Transcriptomic stratification of breast carcinomas with double-equivocal HER2 status

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Osteogenic potential of fast set bioceramic cements: Molecular and in vitro study

Recently, pre-mixed bioceramics in fast set formulations have been increasingly utilized in clinical practice as an alternative to mineral trioxide aggregate (MTA) for their shorter setting time and better handling properties. However, the impact on their osteogenic potential, due to modifications in chemical composition to promote a fast setting, is still unclear. This molecular and in vitro study compared the osteogenic potential of root repairing material putty fast set (FSP) with root-repairing material putty (RRMPU), root-repairing material paste (RRMPA), Biodentine™ and MTA. The null hypothesis tested was that there are no differences among the tricalcium silicate materials in terms of osteogenic potential. Standardized discs were cultured with MG-63 human osteoblastic-like cells to assess biocompatibility, the activity of alkaline phosphatase (ALP) and osteogenic potential. Biocompatibility was evaluated at baseline and after 24 and 48 h. Osteogenic differentiation was assessed after 15 days. Data were analyzed with one-way ANOVAs and Tukey’s post-hoc test (p < 0.05). All materials showed biocompatibility and bioactivity. ALP activity, which induces mineral nodule deposition, increased in all the cements tested, with a significant increase in RRMPU (p < 0.001) and FSP (p < 0.001) samples versus MTA. In vitro mineralization was significantly increased for RRMPU (p < 0.0001), FSP (p = 0.00012) and Biodentine™ (p < 0.0001) versus MTA. The bioceramics tested showed higher levels of biocompatibility and bioactivity than MTA; a higher capacity for mineralization was observed with RRMPU and FSP versus MTA

AR/ER Ratio Correlates with Expression of Proliferation Markers and with Distinct Subset of Breast Tumors

The co-expression of androgen (AR) and estrogen (ER) receptors, in terms of higher AR/ER ratio, has been recently associated with poor outcome in ER-positive (ER+) breast cancer (BC) patients. The aim of this study was to analyze if the biological aggressiveness, underlined in ER+ BC tumors with higher AR/ER ratio, could be due to higher expression of genes related to cell proliferation. On a cohort of 47 ER+ BC patients, the AR/ER ratio was assessed by immunohistochemistry and by mRNA analysis. The expression level of five gene proliferation markers was defined through TaqMan®-qPCR assays. Results were validated using 979 BC cases obtained from gene expression public databases. ER+ BC tumors with ratios of AR/ER ≥ 2 have higher expression levels of cellular proliferation genes than tumors with ratios of AR/ER < 2, in both the 47 ER+ BC patients (P < 0.001) and in the validation cohort (P = 0.005). Moreover, BC cases with ratios of AR/ER ≥ 2 of the validation cohort were mainly assigned to luminal B and HER2-enriched molecular subtypes, typically characterized by higher proliferation and poorer prognosis. These data suggest that joint routine evaluation of AR and ER expression may identify a unique subset of tumors, which show higher levels of cellular proliferation and therefore a more aggressive behavior

Identification of TENM4 as a novel cancer stem cell-associated molecule and potential target in triple negative breast cancer

Triple-negative breast cancer (TNBC) is insensitive to endocrine and Her2-directed therapies, making the development of TNBC-targeted therapies an unmet medical need. Since patients with TNBC frequently show a quicker relapse and metastatic progression compared to other breast cancer subtypes, we hypothesized that cancer stem cells (CSC) could have a role in TNBC. To identify putative TNBC CSC-associated targets, we compared the gene expression profiles of CSC-enriched tumorspheres and their parental cells grown as monolayer. Among the up-regulated genes coding for cell membrane-associated proteins, we selected Teneurin 4 (TENM4), involved in cell differentiation and deregulated in tumors of different histotypes, as the object for this study. Meta-analysis of breast cancer datasets shows that TENM4 mRNA is up-regulated in invasive carcinoma specimens compared to normal breast and that high expression of TENM4 correlates with a shorter relapse-free survival in TNBC patients. TENM4 silencing in mammary cancer cells significantly impaired tumorsphere-forming ability, migratory capacity and Focal Adhesion Kinase (FAK) phosphorylation. Moreover, we found higher levels of TENM4 in plasma from tumor-bearing mice and TNBC patients compared to the healthy controls. Overall, our results indicate that TENM4 may act as a novel biomarker and target for the treatment of TNBC

A collection of primary tissue cultures of tumors from vacuum packed and cooled surgical specimens: a feasibility study

Primary cultures represent an invaluable tool to set up functional experimental conditions; however, creation of tissue cultures from solid tumors is troublesome and often unproductive. Several features can affect the success rate of primary cultures, including technical issues from pre-analytical procedures employed in surgical theaters and pathology laboratories. We have recently introduced a new method of collection, transfer, and preservation of surgical specimens that requires immediate vacuum sealing of excised specimens at surgical theaters, followed by time-controlled transferring at 4°C to the pathology laboratory. Here we investigate the feasibility and performance of short-term primary cell cultures derived from vacuum packed and cooled (VPAC) preserved tissues. Tissue fragments were sampled from 52 surgical specimens of tumors larger than 2 cm for which surgical and VPAC times (the latter corresponding to cold ischemia time) were recorded. Cell viability was determined by trypan blue dye-exclusion assay and hematoxylin and eosin and immunohistochemical stainings were performed to appreciate morphological and immunophenotypical features of cultured cells. Cell viability showed a range of 84-100% in 44 out of 52 (85%) VPAC preserved tissues. Length of both surgical and VPAC times affected cell viability: the critical surgical time was set around 1 hour and 30 minutes, while cells preserved a good viability when kept for about 24 hours of vacuum at 4°C. Cells were maintained in culture for at least three passages. Immunocytochemistry confirmed the phenotype of distinct populations, that is, expression of cytokeratins in epithelioid cells and of vimentin in spindle cells. Our results suggest that VPAC preserved tissues may represent a reliable source for creation of primary cell cultures and that a careful monitoring of surgical and cold ischemia times fosters a good performance of primary tissue cultures

Particles as probes for complex plasmas in front of biased surfaces

An interesting aspect in the research of complex (dusty) plasmas is the experimental study of the interaction of micro-particles with the surrounding plasma for diagnostic purposes. Local electric fields can be determined from the behaviour of particles in the plasma, e.g. particles may serve as electrostatic probes. Since in many cases of applications in plasma technology it is of great interest to describe the electric field conditions in front of floating or biased surfaces, the confinement and behaviour of test particles is studied in front of floating walls inserted into a plasma as well as in front of additionally biased surfaces. For the latter case, the behaviour of particles in front of an adaptive electrode, which allows for an efficient confinement and manipulation of the grains, has been experimentally studied in dependence on the discharge parameters and on different bias conditions of the electrode. The effect of the partially biased surface (dc, rf) on the charged micro-particles has been investigated by particle falling experiments. In addition to the experiments we also investigate the particle behaviour numerically by molecular dynamics, in combination with a fluid and particle-in-cell description of the plasma.Comment: 39 pages, 16 figures, submitted to New J. Phy