Oxidative Stress In Bacteria Research Articles

Investigating the potential use of an oleaginous bacterium, Rhodococcus opacus PD630, for nano-TiO2 remediation.

The occurrence of titanium dioxide nanoparticles (nTiO2), in the effluents released from wastewater treatment plants, has raised concerns. The fate of nTiO2 and their potential impact on organisms from different ecosystems are widely investigated. For the first time, in this work, we report the responses of an oleaginous bacteria Rhodococcus opacus PD630, belonging to an ecologically important genus Rhodococcus to environmentally relevant concentrations of nTiO2, under dark and UV light conditions. We observed a dose-dependent increase in nTiO2 uptake by the bacteria that reached a maximum of 1.4 mg nTiO2 (g cell)-1 under mid-log UV exposure, corresponding to 97% uptake. The nTiO2 induced oxidative stress in bacteria that increased from 25.1 to a maximum of 100.3, 44.1, and 51.7 μmol.OH (g cell)-1 under dark, continuous, and mid-log UV, respectively. However, nTiO2 did not affect bacterial viability. Further, due to oxidative stress, the triacylglycerol (biodiesel) content from bacteria increased from 30% to a maximum of 54% CDW. Based on our findings, we propose an application of R. opacus PD 630 in nTiO2 remediation due to their high nTiO2 uptake and resistance.

A novel role for the glutamate decarboxylase system in Listeria monocytogenes; protection against oxidative stress

The GAD system is widely present in several types of organisms and is known to play an important role in bacterial acid tolerance. There is only one account of this system playing a role in oxidative stress in bacteria and one in yeasts. Here we show for first time that it affects the oxidative stress resistance of a Gram-positive bacterium, (L. monocytogenes, tested in three strains; 10403S, EGD-e, and LO28). We found a statistically significant reduction in survival after H2O2 exposure in ΔgadD3 and ΔgadD2 of EGD-e and in ΔgadD1 of LO28. Furthermore, we observed a lag phase prolongation in ΔgadD3 of 10403S and EGD-e and a larger inhibition zone in disk diffusion assay for ΔgadD1 and ΔgadD3 of EGD-e upon H2O2 exposure. All GAD genes playing a role in oxidative stress resistance are part of GADi system and this occurs partly through catalase activity, while the most potent GADe system plays no role. The latter effects could occur through the GABA shunt, but we show here that mutants in succinate semialdehyde dehydrogenase do not show a phenotype suggesting that either effects are through the GABA transaminase or, this pathway is not involved. Our study highlights for first time the role of the GAD system in oxidative stress resistance of a Gram-positive bacterium, which could be used in Food Hurdle Technology to eliminate pathogens such as L. monocytogenes, while it gives an insight on the general mechanism.

The enhanced photocatalytic properties of MnO2/g-C3N4 heterostructure for rapid sterilization under visible light.

Herein, a heterostructure based on MnO2 and g-C3N4 was constructed on the surface of metallic Ti implants, in which MnO2 favored the transfer and separation of free charges to enhance the photoconversion efficiency of g-C3N4 by 21.11%. Consequently, the yield of ROS was promoted significantly, which denatured protein and damaged DNA to kill bacteria efficiently. In addition, glutathione (GSH, l-γ-glutamyl-l-cysteinyl-glycine) defending oxidative stress in bacteria, was oxidized by MnO2 in the hybrid coating once the bacterial membrane was disrupted by ROS. Hence, after visible light irradiation for 20 min, MnO2/g-C3N4 coating exhibited superior disinfection efficacy of 99.96% and 99.26% against S. aureus and E. coli severally. This work provided a practical sterilization strategy about MnO2/g-C3N4 systems through the synergistic effects of enhanced photodynamic antibacterial therapy and oxidization effect of MnO2 with great biosafety, in which MnO2 enhanced the photocatalyst property of g-C3N4 to generate more ROS and deplete GSH to improve antibacterial efficiency. It will bring more insight into rapid and highly effective disinfection and antibacterial strategy without using traditional high-temperature, ultraviolet ray and antibiotics that cause side-effects.

Coenzyme A: a protective thiol in bacterial antioxidant defence.

Coenzyme A (CoA) is an indispensable cofactor in all living organisms. It is synthesized in an evolutionarily conserved pathway by enzymatic conjugation of cysteine, pantothenate (Vitamin B5), and ATP. This unique chemical structure allows CoA to employ its highly reactive thiol group for diverse biochemical reactions. The involvement of the CoA thiol group in the production of metabolically active CoA thioesters (e.g. acetyl CoA, malonyl CoA, and HMG CoA) and activation of carbonyl-containing compounds has been extensively studied since the discovery of this cofactor in the middle of the last century. We are, however, far behind in understanding the role of CoA as a low-molecular-weight thiol in redox regulation. This review summarizes our current knowledge of CoA function in redox regulation and thiol protection under oxidative stress in bacteria. In this context, I discuss recent findings on a novel mode of redox regulation involving covalent modification of cellular proteins by CoA, termed protein CoAlation.

Preparation and Study of the Antibacterial Applications and Oxidative Stress Induction of Copper Maleamate-Functionalized Mesoporous Silica Nanoparticles.

Mesoporous silica nanoparticles (MSNs) are an interesting class of nanomaterials with potential applications in different therapeutic areas and that have been extensively used as drug carriers in different fields of medicine. The present work is focused on the synthesis of MSNs containing a maleamato ligand (MSN-maleamic) and the subsequent coordination of copper(II) ions (MSN-maleamic-Cu) for the exploration of their potential application as antibacterial agents. The Cu-containing nanomaterials have been characterized by different techniques and the preliminary antibacterial effect of the supported maleamato-copper(II) complexes has been tested against two types of bacteria (Gram positive and Gram negative) in different assays to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The biological results showed a moderate antibacterial activity against Escherichia coli which motivated a more detailed study of the antibacterial mechanism of action of the synthesized maleamate-containing nanosystems and whose findings showed oxidative stress generation in bacterial cells. All the prepared nanomaterials were also tested as catalysts in the “solvent free” selective oxidation of benzyl alcohol, to observe if there is a potential correlation between the catalytic oxidation capacity of the materials and the observed oxidative stress in bacteria. This may help in the future, for a more accurate rational design of antibacterial nanosystems, based on their observed catalytic oxidation activity.

Identification of novel sRNAs involved in oxidative stress response in the fish pathogen Vibrio alginolyticus by transcriptome analysis.

Vibrio alginolyticus as an important pathogen in aquaculture can encounter the oxidative stress produced by the immune system during infection. Previous studies showed that sRNAs have important functions in response to oxidative stress in bacteria; however, less of sRNAs related to oxidative stress response were identified in V.alginolyticus. In this study, a total of 749 novel sRNAs were identified by RNA sequencing; among them, 128 sRNAs were up- or downregulated in response to oxidative stress. In addition, 1,870 genes exhibited variation on mRNA levels in oxidative stress response. By analysing the target genes of the sRNAs, we concluded that these sRNAs could regulate expressions of genes responsible for iron transport, catalase, GSH-dependent defence system, electron transferred and stress response. Moreover, the functions of the sRNAs are also seemed related to the pathogenicity in V.alginolyticus. Based on the results, we constructed the oxidative stress model in V.alginolyticus. This study provides us the first outlook of sRNAs function in oxidative stress response in V.alginolyticus. Furthermore, this study can help us to prevent and control this important opportunistic pathogen in aquaculture.

The MmpS6-MmpL6 Operon Is an Oxidative Stress Response System Providing Selective Advantage to Mycobacterium tuberculosis in Stress.

The stress response adaptability of Mycobacterium tuberculosis (Mtb) is still unresolved. In this study, we ascribe an important function to the MmpS6-MmpL6 (M6) operon in Mtb stress management. By using a novel promoter probe in a high-throughput unbiased screen, we identified several quinones as potent inducers of the M6 operon in addition to triclosan. Triclosan and plumbagin effectively altered the intracellular redox potential in Mtb suggestive of oxidative stress in bacteria. Presence of the functional M6 operon correlated with an enhanced ability of clinical strains to survive in the presence of triclosan. Similar to the addition of a powerful reactive oxygen species-quenching agent such as N-acetyl cysteine in the medium, introduction of the complete M6 operon was sufficient to increase tolerance of the M6- strains to triclosan and plumbagin by effectively ablating the change in intracellular redox potential of Mtb, signifying the importance of this operon in oxidative stress survival in mycobacteria.

Nanosilver and the microbiological activity of the particulate solids versus the leached soluble silver

Nanosilver (Ag NPs) is currently one of the most commercialized antimicrobial nanoparticles with as yet, still unresolved cytotoxicity origins. To date, research efforts have mostly described the antimicrobial contribution from the leaching of soluble silver, while the undissolved solid Ag particulates are often considered as being microbiologically inert, serving only as source of the cytotoxic Ag ions. Here, we show the rapid stimulation of lethal cellular oxidative stress in bacteria by the presence of the undissolved Ag particulates. The cytotoxicity characteristics are distinct from those arising from the leached soluble Ag, the latter being locked in organic complexes. The work also highlights the unique oxidative stress-independent bacterial toxicity of silver salt. Taken together, the findings advocate that future enquiries on the antimicrobial potency and also importantly, the environmental and clinical impact of Ag NPs use, should pay attention to the potential bacterial toxicological responses to the undissolved Ag particulates, rather than just to the leaching of soluble silver. The findings also put into question the common use of silver salt as model material for evaluating bacterial toxicity of Ag NPs.

Oxidative Stress in Bacteria Measured by Flow Cytometry

Oxidative Stress in Bacteria Measured by Flow Cytometry

Mechanisms of antibiotic resistance in bacteria mediated by silver nanoparticles

ABSTRACT Silver nanoparticles (AgNPs) are widely used in industry, consumer products, and medical appliances due to their efficient antimicrobial properties. However, information on environmental toxicity and bacterial impact of these particles is not completely elucidated. Results showed that AgNPs produced growth inhibition and oxidative stress in bacteria Escherichia coli (gram negative) and Staphylococcus aureus (gram positive), with half-maximal inhibitory concentrations (IC50) of 12 and 7 mg/L, respectively. Surprisingly, bacteria pre-exposed to sublethal dose of AgNPs exhibited increased resistance toward antibiotics (ampicillin and Pen-Strep) with IC50 elevated by 3–13-fold. Further, AgNP pre-exposure raised the minimal inhibitory concentration and minimal biocidal concentration by two- to eightfold when cells were challenged with antibiotics with diverse mechanisms of action (penicillin, chloramphenicol, and kanamycin). Interestingly, we found that upon exposure to ampicillin, strains pretreated with AgNPs exhibited lower levels of membrane damage and oxidative stress, together with elevated levels of intracellular ATP relative to untreated cells. Bacterial reverse mutation assay (Ames test) showed that AgNPs are highly mutagenic, consistent with further assays demonstrating abiotic reactive oxygen species (ROS) generation and intrinsic DNA cleavage activity in vitro of AgNPs. Overall, our results suggest that AgNPs enhance bacterial resistance to antibiotics by promoting stress tolerance through induction of intracellular ROS. Our data suggest potential consequences of incidental environmental exposure of bacteria to AgNPs and indicate the need to regulate use and disposal of AgNPs in industry and consumer products.

Involvement of multiple stressors induced by non-thermal plasma-charged aerosols during inactivation of airborne bacteria.

A lab-scale, tunable, single-filament, point-to-point nonthermal dieletric-barrier discharge (DBD) plasma device was built to study the mechanisms of inactivation of aerosolized bacterial pathogens. The system inactivates airborne antibiotic-resistant pathogens efficiently. Nebulization mediated pre-optimized (4 log and 7 log) bacterial loads were challenged to plasma-charged aerosols, and lethal and sublethal doses determined using colony assay, and cell viability assay; and the loss of membrane potential and cellular respiration were determined using cell membrane potential assay and XTT assay. Using the strategies of Escherichia coli wildtype, over-expression mutant, deletion mutants, and peroxide and heat stress scavenging, we analyzed activation of intracellular reactive oxygen species (ROS) and heat shock protein (hsp) chaperons. Superoxide dismutase deletion mutants (ΔsodA, ΔsodB, ΔsodAΔsodB) and catalase mutants ΔkatG and ΔkatEΔkatG did not show significant difference from wildtype strain, and ΔkatE and ΔahpC was found significantly more susceptible to cell death than wildtype. The oxyR regulon was found to mediate plasma-charged aerosol-induced oxidative stress in bacteria. Hsp deficient E. coli (ΔhtpG, ΔgroEL, ΔclpX, ΔgrpE) showed complete inactivation of cells at ambient temperature, and the treatment at cold temperature (4°C) significantly protected hsp deletion mutants and wildtype cells, and indicate a direct involvement of hsp in plasma-charged aerosol mediated E. coli cell death.

Expression of alkyl hydroperoxide reductase is regulated negatively by OxyR1 and positively by RpoE2 sigma factor in Azospirillum brasilense Sp7

OxyR proteins are LysR-type transcriptional regulators, which play an important role in responding to oxidative stress in bacteria. Azospirillum brasilense Sp7 harbours two copies of OxyR. The inactivation of the oxyR1, the gene organized divergently to ahpC in A. brasilense Sp7, led to an increased tolerance to alkyl hydroperoxides, which was corroborated by an increase in alkyl hydroperoxide reductase (AhpC) activity, enhanced expression of ahpC :lacZ fusion and increased synthesis of AhpC protein in the oxyR1::km mutant. The upstream region of ahpC promoter harboured a putative OxyR binding site, T-N11-A. Mutation of T, A or both in the T-N11-Amotif caused derepression of ahpC in A. brasilense suggesting that T-N11-A might be the binding site for a negative regulator. Retardation of the electrophoretic mobility of the T-N11-A motif harbouring oxyR1-ahpC intergenic DNA by recombinant OxyR1, under reducing as well as oxidizing conditions, indicated that OxyR1 acts as a negative regulator of ahpC in A. brasilense. Sequence of the promoter of ahpC, predicted on the basis of transcriptional start site, and an enhanced expression of ahpC:lacZ fusion in chrR2::km mutant background suggested that ahpC promoter was RpoE2 dependent. Thus, this study shows that in A. brasilense Sp7, ahpC expression is regulated negatively by OxyR1 but is regulated positively by RpoE2, an oxidative-stress-responsive sigma factor. It also shows that OxyR1 regulates the expression RpoE1, which is known to play an important role during photooxidative stress in A. brasilense.

Oxidative stress generation of silver nanoparticles in three bacterial genera and its relationship with the antimicrobial activity

Oxidative stress is a condition caused by the high intracellular concentrations of reactive oxygen species (ROS) that includes superoxide anion radicals, hydroxyl radicals and hydrogen peroxide. Nanoparticles could cause rapid generation of free radicals by redox reactions. ROS can react directly with membrane lipids, proteins and DNA and are normally scavenged by antioxidants that are capable of neutralizing; however, elevated concentrations of ROS in bacterial cells can result in oxidative stress. The aim of this work was contribute to the knowledge of action mechanism of silver nanoparticles (Ag-NPs) and their relation to the generation of oxidative stress in bacteria. We demonstrated that Ag-NPs generated oxidative stress in Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa mediated by the increment of ROS and this increase correlated with a better antimicrobial activity. On the other hand, we showed that the oxidative stress caused by the Ag-NPs biosynthesized was associated to a variation in the level of reactive nitrogen intermediates (RNI). Oxidative stress in bacteria can result from disruption of the electronic transport chain due to the high affinity of Ag-NPs for the cell membrane. This imbalance in the oxidative stress was evidentiated by a macromolecular oxidation at level of DNA, lipids and proteins in E. coli exposed to Ag-NPs. The formation of ROS and RNI by Ag-NPs may also be considered to explain the bacterial death.

The Antibacterial Activity of Date Syrup Polyphenols against S. aureus and E. coli.

Plant-derived products such as date syrup (DS) have demonstrated antibacterial activity and can inhibit bacteria through numerous different mechanisms, which may be attributed to bioactive compounds including plant-derived phenolic molecules. DS is rich in polyphenols and this study hypothesized that DS polyphenols demonstrate inherent antimicrobial activity, which cause oxidative damage. This investigation revealed that DS has a high content of total polyphenols (605 mg/100 g), and is rich in tannins (357 mg/100 g), flavonoids (40.5 mg/100 g), and flavanols (31.7 mg/100 g) that are known potent antioxidants. Furthermore, DS, and polyphenols extracted from DS, the most abundant bioactive constituent of DS are bacteriostatic to both Gram positive and Gram negative Escherichia coli and Staphylococcus aureus, respectively. It has further been shown that the extracted polyphenols independently suppress the growth of bacteria at minimum inhibitory concentration (MIC) of 30 and 20 mg/mL for E. coli and S. aureus, and have observed that DS behaves as a prooxidant by generating hydrogen peroxide that mediates bacterial growth inhibition as a result of oxidative stress. At sub-lethal MIC concentrations DS demonstrated antioxidative activity by reducing hydrogen peroxide, and at lethal concentrations DS demonstrated prooxidant activity that inhibited the growth of E. coli and S. aureus. The high sugar content naturally present in DS did not significantly contribute to this effect. These findings highlight that DS’s antimicrobial activity is mediated through hydrogen peroxide generation in inducing oxidative stress in bacteria.

Exploring the redox balance inside gram-negative bacteria with redox-sensitive GFP

Aerobic bacteria are continuously fighting potential oxidative stress due to endogenous and exogenous reactive oxygen species (ROS). To achieve this goal, bacteria possess a wide array of defenses and stress responses including detoxifying enzymes like catalases and peroxidases; however until now, the dynamics of the intra-bacterial redox balance remained poorly understood. Herein, we used redox-sensitive GFP (roGFP2) inside a variety of gram-negative bacteria to study real-time redox dynamics immediately after a challenge with hydrogen peroxide. Using this biosensor, we determined the individual contributions of catalases and peroxidases and found that each enzyme contributes more to rapid detoxification or to prolonged catalytic activity. We also found that the total catalytic power is affected by environmental conditions. Additionally, using a Salmonella strain that is devoid of detoxifying enzymes, we examined endogenous ROS production. By measuring endogenous ROS production, we assessed the role of oxidative stress in toxicity of heavy metals and antibiotics. We found that exposure to nickel induced significant oxidative stress whereas cobalt (which was previously implicated to induce oxidative stress) did not induce ROS formation. Since a turbulent debate evolves around oxidative stress as a general killing mechanism by antibiotics (aminoglycosides, fluoroquinolones and β-lactams), we measured oxidative stress in bacteria that were challenged with these antibiotics. Our results revealed that antibiotics do not induce ROS formation in bacteria thereby disputing a role for oxidative stress as a general killing mechanism. Together, our results expose how the intra-bacterial redox balance in individual microorganisms is affected by environmental conditions and encounters with stress-inducing compounds. These findings demonstrate the significant potential of roGFP2 as a redox biosensor in gram-negative bacteria to investigate redox dynamics under a variety of circumstances.

A Kinetic Platform to Determine the Fate of Hydrogen Peroxide in Escherichia coli.

Hydrogen peroxide (H2O2) is used by phagocytic cells of the innate immune response to kill engulfed bacteria. H2O2 diffuses freely into bacteria, where it can wreak havoc on sensitive biomolecules if it is not rapidly detoxified. Accordingly, bacteria have evolved numerous systems to defend themselves against H2O2, and the importance of these systems to pathogenesis has been substantiated by the many bacteria that require them to establish or sustain infections. The kinetic competition for H2O2 within bacteria is complex, which suggests that quantitative models will improve interpretation and prediction of network behavior. To date, such models have been of limited scope, and this inspired us to construct a quantitative, systems-level model of H2O2 detoxification in Escherichia coli that includes detoxification enzymes, H2O2-dependent transcriptional regulation, enzyme degradation, the Fenton reaction and damage caused by •OH, oxidation of biomolecules by H2O2, and repair processes. After using an iterative computational and experimental procedure to train the model, we leveraged it to predict how H2O2 detoxification would change in response to an environmental perturbation that pathogens encounter within host phagosomes, carbon source deprivation, which leads to translational inhibition and limited availability of NADH. We found that the model accurately predicted that NADH depletion would delay clearance at low H2O2 concentrations and that detoxification at higher concentrations would resemble that of carbon-replete conditions. These results suggest that protein synthesis during bolus H2O2 stress does not affect clearance dynamics and that access to catabolites only matters at low H2O2 concentrations. We anticipate that this model will serve as a computational tool for the quantitative exploration and dissection of oxidative stress in bacteria, and that the model and methods used to develop it will provide important templates for the generation of comparable models for other bacterial species.

Genome-wide Reconstruction of OxyR and SoxRS Transcriptional Regulatory Networks under Oxidative Stress in Escherichia coli K-12 MG1655.

Three transcription factors (TFs), OxyR, SoxR, and SoxS, play a critical role in transcriptional regulation of the defense system for oxidative stress in bacteria. However, their full genome-wide regulatory potential is unknown. Here, we perform a genome-scale reconstruction of the OxyR, SoxR, and SoxS regulons in Escherichia coli K-12 MG1655. Integrative data analysis reveals that a total of 68 genes in 51transcription units (TUs) belong to these regulons. Among them, 48 genes showed more than 2-fold changes in expression level under single-TF-knockout conditions. This reconstruction expands the genome-wide roles of these factors to include direct activation of genes related to amino acid biosynthesis (methionine and aromatic amino acids), cell wall synthesis (lipid A biosynthesis and peptidoglycan growth), and divalent metal ion transport (Mn(2+), Zn(2+), and Mg(2+)). Investigating the co-regulation of these genes with other stress-response TFs reveals that they are independently regulated by stress-specific TFs.

The sRNA SorY confers resistance during photooxidative stress by affecting a metabolite transporter in Rhodobacter sphaeroides

Exposure to oxygen and light generates photooxidative stress by the bacteriochlorophyll a mediated formation of singlet oxygen (1O2) in the facultative photosynthetic bacterium Rhodobacter sphaeroides. We have identified SorY as an sRNA, which is induced under several stress conditions and confers increased resistance against 1O2. SorY by direct interaction affects the takP mRNA, encoding a TRAP-T transporter. We present a model in which SorY reduces the metabolite flux into the tricarboxylic acid cycle (TCA cycle) by reducing malate import through TakP. It was previously shown that oxidative stress in bacteria leads to switch from glycolysis to the pentose phosphate pathway and to reduced activity of the TCA cycle. As a consequence the production of the prooxidant NADH is reduced and production of the protective NADPH is enhanced. In R. sphaeroides enzymes for glycolysis, pentose phosphate pathway, Entner–Doudoroff pathway and gluconeogenesis are induced in response to 1O2 by the alternative sigma factor RpoHII. The same is true for the sRNA SorY. By limiting malate import SorY thus contributes to the balance of the metabolic fluxes under photooxidative stress conditions. This assigns a so far unknown function to an sRNA in oxidative stress response.

Futile cycling increases sensitivity toward oxidative stress in Escherichia coli

Reactive oxygen species (ROS) are toxic molecules utilized by the immune system to combat invading pathogens. Recent evidence suggests that inefficiencies in ATP production or usage can lead to increased endogenous ROS production and sensitivity to oxidative stress in bacteria. With this as inspiration, and knowledge that ATP is required for a number of DNA repair mechanisms, we hypothesized that futile cycling would be an effective way to increase sensitivity to oxidative stress. We developed a mixed integer linear optimization framework to identify experimentally-tractable futile cycles, and confirmed metabolic modeling predictions that futile cycling depresses growth rate, and increases both O2 consumption and ROS production per biomass generated. Further, intracellular ATP was decreased and sensitivity to oxidative stress increased in all actively cycling strains compared to their catalytically inactive controls. This research establishes a fundamental connection between ATP metabolism, endogenous ROS production, and tolerance toward oxidative stress in bacteria.

Structural diversity in dinuclear copper(I) halide complexes of 2,4-dithiouracil: Synthesis, crystal structures, induction of DNA damage and oxidative stress mediated by ROS

2,4-Dithiouracil (dtucH2) reacts with [CuX(PPh3)]4 (X=Cl, Br, I) to give dinuclear complexes with different bridging motifs, namely doubly bridged [Cu2(μ-Cl)(μ-S,S-dtucH)(PPh3)4] (1), single bridged [Cu2Br2(μ-S,S-dtucH2)(PPh3)4] (2) and triply bridged [Cu2(μ-I)2(μ-S,S-dtucH2)(PPh3)2] (3). In the structure of 1 the two copper(I) centers are joined by one bridging chlorine atom as well as a 2,4-dithiouracil anion acting as an S,S-donor, while compound 3 contains two bridging iodine atoms besides the neutral bridging 2,4-dithiouracil ligand. The structure of 2 consists of two CuBr(PPh3)2 sub-units joined by the S-atoms of a neutral dtucH2. The biological properties of the new complexes were evaluated in relevance to their binding with calf-thymus double stranded DNA (dsCTDNA) in vitro monitored by mobility shift experiments in agarose gel electrophoresis, and their ability to induce oxidative stress in bacteria Escherichia coli. The experiments showed that these complexes are able to interact with dsCTDNA and to cause DNA damage, and are also able to induce oxidative stress in bacteria. Because of the relevance of these properties, these complexes are regarded as a very promising starting point for biological studies in living systems.