Authors:Eszter Horváth, Nikoletta Kálmán, M. Pesti, K. Iwata, and S. Kunsági-Máté
The effects of the mycotoxin patulin on the thermodynamics and kinetics of the transition of bovine serum albumin (BSA) in aqueous solution were studied by Differential Scanning Calorimetry and Photoluminescence methods. Results show that in the presence of patulin, the free enthalpy change during the transition of BSA was decreased by an average of ∼ 46 kJ/mol, the free energy change was decreased by ∼ 4 kJ/mol, and the activation energy fell from ∼ 1546 to ∼ 840 kJ/mol. These results indicate that the bioactivity of patulin is based on the kinetic rather than on the thermodynamic properties of the transition. This is the first evidence of the direct interaction of patulin with the free thiol-containing BSA, a process which could contribute to the adverse cyto- and genotoxic effects induced by patulin.
Authors:E. Horváth, G. Papp, Z. Gazdag, J. Belágyi, Á. Blaskó, J. Deli, Cs. Vágvölgyi, and M. Pesti
A carotenoid-less Phaffia rhodozyma mutant (MCP 325) exhibited significantly higher resistance to oxidative stressors such as menadione, H2O2 and K2Cr2O7 than its astaxanthin-producing parental strain (MCP 324). The absence of carotenoids in the mutant did not explain this phenomenon. The cause of the decreased superoxide, hydroxyl radical and glutathione contents, the increased peroxide concentration and the elevated specific activity of catalase under uninduced conditions may be a second mutation. Peroxide treatment induced specific catalase activity in the mutant but not in the parental strain. Regulation of these processes led to the result that, in spite of the mutations, the two strains exhibited the same multiplication rate and generation time.
Authors:Z. Gazdag, Nikoletta Kálmán, Ágnes Blaskó, Eszter Virág, J. Belágyi, and M. Pesti
The one-gene mutation in the tert-butyl hydroperoxide-resistant mutant hyd1-190 of the fission yeast Schizosaccharomyces pombe led to a 4-fold increase in resistance to t-BuOOH and decreased specific concentrations of superoxide and total thiols in comparison with the parental strain hyd+. It suggested an unbalanced redox state of the cells, which induced continuously increased specific activities of glutathione peroxidase, glutathione reductase and glutathione S-transferase and decreased activities of the antioxidant enzymes superoxide dismutases and glucose-6-phosphate dehydrogenase to regulate the redox balance of the mutation-induced permanent, low-level but tolerable internal stress. These results may contribute to the understanding of internal, oxidative stress-related human diseases.
Authors:Eszter Virág, Á. Juhász, R. Kardos, Z. Gazdag, G. Papp, Ágota Pénzes, M. Nyitrai, Cs. Vágvölgyi, and M. Pesti
Interaction of primycin antibiotic with plasma membrane, and its indirect biological effects were investigated in this study. The antifungal activity of primycin against 13 human pathogenic Candida ATCC and CBS reference species and 74 other Candida albicans clinical isolates was investigated with a microdilution technique. No primycin-resistant strain was detected. Direct interaction of primycin with the plasma membrane was demonstrated for the first time by using an ergosterol-producing strain 33erg+ and its ergosterol-less mutant erg-2. In growth inhibition tests, the 33erg+ strain proved to be more sensitive to primycin than its erg-2 mutant, indicating the importance of the plasma membrane composition in primycin-induced processes. The 64 μg ml−1 (56.8 nM) primycin treatment induced an enhanced membrane fluidity and altered plasma membrane dynamics, as measured by steady-state fluorescence anisotropy applying a trimethylammonium-diphenylhexatriene (TMA-DPH) fluorescence polarization probe. The following consequences were detected. The plasma membrane of the cells lost its barrier function, and the efflux of 260-nm-absorbing materials from treated cells of both strains was 1.5–1.8 times more than that for the control. Depending on the primycin concentration, the cells exhibited unipolar budding, pseudohyphae formation, and a rough cell surface visualized by scanning electron microscopy.
Authors:B. Kádár, M. Szász, Katalin Kristóf, Natasa Pesti, G. Krizsán, Julianna Szentandrássy, L. Rókusz, K. Nagy, and Dóra Szabó
The aim of the study was to investigate the biofilm-production of 60 Pseudomonas aeruginosa strains isolated from clinical samples and to examine the effect of different antimicrobials and their combinations with clarithromycin on biofilm-formation.The minimal inhibitory concentrations (MICs), minimal biofilm inhibitory concentrations (MBICs), and antibiotic synergy by calculating the fractional inhibitory concentration (FIC) index were determined for the following antibiotics: ceftazidime, cefepime, piperacillin/tazobactam, imipenem, meropenem, levofloxacin, ciprofloxacin, gentamicin, amikacin, tobramycin, netilmicin and clarithromycin.A total of 14 (23.3%) isolates out of 60 isolates of P. aeruginosa were biofilm positive. Cefepime, imipenem and meropenem had the lowest MIC90 values. Piperacillin/tazobactam and clarithromycin had the highest MIC90 values. Imipenem, meropenem, piperacillin/tazobactam and clarithromycin had the lowest MBIC90 values.For biofilm-forming P. aeruginosa strains 2-fold to 128-fold higher MBIC values than MIC values were obtained for ceftazidime, cefepime, imipenem, amikacin and netilmicin. The MBIC was 2-fold to 512-fold lower then the MIC values in the case of piperacillin/tazobactam, ciprofloxacin, levofloxacin and clarithromycin.Synergy was generally demonstrated for clarithromycin in combination with aminoglycosides, fluoroquinolones or ceftazidime. However, surprisingly it was found that combinations of clarithromycin with carbapenems or cefepime led to an antagonistic interaction: combination of clarithromycin with imipenem, meropenem or ertapenem showed antagonism in 37.5%, 50% and 62.5% of the strains tested whereas its combination with cefepime expressed antagonism in 75% of the strains, respectively. To the best of our knowledge no one has previously described this phenomenon so far.