum cells was compared to the levels of exhaled nitric oxide (NO). Sputum induction and measurement of exhaled NO was performed in 31 healthy nonatopic volunteers. Cell differentials were counted. Concentrations of interleukin (IL)-4, IL-6, tumour necrosis factor (TNF)alpha, eosinophil cationic protein (ECP) were measured in sputum supernatant, and iNOS was determined. Reproducibility of cell counts was high (r=0.836 total cells, r=0.762 neutrophils, r=0.966 eosinophils, r=0.742 macrophages). IL-4 (r=0.398), IL-6 (r=0.566), TNFalpha (r=0.658) and ECP (r=0.501) were also less reproducible in healthy volunteers. Consistent with the low levels of NO in the exhaled air (18.5+/-2.6 ppb and 19.3+/-2.8 parts per billion (ppb) on the two study days, r=0.976, p=0.0000), expression of iNOS was not detected. In conclusion, in healthy subjects, induced sputum cell counts are reproducible. Even though the success rate in nonatopic populations is relatively low, sputum induction appears to be a valid method for detecting inflammatory changes within the airways, when being performed 48 h apart.

Administration, Inhalation↗