Mean (SE) of PETCO₂ variation during the handgrip maneuver (ANOVA p = 0.330).

<p>Mean (SE) of PETCO₂ variation during the handgrip maneuver (ANOVA p = 0.330).</p

Mean (SE) of time-varying ARI at five distinct phases of HG.

<p>Mean (SE) of time-varying ARI at five distinct phases of HG.</p

Representative pattern of the extracted and derived parameters in a 19 year-old female subject.

<p>The grey bar shows the duration of HG. From top to bottom: ABP, CBFV_L (solid line) and CBFV_R (dashed line), CrCP_L (solid line) and CrCP_R (dashed line), RAP_L (solid line) and RAP_R (dashed line), ARI_L (solid line) and ARI_R (dashed line). Subscripts R and L indicate right and left MCA respectively.</p

Parameter values of Baseline and Plateau during Handgrip maneuver.

<p>Values are means (SD). CBFV, cerebral blood flow velocity; ABP, arterial blood pressure; HR, heart rate; CrCP, critical closing pressure; RAP, resistance area product; CVR, cerebrovascular resistance. Subscripts R and L indicate right and left respectively. P-values from repeated measures ANOVA.</p>#<p>p<0.005,</p>*<p>p<0.05 compared to baseline.</p

Population mean values during baseline synchronized by the beginning of recording.

<p>From top to bottom: ABP, CBFV_L (solid line) and CBFV_R (dashed line), ARI_L (solid line) and ARI_R (dashed line). Subscripts R and L indicate right and left MCA respectively. For clarity only the largest ±1SE is represented at the point of occurrence.</p

Population mean values of CrCP_L (solid line) and CrCP_R (dashed line), RAP_L (solid line) and RAP_R (dashed line), autoregulation index (ARI_L: solid line and ARI_R: dashed line).

<p>Subscripts R and L indicate right and left MCA respectively. For clarity only the largest ±1 SE is represented at the point of occurrence.</p

Differential Expression of <i>ID4</i> and Its Association with <i>TP53</i> Mutation, <i>SOX2</i>, <i>SOX4</i> and <i>OCT-4</i> Expression Levels

<div><p>Inhibitor of DNA Binding 4 (ID4) is a member of the helix-loop-helix ID family of transcription factors, mostly present in the central nervous system during embryonic development, that has been associated with <i>TP53</i> mutation and activation of <i>SOX2</i>. Along with other transcription factors, <i>ID4</i> has been implicated in the tumorigenic process of astrocytomas, contributing to cell dedifferentiation, proliferation and chemoresistance. In this study, we aimed to characterize the <i>ID4</i> expression pattern in human diffusely infiltrative astrocytomas of World Health Organization (WHO) grades II to IV of malignancy (AGII-AGIV); to correlate its expression level to that of <i>SOX2</i>, <i>SOX4</i>, <i>OCT-4</i> and <i>NANOG</i>, along with <i>TP53</i> mutational status; and to correlate the results with the clinical end-point of overall survival among glioblastoma patients. Quantitative real time PCR (qRT-PCR) was performed in 130 samples of astrocytomas for relative expression, showing up-regulation of all transcription factors in tumor cases. Positive correlation was found when comparing <i>ID4</i> relative expression of infiltrative astrocytomas with <i>SOX2</i> (<i>r</i> = 0.50; <i>p</i><0.005), <i>SOX4</i> (<i>r</i> = 0.43; <i>p</i><0.005) and <i>OCT-4</i> (<i>r</i> = 0.39; <i>p</i><0.05). The results from <i>TP53</i> coding exon analysis allowed comparisons between wild-type and mutated status only in AGII cases, demonstrating significantly higher levels of <i>ID4</i>, <i>SOX2</i> and <i>SOX4</i> in mutated cases (<i>p</i><0.05). This pattern was maintained in secondary GBM and further confirmed by immunohistochemistry, suggesting a role for <i>ID4</i>, <i>SOX2</i> and <i>SOX4</i> in early astrocytoma tumorigenesis. Combined hyperexpression of <i>ID4</i>, <i>SOX4</i> and <i>OCT-4</i> conferred a much lower (6 months) median survival than did hypoexpression (18 months). Because both ID4 alone and a complex of SOX4 and OCT-4 activate <i>SOX2</i> transcription, it is possible that multiple activation of <i>SOX2</i> impair the prognosis of GBM patients. These observational results of associated expression of <i>ID4</i> with <i>SOX4</i> and <i>OCT-4</i> may be used as a predictive factor of prognosis upon further confirmation in a larger GBM series.</p> </div

Comparison of gene expression levels between the wild-type <i>TP53</i> (<i>WT TP53</i>) and the mutated <i>TP53</i> (<i>Mutated TP53</i>) in AGII cases.

<p>Higher expressions of <i>ID4</i> (A), <i>SOX2</i> (B), <i>SOX4</i> (C) and <i>NANOG</i> (E) were observed on the mutated <i>TP53</i> AGII cases. No difference was found for <i>OCT-4</i> (D) and <i>CD133</i> (F) relative expression between the two groups. White lozenges represent the deceased patients. The <i>p</i> values are: *<i>p</i><0.05 and ** <i>p</i><0.005, Mann-Whitney test.</p

Median of relative expression levels of the analyzed genes in astrocytomas, according to morphology.

a<p>NN, non-neoplastic; AGII, low-grade astrocytoma; AGIII, anaplastic astrocytoma; GBM, glioblastoma.</p

Heatmap displaying the relative gene expressions in low-grade astrocytoma (AGII), anaplastic astrocytoma (AGIII) and GBM cases according to <i>TP53</i> mutation status.

<p>The <i>TP53</i> mutated cases are represented by side dashes. The mutated <i>TP53</i> AGII cases showed more elevated expression levels of <i>ID4</i>, <i>SOX2</i>, <i>SOX4</i> and <i>NANOG</i>. <i>CD133</i> expressions were more heterogeneous among the cases. <i>SOX2</i> and <i>SOX4</i> showed similar expression levels to <i>ID4</i>. Similarly, secondary GBM cases also presented higher <i>ID4</i>, <i>SOX2</i>, <i>SOX4</i> expression levels. <i>OCT-4</i>, <i>NANOG</i> and <i>CD133</i> expression levels were heterogeneous among secondary GBM cases, and <i>OCT-4</i> presented higher mRNA levels in primary GBM.</p