Exhaled breath condensates from healthy children induce cell death of in vitro cultured cells by activation of apoptosis.

INTRODUCTION: Exhaled breath condensate (EBC) is a liquefied air, containing a mixture of non-volatile compounds, reflecting pathophysiological status of the bronchopulmonary system. Therefore, EBC analysis may be useful in diagnostics and monitoring of various respiratory diseases. In previous studies it was found that EBC from asthmatic children contained several regulators of angiogenesis. In vitro experiments with EBCs from children with asthma revealed their weak influence on proliferation of various cells. Surprisingly, EBCs from healthy children led to apoptosis of all tested cells. AIM: To assess the expression of selected apoptosis-related proteins in human and murine cells exposed to EBC from healthy children. MATERIAL AND METHODS: EBCs from healthy children were added to cultures of murine endothelial cells (C166) or human lung fibroblasts (HLF) to induce their apoptosis. For proteome analysis the apoptosis pathway-specific protein microarrays were used. RESULTS: The homogenates from EBC-treated C166 cells contained low amounts of Hsp27, which correlated with their fast death. Contrary to C166, the lysates from EBC-treated fibroblasts displayed increased amounts of Hsp27, which correlated with delayed HLF response to the induction of apoptosis. Except for increased caspase-3 in EBC-treated HLF, none of the other apoptosis regulators revealed any significant changes in that analysis. CONCLUSIONS: The screening of apoptosis pathways with microarray technology allowed identification of two molecules, Hsp27 and caspase-3, involved in cellular response to EBC. However, the factor responsible for induction of the cytotoxic effect of EBC from healthy children still remains unknown.

Influence of exhaled breath condensates from children with asthma on endothelial cells cultured in vitro. Do we really know everything about our breath condensate?

INTRODUCTION: Asthma-associated remodelling involves subepithelial fibrosis and increased vascularization of the bronchial wall. The latter may be associated with excessive production of several angiogenesis regulators which may be found in exhaled breath condensates (EBCs) collected from children with asthma. AIM: To assess the influence of EBC samples of asthmatic children and healthy controls on in vitro cultures of normal human lung microvascular endothelial cells (HLMVEC) and murine endothelial cell line (C-166). Moreover, the proteomic profile of cytokines in EBC samples was analysed. MATERIAL AND METHODS: Breath condensates collected from children with mild asthma (n = 10) and from healthy controls (n = 10) were used for experiments. Colorimetric tetrazolium salt reduction assay was used to evaluate the effect of EBCs on HLMVEC and C-166 cell lines. Furthermore, influence of EBCs on C-166 cell line was assessed using Annexin V-binding assay. The cytokine screening of EBC samples was performed using a proteome microarray system. RESULTS: The EBCs from patients with asthma revealed a weak inhibitory influence on human and murine endothelial cells. Surprisingly, EBCs from healthy children led to cell death, mainly by the induction of apoptosis. There were no statistically significant differences in the cytokine profile between EBC samples from children with asthma and healthy controls. CONCLUSIONS: Our preliminary report shows for the first time that the incubation of EBCs from healthy controls induced apoptosis in endothelial cells. The detailed mechanism responsible for this action remains unknown and requires further research.

Hydrogel chitosan sorbent application for nutrient removal from soilless plant cultivation wastewater.

In this study, we determined the effectiveness of removal of nutrients (nitrates and orthophosphates) from greenhouse wastewaters (GW) using non-cross-linked chitosan (CHs) and chitosan cross-linked with epichlorohydrin (CHs-ECH) in the form of hydrogel beads. GW used in the study had the following parameters: N-NO3 621.1 mg/L, P-PO4 60.8 mg/L, SO42- 605.0 mg/L, Cl- 0.9 mg/L, Ca2+ 545.0 mg/L, Mg2+ 178.0 mg/L, K+ 482.0 mg/L, hardness 113° dH, and pH 6.2. The scope of the study included determination of the effect of pH on wastewater composition and effectiveness of nutrient sorption, analyses of nutrient sorption kinetics, and analyses of the effect of sorbent dose on percentage removal of nutrients from GW. CHs-ECH was able to sorb 79.4% of P-PO4 and 76.7% of N-NO3 from GW, whereas CHs to remove 92.8% of P-PO4 and 53.2% of N-NO3.

Metastasising leiomyoma of the uterus with pulmonary involvement - case report.

Benign metastasising leiomyoma (BML) is characterised by extrauterine smooth muscle tumours in women after surgical treatment for uterine leiomyoma. Usually manifested as solitary or multiple focal lesions in various organs, it imposes a scrutinous diagnostic work-up to exclude a malignant disease and requires confirmation in microscopic examination of the extrauterine focus. The authors present a case of a 56-year-old woman with BML manifesting as bilateral multiple pulmonary nodules, with a tentative diagnosis of a disseminated malignant disease of mesenchymal origin. The patient underwent multiple diagnostic tests, which excluded malignancy. The definite diagnosis was established after the microscopic reevaluation of an excised pulmonary nodule. The patient is monitored with chest magnetic resonance. BML should be considered in the differential diagnosis of multiple pulmonary nodules in asymptomatic women. Patients with BML require long-term monitoring, therefore the selected imaging method should not carry the risk of cumulative side effects.