The Chick Chorioallantoic Membrane Model: A New In Vivo Tool to Evaluate Breast Cancer Stem Cell Activity

The high plasticity of cancer stem-like cells (CSCs) allows them to differentiate and proliferate, specifically when xenotransplanted subcutaneously into immunocompromised mice. CSCs are highly tumorigenic, even when inoculated in small numbers. Thus, in vivo limiting dilution assays (LDA) in mice are the current gold standard method to evaluate CSC enrichment and activity. The chick embryo chorioallantoic membrane (CAM) is a low cost, naturally immune-incompetent and reproducible model widely used to evaluate the spontaneous growth of human tumor cells. Here, we established a CAM-LDA assay able to rapidly reproduce tumor specificities—in particular, the ability of the small population of CSCs to form tumors. We used a panel of organotropic metastatic breast cancer cells, which show an enrichment in a stem cell gene signature, enhanced CD44⁺/CD24^−/low cell surface expression and increased mammosphere-forming efficiency (MFE). The size of CAM-xenografted tumors correlate with the number of inoculated cancer cells, following mice xenograft growth pattern. CAM and mice tumors are histologically comparable, displaying both breast CSC markers CD44 and CD49f. Therefore, we propose a new tool for studying CSC prevalence and function—the chick CAM-LDA—a model with easy handling, accessibility, rapid growth and the absence of ethical and regulatory constraints.     

Target Score—A Proteomics Data Selection Tool Applied to Esophageal Cancer Identifies GLUT1-Sialyl Tn Glycoforms as Biomarkers of Cancer Aggressiveness

Esophageal cancer (EC) is a life-threatening disease, demanding the discovery of new biomarkers and molecular targets for precision oncology. Aberrantly glycosylated proteins hold tremendous potential towards this objective. In the current study, a series of esophageal squamous cell carcinomas (ESCC) and EC-derived circulating tumor cells (CTCs) were screened by immunoassays for the sialyl-Tn (STn) antigen, a glycan rarely expressed in healthy tissues and widely observed in aggressive gastrointestinal cancers. An ESCC cell model was glycoengineered to express STn and characterized in relation to cell proliferation and invasion in vitro. STn was found to be widely present in ESCC (70% of tumors) and in CTCs in 20% of patients, being associated with general recurrence and reduced survival. Furthermore, STn expression in ESCC cells increased invasion in vitro, while reducing cancer cells proliferation. In parallel, an ESCC mass spectrometry-based proteomics dataset, obtained from the PRIDE database, was comprehensively interrogated for abnormally glycosylated proteins. Data integration with the Target Score, an algorithm developed in-house, pinpointed the glucose transporter type 1 (GLUT1) as a biomarker of poor prognosis. GLUT1-STn glycoproteoforms were latter identified in tumor tissues in patients facing worst prognosis. Furthermore, healthy human tissues analysis suggested that STn glycosylation provided cancer specificity to GLUT1. In conclusion, STn is a biomarker of worst prognosis in EC and GLUT1-STn glycoforms may be used to increase its specificity on the stratification and targeting of aggressive ESCC forms.     

Expression and Roles of Individual HIF Prolyl 4-Hydroxylase Isoenzymes in the Regulation of the Hypoxia Response Pathway along the Murine Gastrointestinal Epithelium

The HIF prolyl 4-hydroxylases (HIF-P4H) control hypoxia-inducible factor (HIF), a powerful mechanism regulating cellular adaptation to decreased oxygenation. The gastrointestinal epithelium subsists in “physiological hypoxia” and should therefore have an especially well-designed control over this adaptation. Thus, we assessed the absolute mRNA expression levels of the HIF pathway components, Hif1a, HIF2a, Hif-p4h-1, 2 and 3 and factor inhibiting HIF (Fih1) in murine jejunum, caecum and colon epithelium using droplet digital PCR. We found a higher expression of all these genes towards the distal end of the gastrointestinal tract. We detected mRNA for Hif-p4h-1, 2 and 3 in all parts of the gastrointestinal tract. Hif-p4h-2 had significantly higher expression levels compared to Hif-p4h-1 and 3 in colon and caecum epithelium. To test the roles each HIF-P4H isoform plays in the gut epithelium, we measured the gene expression of classical HIF target genes in Hif-p4h-1^−/−, Hif-p4h-2 hypomorph and Hif-p4h-3^−/− mice. Only Hif-p4h-2 hypomorphism led to an upregulation of HIF target genes, confirming a predominant role of HIF-P4H-2. However, the abundance of Hif-p4h-1 and 3 expression in the gastrointestinal epithelium implies that these isoforms may have specific functions as well. Thus, the development of selective inhibitors might be useful for diverging therapeutic needs.     

Antibacterial properties of polyaniline derivatives

This work presents a detailed study on the effect of various functional groups both at the ortho position of the aromatic ring and in the amino group of PANI on the antibacterial properties of polymers against gram-positive (Bacillus subtilis) and gram-negative (Pseudomonas aureofaciens) microorganisms. It was found that incorporation of methyl, methoxy, or pentyl groups into the ortho-position of PANI did not increase the antibacterial activity but in most cases causes a significant decrease in the antibacterial properties of functionally substituted polyanilines. At the same time, PANI derivatives modified by incorporation of pentyl groups into the amino group were found to be more efficient antibacterial compounds against both gram-positive and gram-negative microorganisms than the original polymer. It was also found that N-substituted PANI derivatives manifest not only bactericidal but also bacteriostatic properties toward the test microorganisms. Varying the nature and position of the substituent allowed us to conclude that the synthesis of various N-substituted PANI derivatives with a high degree of doping is the most promising approach to PANI modification for application in bacterial growth inhibition.     

Ni/Al Intermetallics Plasma Transferred Arc Processing

TAILORING COMPONENT SUFACES for elevatedtemperature operations has been a goal of researchersduring the last century.Process and material selectionare the key factors for this development.In particularlythe latter can be described to have started with thedevelopment of stainless steels.Ni and Co based alloys,the superalloys,attended the increasing technologicaldemands for higher service temperatures(1).Morerecently the higher operating temperatures ofequipments,such as gas turbines,r…     

The Influence of High Temperature Exposure on the Wear Performance of a Ni Based Alloy PTA Coating

Degradation phenomena like wear and corrosion, may have their effects accelerated as components operate at high temperature. The aim of this work is to make an indirect evaluation of the influence of high temperature exposure on wear behavior of Ni coatings. A Ni-Cr-Mo-W alloy, was deposited with Plasma Transferred Arc (PTA) process. An indirect approach based on the Ahrrenius relationship was used to evaluated long time exposures at high temperatures. To simulate longer exposures at lower temperatures, coatings were exposed to higher temperatures for the same time interval. So coatings were soaked in an air furnace at 650℃, 1000℃ and 1200℃. Metal/metal wear behavior was evaluated by pin-on-disc tests. Pins were removed from the coatings by eletrodischarge machining and rubbed against a VC31 quenched and tempered tool steel. Two sets of wear tests were run at 2m/s, for different loads (0.5, 1.0, 2.0 and 3.0Kg) and for different sliding distances (120, 720, 1080, 1440 and 1800m).Coatings characterization was done by scanning electronic microscopy and Vickers microhardness. Results showed as temperature is increased, coatings wear performance is altered,and this is associated with microstructural changes.     

Smart Workplaces for older adults: coping ‘ethically’ with technology pervasiveness

Universal Access in the Information Society - Pervasive technologies such as Artificial Intelligence, Virtual Reality and the Internet of Things, despite their great potential for improved...     

Impact of white-matter mask selection on DTI histogram-based metrics as potential biomarkers in cerebral small vessel disease

Magnetic Resonance Materials in Physics, Biology and Medicine - Histogram-based metrics extracted from diffusion-tensor imaging (DTI) have been suggested as potential biomarkers for cerebral small...