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ObjectivesThe aim of our study was to determine whether there is a correlation between transcription factors expression and Th17/Treg ratio, cytokine profile in the RA phenotype as well as to identify transcription factors that could be a potential biomarker for RA.MethodsThe study was conducted on 45 patients with RA, 27 patients with OA and 46 healthy controls (HCs). Th17 and Treg frequency was determined by flow cytometry (15 patients with RA/OA and 15 subjects of HC). Gene expression was estimated by qPCR, and the serum cytokine levels were determined by ELISA.ResultsThe percentage of Treg (CD4+CD25highCD127-) cells in RA patients was lower than in OA patients or HCs. Proportions of Th17 (CD4+CCR6+CXCR3-) cells were higher in RA and OA in comparison to HCs. STAT5 showed a very high expression in the blood of RA patients compared to healthy subjects. The expression of STAT5 and HELIOS was not detected in Th17 cells. A positive correlation between SMAD3 and STAT3 in RA patients was observed. Negative correlations between HIF-1A and SMAD2 in RA Treg cells and DAS-28 score were observed. The range of serum of IL-17 and IL-21 were higher in RA patients than in OA patients. Concentrations of serum IL-2 and IFN-γ were higher in RA and OA patients than in healthy subjects. Based on the ROC analysis, the diagnostic potential of the combination of HIF1A, SMAD3 and STAT3, was determined at AUC 0.95 for distinguishing RA patients from HCs. For distinguishing RA patients from OA patients the diagnostic potential of the combination of SMAD2, SMAD3, SMAD4 and STAT3, was determined at AUC 0.95.ConclusionBased on our study, we conclude that SMAD3 and STAT3 could be potential diagnostic biomarkers for RA.

PurposeNew-onset diabetes after transplantation (NODAT) is a major complication after kidney transplantation. The risk factors for NODAT include the use of calcineurin inhibitors as part of the immunosuppressive regimen, among which tacrolimus has the most pronounced diabetogenic effect. Both NODAT and type 2 diabetes mellitus (T2DM) share several risk factors. Recent studies have identified a number of common genetic variants associated with increased risk of T2DM. Here we report the results of our study on the potential effect of single nucleotide polymorphisms (SNPs) previously associated with T2DM on the risk of NODAT in kidney transplant patients medicated with tacrolimus.MethodsSeven SNPs in six genes known to increase the risk of T2DM in Caucasians were genotyped by means of TaqMan assays in 235 kidney transplant patients medicated with tacrolimus: rs4402960 and rs1470579 in IGF2BP2; rs1111875 in HHEX; rs10811661 upstream of CDKN2A/B; rs13266634 in SLC30A8; rs1801282 in PPARG; rs5215 in KCNJ11. The TCF7L2 rs7903146 SNP was also included in the multivariate analysis.ResultsNone of the analyzed SNPs was significantly associated with the risk of NODAT. However, the IGF2BP2 rs4402960 T allele was present significantly more frequently among patients diagnosed with NODAT more than 2 weeks after transplantation (p = 0.048). Mean (± standard deviation) number of the analyzed alleles tended to be lower in patients without NODAT (6.19 ± 1.71) than in NODAT patients (6.58 ± 1.1.95; p = 0.09) and significantly lower compared to late-onset NODAT patients (7.03 ± 1.88; p = 0.018). Multivariate analysis confirmed the significance of ‘diabetogenic’ allele number in late-onset NODAT development [odds ratio (OR) 1.37, 95 % confidence interval (CI) 1.05–1.78; p = 0.017]. Additionally, individuals carrying >7 of the analyzed ‘diabetogenic’ alleles were at a significantly higher risk of NODAT (OR 2.17, 95 % CI 1.18–3.99; p = 0.015).ConclusionsComplex analysis of genotypes increasing the risk of diabetes may lead to the identification of NODAT susceptibility predictors.

MicroRNAs regulate gene expression of transcriptional factors, which influence Th17/Treg (regulatory T cells) balance, establishing the molecular mechanism of genetic and epigenetic regulation of Treg and Th17 cells is crucial for understanding rheumatoid arthritis (RA) pathogenesis. The study goal was to understand the potential impact of the selected microRNAs expression profiles on Treg/Th17 cells frequency, RA phenotype, the expression profile of selected microRNAs, and their correlation with the expression profiles of selected transcriptional factors: SOCS1, SMAD3, SMAD4, STAT3, STAT5 in RA; we used osteoarthritis (OA) and healthy controls (HCs) as controls. The study was conducted on 14 RA and 11 OA patients, and 15 HCs. Treg/Th17 frequency was established by flow cytometry. Gene expression analysis was estimated by qPCR. We noticed correlations in RA Th17 cells between miR-26 and SMAD3, STAT3, SOCS1; and miR-155 and STAT3—and in RA Treg cells between miR-26 and SOCS1; miR-31, -155 and SMAD3; and miR-155 and SMAD4. In RA Tregs, we found a negative correlation between miR-26, -126 and STAT5a. The expression level of miR-31 in Th17 cells from RA patients with DAS28 ≤ 5.1 is higher and that for miR-24 is greater in Tregs from patients with DAS28 > 5.1. MiR-146a in Tregs is higher in rheumatoid factor (RF) positive RA patients.

In this contribution we present comparative study on synthesis, bio-characterization and antibacterial properties of mesoporous silica nanospheres modified by titanium dioxide. Mesoporous silica nanospheres functionalized by titania were studied as light activated antibacterial agents. The analysis of the antibacterial effects on E. coli ATCC 25922 shows strong enhancement during the visible and ultraviolet light irradiation in respect to the commercial catalyst and sample free from the nanomaterials. In darkness the mesoporous silica/titania nanostructures revealed low antibacterial activity dependent on the stirring intensity of the suspension containing nanomaterials and bacteria. The nanomaterials toxicity was determined on the amount of lactate dehydrogenase released from mouse fibroblast cells L929 with LDH assay. Sample was characterized in details by means of high resolution transmission electron microscopy (HR-TEM), Raman spectroscopy, XRD and BET Isotherm. photocatalytic efficiency [18-21]. Composite structures of titania with Ag nanoparticles are studied for their photocatalytic activity and have shown the enhanced photodegradation of organic molecules [22-24]. This article describes a simple synthesis methodology of mesoporous silica nanospheres functionalized by TiO 2 with enhanced bactericidal efficiency and photocatalytic properties. The detailed studies on spherical mesoporous structures have been performed. Furthermore, the correlation between cytotoxicity and the antibacterial activity of the samples has been revealed. Materials and Methods Materials Silica (tetraethyl orthosilicate-TEOS) and titania precursor (Titanium(IV) butoxide-TBT) was purchased from Sigma-Aldrich. Ammonium solution, ethanol and n-propanol were provided from Chempure (Poland, Piekary Sląskie) and from Polskie Odczynniki Chemiczne-POCH S.A. (Poland, Gliwice). Synthesis of silica nanospheres with mesoporous shell (mSiO 2) 1.5 mL of the tetraethyl orthosilicate (TEOS) and 50.0 mL of