Effect of phoshpodiesterase 4 (PDE4) inhibibtors on eotaxin expression in humen bronchial epithelial cells.

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.

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.

Effects of the phosphodiestrase-4 inhibitor rolipram on lung resistance and inflammatory reaction in experimental asthma.

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.

Effect of IFN-gamma stimulation on expression of intercellular adhesion molecule-1 (ICAM-1) on alveolar macrophages in patients with non-small cell lung cancer.

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.

Reactivity of alveolar macrophages in lung cancer patients and healthy subjects: surface ICAM-1 after INF-gamma stimulation in vitro.

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.

CD4/CD8 lymphocytes in BALF during the efferent phase of lung delayed-type hypersensitivity reaction induced by single antigen inhalation.

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.

[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.

[The effect of 6 week treatment with theophylline on spirometric parameters, nonspecific bronchial reactivity and the concentration of soluble interleukin 2 receptor in serum of asthmatic patients]. / Wplyw 6-tygodniowego leczenia teofilina o przedluzonym dzialaniu na parametry spirometryczne, nieswoista nadreaktywnosc oskrzeli i stezenie receptora dla interleukiny 2 w surowicy krwi u chorych na astme oskrzelowa.

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.

[Angioedema is caused by a defect in C1-inhibitor synthesis]. / Obrzek naczynioruchowy zalezny od defektu syntezy C1-inhibitora.

Deficit of the first component of complement inhibitor (C1-inhibitor, C1-inh) may clinically be manifested as angioedema. The disease is characterized by episodic swellings of mucosa and subcutaneous tissue at different locations of the body. Laryngeal swelling can be life-threatening. The major mediators of edema are discussed to be bradykinin and C2b derived peptides. These mediators increase capillary permeability. Antifibrinolytic agents (aminocaproic acid, tranexamic acid) and attenuated androgens (danazol or stanazolol) are used for prophylaxis. Prolonged use both of them might result in more or less severe side effects. In experiments in vitro it has been shown that IFN-gamma, IL-1, IL-6 have a stimulatory effect on C1-inh synthesis. We want to verify the practical use of probiotics as natural inductors of IFN-gamma synthesis for elevating C1-inh level.

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.

Serum and bronchoalveolar IgG against A60 and 38 kDa antigens in the diagnosis 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.

[The effect of salmeterol on the number of hypodense eosinophils, eosinophilic cationic protein levels ECP in serum, bronchial hyperresponsiveness and lung function test in patients with bronchial asthma]. / Wplyw salmeterolu na liczbe granulocytów kwasochlonnych hipodensyjnych, stezenie eozynofilowego bialka kationowego ECP w surowicy, badania czynnosciowe ukladu oddechowego i nadreaktywnosc oskrzelowa u chorych na astme oskrzelowa.

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.

[The effect of an isotonic solution of theophylline 300 on eosinophil cationic protein levels]. / Wplyw izotonicznego roztworu teofilinum 300 na stezenie eozynofilowego bialka kationowego.

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.

[Clinical evaluation of the isotonic solution of theophylline for intravenous injection]. / Ocena kliniczna izotonicznego roztworu teofiliny do wlewów dozylnych.

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.

[Clinical, radiologic and functional changes in the respiratory system and changes in cytology of bronchoalveolar lavage fluid of patients with idiopathic pulmonary fibrosis]. / Zmiany kliniczne, radiologiczne i czynnosciowe ukladu oddechowego oraz zmiany w skladzie komórkowym plynu z plukania oskrzelowo-pecherzykowego u chorych na samoistne wlóknienie pluc.

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.

[Effect of 16-week use of salmeterol on ECP levels, pulmonary function tests and bronchial hyperreactivity in patients with chronic obstructive lung disease]. / Wplyw 16-tygodniowego stosowania salmeterolu na stezenie ECP, badania czynnosciowe pluc i nadreaktywnosc oskrzeli u chorych z przewlekla obturacyjna choroba pluc.

The aim of the study was to compare the influence of salmeterol on lung function tests (VC, FEV1), bronchial hyperreactivity (PC20) and eosinophil cationic protein (ECP) level in serum and in sputum in 10 patients with COPD. FEV1 was less than or equal 70% of predicted values but greater than 0.6 L. FEV1%VC less or equal 60% (reversibility after 200 micrograms salbutamol between 5-15%. Patients were treated with 2 x 50 micrograms salmeterol for 16 weeks. Lung function test didn't change during the observation but hyperreactivity had decreased (0.08 + 0.1 microgram/ml before, 0.24 + 0.2 microgram/ml after 4 weeks and 0.4 + 0.16 microgram/ml after 16 weeks) ECP level had decreased in serum (53 +/- 24 micrograms/l before, 32 +/- 12 micrograms/l after 4 weeks and 36 +/- 13 micrograms/l after 16 weeks) and in sputum (2345 +/- 789 micrograms/g before 1342 +/- 894 micrograms/g after 4 weeks, 1742 mu 698 micrograms/g after 16 weeks). We concluded that salmeterol didn't change lung function test but decreased bronchial hyperreactivity and decreased ECP level in serum and sputum.

[Correlation of ECP levels in sputum and blood with parameters of airflow obstruction and hyperreactivity in patients with irreversible bronchial obstruction due to chronic bronchitis]. / Korelacja stezenia ecp w plwocinie i surowicy ze stopniem obturacji dróg oddechowych i stopniem nadreaktywnosci oskrzeli u chorych z nieodwracalna obturacja w przebiegu przewleklego zapalenia oskrzeli.

The aim of this study was to find if there was any correlation between the serum and sputum eosinophil cationic protein (ECP) concentrations with the parameters of airflow obstruction as well as degree of bronchial hyperreactivity. 18 patients aged 40-75 years with irreversible bronchial obstruction (FEV1 > 0.61 or < 70% of the predicted value, FEV1%VC < 60%) without eosinophilia and high hyperreactivity (PC20 0.02 mg/ml -0.4 mg/ml) were examined. The FEV1 reversibility after 400 micrograms MDI salbutamol was 5-10%. ECP was measured by radioimmunosorbent assay using the ECP RIA kit (Pharmacia). The range of the standard curve was 2-100 micrograms ECP/L, while all assays were run in duplicate. The bronchial challenge was performed with histamine. The challenge was stopped when the FEV1 had decreased by > 20%. Serum ECP concentrations ranged from 0.0001-0.063 mg/ml, whereas sputum ECP concentrations were between 0.0002-1.84 mg per g sputum. There was no correlation between sputum and serum ECP concentrations and the degree of hyperreactivity and were not related to the parameters of hyperreactivity and were not related to the parameters of airway obstruction.

In vitro and in vivo effect of salbutamol on histamine release from human basophils.

The effect of salbutamol, a selective beta 2-adrenergic agonist, on immunological release of histamine from leukocytes isolated from blood of 14 allergic, asthmatic patients, was investigated after in vivo or in vitro drug administration. Simultaneously, the effect of this drug on peak expiratory flow rate (PEFR) was estimated. Histamine was assayed by the single isotope-enzymatic method. Obtained results have confirmed that salbutamol is a rather poor inhibitor of histamine release from basophils. Although both administered salbutamol doses (0.5 or 1 mg, i.v.) significantly increased PEFR (p less than 0.01), inhibitory effect on histamine release by allergen was seen only after higher dose administration in vivo (p less than 0.01 against control). Similarly, a small but significant decrease in histamine release by both concanavalin A (by 20%, p less than 0.01) and allergen (by 30%, p less than 0.025) was seen in vitro after incubation of leukocytes with 8 x 10(-6) mol/l salbutamol, whereas 4 x 10(-7) mol/l salbutamol was without effect.

[Lipid proteinosis of the pulmonary alveoli--diagnostic difficulties]. / Lipoproteinoza pecherzyków plucnych--trudnosci diagnostyczne.

It is the first polish case of diagnosis of alveolar lipid proteinosis diagnosed during life is described. The diagnosis was based on biochemical analysis and morphological assessment in electron microscope of the fluid obtained in bronchoalveolar large in a patient aged 18 years with disseminated radiological pulmonary changes. Diagnostic difficulties are discussed comparing them with the cases of this disease reported as yet in Poland. The importance of bronchoalveolar lavage for the diagnosis is emphasized.