DataSheet_1_The relationship between radiomics and pathomics in Glioblastoma patients: Preliminary results from a cross-scale association study.pdf

Glioblastoma multiforme (GBM) typically exhibits substantial intratumoral heterogeneity at both microscopic and radiological resolution scales. Diffusion Weighted Imaging (DWI) and dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) are two functional MRI techniques that are commonly employed in clinic for the assessment of GBM tumor characteristics. This work presents initial results aiming at determining if radiomics features extracted from preoperative ADC maps and post-contrast T1 (T1C) images are associated with pathomic features arising from H&E digitized pathology images. 48 patients from the public available CPTAC-GBM database, for which both radiology and pathology images were available, were involved in the study. 91 radiomics features were extracted from ADC maps and post-contrast T1 images using PyRadiomics. 65 pathomic features were extracted from cell detection measurements from H&E images. Moreover, 91 features were extracted from cell density maps of H&E images at four different resolutions. Radiopathomic associations were evaluated by means of Spearman’s correlation (ρ) and factor analysis. p values were adjusted for multiple correlations by using a false discovery rate adjustment. Significant cross-scale associations were identified between pathomics and ADC, both considering features (n = 186, 0.45 < ρ < 0.74 in absolute value) and factors (n = 5, 0.48 < ρ < 0.54 in absolute value). Significant but fewer ρ values were found concerning the association between pathomics and radiomics features (n = 53, 0.5 < ρ < 0.65 in absolute value) and factors (n = 2, ρ = 0.63 and ρ = 0.53 in absolute value). The results of this study suggest that cross-scale associations may exist between digital pathology and ADC and T1C imaging. This can be useful not only to improve the knowledge concerning GBM intratumoral heterogeneity, but also to strengthen the role of radiomics approach and its validation in clinical practice as “virtual biopsy”, introducing new insights for omics integration toward a personalized medicine approach.</p

Robust SOD3 expression reduced anaplastic thyroid cancer cell growth.

<p>(a) Real time RT-PCR analysis showed a high-level mRNA production of <i>SOD3</i> transgene in anaplastic thyroid cancer 8505c cells. (b,c) Western blotting and related band intensity analysis showed high-level SOD3 production caused by <i>SOD3</i> transgene. (d) The growth curve analysis suggested significantly (p<0.001) decreased cell numbers in cells transfected with <i>SOD3</i>. (e,f) The BrdU DNA incorporation analysis confirmed significantly (p<0.001) decreased cellular growth caused by SOD3. (g) The 3-D gel invasion assay suggested almost complete lack of invasive capacity of 8505c SOD3 cells. (h,i) The soft agar analysis showed significantly (p<0.001) reduced formation of colonies suggesting decreased anchorage independent growth ability for SOD3 cells as compared to 8505c control vector transfected cells. (j) The growth curve analysis of control and SOD3 cells in the presence of 2.5 mM N-Acetyl-cysteine (NAC) showed growth reduction both in 8505c control and SOD3 expressing cells upon the treatment. (k) <i>CATALASE</i> mRNA expression. The quantitative RT-PCR showed no difference between 8505c control and SOD3 cells in <i>CATALASE</i> expression. Data are expressed as mean ±SD. The p-values (* = p<0.05, ** = p<0.01, *** = p<0.001).</p

Extracellular Superoxide Dismutase Regulates the Expression of Small GTPase Regulatory Proteins GEFs, GAPs, and GDI

<div><p>Extracellular superoxide dismutase (SOD3), which catalyzes the dismutation of superoxide anions to hydrogen peroxide at the cell membranes, regulates the cellular growth in a dose-dependent manner. This enzyme induces primary cell proliferation and immortalization at low expression levels whereas it activates cancer barrier signaling through the p53-p21 pathway at high expression levels, causing growth arrest, senescence, and apoptosis. Because previous reports suggested that the SOD3–induced reduction in the rates of cellular growth and migration also occurred in the absence of functional p53 signaling, in the current study we investigated the SOD3-induced growth-suppressive mechanisms in anaplastic thyroid cancer cells. Based on our data, the robust over-expression of SOD3 increased the level of phosphorylation of the EGFR, ERBB2, RYK, ALK, FLT3, and EPHA10 receptor tyrosine kinases with the consequent downstream activation of the SRC, FYN, YES, HCK, and LYN kinases. However, pull-down experiments focusing on the small GTPase RAS, RAC, CDC42, and RHO revealed a reduced level of growth and migration signal transduction, such as the lack of stimulation of the mitogen pathway, in the SOD3 over-expressing cells, which was confirmed by MEK1/2 and ERK1/2 Western blotting analysis. Interestingly, the mRNA expression analyses indicated that SOD3 regulated the expression of guanine nucleotide-exchange factors (<i>RHO GEF16</i>, <i>RAL GEF RGL1</i>), GTPase-activating proteins (<i>ARFGAP ADAP2</i>, <i>RAS GAP RASAL1</i>, <i>RGS4</i>), and a Rho guanine nucleotide-disassociation inhibitor (<i>RHO GDI 2</i>) in a dose dependent manner, thus controlling signaling through the small G protein GTPases. Therefore, our current data may suggest the occurrence of dose-dependent SOD3–driven control of the GTP loading of small G proteins indicating a novel growth regulatory mechanism of this enzyme.</p></div

Growth-associated genes showing SOD3-mediated downregulated expression.

<p>Growth-associated genes showing SOD3-mediated downregulated expression.</p

Reversed growth characteristics by <i>H-RasV12</i> and <i>RHO GEF16</i> over-expression in <i>SOD3</i> transfected cells.

<p>(a,b) The 8505c SOD3 cells were stable transfected with <i>H-RasV12</i> oncogene and analyzed by growth curve and 3-D invasion analysis. Both analysis suggested marked increase in the growth of SOD3 cells after <i>H-RasV12</i> transfection. (c) The 8505c SOD3 cells were stably transfected with <i>RHO GEF16</i>. The real time RT-PCR suggested significant (p<0.01) increase in <i>RHO GEF16</i> mRNA production that was confirmed by Western blotting (lower panel). (d,e) The BrdU DNA incorporation analysis showed significantly (p<0.001) increased nuclear proliferation after <i>RHO GEF16</i> over-expression as compared to 8505c SOD3 cells. (f) The RNAi of <i>RGS4</i> in 8505c SOD3 cells showed significant downregulation of <i>RGS4</i> mRNA production. (g,h) However, the BrdU DNA incorporation analysis failed to show differences between 8505c control and 8505c SOD3/siRGS4 cells. (i) A schematic drawing hypothesizing the SOD3 action in the cells. SOD3 over-expression activates cell membrane RTKs and membrane associated non-receptor tyrosine kinases. Depending on the expression level of <i>SOD3</i> the proliferative and migratory signal is either halted or promoted at the level of small GTPase regulatory genes and small GTPases.</p

SOD3 regulated the expression of small GTPase regulatory genes and affected the activation of small GTPases.

<p>(a-f) Quantitative RT-PCR for GTPase superfamily regulatory genes. The panels show the expression levels of the regulatory genes <i>in vitro</i> cultured cells containing high-level <i>SOD3</i> mRNA and reduced growth abilities. (g-k) Pull-down assay for small GTPase proteins RAS, RAC, CDC42, and RHO. The assay suggested significantly (p<0.001) reduced GTP loading to each tested GTPase therefore being in line with reduced cell proliferation, migration, and growth related signal transduction. Data are expressed as mean ±SD. The p-values (* = p<0.05, ** = p<0.01, *** = p<0.001).</p

MOESM1 of Clinical relevance of thyroid cell models in redox research

Additional file 1: Table S1. PCR primers

Western blotting and DNA damage response analysis.

<p>(a,b) Western blotting analyses supported the protein array data showing the increased activation of RTKs and non-receptor kinases in the <i>SOD3</i>-transfected 8505c cells. Note, MEK1/2 and ERK1/2 phosphorylation was moderately downregulated based on the band density analyses. (c,d) Western blotting and the related band intensity analysis for β-catenin nuclear and cytosolic proteins showed significantly (p<0.001) increased β-catenin production in cytosolic compartment. Tubulin was used to normalize cytosolic proteins and SP1 was used to normalize nuclear proteins. (e,f) γH2AX staining for 8505c cells suggested significantly (p<0.001) increased histone H2AX phosphorylation, suggesting DNA damages in 8505c SOD3 cells, which supported the increased CHK2 phosphorylation observed in protoarray and in the Western blotting. The white arrows in the control and the SOD3 merge panels indicate cells that are magnified to detect the nuclear foci. The white arrows in the control and the SOD3 magnification images indicate the nuclear foci. Data are expressed as mean ±SD. The p-values (* = p<0.05, ** = p<0.01, *** = p<0.001).</p

Time lapses between distribution of influenza vaccines to health authorities and their administration by General Practitioners (GPs) to older adults: a retrospective study over five influenza seasons in Italy

Delays in influenza vaccine delivery and administration can hinder vaccine coverage and protection. This study examines the differentials in distributing and administering adjuvanted trivalent (aTIV) and quadrivalent influenza vaccines (aQIV) to older adults in Italy’s primary care setting and its potential impact on hospitalization risk over 5 epidemic seasons. Using a primary care database, individuals aged ≥ 65 years were selected. The proportion of vaccine distribution to regional authorities and subsequent administration by GPs was estimated using census data. Using quantile (median) regression, we examined the relationship between velocities of vaccine distribution and administration (doses/week) and the incidence of hospitalizations. Over the 5 influenza seasons, the velocity of distribution and administration of aTIV/aQIV ranged 341–833 and 152–270 median doses/week; no trend was yielded for the difference between these velocities (p = 0.189) or vaccine coverage (p = 0.142). An association was observed for each differential dose/week between distributed and administered vaccines and all-cause hospitalizations with a 10% increase in 2017–2018, 54% in 2018–2019, and 12% in 2020–2021 season. These findings highlight the importance of minimizing the time lapse between vaccine distribution and administration to mitigate the impact of influenza and address factors that contribute to vaccination barriers.</p

Semiquantitative visual assessment.

<p>Frequency histogram of the semiquantitative scores for the display of the brain structures of the MNLM-SWI (gray), MNLM-HP-SWI (orange), SWI (green), NLM-SWI (yellow), IR-SWI (cyan) and MIR-SWI (red) images. Score values from 1 to 5 indicate increasing overall image quality (see text).</p