Development and Evaluation of the New Predictive Models in Tuberculous Pleuritis.

Different pleural fluid biomarkers have been found useful in the discrimination between tuberculous pleural effusion (TPE) and non-TPE, with interferon gamma (IFN-γ) showing the highest single marker diagnostic accuracy. The aim of the present study was to develop predictive models based on clinical data and pleural fluid biomarkers, other than IFN-γ, which could be applied in differentiating TPE and non-TPE. Two hundred and forty two patients with newly diagnosed pleural effusion were prospectively enrolled. Upon completion of the diagnostic procedures, the underlying disease was identified in 203 patients (117 men and 86 women, median age 65 years; 44 patients with TPE and 159 with non-TPE) who formed the proper study group. Pleural fluid level of ADA, IFN-γ, IL-2, IL-2sRα, IL-12p40, IL-18, IL-23, IP-10, Fas-ligand, MDC, and TNF-α was measured and then ROC analysis and multivariate logistic regression were used to construct the predictive models. Two predictive models with very high diagnostic accuracy (AUC > 0.95) were developed. The first model included body temperature, white blood cell count, pleural fluid ADA and IP-10. The second model was based on age, sex, body temperature, white blood cell count, pleural fluid lymphocyte percentage, and IP-10 level. We conclude that two new predictive models based on clinical and laboratory data demonstrate very high diagnostic performance and can be potentially used in clinical practice to differentiate between TPE and non-TPE.

Diagnostic performance of different pleural fluid biomarkers in tuberculous pleurisy.

Due to the paucibacillary nature of tuberculous pleural effusion the diagnosis of pleural tuberculosis is challenging. This prospective study was undertaken to evaluate the diagnostic performance of ten different pleural fluid biomarkers in the differentiation between tuberculous and non-tuberculous pleural effusions. Two hundred and three patients with pleural effusion (117 men and 86 women, median age 65 years) were enrolled. Routine diagnostic work-up, including thoracentesis and pleural fluid analysis, was performed to determine the cause of pleural effusion. The following biomarkers were measured in pleural fluid: adenosine deaminase (ADA), interferon gamma (IFN-γ), interleukin 2 soluble receptor (IL-2sRα), sub-unit p40 of interleukin 12b (IL-12p40), interleukin 18 (IL-18), interleukin 23 (IL-23), IFN-γ induced protein 10 kDa (IP-10), Fas-ligand, human macrophage-derived chemokine (MDC) and tumor necrosis factor alfa (TNF-α). There were 44 (21.7%) patients with tuberculous pleural effusion, 88 (43.3%) patients with malignant pleural effusion, 35 (17.2%) with parapneumonic effusion/pleural empyema, 30 (14.8%) with pleural transudates, and 6 (3%) with miscellaneous underlying diseases. Pleural fluid IFN-γ was found the most accurate marker differentiating tuberculous from non-tuberculous pleural effusion, with sensitivity, specificity, PPV, NPV, and AUC 97%, 98%, 95.5%, 99.4%, and 0.99, respectively. Two other biomarkers (IP-10 and Fas ligand) also showed very high diagnostic accuracy with AUC≥0.95. AUC for ADA was 0.92. We conclude that IFN-γ, IP-10, and Fas-ligand in pleural fluid are highly accurate biomarkers differentiating tuberculous from non-tuberculous pleural effusion.

CD4+/CD25+ cells in systemic inflammation in COPD.

The autoimmune reaction is recently suspected to play a role in the pathogenesis of chronic obstructive lung disease (COPD). As COPD is a systemic disease, the elements of an autoimmune response in circulatory system is of interest. It has been shown that regulatory T cells are important in the control of autoimmunity. There are some data on a role of adiponectin in the regulation of immune reactions. The objective of this study was to assess the elements of autoimmune reaction in the peripheral blood (PB) of patients with COPD. Twenty-eight patients with mild/moderate COPD and 20 healthy volunteers were investigated. Flow cytometry method with mixtures of monoclonal antibodies anti: CD14/CD45, CD3/CD19, CD4/CD25/CTLA4 and CD8/CD25 were used. Concentration of adiponectin was measured using ELISA method. We observed significantly lower proportion of CD4+/CD25+ as well as CD4+/CD25+ (high) cells in COPD patients than in healthy controls (15.3 versus 17.8% and 0.79 versus 1.54%, respectively, P < 0.05). The proportion of CTLA4+ cells in CD25+ cells and the mean fluorescence of CTLA4 on CD4+ cells were higher in patients than in healthy controls (10.4 versus 4.7%, P < 0.05, 189% versus 149%, non significant, respectively). We found significantly elevated concentration of adiponectin in patients when compared to healthy subjects (15.4 versus 8.5 µl/ml, P < 0.05). We found that the adiponectin/BMI ratio correlated with the decrease of FEV(1) %. The results of this study support the possible role of CD4/CD25/CTLA4 cells and adiponectin in the systemic inflammation in COPD.

Nontuberculous mycobacterial infections among patients suspected of pulmonary tuberculosis.

The purpose of this study was to present a retrospective analysis of the frequency of nontuberculous mycobacteria (NTM)-related pulmonary infections among the AFB-positive and/or culture-positive patients in the Warsaw region who were suspected of tuberculosis (TB) and hospitalized in the university hospital between 1999 and 2005. All the AFB-positive pulmonary samples were examined with a molecular method using the Amplicor MTB test (Roche) for detection of Mycobacterium tuberculosis complex, and all mycobacterial isolates were speciated by high performance liquid chromatography (HPLC) analysis of mycolic acids. Patients who met clinical, radiological, and bacteriological criteria of mycobacteriosis were classified according to the American Thoracic Society (ATS) guidelines for diagnosis of NTM related disease. Among the 445 smear-positive or/and culture-positive patients, 142 subjects (31.9%) were found to be infected with M. tuberculosis. Among 303 non-TB patients, mycobacteriosis was found in 27 (8.9%) subjects. The frequency of NTM-related lung disease as compared to the bacteriologically-confirmed lung TB was estimated at 1:5. The rapid, precise methods of NTM speciation are necessary for progress in diagnostics of NTM related diseases.

Diagnostic utility of pleural fluid and serum markers in differentiation between malignant and non-malignant pleural effusions.

STUDY OBJECTIVE: To evaluate the diagnostic value of four different tumor markers: cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA 21-1) and neuron specific enolase (NSE) in patients with malignant and non-malignant pleural effusion. MATERIAL AND METHODS: One hundred and two patients with pleural effusion treated in the University Hospital in Warsaw between 2001 and 2003 were studied. They underwent an extensive, diagnostic work-up in order to determine the pleural effusion etiology. Patients with known pleural fluid etiology were labeled as the study group and submitted for further analysis. Pleural fluid and serum samples for CA-125, CEA, CYFRA 21-1 and NSE measurements were collected during the first thoracentesis, centrifuged, and frozen until further use. Pleural fluid and serum concentration of tumor markers were assessed by electrochemiluminescence methods using commercial kits. RESULTS: 74 patients (32 M, 42 F; mean age 65 +/-14 years) composed the final study group. Exudative pleural effusion was found in 62 patients; of these 36 were malignant (48.6% of all effusions), 20 parapneumonic (or pleural empyema), and 6 tuberculous. In 12 patients, pleural transudate was diagnosed. The highest diagnostic sensitivity for malignant pleural effusion was found for NSE (94.4% and 80.6% in the pleural fluid and serum, respectively). However, the specificity of NSE measurement was relatively low (36.1% and 47.4% in pleural fluid and serum, respectively). The most specific markers of malignant pleural fluid etiology were pleural fluid CYFRA 21-1 and CEA levels (92.1% and 92.1%, respectively). CA-125 was found to be the most specific serum marker of pleural malignancies (78.9%). The AUC for combined pleural markers was 0.89, combined serum markers 0.82, combined ratio pleural/serum markers 0.88. CONCLUSIONS: There are significant differences between the diagnostic value of various pleural fluid and serum markers. Overall, pleural fluid markers are superior to serum markers in determining the pleural fluid etiology. A combination of two or more tumor markers may help improve their diagnostic accuracy. Pleural fluid and serum measurements of different tumor markers play a limited role in the differentiation between malignant and non-malignant pleural effusions.

Pleural fluid adenosine deaminase and interferon gamma as diagnostic tools in tuberculosis pleurisy.

Several biological markers have been proposed to improve the efficacy of diagnosing tuberculous pleurisy. The study was undertaken to evaluate the accuracy of pleural fluid adenosine deaminase (ADA) activity and interferon-gamma (IFN-gamma) concentration in differentiating tuberculous pleural effusion (TPE) and nontuberculous pleural effusion (non-TPE). Ninety four patients (50 M and 44 F, mean age 60+/-18, range 18-95 years) with pleural effusion (PE) were studied. TPE was diagnosed in patients with: (i) positive pleural fluid or pleural biopsy culture or (ii) granulomas in the pleural biopsy specimen, after exclusion of other granulomatous diseases. Pleural fluid ADA activity was measured with the colorimetric method of Giusti, while IFN-gamma level was measured with ELISA. TPE was diagnosed in 28 patients. The non-TPE group consisted of 35 patients with malignant PE, 20 patients with parapneumonic effusion/pleural empyema, 5 with pleural transudate, and 6 with miscellaneous PE. The ADA activity and IFN-gamma concentration were significantly higher in TPE than in non-TPE (614.1+/-324.5 vs. 15.1+/-36.0 pg/ml, P<0.0001 and 75.1+/-39.1 vs. 11.0+/-16.6 U/l respectively, P<0.0001). The diagnostic sensitivity and specificity of IFN-gamma measurement (cut-off value of 75.0 pg/ml) were 100% and 98.5% respectively and were similar to those of ADA (100% and 93.9% at the cut-off value of 40.3 U/L). We conclude that pleural fluid ADA activity and IFN-gamma concentration are highly sensitive and specific markers of tuberculous pleurisy.

[Levels of CEA, antibodies against mycobacterial antigens and ACE activity in serum and in BALF in patients with sarcoidosis, tuberculosis and lung cancer--preliminary results]. / Stezenie CEA, przeciwcial skierowanych przeciw antygenom pratkowym oraz aktywnosc ACE w surowicy i w BALF chorych na sarkoidoze, gruzlice i raka pluca--wyniki wstepne.

UNLABELLED: Assessment of CEA concentration, levels of the antibodies against mycobacterial antigens A60 and 38 kDa and ACE activity in serum and in BALF of the patients with sarcoidosis (n = 8), tuberculosis (n = 13) and lung cancer (n = 10) was performed. Nine healthy volunteers were referred to as the control group. Sarcoidosis and lung cancer were confirmed histopathologically. Diagnosis of tuberculosis was stated using the radiometric Bactec 460 system. Serum and BALF CEA concentration and IgG levels against A-60 and 38 kDa were measured by ELISA: IMx, Immunozym and Pathozyme, respectively. ACE activity was assayed by the spectrophotometric method. Statistically significant (p < 0.01) increase in BALF-ACE activity in patients with sarcoidosis compared to each group was observed. Serum level of antibodies against 38 kDa and serum ACE activity were specific for tuberculosis and sarcoidosis respectively, but their sensitivity were very poor. Serum CEA concentration was fairly specific for lung cancer but its sensitivity was not satisfactory. CONCLUSIONS: 1. Determination of BALF-ACE activity may be helpful in diagnosis of sarcoidosis but it is recommended to continue the study on more numerous groups of patients taking into consideration polymorphism in the ACE gene. 2. High level of antibodies against 38 kDa and ACE activity in serum are specific for tuberculosis and sarcoidosis respectively but both tests have restricted application because of their poor sensitivity. 3. Serum CEA concentration could be considered only as an auxiliary test in the diagnosis of lung cancer.

Bioavailability of rifampicin, isoniazid and pyrazinamide from fixed-dose combination capsules.

SETTING: The absorption of rifampicin, isoniazid and pyrazinamide was tested after administration of each drug in free combinations and in a fixed-dose combination of the three drugs, known as Trifazid. OBJECTIVE: To examine the relative bioavailability of rifampicin, isoniazid and pyrazinamide after oral administration of the drugs given alone in comparison to that of the same drugs after administration of Trifazid. DESIGN: An open, randomized, cross-over study comprising 16 healthy volunteers. RESULTS: The pattern of absorption, plasma concentrations and pharmacokinetic parameters were very similar after administration of the drugs in free and fixed combinations. CONCLUSION: The triple combination of antituberculosis drugs could replace the separate drugs in the treatment of tuberculosis.

Isolation and characterization of an iron-containing superoxide dismutase from tomato leaves, Lycopersicon esculentum.

A cyanide-insensitive superoxide dismutase was purified from tomato leaves (Lycopersicon esculentum, Mill., var. Venture) to apparent homogeneity. The enzyme had twofold higher specific activity (about 4000 standard units) than ferric superoxide dismutases purified from Brassica campestris [Salin, M. L. and Bridges, S. M. (1980) Arch. Biochem. Biophys. 201, 369-374] and Nuphar luteum [Salin, M.L. and Bridges, S. M. (1982) Plant Physiol. 69, 161-165]. The protein had a relative molecular mass of about 42000 and was composed of two equal subunits noncovalently joined. It was negatively charged (pI = 4.6) and contained about 1.45 mol Fe/mol dimer and negligible amounts of Mn, Cu and Zn. Absorption spectrum and sensitivity to NaN3, H2O2 and temperature are also reminiscent of other ferric superoxide dismutases. Comparison of amino acid composition indicated, however, a closer relationship to the Mn-containing enzymes rather than to other Fe-containing superoxide dismutases. Two possible ways of Fe-containing superoxide dismutase acquisition by vascular plants were suggested.