Molecular Beacon for Detection miRNA-21 as a Biomarker of Lung Cancer.

Lung cancer (LC) is the leading cause of cancer-related death worldwide. Although the diagnosis and treatment of non-small cell lung cancer (NSCLC), which accounts for approximately 80% of LC cases, have greatly improved in the past decade, there is still an urgent need to find more sensitive and specific screening methods. Recently, new molecular biomarkers are emerging as potential non-invasive diagnostic agents to screen NSCLC, including multiple microRNAs (miRNAs) that show an unusual expression profile. Moreover, peripheral blood mononuclear cells' (PBMCs) miRNA profile could be linked with NSCLC and used for diagnosis. We developed a molecular beacon (MB)-based miRNA detection strategy for NSCLC. Following PBMCs isolation and screening of the expression profile of a panel of miRNA by RT-qPCR, we designed a MB targeting of up-regulated miR-21-5p. This MB 21-5p was characterized by FRET-melting, CD, NMR and native PAGE, allowing the optimization of an in-situ approach involving miR-21-5p detection in PBMCs via MB. Data show the developed MB approach potential for miR-21-5p detection in PBMCs from clinical samples towards NSCLC.

Inhaler training and asthma knowledge are associated with a higher proportion of patients with correct inhaler technique in young but not in elderly asthmatic patients.

Objective: Incorrect inhaler usage is frequent, particularly in elderly asthmatic patients. This study aimed at comparing inhaler technique errors and their determinants, as well inhaler technique self-perception versus real performance, between elderly and non-elderly asthmatics. Methods: Cross-sectional assessment of 92 elderly and 100 non-elderly asthmatics followed at specialty clinics. A standardized questionnaire was applied and inhaler technique demonstration was requested. Errors were assessed using checklists based on manufacturers' instructions and inhaler technique was graded as correct, acceptable or incorrect. Chi-Square Test and Fischer's Exact Test were used for comparative analysis of nominal variables. A p value equal to or less than 0.05 was considered statistically significant. Results: Inhaler technique was correct in a minority of elderly and young patients, without significant differences between study groups. Only 11.1% of the elderly who classified their inhaler as easy and 12.7% who stated their technique was correct had no errors. Previous regular inhaler training was associated with better actual performance in young but not in elderly patients. Conclusion: Our study showed that in spite of regular follow up at specialized outpatient clinics, inhaler devices are associated with a high frequency of errors in elderly and non-elderly asthmatics. In addition, most patients tend to overestimate their technique as correct. Finally, previous, frequent training was associated with a significantly higher percentage of patients showing correct or acceptable technique but only in non-elderly asthmatics, which suggests that elderly asthmatics may need specifically tailored inhaler education programs.

Factors Affecting Usage of a Digital Asthma Monitoring Application by Old-Age Asthmatics Living in Inner Central Portugal.

Purpose: To analyse factors affecting the ability to use the digital asthma monitoring application Mask-Air® in old-age individuals living in inland Portugal. Patients and Methods: In this observational study, patients with medically confirmed asthma who agreed to participate were interviewed and subdivided into Non-users Group: those who could not use the application and Users Group: those who could. Sociodemographic and psychological data, comorbidities, and asthma status were compared between groups. Assessment of reasons for refusal was based on a 6-item questionnaire. Results: Among the 72 sequentially recruited patients (mean age±SD 73.26±5.43 yrs; 61 women; 11 men), 44 (61.1%; mean age±SD 74.64±5.68 yrs; 38 women; 6 men)) were included in Non-users Group and 28 (38.9%; mean age±SD 71.11±4.26 yrs; 23 women; 5 men) in Users Group. Non-users Group patients were significantly older, had lower socioeconomic level, and more frequently had severe asthma (25% vs 3.6%; Odds ratio=0.08 (95% CI=0.01-0.81; p=0.033)) and diabetes (32.6% vs 7.4%; Odds ratio=0.17 (95% CI=0.03-0.80; p=0.025)) than Users Group. The main reasons for not using the App were "Lack of required hardware" (n=35) and "Digital illiteracy" (n=26), but lack of interest to use the App among those who had conditions to use it was uncommon. Conclusion: Most old-age asthmatics living in Beira Interior either lack a smartphone or digital skills, which are significant obstacles to implementing app-based monitoring studies.

This study was done to see whether it was possible to use a mobile phone application (App) to help old-age asthmatics living in inner Central Portugal better monitor and self-manage their disease. The researchers interviewed a group of 72 patients with proven asthma who agreed to participate in the study. This group was subdivided into two subgroups: Non-users Group (44 patients) included those who could not use the App because they did not have a smartphone; Users Group (28 patients) included those who had all the conditions to use the App. Patients were helped to download the App (called MASK-Air), were given a thorough explanation about it, and about how it should be used on a daily basis to monitor their asthma symptoms. The researchers found that patients in Non-users Group were significantly older, had worse socioeconomic conditions, and more often had severe asthma and diabetes. They also discovered that the main reasons for not using the App were lack of a smartphone and not knowing how to use a smartphone. These results show that lacking a smartphone and not knowing how to use digital tools are frequent situations in old-age asthmatics living in inner Central Portugal, and these may be obstacles for patients in monitoring their own asthma symptoms.

Targeting nucleolin by RNA G-quadruplex-forming motif.

A high level of nucleolin (NCL) expression is often associated with a poor prognosis of patients with lung cancer (LC), suggesting that NCL can be used as a possible biomarker. NCL has been shown to display a marked preference for the binding to G-quadruplexes (G4). Here, we investigate the formation of an RNA quadruplex structure in a sequence found in the human precursor pre-MIR150 with the potential to recognize NCL. Circular dichroism (CD) spectra of pre-MIR150 G4-forming sequence (designated by rG4) indicate the formation of a parallel quadruplex structure in KCl or when complexed with the well-known G4 ligand PhenDC3. The thermal stability of rG4 is very high, and further increases in the presence of PhenDC3. The binding affinities of rG4 to PhenDC3 and NCL RBD1,2 are similar with KD values in the nanomolar range. PAGE results suggest the formation of a ternary quadruplex-ligand-protein complex (rG4-PhenDC3-NCL RBD1,2), indicative that PhenDC3 does not prevent the binding of rG4 to NCL RBD1,2. Finally, rG4 can recognize NCL-positive cells and, when fluorescently labeled, can be used as a probe for this protein. ELISA experiments indicate altered NCL expression patterns in liquid biopsies of LC patients in a non-invasive manner, potentially helping the diagnosis, prognosis, and patient response to treatment. Hence, labeled rG4 could be used as a detection probe of LC in liquid biopsies.