Urine cultures.

<p>Urine cultures.</p

Changes in plasma ammonia levels and level of consciousness.

<p>The case 5 patient died shortly after admission due to complicated aspiration pneumonia.</p><p>Changes in plasma ammonia levels and level of consciousness.</p

Clinical characteristics of five cases with high plasma ammonia levels.

<p>Clinical characteristics of five cases with high plasma ammonia levels.</p

Eastern Cooperative Oncology Group performance status.

<p>Eastern Cooperative Oncology Group performance status.</p

Schematic flow of the discovery and validation of the optimal preanalytical conditions.

The effects of different blood collection tubes and sample storage periods were examined during the discovery stage. The library concentrations, identification rate, UMI ratios, and expression profiles were used to identify optimal conditions. During the validation stage, 60 blood samples from cancer patients were collected from three separate facilities to assess the effects of the different conditions. The library concentrations, identification rate, UMI ratios, and expression profiles were assessed to determine whether these measurements were within the assumed range and to validate the preanalytical conditions.</p

QC data from NGS measurement for comparison of variances among samples subjected to different conditions.

(XLSX)</p

RNA concentrations in the purified RNA samples.

Concentration of RNAs obtained from samples isolated using different blood collection tube conditions (A). The crossbar indicates the mean, and the error bar indicates the standard deviation (SD). Significant differences were identified using one-way ANOVA with a post hoc Tukey test and are indicated as * p (TIFF)</p

MiRNAs with higher expression at 2 days and 3 days than at 1 hour.

MiRNAs with higher expression at 2 days and 3 days than at 1 hour.</p

Assessment of estimated conditions using clinical samples from facilities A, B and C.

The library concentrations (A), identification rate (B) and UMI ratios (C) were compared among samples obtained from different facilities. The expression levels of hsa-miR-451a (D), an indicator of hemolysis, and hsa-miR-126-3p (E), an indicator of platelet contamination, are shown. The crossbar indicates the mean, and the error bar indicates the standard deviation (SD). Significant differences were identified using one-way ANOVA with a post hoc Tukey test and are indicated as ** p<0.01.</p

Stability of plasma in storage periods up to 30 days at -20°C.

The library concentrations (A), identification rate (B) and UMI ratios (C) were compared between samples stored for 1 hour and 1 day, 3 days, 5 days, 7 days and 30 days after blood collection from five healthy volunteers. The mean miRNA expression profiles of samples stored for 1 hour vs. 1 day (D), 1 hour vs. 3 days (E), 1 hour vs. 5 days (F), 1 hour vs. 7 days (G), and 1 hour vs. 30 days (H) were compared. MiRNAs with levels that exhibited a fold change greater than 2-fold or less than half-fold are indicated outside of the dashed lines. The X-axis or Y-axis of the scatter plot indicates log2(miRNA expression counts). Significant differences were identified using one-way ANOVA with a post hoc Dunnett’s test.</p