IMPORTANCEThe optimal first-line mode of noninvasive respiratory support for acutely ill children is not known.OBJECTIVE To evaluate the noninferiority of high-flow nasal cannula therapy (HFNC) as the first-line mode of noninvasive respiratory support for acute illness, compared with continuous positive airway pressure (CPAP), for time to liberation from all forms of respiratory support. DESIGN, SETTING, AND PARTICIPANTS Pragmatic, multicenter, randomized noninferiority clinical trial conducted in 24 pediatric critical care units in the United Kingdom among 600 acutely ill children aged 0 to 15 years who were clinically assessed to require noninvasive respiratory support, recruited between August 2019 and November 2021, with last follow-up completed in March 2022. INTERVENTIONS Patients were randomized 1:1 to commence either HFNC at a flow rate based on patient weight (n = 301) or CPAP of 7 to 8 cm H 2 O (n = 299). MAIN OUTCOMES AND MEASURESThe primary outcome was time from randomization to liberation from respiratory support, defined as the start of a 48-hour period during which a participant was free from all forms of respiratory support (invasive or noninvasive), assessed against a noninferiority margin of an adjusted hazard ratio of 0.75. Seven secondary outcomes were assessed, including mortality at critical care unit discharge, intubation within 48 hours, and use of sedation. RESULTSOf the 600 randomized children, consent was not obtained for 5 (HFNC: 1; CPAP: 4) and respiratory support was not started in 22 (HFNC: 5; CPAP: 17); 573 children (HFNC: 295; CPAP: 278) were included in the primary analysis (median age, 9 months; 226 girls [39%]). The median time to liberation in the HFNC group was 52.9 hours (95% CI, 46.0-60.9 hours) vs 47.9 hours (95% CI, 40.5-55.7 hours) in the CPAP group (absolute difference, 5.0 hours [95% CI -10.1 to 17.4 hours]; adjusted hazard ratio 1.03 [1-sided 97.5% CI, 0.86-ϱ]). This met the criterion for noninferiority. Of the 7 prespecified secondary outcomes, 3 were significantly lower in the HFNC group: use of sedation (27.7% vs 37%; adjusted odds ratio, 0.59 [95% CI, 0.39-0.88]); mean duration of critical care stay (5 days vs 7.4 days; adjusted mean difference, −3 days [95% CI, −5.1 to −1 days]); and mean duration of acute hospital stay (13.8 days vs 19.5 days; adjusted mean difference, −7.6 days [95% CI, −13.2 to −1.9 days]). The most common adverse event was nasal trauma (HFNC: 6/295 [2.0%]; CPAP: 18/278 [6.5%]).CONCLUSIONS AND RELEVANCE Among acutely ill children clinically assessed to require noninvasive respiratory support in a pediatric critical care unit, HFNC compared with CPAP met the criterion for noninferiority for time to liberation from respiratory support.

The genetic alphabet consists of the four letters: C, A, G, and T in DNA and C,A,G, and U in RNA. Triplets of these four letters jointly encode 20 different amino acids out of which proteins of all organisms are built. This system is universal and is found in all kingdoms of life. However, bases in DNA and RNA can be chemically modified. In DNA, around 10 different modifications are known, and those have been studied intensively over the past 20 years. Scientific studies on DNA modifications and proteins that recognize them gave rise to the large field of epigenetic and epigenomic research. The outcome of this intense research field is the discovery that development, ageing, and stem-cell dependent regeneration but also several diseases including cancer are largely controlled by the epigenetic state of cells. Consequently, this research has already led to the first FDA approved drugs that exploit the gained knowledge to combat disease. In recent years, the ~150 modifications found in RNA have come to the focus of intense research. Here we provide a perspective on necessary and expected developments in the fast expanding area of RNA modifications, termed epitranscriptomics.

Neuroblastoma (NB) is the most common occurring solid paediatric cancer in children under the age of five years. Whether of familial or sporadic origin, chromosome abnormalities contribute to the development of NB and cause dysregulation of microRNAs (miRNAs). MiRNAs are small non-coding, single stranded RNAs that target messenger RNAs at the post-transcriptional levels by repressing translation within all facets of human physiology. Such gene ‘silencing’ activities by miRNAs allows the development of regulatory feedback loops affecting multiple functions within the cell, including the possible differentiation of neural stem cell (NSC) lineage selection. Neurogenesis includes stages of self-renewal and fate specification of NSCs, migration and maturation of young neurones, and functional integration of new neurones into the neural circuitry, all of which are regulated by miRNAs. The role of miRNAs and their interaction in cellular processes are recognised aspects of cancer genetics, and miRNAs are currently employed as biomarkers for prognosis and tumour characterisation in multiple cancer models. Consequently, thorough understanding of the mechanisms of how these miRNAs interplay at the transcriptomic level will definitely lead to the development of novel, bespoke and efficient therapeutic measures, with this review focusing on the influences of miRNAs on neuroblast modulations leading to neuroblastoma.

Mesenchymal Stem Cells (MSCs) are widely used in therapeutic applications. Their plasticity and predisposition to differentiate into a variety of cell types, including those of the neuronal lineage, makes them ideal to study whether a selection of miRNAs may direct the differentiation of MSCs into neuroblasts or neuroblastoma to mature neurons. Following a short-listing, miR-107, 124 and 381 were selected as the most promising candidates for this differentiation. MSCs differentiated into cells of the neural lineage (Conditioned Cells) upon addition of conditioned medium (rich in microvesicles containing miRNAs) obtained from cultured SH-SY5Y neuroblastoma cells. Characterisation of stemness (including SOX2, OCT4, Nanog and HCG) and neural markers (including Nestin, MASH1, TUBB3 and NeuN1) provided insight regarding the neuronal state of each cell type. This was followed by transfection of the three miRNA antagonists and mimics, and quantification of their respective target genes. MiRNA target gene expression following transfection of MSCs with miRNA inhibitors and mimics demonstrated that these three miRNAs were not sufficient to induce differentiation. In conditioned cells the marginal changes in the miRNA target expression levels reflected potential for the modulation of intermediate neural progenitors and immature neuron cell types. Transfection of various combinations of miRNA inhibitors and/or mimics revealed more promise. Undoubtedly, a mix of biomolecules is being released by the SH-SY5Y in culture that induce MSCs to differentiate. Screening for those biomolecules acting synergistically with specific miRNAs will allow further combinatorial testing to elucidate the role of miRNA modulation.

Aims: The necessity for Platelet Concentrates (PCs) in transfusion has increased, and several measures have been undertaken to try and increase the shelf-life of this product. Increasing the shelf-life to 10 days from the current 5 days will increase availability and reduce waste and cost. However, the PCs need to maintain their safety and therapeutic efficacy over the extended storage time. To ensure the safety of this product, the bag should be sterile, with low concentrations of cytokines to prevent transfusion reactions, and the platelet indices should be within the therapeutic range to ensure efficacy. The aim of this study is to determine whether PCs retain quality standards and thus, remain fit for transfusion after being stored for 10 days in SSP+ at 22±2°C with constant agitation. Study Design:  Qualitative study. Place and Duration of Study: National Blood Transfusion Service Malta, between March 2021 and June 2022. Methodology: In this study, 20 PCs were analysed by sampling on Day 5 and Day 10 of storage and tested for platelet count and indices, pH, sterility of the product and cytokine concentrations. Results: The results obtained showed a significant differences in the platelet count and platelet indices and pH. The difference in cytokine concentration of Interleukin-6 (IL-6), Tumour necrosis factor-α (TNF—α) and transforming growth factor-β (TGF-β) were not found to be statistically different between Day 5 and Day 10. Furthermore, 1 PC unit was positive during testing for bacteria detection. Conclusion: These results showed that the difference between Day 5 and Day 10 is not alarming, and further studies should be made to prolong the storage life to 10 days.

Storage lesions of blood products intended for transfusion have been shown to give rise to the accumulation of proin flammatory cytokines, which are known to cause transfusion associated adverse events and immunomodulatory effects. Regulations which determine the eligibility of blood donors do not take into consideration metabolic disorders such as Type 2 Diabetes Mellitus (T2DM), unless complications arise from said disorder. However, the increased levels of cytokines in T2DM classifies this condition as an inflammatory condition. This outcome dictates the need to investigate whether detected levels of pro-inflammatory cytokines due to inflammation combined with those present in red cell concentrates as a result of storage lesions may lead to undesirable outcomes in recipients who already have underlying inflammatory conditions. Due to the largely unavailable information on the subject, a few important considerations for undertaking such investigations have been summarised.

Background and Aim: Whole blood is processed to derive a red cell concentrate, plasma, and buffy coat (BC) (from which platelets can be further extracted). Unused plasma and BCs are common in most blood establishments and considered a liability. The redirection of these products to xeno-free applications is not complicated or time-consuming and cannot only benefit the research recipients but also the blood establishment suppliers interested in research collaboration. The aim of this study is to describe a diverse yet by no means an exhaustive list of options for preparing blood products for research applications. Materials and Methods: Plasma and BCs from healthy donors were processed using basic laboratory techniques under aseptic conditions and tested for their ability to support the culture of mesenchymal stem cells in both 2D and 3D cultures using fibrin clots. The white blood cells (WBC) from the BCs were induced by phytohemagglutinin and CD marker expression was monitored using quantitative polymerase chain reaction. Results: All the methods tested for preparing blood products were successful but the applicability to different settings varied greatly with the most successful being the supplementation of Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 with 20% cryodepleted plasma and 0.1 mg/mL platelet lysate, the formation of fibrin clots using a ratio of 3:1 (medium: plasma) and the culturing of WBCs with 5 µg/mL phytohemagglutinin. Conclusions: Using the wastes and by-products of blood establishments for xeno-free cell culturing of stem cells will reduce the reliance on commercially available, ready-made products, and increasing the potential for therapeutic stem cell research. Despite the benefits presented both in terms of cost and applications, further characterization and optimization of each blood product for reproducibility of results is required. Relevance for Patients: The availability of low-cost xeno-free reagents will speed up therapeutic stem cell research and allow patients to receive treatments of the expected high standards at lower costs.

BackgroundHemoglobin (Hb) evaluation by point-of-care testing (POCT) identifies borderline or anaemic asymptomatic blood donors. Although quality control checks confirm that this device is fit for use, it is still not clear whether the analyser is performing effectively. A protocol comparing the POCT EKF Diagnostics with the Sysmex XN-550 automated cell counter (ACC) has been designed.MethodsVarious scenarios of Hb measurements from the ACC and the POCT device are compared using the Spearman correlation and Intraclass correlation. The Bland-Altman method was used to analyse the level of agreement between the two devices.ResultsCorrelation between the two devices was best observed in the venous vs venous blood scenario.ConclusionThe POCT device overestimates the Hb levels in capillary blood, meaning that Hb requirements should be adjusted and when feasible testing repeated on venous blood using an ACC. Furthermore, it is suggested thar each Facility determine their own Hb threshold.