Clarithromycin Decreases IL-6 Concentration in Serum and BAL Fluid in Patients with Cryptogenic Organizing Pneumonia.

BACKGROUND: Inflammatory cytokines are involved in the development of cryptogenic organizing pneumonia (COP). It has been shown that macrolides inhibit cytokine production in the alveolar macrophages of COP patients. OBJECTIVES: The aim of the study was to assess the concentrations of interleukin 1ß (IL-1ß), IL-6, IL-8 and transforming growth factor ß (TGF-ß) in serum and in bronchoalveolar lavage fluid (BAL-f) in COP patients treated with clarithromycin (CAM). MATERIAL AND METHODS: The study involved 26 patients (18 women and 8 men, mean age 56.46 ± 8.83 years) with biopsy-proven COP. After being treated with CAM, a complete recovery was achieved in 22 patients, while four patients did not respond to the treatment. The ELISA method was used to measure the serum and BAL-f concentrations of IL-1ß, IL-6, IL-8 and TGF-ß. RESULTS: Before treatment, the serum IL-1ß1, IL-6, IL-8 and TGF-ß1 concentrations were similar in responders and non-responders. Significant decreases in serum concentrations of IL-6 (8.98 ± 13.26 pg/mL vs. 3.1 ± 6.95 pg/mL; p = 0.005), IL-8 (20.14 ± 25.72 pg/mL vs. 10.14 ± 6.8 pg/mL; p = 0.007) and TGF-ß1 (37.89 ± 12.49 ng/mL vs. 26.49 ± 12.45 ng/mL; p = 0.001) were found after treatment, as well as a significant decrease in the BAL-f concentration of IL-6 (30.56 ± 56.78 pg/mL vs. 4.53 ± 5.84 pg/mL; p = 0.036). Clarithromycin treatment resulted in a significantly lower mean value of serum IL-6 responders than non-responders. CONCLUSIONS: In COP patients, response to clarithromycin treatment was associated with decreases in serum concentrations of IL-6, IL-8 and TGF-ß, and of rations, and of the BAL-f concentration of IL-6.

Correlation of serum vascular endothelial growth factor-D concentration with clinical presentation and course of lymphangioleiomyomatosis.

BACKGROUND: Increased serum vascular endothelial growth factor D (VEGF-D) concentration has been accepted as a diagnostic marker in lymphangioleiomyomatosis (LAM). The study was performed to evaluate the correlation of VEGF-D with clinical presentation and course of LAM. MATERIAL: The study group comprised of 48 women with LAM (27 with sLAM, 9 with sLAM and lymphangioma (sLAM-LYM) and 12 patients with TSC/LAM). Patients were assessed at the time of VEGF-D examination, and pulmonary function parameters were compared with those, obtained one year before. VEGF-D serum concentration was measured by ELISA method. RESULTS: Patients with TSC/LAM and sLAM-LYM displayed higher concentrations of VEGF-D than patients with sLAM (2682 ± 1347 pg/mL and 2223 ± 1184 pg/mL vs.1281 ± 791 pg/mL; p = 0.0002, p = 0.009) respectively. Patients with sLAM and VEGF-D concentration <800 pg/mL (sLAM-L) had better lung function as assessed by FEV1 (2.38 ± 0.88 L vs. 1.75 ± 0.8 L; p < 0.015) and DL,CO (5.8 ± 2.25 vs. 3.93 ± 1.74 mL/min/mmHg; p < 0.028), had higher blood oxygenation, then those with VEGF-D >800 pg/mL (sLAM-H). Significant yearly increase of TLC (390 ± 700 mL; p < 0.021) and RV (340 ± 790 mL; p < 0.03), and decrease of distance in 6MWT (-30 ± 50 m; p = 0.04) were observed in sLAM-H group. Lung function parameters remained constant in sLAM-L patients. Patients with sLAM-H displayed higher yearly decline of FVC (120 vs. 50 mL; p = 0.035) and increase of TLC (390 vs. -80 mL; p = 0.038) and RV (340 vs. 90 mL; p = 0.045) than sLAM-L patients. Negative correlations between VEGF-D concentration and DL,CO, PaO2, PaCO2, and positive with HRCT grading, and desaturation in 6MWT were noticed in sLAM patients without lymphangioma. CONCLUSIONS: Serum VEGF-D is the useful biomarker of LAM extension, and might also prove predictive towards therapeutic decision.

The efficacy of EGFR gene mutation testing in various samples from non-small cell lung cancer patients: a multicenter retrospective study.

INTRODUCTION: Testing for the epidermal growth factor receptor (EGFR) gene mutations requires considerable multidisciplinary experience of clinicians (for appropriate patient selection), pathologists (for selection of appropriate cytological or histological material) and geneticists (for performing and reporting reliable molecular tests). We present our experience on the efficacy of routine EGFR testing in various types of tumor samples and the frequency of EGFR mutations in a large series of Polish non-small cell lung cancer (NSCLC) patients. METHODS: Deletions in exon 19 and substitution L858R in exon 21 of EGFR gene were assessed using real-time PCR techniques in 1,138 small biopsies or cytological specimens and in 1,312 surgical samples. RESULTS: Out of 2,450 diagnostic samples (containing >10% of tumor cells), the occurrence of EGFR gene mutations was 9%; more frequently in women (13.9%) and adenocarcinoma patients (10%), particularly with accompanying expression of TTF1 (13.0%). The frequency of EGFR gene mutations was similar in cytological and histological specimens, and in primary and metastatic lesions, and did not depend on the percentage of tumor cells and quality of isolated DNA. Cytological or small biopsy, compared to surgical specimens showed lower percentage of tumor cells, with no impact on the quality of real-time PCR assay. CONCLUSION: Cytological and small biopsy samples with low (10-20%) content of tumor cells and specimens from metastatic lesions are a sufficient source for EGFR mutation testing in NSCLC patients. The incidence of EGFR gene mutations in examined population was similar to those reported in other Caucasian populations.

Role of serum vascular endothelial growth factor D in discrimination of patients with polycystic lung diseases.

INTRODUCTION: Polycystic lung diseases (PLDs) include numerous rare diseases including lymphangioleiomyomatosis (LAM), pulmonary Langerhans cell histiocytosis (PLCH), and lymphocytic interstitial pneumonia. In these cases, diagnosis is based on a histological examination of open lung biopsy samples; however, it is not always possible to perform this procedure. Serum markers characteristic for a given entity are still being sought. OBJECTIVES: The aim of the study was to determine the usefulness of assessing serum vascular endothelial growth factor D (VEGF­D) concentration in the differential diagnosis of LAM and other PLDs (OPLDs). PATIENTS AND METHODS: Serum VEGF­D levels were measured by an enzyme­linked immunosorbent assay in 75 patients with PLDs including 29 women with LAM and 46 patients with OPLDs (28 women and 18 men). RESULTS: Serum VEGF­D levels were significantly higher in patients with LAM (median, 1557 pg/ml; interquartile range [IQR], 636-2593 pg/ml) than in all patients with OPLDs (median, 292 pg/ml; IQR, 233-405 pg/ml, P <0.0001) or than in women with OPLDs (median, 344 pg/ml; IQR, 243-452 pg/ml, P <0.0001). The serum VEGF­D level exceeding 468 pg/ml identified LAM patients with the specificity of 90% and sensitivity of 87% (area under the curve of 0.908; 95% confidence interval, 0.820-0.996). In none of the patients with OPLDs serum VEGF­D concentrations exceeded 800 pg/ml. CONCLUSIONS: An increased serum VEGF­D level is a highly specific biomarker useful in a differential diagnosis of LAM and OPLDs.

[Pneumocystosis in a patient with lymphangioleiomyomatosis]. / Pneumocystozowe zapalenie pluc u chorej na limfangioleiomiomatoze

A 47-year-old, non-smoking woman was admitted to the National Tuberculosis and Lung Diseases Research Institute for diagnosis of progressive exertional dyspnoea and numerous small thin-walled, air-filled cysts equally distributed throughout both lungs revealed in HRCT (high resolution computed tomography) examination. Histological assessment of specimens obtained by open lung biopsy revealed proliferation of immature smooth muscle, showing the expression of the antigen HMB45. On this basis, diagnosis of lymphangioleiomyomatosis was established. The disease caused essential ventilation damage of the lungs (FEV1 1.34 L; 39.71% pred, VC 4.02 L; 94.96% pred, FEV1/ /VC 0.33-4 1.81% pred, DLCO 3.65 mmol/min/Kpa 38.35% pred).During the observation, despite the lack of immunological disorders, the patient developed Pneumocystis jiroveci pneumonia (PCP) that was treated with trimethoprimsulfamethoxazole. Lymphangioleiomyomatosis is a rare disease which results from a defect of TSC genes. The disease is not related to immunological defects or disorders. However, the considerable cystic destruction of the lungs can predispose the patient to opportunistic infections such as the one in the presented case.

[Probiotics in the prevention and treatment of allergic diseases]. / Rola probiotyków w profilaktyce i leczeniu chorób alergicznych.

Physiological gastrointestinal microflora dominated by lactic acid bacteria is crucial for the maturation and proper functioning of human immune system. Thus, lactic acid bacteria eradication followed by intestinal colonization by other anaerobes seems to play an important role in the pathogenesis of numerous diseases, including allergy. This paper discusses the effect of physiological intestinal microflora on the physiological immune reactivity as well as its immunomodulatory potential. The critical review of current research on the effectiveness of probiotic dietary supplementation in the prevention and treatment of allergic diseases is provided.

Simple flow cytometric protocol of CD4+/CD8+ lymphocyte ratio assessment in bronchoalveolar lavage fluids from patients with interstitial lung diseases.

OBJECTIVE: To validate the fast and accurate flow cytometric (FCM) protocol using blood-standardized antibodies for alveolar lymphocyte subtyping with respect to standard immunocytochemistry (IC). STUDY DESIGN: FCM and IC were applied to immunophenotype T cell subsets in bronchoalveolar lavage (BAL) fluids from patients with interstitial lung diseases. Diagnostic BAL specimens from 50 patients with suspected sarcoidosis, idiopathic pulmonary fibrosis, and hypersensitivity pneumonitis were evaluated by both IC and FCM. In FCM, CD4+ and CD8+ T cells were identified by light scatter gating with CD3 selection using basic tricolor cytometer. RESULTS: Relative amounts of CD4+, CD8+ T cells, and CD4+/CD8+ ratios demonstrated by the FCM showed excellent, significant correlations with IC results. FCM values did not differ significantly from IC results. However, the sensitivity and specificity of conventional IC staining were not sufficient to assess CD4+/ CD8+ ratio in most idiopathic pulmonary fibrosis cases. Additionally, performing IC immunophenotyping in BAL samples with low lymphocyte content introduced a remarkable error into CD4+/CD8+ ratio assessment. CONCLUSION: FCM allowed reliable, precise, and fast T-cell subset measurement in all BAL samples, overcoming the IC disadvantages. Our validated FCM protocol provides diagnostically relevant CD4+/CD8+ ratio determination by simple light scatter gating strategy with CD3 selection.

[Cell phenotype determines PAI-1 antiproliferative effect - suppressed proliferation of the lung cancer but not prostate cancer cells]. / Fenotyp komórek warunkuje antyproliferacyjne dzialanie PAI-1 -hamujacy wplyw na aktywnosc proliferacyjna komórek raka pluca.

INTRODUCTION: Plasminogen inhibitor activator type 1 (PAI-1) is an important regulator of tumor growth and metastasis formation acting directly via specific urokinase complexing or indirectly due to its affinity to vitronectin. We have shown previously that PAI-1 modifies angiogenic activity of endothelial cells in a dose-dependent manner but also in close relationship to the cell phenotype. Present study aimed on evaluating the PAI-1 effect on the proliferative activity of lung cancer cells (A549), prostate cancer cells (DU145) as well as endothelial cells (HUVEC). RESULTS: Mutated PAI-1 (1, 10, 100 microg/mL) characterized by the prolonged antifibrinolytic activity (T1/2 approximately 7000 h) inhibited proliferation of lung cancer A549 cells in a dose-dependent (p < 0.001) and time-dependent (p < 0.001) manner. No significant effect on the DU145 prostate cancer cells has been observed except of the 72 h cultures with highest PAI-1 concentration (100 microg/ml) (p < 0.001). Proliferative activity of endothelial cells (HUVEC) was affected by 100 microg/ml PAI-1 only, and independent of the culture period (24, 48 and 72 h, p < 0.001). CONCLUSION: Plasminogen inhibitor activator type 1 modulates cell proliferation via antifibrynolitic mechanizm time- and dose-dependently, however final outcome is strongly affected by the cell phenotype.

[Diagnostics of selected respiratory virus infections]. / Diagnostyka wybranych zakazen wirusowych ukladu oddechowego.

Viral infections are the most common infectious diseases of the respiratory tract characterized by the considerable mortality (especially among children and elderly people) and considered as the significant economic burden. It has been demonstrated that implementation of rapid diagnostic methods enabled more appropriate treatment of respiratory viral infections, reduced mean hospitalization time and cost, as well as resulted in the significantly decreased mortality. Modern diagnostic methods effectively identify respiratory virus, its antigen or nucleic acids in biological samples by means of the immunological and molecular techniques. This article presents critical overview of those methods with particular emphasis on their clinical usefulness and clinical reliability.