MicroRNAs are small non-coding RNAs that regulate immune response and inflammation. We assumed that miRNAs may be involved in the immune response during cystic fibrosis pulmonary exacerbations (CFPE) and that altered expression profile in the airways and blood may underlie clinical outcomes in CF pediatric patients. Methods: We included 30 pediatric patients diagnosed with cystic fibrosis. The biologic material (blood, sputum, exhaled breath condensate) was collected during pulmonary exacerbation and in stable condition. The miRNA expression profile from blood and sputum (n = 6) was done using the next-generation sequencing. For validation, selected four miRNAs were analyzed by qPCR in exosomes from sputum supernatant and exhaled breath condensate (n = 24). NGS analysis was done in Base Space, correlations of gene expression with clinical data were done in Statistica. Results: The miRNA profiling showed that four miRNAs (miR-223, miR-451a, miR-27b-3p, miR-486-5p) were significantly altered during pulmonary exacerbation in CF patients in sputum but did not differ significantly in blood. MiRNA differently expressed in exhaled breath condensate (EBC) and sputum showed correlation with clinical parameters in CFPE. Conclusion: MiRNA expression profile changes in the airways during pulmonary exacerbation in CF pediatric patients. We suggest that miRNA alterations during CFPE are restricted to the airways and strongly correlate with clinical outcome.
Introduction Bronchial asthma is a chronic respiratory disease characterized by airway inflammation, allergen‐induced hypersensitivity and dyspnea. Most asthmatic patients demonstrate oscillations of disease symptoms within 24 hours regulated by circadian clock genes. We hypothesized that these genes may be regulators of childhood asthma risk. Objectives The aim was to investigate whether single‐nucleotide polymorphisms (SNPs) in the circadian clock genes are associated with childhood asthma risk. We also aimed to analyze the mRNA level of clock genes in the blood of asthmatic children and NHBE cells stimulated with IL‐13. Materials and Methods Peripheral blood was collected from 165 asthmatic and 138 healthy Polish children. NHBE cells were culture at the air‐liquid interface (ALI) with IL‐13 as an in vitro model of allergic inflammation. Using TaqMan probes, we genotyped 32 SNPs in: CLOCK, BMAL1, PER3 and TIMELESS. Expression analysis for TIMELESS was performed using real‐time PCR with SYBR Green. For haplotype and genotype statistical analysis we used Haploview 4.2 and STATISTICA version 12, respectively. Gene expression analysis was performed in DataAssist v3.01. Results We found that three polymorphisms in TIMELESS (rs2291739, rs10876890, rs11171856) and two haplotypes (TTTT and CTAC) were associated with asthma risk. We also found significantly decreased expression of TIMELESS in the blood of asthmatic children as compared to the healthy children (P = 0.0289) and in NHBE cells stimulated with IL‐13 (P = 0.0302). Conclusions In our study, we showed for the first time that TIMELESS variants and expression may be associated with childhood asthma.
Immunization is a very effective health intervention. Moreover, the global vaccines market has been growing rapidly and costs of full children immunization are dramatically higher nowadays than they were 30 years ago. High vaccine prices may limit affordability of vaccination which is why we attempted an evaluation of availability and affordability of 7 and non-reimbursed vaccines mostly used in children in Poland. The study was conducted between October 2016 and October 2017 using a specially designed anonymous questionnaire comprising three closed-ended questions. The study tool was distributed by direct contact or via the Internet. Eventually, answers from 505 pharmacies from the Greater Poland region and 10 primary care clinics were included. 5 out of 7 vaccines were available in all types of facilities. There were some issues, however, with availability of BexseroAE and NimenrixAE. Considering prices, the highest difference (of more than 100%) was found for Infanrix hexaAE and the lowest (38.5%), for Infanrix IPV+HIBAE. In 88.17% of pharmacies included, patients were informed about a thermo-insulating package. 48.39% of respondents indicated that such package is free of charge, while in other pharmacies an average price for the package was EUR 0.48. Although availability and affordability of medicines are crucial objectives of the public health policy, it seems that access to vaccines in Poland might be an area for improvement. Thus, prices of non-reimbursed vaccines could be regulated in Poland nationwide. Moreover, to provide trustworthy information concerning vaccination, healthcare decision makers should consider social education about immunization as an important issue.
Due to their structural conservation and functional role in critical signalling pathways, non-coding RNA (ncRNA) is a promising biomarker and modulator of pathological conditions. Most research has focussed on the role of microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs). These molecules have been investigated both in a cellular and an extracellular context. Sources of ncRNAs may include organ-specific body fluids. Therefore, studies on ncRNAs in respiratory diseases include those on sputum, bronchoalveolar lavage fluid (BALF) and exhaled breath condensate (EBC). It is worth identifying the limitations of these biosamples in terms of ncRNA abundance, processing and diagnostic potential. This review describes the progress in the literature on the role of ncRNAs in the pathogenesis and progression of severe respiratory diseases, including cystic fibrosis, asthma and interstitial lung disease. We showed that there is a deficit of information on lncRNAs and circRNAs in selected diseases, despite attempts to functionally bind them to miRNAs. miRNAs remain the most well-studied, but only a few investigations have been conducted on the least invasive biosample material, i.e., EBC. To summarise the studies conducted to date, we also performed a preliminary in silico analysis of the reported miRNAs, demonstrating the complexity of their role and interactions in selected respiratory diseases.
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