This study extends current knowledge of miRNA-mediated gene expression regulation in mTLE+HS by identifying miRNAs with altered expression in mTLE+HS, including nine novel abnormally expressed miRNAs and their putative targets. These observations further encourage the potential of microRNA-based biomarkers or therapies.

Abstract:We report on improvement of the adsorptive transfer stripping technique (AdTS) coupled with the differential pulse voltammetry Brdicka reaction to determine a thiol-protein. The current technique has been unable to generate reproducible results when analyzing very low sample volumes (nanolitres). This obstacle can be overcome technically by modifying the current transfer technique including cooling step of the adsorbed analyte. We tested the technique on determination of a promising tumour disease marker protein called metallothionein (MT). The detection limit (3 S/N) of MT was evaluated as 500 zeptomoles per 500 nL (1 pM) and the quantification limit (10 S/N) as 1,500 zeptomoles per 500 nL (3 pM). Further, the improved AdTS technique was utilized to analyze blood serum samples from patients with breast cancer. Based on the results obtained it can be concluded that the improved technique can be used to detect a thiolprotein in very low sample volumes and can also prevent interferences during the washing and transferring step.

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Due to unique physico-chemical properties of -SH moiety thiols comprise wide group of biologically important compounds. A review devoted to biological functions of glutathione and phytochelatins with literature survey of methods used to analysis of these compounds and their interactions with cadmium(II) ions and Murashige-Skoog medium is presented. For these purposes electrochemical techniques are used. Moreover, we revealed the effect of three different cadmium concentrations (0, 10 and 100 μM) on cadmium uptake and thiols content in maize plants during 192 hours long experiments using differential pulse anodic stripping voltammetry to detect cadmium(II) ions and high performance liquid chromatography with electrochemical detection to determine glutathione. Cadmium concentration determined in tissues of the plants cultivated in nutrient solution containing 10 μM Cd was very low up to 96 hours long exposition and then the concentration of Cd markedly increased. On the contrary, the addition of 100 μM Cd caused an immediate sharp increase in all maize plant parts to 96 hours Cd exposition but subsequently the Cd concentration increased more slowly. A high performance liquid chromatography with electrochemical detection was used for glutathione determination in treated maize plants after 96 and 192 hours of treatment. The highest total content of Sensors 2007, 7 933 glutathione per one plant was 6 μg (96 h, 10 μM Cd) in comparison with non-treated plant (control) where glutathione content was 1.5 μg. It can be concluded that electrochemical techniques have proved to be useful to analyse plant thiols.

Abstract:The main aim of this work was to use multi-instrumental analytical apparatus to investigate the effects of treatment with cadmium(II) and/or lead(II) ions (50, 250 and 500 µM) for twelve days on early somatic spruce embryos (ESEs). Primarily we used image analysis for estimation of growth and a fluorimetric sensor for enzymatic detection of viability of the treated ESEs. It follows from the obtained results that Cd caused higher toxicity to ESEs than Pb. Besides this fundamental finding, we observed that ESEs grew and developed better in the presence of 500 µM of the metal ions than in the presence of 250 µM. Based on the results obtained using nuclear magnetic resonance this phenomenon was related to an increase of the area of ESE clusters by intensive uptake of water from the cultivation medium, due to dilution of the heavy metal concentration inside the cluster. In addition we studied the glutathione content in treated ESEs by the adsorptive transfer stripping technique coupled with the differential pulse voltammetry Brdicka reaction. GSH contents increased up to 148 ng/mg (clone 2/32) and 158 ng/mg (clone PE 14) after twelve Sensors 2007, 7 744 day long treatment with Cd-EDTA ions. The GSH content was about 150 and 160 % higher in comparison with the ESEs treated with Pb-EDTA ions, respectively. The difference between GSH contents determined in ESEs treated with Pb-EDTA and Cd-EDTA ions correlates with the higher toxicity of cadmium(II) ions.

S100A11 (calgizzarin), a member of S100 family, is associated with several autoimmune diseases, including rheumatoid arthritis (RA). Neutrophil extracellular traps (NETs) are implicated in the pathogenesis of RA and in the externalization of some S100 family members. Therefore, we aimed to determine the association between S100A11 and NETs in RA. For this purpose, the levels of S100A11 and NETosis markers were detected in the RA synovial fluid by immunoassays. The expression of S100A11 by neutrophils in the RA synovial tissue was assessed. Neutrophils isolated from peripheral blood were exposed to S100A11 or stimulated to release NETs. The levels of NETosis- and inflammation-associated proteins were analysed by immunoassays. NETs were visualized by immunofluorescence. We showed that S100A11 was expressed by the neutrophils in the RA synovial tissue. Moreover, S100A11 in the RA synovial fluid correlated with several NETosis markers. In vitro, S100A11 was abundantly released by neutrophils undergoing NETosis compared to untreated cells (p < 0.001). Extracellular S100A11 increased the secretion of IL-6 (p < 0.05) and TNF (p < 0.05) by neutrophils but did not induce NETosis. This study demonstrates, for the first time, that the release of S100A11 is dependent on NETosis and that extracellular S100A11 augments the inflammatory response by inducing pro-inflammatory cytokines in neutrophils.

In the present paper potato plants were cultivated in the presence of ammonium sulphate or elemental sulphur supplementation into the soil to reveal the effects of different sulphur forms on content of nitrogen, phosphorus, potassium, calcium, magnesium and sulphur, and yield of tubers. During the investigation of the influence of different sulphur forms on yield of potato tubers we did not observe significant changes. Average weight of tubers of control plants per one experimental pot was 355 g. Application of sulphur in both forms resulted in moderate potato tubers weight reduction per one experimental pot compared to control group; average value ranged from 320 to 350 g per one experimental pot. Further we treated the plants with two different supplementation of sulphur with cadmium(II) ions (4 mg of cadmium(II) acetate per kilogram of the soil). The significantly lowest cadmium content (p < 0.05) was determined in tissues of plants treated with the highest dosage of elemental sulphur (0.64 mg Cd/kg) compared to control plants (0.82 mg Cd/kg). We also aimed our attention on the cadmium content in proteins, lipids or soluble carbohydrates and ash. Application of sulphate as well as elemental sulphur resulted in significant cadmium content reduction in lipid fraction compared to control plants. In addition to this we quantified content of low molecular mass thiols in potatoes tissues. To determine the thiols content we employed differential pulse voltammetry Brdicka reaction. After twelve days of the treatment enhancing of thiols level was observed in all experimental groups regardless to applied sulphur form and its concentration. Finally we evaluated the effect of sulphur supplementation on Phytophora infestans infection of potato plants.

Temporal lobe epilepsy (TLE) is a severe neurological disorder accompanied by recurrent spontaneous seizures. Although the knowledge of TLE onset is still incomplete, TLE pathogenesis most likely involves the aberrant expression of microRNAs (miRNAs). miRNAs play an essential role in organism homeostasis and are widely studied in TLE as potential therapeutics and biomarkers. However, many discrepancies in discovered miRNAs occur among TLE studies due to model-specific miRNA expression, different onset ages of epilepsy among patients, or technology-related bias. We employed a massive parallel sequencing approach to analyze brain tissues from 16 adult mesial TLE (mTLE)/hippocampal sclerosis (HS) patients, 8 controls and 20 rats with TLElike syndrome, and 20 controls using the same workflow and categorized these subjects based on the age of epilepsy onset. All categories were compared to discover overlapping miRNAs with an aberrant expression, which could be involved in TLE. Our cross-comparative analyses showed distinct miRNA profiles across the age of epilepsy onset and found that the miRNA profile in rats with adult-onset TLE shows the closest resemblance to the profile in mTLE/HS patients. Additionally, this analysis revealed overlapping miRNAs between patients and the rat model, which should participate in epileptogenesis and ictogenesis. Among the overlapping miRNAs stand out miR-142-5p and miR-142-3p, which regulate immunomodulatory agents with pro-convulsive effects and suppress neuronal growth. Our cross-comparison study enhanced the insight into the effect of the age of epilepsy onset on miRNA expression and deepened the knowledge of epileptogenesis. We employed the same methodological workflow in both patients and the rat model, thus improving the reliability and accuracy of our results.