Correlates and associations of pulse pressure with kidney function and hemolysis.

<p><b>A</b>, Pulse pressure has a significant positive correlation with the hemolytic component in HbSS patients, but not in HbSC patients. <b>B</b>, Pulse pressure has a significant positive correlation with serum creatinine in both HbSS and HbSC patients. <b>C</b>, Elevated pulse pressure is significantly associated with presence of proteinuria in HbSS patients, while the association is not significant in HbSC patients.</p

Correlations of pulse pressure with clinical and laboratory characteristics by hemoglobin genotype.

<p>*p values <0.002 remained significant after Bonferroni's adjustment for multiple comparisons.</p>†<p>N-terminal prohormone of brain natriuretic peptide.</p><p>Correlations of pulse pressure with clinical and laboratory characteristics by hemoglobin genotype.</p

Independent predictors of pulse pressure.

<p>*transformed using the natural log function.</p><p>Independent predictors of pulse pressure.</p

Twenty most differentially expressed (A) up- and (B) down-regulated microRNAs (P<0.05, fold change>2) between sickle cell disease (N = 26) and controls (N = 17) ranked according to p-value (low to high).

<p>Twenty most differentially expressed (A) up- and (B) down-regulated microRNAs (P<0.05, fold change>2) between sickle cell disease (N = 26) and controls (N = 17) ranked according to p-value (low to high).</p

Most significant predicted canonical pathways regulated by the target genes of the 3 miRNAs: 376a, 409-3p and 1225-3p (p-value<0.01).

<p>Orange squares depict ratio which is the number of target genes associated with each pathway. The vertical line across the bars represents the cut-off of the p-value.</p

Characteristics of Study Participants.^*

*<p>Age and Laboratory values indicate median (range).</p

MiRNAs in platelets of SCD patients are differentially expressed.

<p>(<b>A</b>) Characterization of purified platelet preparation by flow cytometry. (<b>B</b>) Bioanalyzer assessment of RNA samples from purified platelet preps showing good quality with RIN of 7.8. (<b>C</b>) Venn diagram comparing differentially expressed miRNAs from two independent study cohorts. Comparison of 259 differentially expressed miRNAs from NHLBI samples and 67 differentially expressed miRNAs from UPMC samples. In total, there are 40 miRNAs overlapping between the two cohorts. (<b>D</b>) The heatmap depicts the 40 statistically significant (<i>p</i>-value<0.05, FC>2) differentially expressed miRNAs between SCD and controls that were common between the two cohorts. Columns represent individual samples and each represents a miRNA. Upregulated miRNAs expression levels are shown in progressively brighter shades of yellow, depending on the fold difference. Downregulated miRNAs are shown in progressively brighter shades of purple. No difference is represented as grey. The names of the miRNAs are displayed to the right of the heatmap.</p

miR-1225-3p regulates gene expression after transfection in MEG-01 cells.

<p>(<b>A</b>) Expression of the flourescent marker pmaxGFP plasmid as determined by flow cytometry demonstrates 76% GFP expressing cells out of the viable cell population. (<b>B</b>) qRT-PCR assay confirms overexpression of miR-1225-3p in MEG-01 cells (n = 5) vs scrambled (scr) (n = 5). (<b>C</b>) MT1X, PTPN6, IFI6, FCER1G and RAP2A are significantly downregulated in MEG-01 transfected cells as validated by qRT-PCR. (<b>D</b>) Schematic representation of overlap in differentially expressed genes between miR-1225-3p transfected MEG-01 cells and platelets from SCD patients. The numbers in the circles denote the differentially expressed genes with the leftmost figure representing genes using a FDR of 5% and the rightmost figure after applying a stringent statistical filter of FC>2. Further overlap with ComiR predicted target list of the 40 differentially expressed miRNAs resulted in a list of 7 genes potentially regulated by miR-1225-3p. (<b>E</b>) Out of the 7 genes, PLAGL2 and PBXIP1 genes are significantly downregulated and PHF20L1 is significantly upregulated in miR-1225-3p transfected cells vs scrambled. In the qRT-PCR figures (B, C and D), for each gene, the blue bar represents the cells transfected with scrambled RNA (n = 5) and the red bar represents the miR-1225-3p transfected cells (n = 5). Y-axis shows fold change of miR-1225-3p in transfected cells with the expression level in scrambled set to 1. Error bars are based on standard deviation. *denotes p-value<0.05.</p

List of novel putative target genes of miR-1225-3p from the differentially expressed genes (FDR<5%, FC>2).

<p>List of novel putative target genes of miR-1225-3p from the differentially expressed genes (FDR<5%, FC>2).</p

Functional annotation clustering of predicted target genes to the differentially expressed microRNAs belonging to three distinct families located at chromosome 14q32.31.

<p>Functional annotation clustering of predicted target genes to the differentially expressed microRNAs belonging to three distinct families located at chromosome 14q32.31.</p