Absolute and Relative Quantification of Placenta-Specific MicroRNAs in Maternal Circulation with Placental Insufficiency–Related Complications  Ilona Hromadnikova, Katerina Kotlabova, Jindrich Doucha, Klara Dlouha, Ladislav Krofta  The Journal of Molecular Diagnostics  Volume 14, Issue 2, Pages 160-167 (March 2012) DOI: 10.1016/j.jmoldx.2011.11.003 Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Absolute quantification of extracellular miRNAs in maternal circulation in women with normally progressing pregnancies and those with PIRCs. Although no differences between the levels of ubiquitous miR-16 and let-7d were found over time in those with normally progressing pregnancies, a statistically significant difference was observed between the first and the third trimesters of gestation in the cohort of women with normally progressing pregnancies for all placenta-specific miRNAs in maternal circulation. No difference in the levels of extracellular miRNAs was observed between those with normally progressing pregnancies and those with PIRCs. The Journal of Molecular Diagnostics 2012 14, 160-167DOI: (10.1016/j.jmoldx.2011.11.003) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 Relative quantification of extracellular placenta-specific miRNAs in maternal circulation in those with normally progressing pregnancies and those with PIRCs. Significantly higher expression of all tested placenta-specific miRNAs was observed in the third trimester of gestation compared with the first trimester of gestation in the cohort of those with normally progressing pregnancies using the comparative CT method relative to the normalization factor (geometric mean of miR-16 and let-7d). PIRCs showed no difference in placenta-specific miRNA expression profile in maternal plasma samples when compared with those with normally progressing pregnancies. The Journal of Molecular Diagnostics 2012 14, 160-167DOI: (10.1016/j.jmoldx.2011.11.003) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 Demonstration of quantitative aberrations of extracellular placenta-specific miRNAs in maternal circulation throughout gestation in patients who later developed preeclampsia and/or IUGR. Significant elevation of the concentrations (A) and expression (B) of extracellular placenta-specific miRNAs was observed during early gestation (within the 12th to 16th weeks of gestation) in pregnant women who later developed preeclampsia and/or IUGR. The mean ± 2 SDs for extracellular miRNAs was used as the cutoff to differentiate between those with PIRCs and those with normally progressing pregnancies. This cutoff showed the maximum feasible specificity (the percentage of those with normally progressing pregnancies identified as not having the condition) for each miRNA assay. The Journal of Molecular Diagnostics 2012 14, 160-167DOI: (10.1016/j.jmoldx.2011.11.003) Copyright © 2012 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions