Validation of microarray data by RT-PCR.

<p>Validation of microarray data by RT-PCR.</p

Venn diagram showing the relationship of present calls between PBMC, PAX and PAX-GR in SCD samples.

<p>Venn diagram showing the relationship of present calls between PBMC, PAX and PAX-GR in SCD samples.</p

Figure 2

<p>A. Expression of 18s, globin transcripts α, β by RT-PCR in PAX and PAX-GR from control and SCD samples. Ct values in the expression of globins are given as mean±SD for each group. Red bar - PAX-Control; Green bar - PAX-GR Control; Yellow bar - PAX-SCD; Blue bar - PAX-GR-SCD. B. Normalized signal intensities for β globin probe sets from microarrays for control and SCD samples. Values are given as mean±SD. Grey bar - PAX; Black bar - PAX-GR. C. Normalized signal intensities for α globin probesets from microarrays for control and SCD samples. Values are given as mean±SD. Grey bar - PAX; Black bar - PAX-GR.</p

Heat Map of differential gene expression in PAX-GR in sickle cell disease (n = 5) in comparison to control subjects (n = 5).

<p>Cluster analysis was applied to gene expression data derived from all probes on HG-U133 plus 2.0 at FDR 10% and FC >5.0. The level of expression of each gene in each sample relative to the median level of expression of that gene across all samples is represented using a red, black and green color scale (green – below median; black – equal to median; red - above median). The dendrogram displays the unsupervised clustering of patients and control subjects using the differentially expressed gene list.</p

List of selected genes that overlap between PAX and PAX-GR.

<p>List of selected genes that overlap between PAX and PAX-GR.</p

Detection of expressed transcripts in PBMC, PAX and PAX-GR in control (Grey bars) and SCD (Black bars) as determined by the present calls for the samples (n = 5) analyzed by microarrays.

<p>Values are given as mean±SD for the number of present transcripts in each of the groups in each sample type.</p

Gene ontology classification for genes common to PAX and PAX – GR.

<p>Many genes have multiple gene ontology descriptions.</p

Exploring the Mechanisms of the Reductase Activity of Neuroglobin by Site-Directed Mutagenesis of the Heme Distal Pocket

Neuroglobin (Ngb) is a six-coordinate globin that can catalyze the reduction of nitrite to nitric oxide. Although this reaction is common to heme proteins, the molecular interactions in the heme pocket that regulate this reaction are largely unknown. We have shown that the H64L Ngb mutation increases the rate of nitrite reduction by 2000-fold compared to that of wild-type Ngb [Tiso, M., et al. (2011) <i>J. Biol. Chem. 286</i>, 18277–18289]. Here we explore the effect of distal heme pocket mutations on nitrite reduction. For this purpose, we have generated mutations of Ngb residues Phe28­(B10), His64­(E7), and Val68­(E11). Our results indicate a dichotomy in the reactivity of deoxy five- and six-coordinate globins toward nitrite. In hemoglobin and myoglobin, there is a correlation between faster rates and more negative potentials. However, in Ngb, reaction rates are apparently related to the distal pocket volume, and redox potential shows a poor relationship with the rate constants. This suggests a relationship between the nitrite reduction rate and heme accessibility in Ngb, particularly marked for His64­(E7) mutants. In five-coordinate globins, His­(E7) facilitates nitrite reduction, likely through proton donation. Conversely, in Ngb, the reduction mechanism does not rely on the delivery of a proton from the histidine side chain, as His64 mutants show the fastest reduction rates. In fact, the rate observed for H64A Ngb (1120 M^–1 s^–1) is to the best of our knowledge the fastest reported for a heme nitrite reductase. These differences may be related to a differential stabilization of the iron–nitrite complexes in five- and six-coordinate globins

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