Preanalytical considerations in quantifying circulating miRNAs that predict end-stage kidney disease in diabetes.

Our previous study identified 8 risk and 9 protective plasma miRNAs associated with progression to end-stage kidney disease (ESKD) in diabetes. This study aimed to elucidate preanalytical factors that influence the quantification of circulating miRNAs. Using the EdgeSeq platform, which quantifies 2,002 miRNAs in plasma, including ESKD-associated miRNAs, we compared miRNA profiles in whole plasma versus miRNA profiles in RNA extracted from the same plasma specimens. Less than half of the miRNAs were detected in standard RNA extraction from plasma. Detection of individual and concentrations of miRNAs were much lower when RNA extracted from plasma was quantified by RNA sequencing (RNA-Seq) or quantitative reverse transcription PCR (qRT-PCR) platforms compared with EdgeSeq. Plasma profiles of miRNAs determined by the EdgeSeq platform had excellent reproducibility in assessment and had no variation with age, sex, hemoglobin A1c, BMI, and cryostorage time. The risk ESKD-associated miRNAs were detected and measured accurately only in whole plasma and using the EdgeSeq platform. Protective ESKD-associated miRNAs were detected by all platforms except qRT-PCR; however, correlations among concentrations obtained with different platforms were weak or nonexistent. In conclusion, preanalytical factors have a profound effect on detection and quantification of circulating miRNAs in ESKD in diabetes. Quantification of miRNAs in whole plasma and using the EdgeSeq platform may be the preferable method to study profiles of circulating cell-free miRNAs associated with ESKD and possibly other diseases.

The Addition of High-Technology Into the Stop the Bleed Program Among School Personnel Improves Short-Term Skill Application, Not Long-Term Retention.

INTRODUCTION: The Stop the Bleed (STB) program trains the general public on identifying and treating life-threatening bleeding. Data on efficacy and retention of skills taught through this program are limited, with the role of high-technology modalities to augment the program, such as simulation and feedback devices, untested. METHODS: A convenience sample of 66 school personnel participated in an open-label observational study from January to August 2019. The control group received the standard bleeding control course, while the intervention group received the bleeding control course with addition of a simulation and a feedback device for wound packing. Assessment was performed by STB instructors using performance metrics from prior studies as well as a feedback device. Retention testing was performed 2 to 8 months after intervention. The study was approved by the hospital's institutional review board. RESULTS: The intervention group performed better than the control group on correct tourniquet application [90.3% vs. 71.0%; odds ratio (OR) = 11.28; P = 0.015; 95% confidence interval (CI) = 1.86 to 104.67] wound packing scores (59.5% vs. 29.6%; OR = 0.33; P = 0.007; 95% CI = 9.36 to 56.00) and were more likely to assess their safety (OR = 5.49; P = 0.034; 95% CI = 1.28 to 27.66), and reported higher comfort scores on stepping into an emergency scenario (OR = 11.19; P = 0.004; 95% CI = 2.51 to 63.11), wound packing (OR = 5.16; P = 0.025; 95% CI = 1.35 to 22.46), and using a tourniquet (OR = 11.41; P = 0.003; 95% CI = 2.57 to 67.59). Thirty-one participants (46.9%) were assessed again at retention 2 to 8 months later where scores for tourniquet placement and wound packing were not significantly different in the two groups. CONCLUSIONS: Augmenting STB with simulation and feedback improved both self-reported comfort level and skill set of participants, but the retention of skills was poor in both groups.