imagine (Institute of molecular genetics and genetic engineering)
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The role of microRNAs involved in regulation TGFβ signaling pathway in metastatic colorectal cancer - functional analysis and response to therapy
Get PDFAntimicrobial Activity of Rhenium Di- and Tricarbonyl Diimine Complexes: Insights on Membrane-Bound S. aureus Protein Binding
Get PDFAntimicrobial resistance is one of the major human health threats, with significant impacts on the global economy. Antibiotics are becoming increasingly ineffective as drug-resistance spreads, imposing an urgent need for new and innovative antimicrobial agents. Metal complexes are an untapped source of antimicrobial potential. Rhenium complexes, amongst others, are particularly attractive due to their low in vivo toxicity and high antimicrobial activity, but little is known about their targets and mechanism of action. In this study, a series of rhenium di- and tricarbonyl diimine complexes were prepared and evaluated for their antimicrobial potential against eight different microorganisms comprising Gram-negative and -positive bacteria. Our data showed that none of the Re dicarbonyl or neutral tricarbonyl species have either bactericidal or bacteriostatic potential. In order to identify possible targets of the molecules, and thus possibly understand the observed differences in the antimicrobial efficacy of the molecules, we computationally evaluated the binding affinity of active and inactive complexes against structurally characterized membrane-bound S. aureus proteins. The computational analysis indicates two possible major targets for this class of compounds, namely lipoteichoic acids flippase (LtaA) and lipoprotein signal peptidase II (LspA). Our results, consistent with the published in vitro studies, will be useful for the future design of rhenium tricarbonyl diimine-based antibiotics
Coriolus versicolor Mushroom Grown on Selenium-Rich Zeolitic Tuff as a Potential Novel Food Supplement
Get PDFResearch background. In the recent years, considerable attention has been given to selenium status since its deficiency is linked with various disorders and affects at least 13 % of world population. Additionally, mushrooms are known to possess pronounced capacity for absorption of various micronutrients, including Se, from soil/substrate. Here, we investigate the possibility of using Se-rich zeolitic tuff as a supplement for production of selenized mushroom. Furthermore, the impact of the enrichment on the activity of antioxidant enzymes and biological potential of Coriolus versicolor medicinal mushroom is studied. Experimental approach. Se(IV)- and Se(VI)-modified natural zeolitic tuff from the Serbian deposit Zlatokop was used as supplement for mushroom cultivation. To examine the effectiveness of selenium enrichment, we determined total selenium with inductively coupled plasma mass spectrometry (ICP-MS), together with the activity of antioxidant enzymes in fresh fruiting bodies and biological potential of methanolic extracts. Antioxidant activity was evaluated using the appropriate tests for: inhibition of lipid peroxidation, DPPH free radical scavenging assay, Fe(III)-reducing antioxidant power assay and ability of chelating Fe2+ ions. The antibacterial activity against foodborne pathogens was measured by broth microdilution assay. Additionally, chemical composition of the prepared extracts was studied using UV-Vis and Fourier transform infrared (FTIR) spectroscopy. Results and conclusions. Content of selenium detected in biofortified C. versicolor was even 470 times higher than in control on dry mass basis ((140.7 +/- 3.8) vs (0.3 +/- 0.1) mu g/g), proving that Se-rich zeolitic tuff is an excellent supplement for mushroom production. Furthermore, the results of monitoring the activity of antioxidant enzymes revealed that most of the Se-enriched mushrooms exhibited higher superoxide dismutase (SOD) and cata lase (CAT) and lower glutathione peroxidase (GSH-Px) activities than control. Due to higher amounts of enzymes, which can quickly catalyze the reduction of superoxide radicals, the quality of selenium-enriched mushrooms is preserved for a longer period of time. Investigation of biological potential indicated that Se-enriched mushroom methanolic extracts, generally, expressed enhanced antioxidant properties. Additionally, extracts showed antibacterial activity against all tested pathogenic microorganisms. Novelty and scientific contribution. Cultivation of mushrooms on Se-enriched zeolitic tuff is a new technological approach for obtaining Se-fortified food/supplements with enhanced antioxidant and antibacterial activities
Electronic cigarette liquids impair metabolic cooperation and alter proteomic profiles in V79 cells
Get PDFBackground Although still considered a safer alternative to classical cigarettes, growing body of work points to harmful effects of electronic cigarettes (e-cigarettes) affecting a range of cellular processes. The biological effect of e-cigarettes needs to be investigated in more detail considering their widespread use. Methods In this study, we treated V79 lung fibroblasts with sub-cytotoxic concentration of e-cigarette liquids, with and without nicotine. Mutagenicity was evaluated by HPRT assay, genotoxicity by comet assay and the effect on cellular communication by metabolic cooperation assay. Additionally, comprehensive proteome analysis was performed via high resolution, parallel accumulation serial fragmentation-PASEF mass spectrometry. Results E-cigarette liquid concentration used in this study showed no mutagenic or genotoxic effect, however it negatively impacted metabolic cooperation between V79 cells. Both e-cigarette liquids induced significant depletion in total number of proteins and impairment of mitochondrial function in treated cells. The focal adhesion proteins were upregulated, which is in accordance with the results of metabolic cooperation assay. Increased presence of posttranslational modifications (PTMs), including carbonylation and direct oxidative modifications, was observed. Data are available via ProteomeXchange with identifier PXD032071. Conclusions Our study revealed impairment of metabolic cooperation as well as significant proteome and PTMs alterations in V79 cells treated with e-cigarette liquid warranting future studies on e-cigarettes health impact
Novel Quorum Quenching YtnP Lactonase From Bacillus paralicheniformis Reduces Pseudomonas aeruginosa Virulence and Increases Antibiotic Efficacy in vivo
Get PDFBacterial infections have become increasingly difficult to treat due to the occurrence of antibiotic-resistant strains. A promising strategy to increase the efficacy of therapy is to combine antibacterials with agents that decrease pathogen virulence via the modulation of the quorum sensing (QS). Lactonases inhibit acylated homoserine lactone (AHL)-mediated QS in Gram-negative bacteria, including the leading nosocomial pathogen Pseudomonas aeruginosa. Here we describe the characteristics of heterologously expressed YtnP lactonase from Bacillus paralicheniformis ZP1 (YtnP-ZP1) isolated from agricultural soil using the culture enrichment method. Purified YtnP-ZP1 hydrolyzed different AHLs with preference to substrates with long acyl residues as evaluated in assays with biosensors and HPLC. The enzyme showed good thermostability and activity in a wide temperature range. YtnP-ZP1 in 50 mu g mL(-1) concentration reduced the amount of P. aeruginosa-produced long-chain AHLs by 85%, while it hydrolyzed 50% of short-chain AHLs. Incubation of P. aeruginosa PAO1 with YtnP-ZP1 reduced its swarming motility and elastolytic activity without bactericidal effect. YtnP-ZP1 caused the inhibition of biofilm formation and disintegration of mature biofilms in P. aeruginosa PAO1 and multiresistant clinical strain BR5H that was visualized by crystal violet staining. The treatment with YtnP-ZP1 in concentrations higher than 25 mu g mL(-1) improved the survival of P. aeruginosa PAO1-infected zebrafish (Danio rerio), rescuing 80% of embryos, while in combination with tobramycin or gentamicin survival rate increased to 100%. The treatment of P. aeruginosa PAO1 biofilms on infected zebrafish tail wounds with 50 mu g mL(-1) YtnP-ZP1 and 2 x MIC tobramycin led to infection clearing in 2 days. The extensive toxicity studies proved YtnP-ZP1 was non-toxic to human cells and zebrafish. In conclusion, novel YtnP-ZP1 lactonase with its effective anti-virulence activity could be used to increase the efficacy of clinically approved antibiotics in clearing both systemic and biofilm-associated P. aeruginosa infections
Interpretative cut-off for alpha-1-antitrypsin concentration in detection of alpha-1-antitrypsin deficiency among adults - a pilot study in the Republic of Serbia
No full textStructural characterization and antimicrobial evaluation of chromium(III) and cobalt(III) complexes with 2,2-diMe-1,3-pdta: Tuning dimensionality of coordination polymer and the water content by alkyl substitution
No full textHexadentate 2,2-dimethyl-1,3-propanediamine-N,N,N',N'-tetraacetate (2,2-diMe-1,3-pdta) ligand, containing two methyl substituents at the central carbon atom of a 1,3-propanediamine, has been prepared and used for the synthesis of Na[Cr(2,2-diMe-1,3-pdta)].3.75H2O (1) and Na[Co(2,2-diMe-1,3-pdta)].3.88H(2)O (2) complexes. These complexes were characterized by IR and electronic absorption spectroscopy, and single-crystal X-ray diffraction analysis. NMR (H-1 and C-13) spectroscopy was additionally applied for the characterization of complex 2. Crystallographic data indicate that the two investigated crystals are isostructural and contain 2,2-diMe-1,3-pdta ligand coordinated to metal ion through 2N and 4O atoms forming an octahedral complex in which the six-membered 1,3-propanediamine chelate ring adopts a twist-boat conformation. There are four such complex anions in the symmetry independent part of the unit cell. Each complex anion is further connected to the sodium counterion(s) via the bridging carboxylate group(s). Structural changes in 2,2-diMe-1,3-pdta-Cr(III) complex stimulated solely by the presence of alkyl side groups are discussed. The present study shows that in 1,3-pdtatype complexes of Cr(III) and Co(III), the environment at coordination centre can be modified by introducing substitution in one of the carbon atoms of the diamine and the resulting difference in the subunit structure can bring about noticeable change in molecular and crystal structure. The examples illustrate the importance of the steric effect for the fine tuning of the dimensionality of the resulting coordination polymer and the water content. The antimicrobial activity of complexes 1 and 2 was evaluated against different bacterial and Candida spp., while their cytotoxic effects were tested on the normal human lung fibroblast cell line (MRC-5)
Autophagy Inhibition Enhances Anti-Glioblastoma Effects of Pyrazolo[3,4-d]pyrimidine Tyrosine Kinase Inhibitors
Get PDFDrug resistance presents a major obstacle to the successful treatment of glioblastoma. Autophagy plays a key role in drug resistance, particularly in relation to targeted therapy, which has prompted the use of autophagy inhibitors to increase the effectiveness of targeted therapeutics. The ability of two Src tyrosine kinase inhibitors, Si306 and its prodrug pro-Si306, to induce autophagy was evaluated in the human glioblastoma cell line U87 and its multidrug-resistant counterpart U87-TxR. Autophagy markers were assessed by flow cytometry, microscopy, and Western blot, and induction of autophagy by these compounds was demonstrated after 3 h as well as 48 h. The effects of Si306 and pro-Si306 on cell proliferation and cell death were examined in the presence or absence of autophagy inhibition by bafilomycin A1. Combined treatments of Si306 and pro-Si306 with bafilomycin A1 were synergistic in nature, and the inhibition of autophagy sensitized glioblastoma cells to Src tyrosine kinase inhibitors. Si306 and pro-Si306 more strongly inhibited cell proliferation and triggered necrosis in combination with bafilomycin A1. Our findings suggest that modulation of Si306- and pro-Si306-induced autophagy can be used to enhance the anticancer effects of these Src tyrosine kinase inhibitors and overcome the drug-resistant phenotype in glioblastoma cells
Techno-Functional Performance of Emmer, Spelt and Khorasan in Spontaneously Fermented Sourdough Bread
Get PDFThe aim of this study was to test the suitability of three different ancient wheat varieties (emmer, spelt and khorasan) to produce spontaneously fermented sourdough bread and to evaluate the impact on the dough rheological properties, ultrastructure and baking quality. Modern wheat sourdough bread and bakery yeast fermented bread were used as controls. Sourdoughs produced from modern and ancient wheats exerted different effects on dough viscoelastic properties, bread specific volume, texture, firming rate, colour and sensory properties, while there was no influence on bread water activity. Both khorasan sourdough, being characterised with the highest dough strength and dense gluten protein matrix, and emmer sourdough, with loose and thin gluten strands of low strength, yielded breads characterised by low specific volume and hard crumb texture. Spelt and modern wheat sourdough were characterised by foam-like dough structures with entrapped gas cells leading to breads of similar specific volume and texture. Although the yeast-fermented wheat flour exerted a higher specific volume and the lowest firmness, the sourdough wheat flour bread had a lower firming rate. A comparison of sourdough bread prepared with modern and ancient wheats revealed that breads based on ancient varieties possess a less noticeable sour taste, odour and flavour, thus contributing to more sensory-appealing sourdough bread
Long non-coding RNA H19 expression in rectal cancer and therapy response
Get PDFBackground & objectives: Long non coding RNA, H19 is an imprinted, maternally expressed gene, usually deregulated in diferent cancer types, including rectal cancer. This study aimed to investigate H19 role as a potential biomarker to predict therapy response in rectal cancer patients. Methods: The study included 14 patients diagnosed with rectal cancer, treated with neoadjuvant chemoradio therapy (nCRT). RNA was isolated by TRIzol reagent from samples of rectal cancer tissue before and after nCRT. Relative expression of H19 was normal- ized to housekeeping GAPDH gene, and expression was analysed by quantitative real-time PCR. Relative expression of H19 was calculated by 2-ΔCt method. Results: Relative expression of H19 was significantly increased in rectal cancer tissue after nCRT (0.244±0.408) compared to the tissue before nCRT (0.043±0.055), p=0.004, Wilcoxon test. According to tumour regression grade (TRG), 85.71% (12/14) of patients did not respond, while 14.28% (2/14) responded to pre- operative CRT. Responders (TRG1, TRG2) and non-responders (TRG3, TRG4) did not differ in H19 expression in tumour tissue before (p=0.659, Mann-Whitney U test) as well as after nCRT (p=0.999, Mann-Whitney U test). Receiver operating curve analy- sis indicates that H19 expression in colorectal tissue before nCRT can not be used as a biomarker for distinguishing responders from non-responders (AUC=0.625, 95%CI=0.257-0.992, p=0.583). Conclusion: Our study suggests H19 upregulation upon neoadju- vant chemoradiotherapy in rectal cancer. The potential predictive value of H19 as a biomarker of therapy response should be studied in a larger group of patients