The effect of DMSO on Saccharomyces cerevisiae yeast with different energy metabolism and antioxidant status.

We studied the effect of dimethyl sulfoxide (DMSO) on the biochemical and physiological parameters of S. cerevisiae yeast cells with varied energy metabolism and antioxidant status. The wild-type cells of varied genetic backgrounds and their isogenic mutants with impaired antioxidant defences (Δsod mutants) or response to environmental stress (ESR) (Δmsn2, Δmsn4 and double Δmsn2msn4 mutants) were used. Short-term exposure to DMSO even at a wide range of concentrations (2-20%) had little effect on the metabolic activity of the yeast cells and the stability of their cell membranes, but induced free radicals production and clearly altered their proliferative activity. Cells of the Δsod1 mutant showed greater sensitivity to DMSO in these conditions. DMSO at concentrations from 4 to 10-14% (depending on the strain and genetic background) activated the ESR programme. The effects of long-term exposure to DMSO were mainly depended on the type of energy metabolism and antioxidant system efficiency. Yeast cells with reduced antioxidant system efficiency and/or aerobic respiration were more susceptible to the toxic effects of DMSO than cells with a wild-type phenotype and respiro-fermentative or fully fermentative metabolism. These studies suggest a key role of stress response programs in both the processes of cell adaptation to small doses of this xenobiotic and the processes related to its toxicity resulting from large doses or chronic exposure to DMSO.

Effect of Sublethal Concentrations of Zinc Oxide Nanoparticles on Bacillus cereus.

Zinc oxide nanoparticles (ZnONPs), which are produced on a large scale, pose a potential threat to various environments because they can interact with the microbial populations found in them. Bacteria that are widespread in soil, water, and plant material include the Bacillus cereus group, which plays an important role in biodegradation and the nutrient cycle and is a major factor determining ecological balance. This group includes, among others, the foodborne pathogen B. cereus sensu stricto (herein referred to as B. cereus). The aim of this study was a comprehensive assessment of the effects of commercially available ZnONPs on B. cereus. The MIC (minimum inhibitory concentration) for B. cereus was 1.6 mg/mL, and the MBC (minimum bactericidal concentration) was 1.8 mg/mL. Growth of B. cereus was inhibited by a concentration of ZnONPs lower than or equal to MIC50. Concentrations from 0.2 to 0.8 mg/mL inhibited the growth of these bacteria in liquid media, induced symptoms of oxidative stress, and stimulated an environmental stress response in the form of biofilm and endospore formation. In addition, ZnONPs negatively affected the ability of the bacteria to break down the azo dye Evans Blue but enhanced the antimicrobial properties of phenolic compounds. Sublethal concentrations of ZnONPs generally decreased the activity of B. cereus cells, especially in the presence of phenolics, which indicates their potential toxicological impact, but at the same time they induced universal defence responses in these cells, which in the case of potential pathogens can hinder their removal.

Model Study for Interaction of Sublethal Doses of Zinc Oxide Nanoparticles with Environmentally Beneficial Bacteria Bacillus thuringiensis and Bacillus megaterium.

Zinc oxide nanoparticles (ZnO NPs), due to their antibacterial effects, are commonly used in various branches of the economy and can affect rhizobacteria that promote plant growth. We describe the effect of ZnO NPs on two model bacteria strains, B. thuringiensis and B. megaterium, that play an important role in the environment. The MIC (minimum inhibitory concentration) value determined after 48 h of incubation with ZnO NPs was more than 1.6 mg/mL for both strains tested, while the MBC (minimum bactericidal concentration) was above 1.8 mg/mL. We tested the effect of ZnO NPs at concentrations below the MIC (0.8 mg/mL, 0.4 mg/mL and 0.2 mg/mL (equal to 50%, 25% and 12,5% MIC, respectively) in order to identify the mechanisms activated by Bacillus species in the presence of these nanoparticles. ZnO NPs in sublethal concentrations inhibited planktonic cell growth, stimulated endospore formation and reduced decolorization of Evans blue. The addition of ZnO NPs caused oxidative stress, measured using nitroblue tetrazolium (NBT), and reduced the activity of catalase. It was confirmed that zinc oxide nanoparticles in sublethal concentrations change metabolic processes in Bacillus bacteria that are important for their effects on the environment. B. thuringiensis after treatment with ZnO NPs decreased indole acetic acid (IAA) production and increased biofilm formation, whereas B. megaterium decreased IAA production but, inversely, increased biofilm formation. Comparison of different Bacillus species in a single experiment made it possible to better understand the mechanisms of toxicity of zinc oxide nanoparticles and the individual reactions of closely related bacterial species.

Quality of Rye Plants (Secale cereale) as Affected by Agronomic Biofortification with Iodine.

This study assessed the possibility of using iodine-containing fertilizers for agronomic biofortification of rye biomass used as fodder for cows, and establish the best application method and form and the optimal dose of iodine (I) under field conditions. The impact of iodine fertilization on grain iodine content was not studied. Results showed that agronomic biofortification of rye plants with iodine, influenced by its dose, form, and method of application was highly effective in increasing I shoot contents. Plant I-enrichment via foliar and soil application significantly affected I concentration in plant biomass even at a low dose (2.5 kg ha-1). Soil I application as KI appeared optimal for rye plants used as fodder for cows, especially cropped under the soil with a neutral reaction. Iodine application improved the biological quality of rye plants by increasing concentrations of sugar, chlorophylls, and at a low rate, protein and total antioxidant capacity.

Chemical Composition, Antioxidant and Antimicrobial Activity of Raspberry, Blackberry and Raspberry-Blackberry Hybrid Leaf Buds.

In our investigation, the chemical composition and bioactive potential of leaf buds of raspberry, blackberry, and a raspberry-blackberry hybrid were determined. Antioxidant and antimicrobial properties were tested in water (W), ethanol-water (EW), and glycerol-water (GW) extracts from the buds. These plant organs contain relatively large amounts of minerals, especially Fe. The total antioxidant capacity (TAC) measured by the ABTS and DPPH methods ranged from 2.86 to 12.19 and 6.75 to 24.26 mmol per 100 g fresh weight (FW) of buds, respectively. TAC values were generally higher in the raspberry than in the case of blackberry and raspberry-blackberry hybrid extracts. The antioxidant properties of the extracts were strongly positively correlated with their content of total phenolic (TP). No such relationship was noted for ascorbic acid (AA), whose concentration in all extracts was at a similarly low level. Antioxidant properties determined in vitro were confirmed for the GW extract from raspberry leaf buds in biological test based on the growth parameters of Δsod1Saccharomyces cerevisiae mutant cells in hypertonic medium. The extracts also exhibited strong antibacterial properties against Staphylococcus aureus and Enterococcus faecalis and weaker against Enterobacter aerogenes. The studied leaf buds could be therefore an unconventional source of minerals, natural antioxidants and antibacterial compounds with potential applications in the food, pharmaceutical, and cosmetics industries.

Yeast as a biosensor for antioxidants: simple growth tests employing a Saccharomyces cerevisiae mutant defective in superoxide dismutase.

Mutants of Saccharomyces cerevisiae devoid of Cu,Zn-superoxide dismutase are hypersensitive to a range of oxidants, hyperbaric oxygen and hyperosmotic media, show lysine and methionine auxotrophy when grown under the atmosphere of air and have a shortened replicative life span when compared to the wild-type strain. Ascorbate and other antioxidants can ameliorate these defects, which may be a basis of simple tests sensing the presence of antioxidants. In particular, tests of growth on solid medium (colony formation) in the absence of methionine and/or lysine, or in the presence of 0.8 M NaCl can be useful for detection and semiquantitative estimation of compounds of antioxidant properties. Hypoxic atmosphere was found to increase the sensitivity of detection of antioxidants. The test of abolishment of lysine auxotrophy showed a concentration dependence of the antioxidant effects of cysteine and N-acetylcysteine which, however, lost their protective action at high concentration, in contrast to glutathione which was effective also at higher concentrations.

Ascorbate abolishes auxotrophy caused by the lack of superoxide dismutase in Saccharomyces cerevisiae. Yeast can be a biosensor for antioxidants.

Yeast (Saccharomyces cerevisiae) mutants lacking cytoplasmic superoxide dismutase (CuZnSOD) show Lys and Met auxotrophy under aerobic conditions. This metabolic defect can be ameliorated by exogenous ascorbate as well as other antioxidants (glutathione, cysteine and N-acetylcysteine). Restoration of growth of CuZnSOD- yeast mutants on media devoid of Met and/or Lys may therefore be a simple and useful means to detect and quantify antioxidants. The protective effect of antioxidants is oxygen-dependent: the lower the oxygen content of the atmosphere, the lower antioxidant concentrations are required to restore prototrophy. Therefore, the sensitivity of the test can be augmented by growing the yeast under lowered partial oxygen pressure. While 6 mM, 10 mM and 30 mM ascorbate was necessary to restore the growth in the absence of Met, in the absence of Lys, and in the absence of Lys and Met, respectively, under 21% oxygen, 3 mM and 6 mM ascorbate was sufficient for growth restoration in the absence of Lys and in the absence of Lys and Met, respectively, under 3% oxygen. The protective effects of cysteine and N-acetylcysteine peaked at 0.5 mM and 6 mM, respectively, disappearing at higher concentrations of these compounds, pointing to the detection of not only protective but also toxic cellular effects of the compounds studied by the test proposed.

A novel test for identifying genes involved in aldehyde detoxification in the yeast. Increased sensitivity of superoxide-deficient yeast to aldehydes and their metabolic precursors.

A novel test for the identification of genes involved in aldehyde metabolism is proposed, based on detection of altered sensitivity of the yeast to corresponding alcohols, metabolic precursors of the aldehydes. This attitude enabled to an unexpected detection increased sensitivity of mutants devoid of CuZn-superoxide dismutase (CuZnSOD) to allyl alcohol (precursor of acrolein) and nonenol. We interpret this finding as due to inactivation of some important element of aldehyde detoxification by increased flux of superoxide in DeltaCuZnSOD mutants.

Ascorbate restores lifespan of superoxide-dismutase deficient yeast.

Yeast (Saccharomyces cerevisiae) mutants lacking CuZn-superoxide dismutase (CuZnSOD) are hypersensitive to oxygen and have significantly decreased replicative life span. Both these defects can be ameliorated by exogenous ascorbate. The effect of ascorbate on life span is complicated by auto-oxidation of its compound in the medium. If negative effects of auto-oxidation are prevented by exchange of the medium, ascorbate prolongs not only mean but also maximal replicative life span of the yeast in the atmosphere of air and of pure oxygen. These results demonstrate that life span shortening due to the lack of a vital antioxidant enzyme can be ameliorated by a low-molecular weight antioxidant.