Inflammatory reactions to ssRNA viruses are induced by the endosomal Toll-like receptors (TLRs) 7 and 8. TLR7/8-mediated inflammatory reaction results in activation of the Nalp3 inflammasome via an unknown mechanism. Here we report for the first time that TLR7/8 mediate activation of xanthine oxidase (XOD) in an HIF-1α-dependent manner. XOD produces uric acid and reactive oxygen species, which could activate Nalp3 and therefore induce activation of caspase 1, known to convert inactive pro-IL-1β into active IL-1β. Specific inhibition of the XOD activity attenuates TLR7/8-mediated activation of caspase 1 and IL-1β release. These results were obtained using human THP-1 myeloid macrophages. The findings were verified by conducting in vivo experiments on mice.

DNA methylation patterns have been recognised as cancer-specific markers with high potential for clinical applications. We aimed at identifying methylation variations that differentiate between breast cancers and other breast tissue entities to establish a signature for diagnosis. Candidate genomic loci were analysed in 117 fresh-frozen breast specimens, which included cancer, benign and normal breast tissues from patients as well as material from healthy individuals. A cancer-specific DNA methylation signature was identified by microarray analysis in a test set of samples (n = 52, p < 2.1 × 10(-4)) and its performance was assessed through bisulphite pyrosequencing in an independent validation set (n = 65, p < 1.9 × 10(-7)). The signature is associated with SFRP2 and GHSR genes, and exhibited significant hypermethylation in cancers. Normal-appearing breast tissues from cancer patients were also methylated at these loci but to a markedly lower extent. This occurrence of methylated DNA in normal breast tissue of cancer patients is indicative of an epigenetic field defect. Concerning diagnosis, receiver operating characteristic curves and the corresponding area under the curve (AUC) analysis demonstrated a very high sensitivity and specificity of 89.3 and 100 %, respectively, for the GHSR methylation pattern (AUC >0.99). To date, this represents the DNA methylation marker of the highest sensitivity and specificity for breast cancer diagnosis. Functionally, ectopic expression of GHSR in a cell line model reduced breast cancer cell invasion without affecting cell viability upon stimulation of cells with ghrelin. Our data suggest a link between epigenetic down-regulation of GHSR and breast cancer cell invasion.

The hyperplastic processes of the endometrium can arise not only against the background of excessive influence of estrogen, but also against the background of epigenetic damages that affect apoptosis, cell proliferation, differentiation, and adhesion, and DNA reparation. The aim of our study was to investigate and analyze the status of methylation of the promoter of SFRP2 gene in patients with hyperplastic processes of the endometrium. Materials and Methods: The study groups were the following: I — patients with endometrial hyperplasia (EH, n = 9); II — patients with endometrial intraepithelial neoplasia (EIN, n = 10), III — control groups: 1) with endometrial cancer (EC, n = 4), and 2) healthy women (n = 4). Determination of promoter methylation of SFRP2 gene was carried out by the semiquantitative method of methylation-specific PCR assay. Results: The maximum level of methylation of SFRP2 gene promoter had been revealed in patients with EC — 42.80 ± 3.55% (р < 0.05). The patients of the I group had the lowest values of methylation of SFRP2 gene promoter — 10.66 ± 0.85%, while in patients of the II group this indicator was higher — 20.60 ± 0.95% (р < 0.05). In healthy women of the control group, methylation of SFRP2 gene promoter was detected in none of the samples. Conclusion: The content of the methylated SFRP2 gene in endometrial tissue of patients with hyperplastic processes higher than 20–25% allows relate these women to the risk group of EC development and dictates the need of intensive observation of such patients.

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