Inter-society consensus for the use of inhaled corticosteroids in infants, children and adolescents with airway diseases

Background: In 2019, a multidisciplinary panel of experts from eight Italian scientific paediatric societies developed a consensus document for the use of inhaled corticosteroids in the management and prevention of the most common paediatric airways disorders. The aim is to provide healthcare providers with a multidisciplinary document including indications useful in the clinical practice. The consensus document was intended to be addressed to paediatricians who work in the Paediatric Divisions, the Primary Care Services and the Emergency Departments, as well as to Residents or PhD students, paediatric nurses and specialists or consultants in paediatric pulmonology, allergy, infectious diseases, and ear, nose, and throat medicine. Methods: Clinical questions identifying Population, Intervention(s), Comparison and Outcome(s) were addressed by methodologists and a general agreement on the topics and the strength of the recommendations (according to the GRADE system) was obtained following the Delphi method. The literature selection included secondary sources such as evidence-based guidelines and systematic reviews and was integrated with primary studies subsequently published. Results: The expert panel provided a number of recommendations on the use of inhaled corticosteroids in preschool wheezing, bronchial asthma, allergic and non-allergic rhinitis, acute and chronic rhinosinusitis, adenoid hypertrophy, laryngitis and laryngospasm. Conclusions: We provided a multidisciplinary update on the current recommendations for the management and prevention of the most common paediatric airways disorders requiring inhaled corticosteroids, in order to share useful indications, identify gaps in knowledge and drive future research

Post-stroke delirium risk factors, signs and symptoms of onset and outcomes as perceived by expert nurses: A focus group study

Higher rates of delirium have been reported among patients with acute stroke. However, poorly modifiable risk factors have been documented to date while sign and symptoms capable of early detecting its onset and outcomes in this specific population have been largely neglected. The aim of this study was to emerge nurses' clinical knowledge and experiences regarding post-stroke delirium (a) risk factors, (b) signs and symptoms of delirium onset, and (c) outcomes

Isolation of Urinary Exosomal miRNAs: Comparative analysis of different methods

Exosomes can be detected in urine and are released from every segment of the nephron. Urinary exosomes harbor unique subset of proteins, reflecting their cellular source, and constitutively contain RNA. The presence of large amounts of small RNAs in exosomes suggests that exosomes may contain specific microRNAs that provide valuable information as noninvasive clinical biomarkers in both diagnostic and prognostic areas for patients with renal pathologies [1]. We report a comparative analysis of different methods for the isolation of urinary exosomes and the analysis of their microRNA content. We applied and compared several methodologies, including nanomembrane concentrators and Exoquick-TCTM precipitation reagent [2] for exosome purification; TRI reagent and various commercial kits (Qiagen, Ambion, miRcury and Seramir) for exosomal RNAs extraction. Purified RNA was subjected to standard validation such as nanodrop quantification, ribo-pico green measurements and bioanalyser analysis. Moreover all the samples were checked for microRNAs/mRNA/small RNAs content by RT-qPCR specific assays. Based on these results we selected the most reliable and convenient method for microRNAs extraction from urinary exosomes. The selection of the appropriate exosomal miRNA isolation method was dependent on the validation results in terms of RNA yield and the absence of contaminants

Urinary Exosomal miRNAs: Best Strategies for Isolation and Analysis

Background & Aims: Urinary exosomes are released from every segment of the nephron and harbor unique subset of proteins and RNA. The abundance of small RNAs in exosomes provides a beneficial \u201ctool\u201d to explore specific microRNAs which, in turn could be helpful as noninvasive clinical biomarkers in cardiovascular pathologies [1]. MicroRNAs are endogenous, small (20-22 nucleotides), non-coding RNAs that negatively regulate gene expression via degradation or translational inhibition of their target mRNAs [2]. The aim of this study was to compare in detail the available strategies for isolation of urinary exosomal microRNAs in combination with different methods for RNA purification. Methods: Urinary Exosomes were isolated using Ultrafiltration (Nanomembrane Concentrators), and Exoquick-TCTM precipitation reagent [3]. Exosomal RNA was isolated using TRI reagent and various commercial kits (Qiagen, Ambion, miRcury and Seramir). Purified total RNA was quantified using Ribogreen, small RNA quantification was obtained using Agilent Bioanalyzer and target miRNAs/mRNAs validation was done by RT-qPCR specific assays. Results: The combination of ultrafiltration for exosomes isolation; and Trizol-miRNeasy for exosomal RNA proved to be efficient compared with all the other methods. Conclusions: The selection of the appropriate urinary exosomal microRNAs isolation method was dependent on the total RNA yield, RNA purity and microRNA abundance. Further analysis in larger groups is required to reconfirm the efficiency of the developed method

Optimizing the purification and analysis of miRNAs from urinary exosomes.

Background: Exosomes are cytoplasm containing vesicles released by many cells that can be found in several biological fluids including urine. Urinary exosomes are released from every segment of the nephron, are detectable in urine, constitutively contain RNA (small RNAs and mRNAs) and harbour unique subset of proteins, reflecting their cellular source. Methods: With the aim to establish the optimal protocol for high throughput analysis of exosomal miRNAs, we compared three different urinary exosomes isolation methods and six RNA extraction techniques. Exosomal RNA yield, size and quality were assessed respectively by specific staining with fluorescent dyes, analysis of spectrophotometric parameters and capillary electrophoresis. MiRNAs detection and abundance was determined by RT-qPCR. Results: Among the exosomes isolation methods, Ultrafiltration resulted to be the most suited. The highest exosomal RNA yield quantified by RiboGreen® staining was obtained with the combination of TRI ReagentTM with miRNeasy®, followed by TRI ReagentTM, SeraMirTM, miRCURYTM, mirVanaTM and miRNeasy®; but after a multivariate analysis, SeraMirTM scored as the method of choice in terms of miRNA yield, purity and RT-qPCR miRNAs quantification accuracy. Storage conditions were also analysed, showing that the relative abundance of urinary exosomal miRNAs is not influenced by urine freezing. Conclusions: The selection of appropriate urinary exosomal miRNAs isolation method was dependent on various validation results. Ultrafiltration in combination with SeraMir™ exoRNA columns represents the optimal procedure for a rapid, cost-effective and efficient purification of miRNAs from urinary exosomes, perfectly suited for further applicative research in the field of miRNAs in kidney physiology and pathology

miRNAs purification and analysis from urinary exosomes.

Urinary exosomes are 40-100 nm membrane vesicles deriving directly from the kidney and the urogenital tract potentially via each epithelial cell facing the renal tubule lumen. They are released into urine and account for 3% of the total urinary proteome and may also be enriched in specific miRNAs related to renal homeostasis and pathologies. In this context urine can be very helpful as non-invasive source of selected population of exosomal miRNAs. We compared three different urinary exosomes isolation methods and six RNA extraction techniques aiming to establish a reliable method both for urinary exosomes isolation [2] and for the subsequent miRNA profiling [1]. Exosomal RNA yield, size, quality and miRNA detection abundance by Real Time qPCR were assessed. Finally, we could obtain a procedure allowing rapid, accurate and efficient purification and detection of miRNA from urinary exosomes, which is best suited for miRNA rather than large RNA detection. Additionally, the advantages and drawbacks of each different purification strategy were also highlighted. Following our experimental design we ended up with a reliable protocol for miRNAs isolation from urinary exosomes, which should be highly beneficial for further applicative research in the field of kidney biology and diseases

Isolation of Urinary Exosomal miRNAs using various methods

Exosomes are involved in a wide spectrum of physiological mechanisms such as immune system modulation, paracrine functions and cell to cell communications. They can be detected in urine being released from every segment of the nephron. Urinary exosomes (UEs) constitutively contain RNA (microRNAs/mRNA/small RNAs) and harbour unique subset of proteins, reflecting their cellular source. With the aim to establish the best method both for urinary exosomes isolation and for the subsequent RNA profiling analysis, we compared three different UEs isolation methods and six RNA extraction techniques. Exosomal RNA yield, quality and size were assessed respectively by specific staining with fluorescent dyes (Ribogreen\uae, RNA specific quantification kit), spectrophotometric quantification (Nanodrop\uae ND \u2013 1000 spectrophotometer) and capillary electrophoresis (Agilent 2100 Bioanalyzer). All the samples were analysed for detection of selected miRNAs and mRNAs by Real Time PCR specific assays. Based on these results the most reliable and convenient method for UEs miRNAs extraction and analysis was selected. Advantages and drawbacks of each methodology were also discussed

Fundamentals of care: revisione narrativa della letteratura

ABSTRACT Introduction Over the last decades, several research lines have been carried out to investigate factors hindering or promoting patient\u2019s safety and quality of care. Among them, the Fundamentals of Care (FOC) framework has reached an increased relevance worldwide. The aim of this narrative review is to summarize the debate on FOC\u2019s concept, as well as its commonalities and differences with other relevant theoretical frameworks, and to underline some practical implications. Methods A narrative literature review has been performed in 2020. Studies have been identified in Medline (via PubMed) and Cumulative Index to Nursing and Allied Health Literature (CINAHL) databases, and the International Learning Collaborative website by using the following keywords: \u2018Fundamentals of Care\u2019, \u2018Fundamental of Care\u2019, \u2018Nursing\u2019; publications performed by the eminent authors in the field (prof. Kitson A. & Feo R) have also been retrieved. Results The nurse-patient relationship is a crucial component to meet the fundamental care needs, articulated in physical, relational, and socio-psychological dimensions. Commonalities with the patient-centred care movement and with that of compassionate and unfinished nursing care have been summarized. Moreover, models of care delivery, studies highlighting the perspective of patients with regards to the FOC and the relevance of nursing minimum data set have been highlighted. Conclusions It is important to continue to build a professional and scientific dialogue on fundamentals of care by including them in the professional and scientific agenda of prioritie

Missed nursing education: Findings from a qualitative study

To understand what nursing education activities are missed in the daily life of nursing programmes, by also identifying antecedents and consequences of missed educational activities