Fig 1.
Overall time course of autoptic tissue samples.
PMI, postmortem interval; RT, room temperature; FFPE, formalin-fixed paraffin-embedded.
Table 1.
Control and biomarker candidate specifications.
Fig 2.
Integrity analysis of smRNAs in autoptic tissues.
Representative electropherograms of frozen tissues (A–C) and FFPE tissues (E–G). Green solid lines indicate the area (10–40 nt) containing miRNA peaks, and green dashed lines indicate the area containing smRNA peaks (0–200 nt). The ratio of smRNAs to miRNAs was evaluated using 11 frozen samples (D), and 17 FFPE samples (H). PMI, postmortem interval; RT, room temperature; FF, formalin-fixed period.
Table 2.
Amplification efficiency of target genes.
Fig 3.
Detection of smRNAs in frozen tissues.
Average Cq values (Y-axis) of 8 smRNAs with various postmortem intervals (X-axis) are shown. Cq values for the miRNAs (dashed lines) appear to be more stable than those of other classes of smRNAs (solid lines). The mean value and standard deviation for each candidate smRNA are shown in Table 3.
Table 3.
Candidate smRNAs in frozen tissues with PMIs up to one week.
Fig 4.
Detection of smRNAs in FFPE tissues.
Average Cq values (Y-axis) of 8 smRNAs with various formalin-fixation periods (FF; X-axis) are shown. The detection of every smRNA candidate was reduced as the duration of fixation increased. The rates of degradation for each miRNA (dashed lines) were less than those observed for the other classes of smRNAs (solid lines) analyzed. The regression formula for each smRNA is shown in Table 4.
Table 4.
Rates of smRNA degradation in FFPE tissues with prolonged fixation up to three years.
Fig 5.
Quantified miRNA biomarker expression in postmortem AMI and control tissues.
Histopathological images of human cardiac tissue stained with Masson trichrome highlighting the dominant contraction band necrosis and low levels of neutrophil infiltration in the early phase of AMI (A) and the regular alignment of muscular fibers in the control tissue (B). Scale bar = 50 μm. Relative expression of miR-1 (C), miR-208b (D), and miR-499a (E) in four AMI and seven control cases after normalization to miR-26b and miR-191. *P < 0.05.