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Airway epithelium acts as multifunctional site of response in the respiratory tract. Epithelial activity plays an important part in the pathophysiology of obstructive lung disease. In this study, we compare normal human epithelial cells from various levels of the respiratory tract in terms of their reactivity to pro-allergic and pro-inflammatory stimulation. Normal human nasal, bronchial and small airway epithelial cells were stimulated with IL-4 and IL-13. The expressions of the eotaxins IL-6 and CXCL8 were evaluated at the mRNA and protein levels. The effects of pre-treatment with IFN-γ on the cell reactivity were measured, and the responses to TNF-α, LPS and IFN-γ were evaluated. All of the studied primary cells expressed CCL26, IL-6 and IL-8 after IL-4 or IL-13 stimulation. IFN-γ pre-treatment resulted in decreased CCL26 and increased IL-6 expression in the nasal and small airway cells, but this effect was not observed in the bronchial cells. IL-6 and CXCL8 were produced in varying degrees by all of the epithelial primary cells in cultures stimulated with TNF-α, LPS or IFN-γ. We showed that epithelial cells from the various levels of the respiratory tract act in a united way, responding in a similar manner to stimulation with IL-4 and IL-13, showing similar reactivity to TNF-α and LPS, and giving an almost unified response to IFN-γ pre-stimulation.

Background: Asthma and COPD are non-infectious inflammatory diseases of the respiratory tract. Allergic rhinitis can be assumed as an intermediate condition between healthy and asthmatic state. Eotaxins are important indicators of allergic reaction. They are strong chemoattractants mainly for eosinophils but also for other cells., Objective: We measured the level of eotaxin expression and inflammatory cell count in the material from nasal brushing in healthy controls and in patients with allergic rhinitis, asthma, and COPD. We studied the correlation between the eotaxin gene expression level in the material from nasal brushing and respiratory tests in asthma and COPD patients., Methods: Expression of eotaxins was measured using quantitative RT-PCR. Number of eotaxin transcript copies was evaluated using real time PCR standard curve method., Results: Of all eotaxins CCL24 had the highest expression in the material from nasal brushing, and its level was increased in allergic asthma. CCL11 was significantly increased in the material from nasal brushing of COPD patients. Increased levels of all three eotaxins were observed in the material from nasal brushing of patients with allergic rhinitis in season. The levels of CCL26 expression and FEV1/FVC factor were correlated negatively in the asthma group and positively in the COPD group., Conclusions: Eotaxins are crucial factors of allergic, asthmatic and also COPD inflammatory reactions. Our results suggest a dual role of CCL26 - it can act as a negative regulator for neutrophils in COPD, while in asthma it may act as a chemoatractant of eosinophils and other cells into the lung., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Bronchoalveolar lavage (BAL) is a useful technique for differential diagnosis of various interstitial lung diseases (ILDs) and is usually realized by analysis of the differential cell count. This study was conducted to estimate the value of bronchoalveolar lavage fluid (BALF) total cell count (TCC) in the diagnosis of ILD. We analyzed 237 BAL samples from patients with ILD: sarcoidosis (SA), idiopathic pulmonary fibrosis (IPF), cryptogenic organizing pneumonia (COP), hypersensitivity pneumonitis (HP), chronic eosinophilic pneumonia (CEP), and smoking-related ILD (sr-ILD). The control group consisted of 30 healthy volunteers. The statistical analysis revealed significant differences in the BALF TCC between healthy controls and patients with SA, IPF, HP, COP, sr-ILD, and eosinophilic disorders (mean values 6.9 vs. 14.5, 22.5, 22.8, 20.7, 64.5, and 27.3 × 10(6), respectively). Logistic regression revealed a significant relation between the TCC and ILD diagnosis. We conclude that the TCC, as well as the value of total number of inflammatory cells, should be reported in the description of BAL.

The bronchial epithelium is a very important factor during the inflammatory response, it produces many key regulators involved in the pathophysiology of asthma and COPD. Local influx of eosinophils, basophils, Th2 lymphocytes and macrophages is the source of many cytotoxic proteins, cytokines and other mediators of inflammation. These cells are attracted by eotaxins (eotaxin-1/CCL11, eotaxin-2/CCL24, eotaxin-3/CCL26). Inhibitors of phosphodiesterase 4 (PDE4) are new anti-inflammatory drugs which cause cAMP accumulation in the cell and inhibit numerous stages of allergic inflammation. The aim of our study was to evaluate the influence of PDE4 inhibitors: rolipram and RO-20-1724 on the expression of eotaxins in human primary bronchial epithelial cells. Cells were preincubated with PDE4 inhibitors for 1 hour and then stimulated with IL-4 or IL-13 alone or in combination with TNF-α. After 48 hours, eotaxin protein level was measured by ELISA and mRNA level by real time PCR. These cells produce CCL24 and CCL26. PDE4 inhibitors increased CCL24 and CCL26 mRNA level irrespectively of the used stimulators. Rolipram and RO-20-1724 had no effect on eotaxin protein production in our experimental conditions. Thus PDE4 inhibitors have no effect on eotaxin protein expression in human primary bronchial epithelial cells. In vitro experiments should be performed using a primary cell model rather than immortalized lines.

Background: IL-6 is strongly implicated in the development of chronic obstructive pulmonary disease (COPD). IL-13 is the well-documented central mediator in allergic asthma. IL-6 is attributed to the proinflammatory activities in COPD as well as asthma. In COPD patients exacerbation is increased by serum IL-6. The association of IL-13 as well as IL-6 with the impaired respiratory function of asthma patients remains controversial., Objectives: The aim of this study was to compare the concentration of IL-6 and IL-13 in the induced sputum of asthma and COPD patients, and to assess the possible association of these cytokines with the impairment of lung function., Methods: Twenty-six subjects with COPD and 18 subjects with asthma were enrolled in this study. IL-6 and IL-13 levels were measured in induced sputum by ELISA and correlated with the results of respiratory tests., Results: The induced sputum of COPD patients had a significantly higher IL-6 level than the sputum of asthma subjects while no significant differences were found in the levels of IL-13. There was a statistically significant negative correlation between IL-6 level and FEV(1) or FEV(1)/FVC in asthma patients (r = -0.59 and -0.54, respectively) and a negative correlation that did not reach statistical significance between IL-6 level and FEV(1), FEV(1)% or FVC in COPD subjects (r = -0.30, -0.30 and -0.38, respectively). There was no relationship between concentrations of IL-13 and impaired respiratory function., Conclusions: Our results confirmed that IL-6, but not of IL-13, is associated with respiratory disorders in both asthma and COPD patients., (Copyright © 2012 S. Karger AG, Basel.)

Introduction: This is the retrospective study on diagnostic effectiveness of two nucleic acid amplification tests: AMPLICOR Mycobacterium tuberculosis (MTB) and Xpert MTB/RIF. The first one was included into diagnostic procedure for tuberculosis from 1999 to 2009 and the second one has been used from November 2009., Material and Methods: Study groups comprised 1875 samples for AMPLICOR MTB (104 were inhibited), and 213 samples for Xpert MTB/RIF., Results: The assay sensitivity was 81.9% for AMPLICOR MTB and 81.8% for Xpert MTB/RIF, and specificity was 97.2% and 99.5% respectively, as compared to the culture on Loewenstein-Jensen medium. However, sensitivity of each test correlated with Ziehl-Neelsen staining. For AFB+ samples, assay sensitivity was 97.8% for AMPLICOR MTB and 100% for Xpert MTB/RIF and for AFB- samples it was 58.1% and 50% respectively., Conclusions: The reported results show a high diagnostic usefulness of Xpert MTB/RIF test for samples, which are positive for Ziehl-Neelsen staining.

Unlabelled: The increasing number of eosinophils into bronchoaelvolar space is observed during noninfectious inflammatory lung diseases. Eotaxins (eotaxin-1/CCL11, eotaxin-2/CCL24, eotaxin-3/CCL26) are the strongest chemotactic agents for eosinophils. Inhibitors of phosphodiesterase 4 (PDE4), the enzyme decomposing cAMP, are anti-inflammatory agents which act through cAMP elevation and inhibit numerous steps of allergic inflammation. The effect of PDE4 inhibitors on eotaxin expression is not known in details. The aim of our study was to evaluate the influence of PDE4 inhibitors: rolipram and RO-20-1724 on expression of eotaxins in bronchial epithelial cell line BEAS-2B. Cells were preincubated with PDE4 inhibitors or dexamethasone for 1 hour and then stimulated with IL-4 or IL-13 alone or in combination with TNF-α. After 48 hours eotaxin protein level was measured by ELISA and mRNA level by real time PCR., Results: PDE4 inhibitors decreased CCL11 and CCL26 expression only in cultures co-stimulated with TNF-α. In cultures stimulated with IL-4 and TNF-α rolipram and RO-20-1724 diminished CCL11 mRNA expression by 34 and 37%, respectively, and CCL26 by 43 and 47%. In cultures stimulated with IL-13 and TNF-α rolipram and RO-20-1724 decreased expression of both eotaxins by about 50%. These results were confirmed at the protein level. The effect of PDE4 inhibitors on eotaxin expression in BEAS-2B cells, in our experimental conditions, depends on TNF-α contribution.

Introduction: The GenoType Mycobacterium CM and the GenoType Mycobacterium AS (HAIN Lifescience, Germany) were evaluated for the ability to differentiate mycobacterial species of clinical isolates. Serial use of the both assays is aimed to identify 38 different molecular patterns, of which 24 patterns can be assigned to single species, 10 patterns are allocated to two or more Mycobacterium species, and 4 patterns correspond to Mycobacterium species and gram-positive bacteria with a high G + C content. The analysis of mycolic acids by high pressure liquid chromatography (HPLC) was the reference method., Material and Methods: A set of 127 nontuberculous mycobacterial isolates on Loewenstein-Jensen slants, derived from different patients between 1999 and 2007, was analyzed. The strains were primary classified by HPLC following the diagnostic procedure, and retyped by GenoType Mycobacterium CM/AS., Results: In total, results obtained by both methods were interpretable for 113 strains. Concordant results were obtained for 105 (93%) mycobacterial strains. One out of 8 inconcordant classified strains, which was classified as M. abscessus/M. chelonae by HPLC, displayed a pattern of M. tuberculosis complex by a molecular method. Eleven clinical strains were differentiated only by one of used methods, either by HPLC (6 strains) or by GenoType CM/ AS (5 strains). Three strains were not classified at all., Conclusions: Our results show that the GenoType Mycobacterium CM/AS system represents a useful tool to identify mycobacterial clinical isolates. The molecular system is as rapid and reliable as the HPLC, but much easier to perform and more friendly for the environment.

Introduction: The GenoType system (HAIN Lifescience, Germany) offers new perspectives of detecting the tuberculous and non-tuberculous mycobacteria at the molecular level. The system compromises five independent tests that could be performed either on direct specimens or isolated strains, to identify the strains and test the resistance against rifampin and isoniazid. Up to now, non GenoType test was applied in Poland. The aim of the study was an evaluation the accuracy of GenoType MTBC test in speciation of the clinical isolates, previously classified as M. tuberculosis complex by HPLC analyze of mycolic acids., Material and Methods: 161 clinical isolates, derived from the TB patients hospitalized in the Warsaw Medical University Hospital between 1999 and 2007 were assayed., Results: On the basis of the hybridization patterns, all 161 studied strains were identified as M. tuberculosis/M. canettii., Conclusions: 1. The GenoType MTBC test (HAIN Lifescience, Germany) precisely recognizes M. tuberculosis complex. The 100% accordance in speciation of M. tuberculosis by the GenoType MTBC test as compared to HPLC method was demonstrated. The GenoType MTBC test can replace HPLC in detection of tuberculous mycobacteria in clinical isolates. 2. As the GenoType MTBC test performs well, the other tests of GenoType system may be considered to be verified in diagnostic procedure of mycobacterial infection.

Asthma is associated with eosinophilic airway inflammation and eosinophils are believed to be important in the pathogenesis of asthma. IL-5 has been considered the central mediator for eosinophilic proliferation, differentiation and eosinophilic inflammation, but results of recent studies suggest that besides IL-5, eotaxin may contribute to the pathogenesis of asthma. Eotaxin is CC chemokine first isolated from guinea pig bronchoalveolar lavage. It selectively binds to a specific receptor (CCR3) highly expressed on eosinophils, basophils, and mast cells being important in the pathogenesis of asthma. Eotaxin is produced mainly by epithelial cells of lung and gut, to mediate organ preferential attraction of eosinophils. Production of eotaxin is stimulated by IL-4, IL-13, TNF(-alpha). Human eotaxin family includes: eotaxin-1 (CCL11), eotaxin-2 (CCL24) and eotaxin-3 (CCL26). It seems that eotaxin-3 may be expressed following allergen challenge. Studies with glucocorticosteroids have shown some inhibitory effect on eotaxin production in cell culture in vitro however, very little in vivo data exists in humans relating to corticosteroid effects on chemokine levels. CCR3 receptor is considered as the possible therapeutic target in asthma treatment.

The aim of this study was to evaluate the influence of pretreatment with the phosphodiesterase-4 inhibitor rolipram on pulmonary resistance, influx of inflammatory cells, and histamine concentration in bronchoalveolar lavage fluid (BALF) during an experimental asthmatic reaction induced in ovalbumin (OA)-sensitized guinea pigs, challenged with OA inhalation. The experiment was performed in three groups of guinea pigs: two experimental groups, pretreated with rolipram or dexamethasone, and a control group without any pretreatment. Lung resistance (LR) was continuously recorded under suppression of spontaneous breathing during early asthmatic reaction. BALF was obtained before and at three time points up to 24 hr after the challenge. In the untreated, control animals a transient, significant increase in neutrophils, total and CD4+ lymphocytes, macrophages, eosinophils, and in histamine concentration in BALF was noted. Pretreatment with rolipram significantly reduced LR, eosinophils infiltration, and histamine release into the bronchoalveolar space during the early asthmatic reaction. These effects were generally comparable with those of dexamethasone, except that dexamethasone also reduced the influx of neutrophils into BALF.

An impairment of in vitro cytotoxicity and tumoricidal function of alveolar macrophages (AMs) in patients with lung cancer was reported in a number of studies. The aim of our study was to evaluate the expression of intercellular adhesion molecule-1 (ICAM-1) on AMs after stimulation with interferon-gamma (IFN-gamma) in patients with non-small cell lung cancer (NSCLC). The study was performed in 13 patients with NSCLC, 6 patients with various nonmalignant pulmonary diseases, and 6 healthy volunteers. AMs were isolated from bronchoalveolar lavage fluid (BALF) by adherence and then cultured with or without IFN-gamma for 24 h. The expression of ICAM-1 on AMs was analyzed by flow cytometry. Stimulation with IFN-gamma caused increased expression of ICAM-1 on AMs in all studied groups (p < 0.05). The degree of the increase in ICAM-1 expression on AMs after IFN-gamma stimulation was significantly lower in patients with NSCLC compared with healthy volunteers (p = 0.002) or the other patients (p = 0.022). The results suggest impaired reactivity of ICAM-1 expression on AMs after stimulation with IFN-gamma in patients with NSCLC, which might be involved in functional defects of AMs in patients with NSCLC.

Introduction: Transforming growth factor (TGF)-beta is one of numerous inhibitory factors produced by cancer cells that regulate antitumor immunity. The aim of this study was to evaluate TGF-beta1 levels and lymphocyte subsets in the broncholaveolar lavage fluid (BALF) of patients with primary lung cancer and to analyze the interdependence of these parameters., Materials and Methods: BALF samples were collected from 38 patients with primary lung cancer prior to treatment and from 23 healthy volunteers. Concentrations of TGF-beta1 were measured in two independent lots of samples using a commercially available sandwich ELISA kit after concentration of the supernatants. Differential cell counts in the BALF were performed on slides stained with the May Grünwald Giemsa method. Flow cytometry with monoclonal antibodies was applied for lymphocyte phenotyping., Results: A higher level of TGF-beta1 in the BALF of patients compared with the healthy subjects was observed in both lots of samples (3.23+/-2.96 pg/ml vs. 1.05+/-0.95 pg/ml, p<0.05, and 16.1+/-19.3 pg/ml vs. 10.1+/-11.1 pg/m,, respectively, difference not significant). There was significant positive correlation of the TGF-beta1 level with the proportion of lymphocytes and negative correlation with both the proportion of macrophages and the percentage of cytotoxic and activated T lymphocytes., Conclusions: Our findings confirmed that TGF-beta takes part in the local response in the course of primary lung cancer.

The polymorphism of the short fragment of the heat shock protein 65 encoding gene was evaluated by the PCR - RFLP technique described by Telenti and further developed by Devallois for identification of mycobacterial species in routine laboratory work. We analysed 58 strains representing 25 different mycobacterial species (24 reference strains and 34 clinical isolates). The results obtained by PCR-RFLP and HPLC identification techniques were highly concordant The results were compatible for 87,5% (21 / 24) reference strains and for 97,1% (33/34) clinical isolates. The PCR - RFLP method allowed for accurate identification mycobacterial species, especially pathogenic strains. Restriction patterns obtained for 25 species of Mycobacteriaceae genus could help in constructing the data base and algorithms used in routine laboratory practice.

Selective phosphodiesterases (PDE) inhibitors are the new group of antiasthmatic drugs, which integrate antiinflammatory activity with bronchoconstriction counteraction. Selective inhibitors of phosphodiesterase type 4 are used as alternative or assist drugs in treatment of respiratory system diseases. So far glucocorticosteroids remain the most efficient and widely used medicine in the treatment of asthma. However application of glucocorticosteroid is greatly limited because of numerous side effects, what induce to permanent search for new antiasthmatic drugs. Examination new substances are executed on animal models. Guinea pig model is widely used to research course of asthmatic reaction. This model is especially convenient on the ground of that: lung is major shock organ, airway respond to histamine, animals demonstrated early asthmatic reaction (EAR) and late asthmatic reaction (LAR), eosinophils flow in bronchoalveolar space during LAR. In ovalbumin (OA) sensitized guinea pigs hypersensitivity reaction breaks out as a result of OA provocation. Aims of our experiments, execute on guinea pig model were to determine the influence of rolipram (PDE 4 inhibitor) on modulation experimental asthmatic reaction and comparison activity of rolipram versus dexamethasone in attribution to chosen parameters of allergic reaction such as: lung resistance, influx of protein and inflammatory cells in airways, and mastocytes degranulation. Experiments were made on guinea pigs sensitized and provoked with ovalbumin The obtain data indicate that rolipram was effective in reduction the rise of lung resistance during EAR, restricted influx of eosinophils to bronchoalveolar space between 1,5 and 24 hours after provocation, and reduced increase of histamine concentration in bronchoalveolar lavage fluid (BALf). Rolipram had no influence on number of neutrophils present in BALf. Dexamethasone in double dose of 1,2mg/kg effectively bordered the growth of lung resistance during EAR, and broke influx of eosinophils and neutrophils to bronchoalveolar space.

Unlabelled: The diagnosis of NTM-related pulmonary disease is based on clinical symptoms, radiological features and several positive cultures of one and the same NTM species from samples obtained from the respiratory tract. Short hospitalization usually does not enable sufficient diagnostic procedures to meet the diagnostic criteria, and this may lead to the reduction of diagnostic sensitivity. The aim of the study was to draw attention to NTM-related pulmonary disease, to share the authors' experience in the diagnosing of pulmonary mycobacteriosis and to indicate the possibilities of improving the diagnostic accuracy in this disease. A group of 31 patients with sputum, bronchial washing and/or bronchoalveolar lavage fluid (BALF) NTM-positive cultures was selected from a cohort of 245 patients evaluated for tuberculous and nontuberculous mycobacterial diseases (total number of 1277 specimens were invastigated). In two of them NTM related pulmonary disease was diagnosed (caused by M. kansasii and M. avium) at the course of initial evaluation. In the remaining 29 patients the microbiological data did not allow to establish the diagnosis of mycobacterial lung disease mainly due to a small number of samples from the respiratory tract. From this group 13 patients were reevaluated within 3 - 6 months from the initial investigation. This allowed to identify two new cases of mycobacteriosis (M. kansasii and M. avium). Thus among 31 patients with NTM positive cultures from respiratory tract specimens 4 patients (4/31, 12,9%) met the diagnostic criteria for mycobacterial disaease., Conclusion: Microbiological analysis of an adequate number of samples in symptomatic patients with radiological features suggestive for NTM-related pulmonary disease increses the diagnostic sensitivity in pulmonary mycobacteriosis. Identification of the species in positive cultures is of great importance.

The aim of this study was to evaluate the utility of the quantitative analysis of mycolic acids by HPLC technique in drug susceptibility testing of the M. tuberculosis isolates to the first-line antituberculous drugs: isoniazid and rifampicin. Drug susceptibility of the 30 clinical M.tbc isolates was examined by the mycolic acids analysis with HPLC technique and results were compared to the proportion method on solid L-J medium and liquid medium in MGIT system. In HPLC method drug susceptibility of M. tuberculosis strains was described by TAMA index defined as the ratio of the total area under mycolic acids peaks (TAMA) from cultures with drug to the TAMA of control. At critical concentrations of drugs, TAMA indexes of resistant strains were >0.5, and TAMA indexes of susceptible strains were <0.05. The average error of the TAMA analysis was +/- 9.5% The quantitative analysis of mycolic acids by HPLC gives results compatible with standard proportion method and is a reliable method for determination of drug susceptibility of M. tuberculosis.

Numerous studies have been performed to determine diagnostic or prognostic utility of tumor markers in patients with lung cancer. The aim of the study was to evaluate the diagnostic usefulness of the tumor markers CA 125, CEA and CYFRA 21-1 in bronchoalveolar lavage fluid (BALF) in patients with non-small cell lung cancer (NSCLC). BAL was performed in 13 patients with NSCLC during diagnostic bronchofibroscopy. The control group consisted of 12 patients with sarcoidosis and 13 healthy volunteers. Tumor markers were determined in BALF supernatants using electrochemiluminescence technique (Elecsys 1010, Roche). To determine optimal cut-off values of tumor markers in BALF ROC curve was used. CEA and CA 125 concentration in BALF were significantly higher in NSCLC patients than in healthy volunteers and patients with sarcoidosis. CYFRA 21-1 in BALF was higher in NSCLC patients than in healthy volunteers, but no significant difference was found between NSCLC and sarcoidosis patients. The cut-off values of BALF concentration of CA 125, CEA and CYFRA 21-1 were 95 IU/mL, 3 ng/ml and 3 ng/ml, respectively. The sensitivity and specificity of CEA and CA 125 in BALF were 100%, 84% and 92%, 80%, respectively. In conclusion, we suggest that among the chosen markers, determination of CEA in BALF is the most useful in diagnosis of NSCLC. It may be a complementary method in diagnosing of patients in whom tumor cannot be visualized by bronchofibroscopy. These results need confirmation in larger groups of patients.

Over the last decades the incidence of infections with non-tuberculous mycobacteria (NTM) has increased. It has been noted in various regions of the world and it seems to concern also our country. The aim of this review is to call attention to clinical presentation and diagnostic criteria of mycobacterioses. The environmental sources of NTM, predispositions to dissemination of NTM infection and current epidemiological data on mycobacterioses are discussed. Since the accuracy of mycobacterial strain identification is most important for microbiological diagnosis of disease, it is suggested to perform it according to modern requirements, exclusively in selected, well-equipped laboratories.

The linearity of ICAM- I expression on alveolar macrophages (AM) before and after INF-gamma stimulation in healthy and lung cancer subjects were compared. AM were collected by bronchoalveolar lavage and incubated with/without INF-gamma according to standard procedures. The harvested cells were analyzed by flow cytometry using monoclonal antibodies against leucocytes and macrophages. Only viable cells were analyzed. Stimulation with INF-gamma revealed two AM subpopulations of similar size differentiated in the intensity of ICAM-1 expression. They were not distinctly marked in every studied case. Our preliminary results did not confirm the previously reported decreasing reactivity of AMs after INF-gamma stimulation in lung cancer patients.

The aim of the study was to estimate the utility of the HPLC-based method of mycolic acids analysis to classify Mycobacterium species in routine diagnostic procedure on the basis of own three-year experience. 2142 patients' specimens were examined. 141 AFB were cultured. 36.2% strains were classified as M. tuberculosis complex by HPLC. The identification was confirmed by AMPLICOR MTB (Roche diagnostic, USA). M. xenopi (17.0%), M. kansasii (14.2%) and M. gordonae (14.2%) were the most frequent identified out of nontuberculous mycobacteria. Four mycobacteriosis cases were suspected because of repeated identification of the isolated strains. 136 strains on L-J slant shipped from other centres were identified. We confirm that the HPLC method is highly effective and specific for Mycobacterium species classification, which can be performed in no more than a couple of hours. In our opinion it is a very helpful tool, hard to replace in diagnostic procedure of tuberculosis and mycobacteriosis.

Unlabelled: ECP released from the granules of activated eosinophils is regarded to be a marker of airway inflammation in asthma. The study was performed to compare the usefulness of measuring serum and sputum ECP for monitoring the asthma treatment. 29 subjects with mild to moderate asthma (mean age 41 +/- 17) were admitted in exacerbation (FEV1 55.54 +/- 87.49% N). 10 subjects with grass pollen asymptomatic asthma and 10 healthy subjects were also enrolled in the study. Patients with symptomatic asthma were ordered 30 mg prednisone for 2 weeks and they continued during next 2 weeks inhaled budesonide therapy. The concentrations of ECP (mcg/L) were determined by CAP-system (Pharmacia). The total eosinophil count and serum ECP in all subjects treated orally and next by inhaled GKS didn't differ statistically. The highest sputum ECP concentration was determined in exacerbation of asthma 84.5 +/- 78 mcg/L and statistically were reduced after 2-weeks of prednisone treatment 24.4 +/- 12.1 mcg/L (p = 0.05). In following 2 weeks of budesonide treatment sputum ECP concentration was statistically negligible in relation to previous treatment in spite of increasing tendency (50 +/- 61.3 mcg/L (p = 0.2394). In asymptomatic grass pollen asthma sputum ECP concentration was 19.7 +/- 9.4 mcg/L, higher than in controls 12 +/- 5.8 mcg/L (p = 0.04). There were a significant correlations between total eosinophil count and serum (r = 0.6396) and sputum ECP(r = 0.4683) in exacerbation., Conclusions: 1. In asthma exacerbation elevated sputum ECP concentration was observed. 2. In consequence of prednisone treatment the sputum ECP concentration was reduced. 3. Sputum ECP measurement is more accurate than serum ECP for monitoring the effectiveness of treatment. 4. Sputum ECP concentration is a sensitive parameter which discriminate asymptomatic patients with asthma from healthy subjects.

HPLC is the most useful method to analyze various species of mycobacteria by using mycolic acids. The purpose was to prepare a library containing chromatographic patterns of mycolic acids derived from reference species Mycobacterium, which had been cultivated in standard conditions. 28 reference strains (27 ones from American Type Culture Collection and one cultivated from the vaccine M. bovis BCG) were used. The analysis of mycolic acids involved chromatographic separation of their bromophenacyl derivatives according to Centers for Disease Control recommendation. Mycolic acids profiles formed by HLPC were reproducible for all reference species in this study. Standard deviation of relative retention time of every peak did not exceed 2.5%. The species included into M. tuberculosis complex beyond M. bovis BCG shared the same mycolic acids pattern. HPLC is the only mean to distinguish M. tuberculosis from M. bovis BCG. The other studied stains had species specific patterns which differed from M. tuberculosis complex and M. bovis BCG. The prepared library comprising 28 reference elution profiles of mycolic acids from known mycobacteria species can be applied in diagnostic procedure of tuberculosis and mycobacteriosis.

Background: The precise mechanisms involved in the pathogenesis of hypersensitivity pneumonitis (HP) have not been identified. HP is characterized by inflammatory lymphocytic alveolitis and a remarkable increase in T-lymphocytes detected in bronchoalveolar lavage fluid (BALF). It is suggested that both CD4+ and CD8+ T cells may contribute to the pathogenesis of HP. Experiments on animal models suggest that cell mediated immunity (CMI) is more important for the pathogenesis of HP than complex-mediated immunity, but the relationship between the subsets of BALF lymphocytes and humoral or cell-mediated allergic reactions is still not clear. The aim of our study was distinguish CD4+ and CD8+ T cells in BALF lymphocytes during a delayed-type hypersensitivity (DTH) reaction in the lung., Material and Methods: The experiment was performed on guinea pigs sensitized with BCG vaccine and subjected to a single inhalation of tubercle bacilli antigens (tuberculin). 24 hours after tuberculin provocation (at the time of maximum lymphocyte infiltration), bronchoalveolar lavage was performed on both sensitized and non-sensitized (control) animals. The total cell count was estimated, and a differential microscopical examination of BAL-fluid cells was performed, along with the phenotyping of BALF lymphocytes (by flow cytometry)., Results: In the BALF of the sensitized animals, as compared to the controls, there was a statistically significant increase in the percentage and absolute count of T-lymphocytes, CD4+ and CD8+. The CD4 / CD8 ratio in both groups did not differ significantly and was individually variable (2.94I0.72 SEM in the experimental group, vs 4.41I1.29 SEM in the control group)., Conclusions: Both CD4+ and CD8+ lymphocytes (with some predominance of helper cells) participate in the efferent phase of the delayed type hypersensitivity reaction in the lung induced by antigen inhalation.

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.

This study allowed to investigate the effect of theophylline (Theo-Dur Astra) given 1 tablet á 200 mg per day in the evening for six weeks on histamine-induced airways responsiveness++ (PC20H), spirometric parameters and the concentration of soluble interleukin 2 receptor (sIL-2R) in serum. 21 patients with mild asthma (6 women, 15 men), average age 34.8 +/- 11.4 years were qualified for this study. PC20H was estimated according to Cockroft's method using pneumatic inhalator (Voyage-Secura Nova). Concentration of sIL-2R in serum was measured by Predicta Interleukin 2 Receptor Kit (Genzyme). Lung function test was measured by means of ABC PNEUMO (abc MED). Six week treatment with theophylline significantly increased PC20H in comparison with basal results (0.74 +/- 0.68 mg/ml before and 0.89 +/- 0.71 mg/ml after p < 0.04). The concentration of sIL-2R in serum did not change significantly during theophylline therapy. Treatment with theophylline significantly increased FEV1 (3.64 l/sec before and 3.84 l/sec after p < 0.001). The anti-inflammatory effect of theophilline is more clearly shown in the decrease of histamine-inducted airways responsiveness than in the decreased concentration of sIL-2R in serum.

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.

Setting: The ELISA test is generally accepted for serodiagnosis of tuberculosis. Its sensitivity and specificity depend on the antigen used and data are not always concordant. In some patients suspected of tuberculosis, sarcoidosis or lung cancer, bronchoalveolar lavage (BAL) is performed. BAL fluid (BALF) contains immunoglobulins--their source is transudation from the serum but local IgG synthesis of IgG is not excluded., Objective: The comparison of BALF versus serum in the serodiagnosis of pulmonary tuberculosis, and its differentiation from sarcoidosis and lung cancer., Design: The IgG against A60 and 38 kDa antigens in serum and BALF were measured using the commercially available IgG EIA kits. The study included 13 tuberculous patients, 8 patients with sarcoidosis, 10 patients with lung cancer and 9 healthy volunteers., Results: In the serum studies the sensitivity of the assays did not exceed 40% and 60% in the estimation of IgG 38 kDa and IgG A60, respectively. The specificity of both assays was similar: 81% (IgG 38 kDa) and 69-75% (IgG A60). In BALF studies the sensitivity of the IgG 38 kDa assay was 50% and its specificity was 44%. The ranges of the IgG A60 concentrations in BALF were similar in all subject groups., Conclusion: In differentiation between tuberculosis, sarcoidosis and lung cancer, the measurement of IgG against A60 and 38 kDa antigens in serum is more useful than that in BALF.

Unlabelled: The aim of the study was to compare the influence of salmeterol xinofoate on blood hypodense eosinophil numbers ECP level in serum and sputum, and on lung function tests (VC, FEV1), bronchial hyperresponsiveness (PC20) Lung function tests (FEV1, VC), bronchial histamine challenge (PC20) and measurements of hypodense eosinophils number and ECP level in sputum and serum were done before two and six weeks after treatment with 50 micrograms salmeterol twice daily. Hypodense eosinophils were measured by Kuo et all, ECP by radioimmunosorbent assay using ECP RIA kit Pharmacia. Normal range of ECP is 2.3-15 micrograms/L Hyperreactivity by standard bronchial provocation techniques. The lung function tests (VC, FEV1, FEV1%VC) were measured with Pneumoscreen., Result: Eosinophil number decrease after treatment: 42.7 +/- 18.3 before vs 23.4 +/- 8.7 after. ECP level had decreased in serum 53 +/- 24 micrograms/L before and 32 +/- 12 micrograms/L after 2 weeks and 36 +/- 13 micrograms/L after 6 weeks: in sputum 2345 +/- 489 micrograms/g before and 1342 +/- 894 micrograms/g after 2 weeks 1741 micrograms/g after 6 weeks. Lung function tests had improved during the observation and hyperresponsiveness had decreased (0.08 +/- 0.1 microgram/ml before 0.24 +/- 0.2 microgram/ml after 2 week sand 0.4 +/- 0.16 after 6 weeks., Conclusion: ECP in serum and sputum in asthmatic patients is higher then in normal patients. Salmeterol improved lung function tests, decreased bronchial responsiveness, decreased hypodense eosinophils number and ECP concentration in serum and sputum., Clinical Implications: This date suggested that salmeterol modified bronchial responsiveness and protected the bronchial inflammation.

Unlabelled: The aim of the study was to compare the influence of theophylline 300 on eosinophil cationic protein (ECP)level in serum in asthmatic patients Theophylline was given twice a day. Measurements of ECP in serum and lung function tests (PEF) were done before and at the first and fifth day after treatment Theophylline concentration were measured before and after treatment on the first injection and second injection at the first and the fifth day. ECP in serum was measured by radioimmunosorbent assay using ECP RIA kit Pharmacia. Concentration of theophylline by Emit Theophylline Assay Syva Company. The lung function test (PEF) were measured with Wright Peak flowmeter, Result: ECP level had decreased in serum 37 + 15 micrograms/l before vs 30 + 12 micrograms/l on the first day and 17 + 9 micrograms/l on the fifth day of observation Lung function tests had improved during the observation.

The safety and tolerance of intravenous theophylline in asthmatic patients was investigated. The effect upon the lung function tests and frequency of cardiac arrhythmias were evaluated in 15 patients after five days treatment with theophylline 300 mg twice a day. Measurements of the peak expiratory flow (PEF) were obtained prior to theophylline administration, immediately after first intravenous infusion and 1.3 and 6 hours later and before and after second infusion, while serum theophylline levels were determined at the same time by enzyme immunoassay (Diagnostica Merc). The same measurements were made at the fifth day of observation. Continuous 24 hour Holter monitoring of the electrocardiogram before the study and at the fifth day of the treatment was performed. Biochemical investigations included aspartate and alanine aminotransferase (GGTPi GOT), sodium and potassium ion concentrations, serum levels of creatinine, glucose and bilirubin hematocrit, hemoglobin concentration, MCHC, red blood cells count and white blood cell differential count were determined. The therapeutic concentration of theophylline in serum were found in all subject treated with theophylline 300 mg twice a day. The improvement in lung function and no changes in cardiac rhythm were observed the biochemical findings did not demonstrate any significant differences after five days of the study. The study confirm safety and good tolerance of intravenous theophylline used in the doses 300 mg twice a day.

The course of the pulmonary fibrosis is difficult to estimate as there are no diagnostic tests specific and sensitive enough to assess disease activity. Together 33 patients with pulmonary fibrosis have been studied. They have been divided into 2 subgroups, depending on the intensity of clinical, radiologic, and functional disorders. In all patients bronchoalveolar lavage has been carried out, and the obtained results have been compared with those in 18 healthy individuals. Changes in the cellular composition of BAL fluid had polymorphic character. In the early phase of the disease, only percentage of lymphocytes in BAL fluid has been increased significantly whereas in the more advanced stage percentage of both neutrophils and eosinophils has also been significantly increased. The use of several parameters simultaneously helps to evaluate pulmonary fibrosis.