Plasmid DNA Damage Research Articles

Exopolysaccharide from Leuconostoc mesenteroides XR1: Yield optimization, partial characterization and properties

The production conditions of exopolysaccharide (EPS) from Leuconostoc mesenteroides XR1 were optimized by response surface methodology (RSM). Maximum EPS yield was 56.59 ± 0.51 g/L under fermentation conditions with 2.6 g/L ammonium citrate, initial pH 6.5 and temperature 23 °C, which was 6.21-fold greater than the EPS yield before optimization. Characterization of the chain conformation using Congo red test and circular dichroism (CD) showed that EPS exhibited a random coil structure in aqueous solution. The CD results revealed that the EPS concentration altered its hydrogen-bond interactions and chirality, but did not change its chain conformation. The average polydispersity index (PDI) of the EPS solution was only 27.16 %, indicating that it was uniformly distributed in the aqueous solution with high stability. The degradation temperature of EPS was 253.11 °C, indicating high thermal stability. EPS possessed the ability to scavenge activities of free radicals and was protective against oxidative stress-induced plasmid DNA damage. In addition, stable hydrogels could be formed at EPS concentrations above 5 % (w/v). These results collectively showed that EPS can be used commercially as an antioxidant and drug delivery carrier.

Dendritic, Water‐Soluble, and Nonaggregated Axially Substituted Silicon Phthalocyanine as Potential Endometrial Anticancer Agent

ABSTRACTIn this work, dendritic, water‐soluble, and nonaggregated axially substituted silicon phthalocyanines (EA‐TD‐Si and EA‐TD‐SiQ) were synthesized and characterized. Then, the supercoiled pBR322 plasmid DNA damage properties of EA‐TD‐Si and EA‐TD‐SiQ were examined using agarose gel electrophoresis without light irradiation. In addition, the cytotoxicity and phototoxicity of EA‐TD‐Si and EA‐TD‐SiQ on human endometrium adenocarcinoma (HEC‐1B) cells were investigated using an MTT assay. The results indicated that EA‐TD‐SiQ had high photodamage properties via singlet oxygen depending on the light dose, while EA‐TD‐Si did not show any DNA damage without light. In addition, EA‐TD‐SiQ had higher phototoxic effects than EA‐TD‐Si on HEC‐1B cells according to the MTT assay. These results revealed the potential of EA‐TD‐SiQ to be used as a PDT agent in endometrial cancer.

Assessment of cytoprotective and genoprotective effects of Moringa oleifera and Tinospora cordifolia extracts in vitro.

Moringa oleifera and Tinospora cordifolia is extensively used as an ingredient of food and in traditional medicine for the management of a variety of diseases. The extracts of leaf of Moringa oleifera and stem of Tinospora cordifolia were assessed to examine their ability to inhibit the oxidative DNA damage (by DNA protection assay), cytoprotective and genoprotective potential (by Comet assay) in V79 cells individually and in combinations. It was found that these extracts could significantly inhibit the OH-dependent damage of pUC18 plasmid DNA. M. oleifera extract (160 and 320μg/mL) and Tinospora cordifolia extract (640, 1,280 and 2,560μg/mL) individually showed higher DNA protection activity. M. oleifera (1,280μg/mL) combined with Tinospora cordifolia (640μg/mL) showed best cytoprotective and genoprotective activities among different concentration combinations and various concentrations of individual plants in V79 cell line against hydrogen peroxide induced cytotoxicity and genotoxicity. This study demonstrates the cytoprotective and genoprotective activity of M. oleifera and Tinospora cordifolia individually or in combination.

Products of Selenite/Thiols Interaction Have Reducing Properties, Cleave Plasmid DNA and Decrease Rat Blood Pressure and Tension of Rat Mesenteric Artery.

Selenium compounds exert their antioxidant activity mostly when the selenium atom is incorporated into selenoproteins. In our work, we tested the possibility that selenite itself interacts with thiols to form active species that have reducing properties. Therefore, we studied the reduction of 2-(4-carboxyphenyl)-4,5-dihydro-4,4,5,5-tetramethyl-1H-imidazol-1-yloxy-3-oxide radical (•cPTIO), damage of plasmid DNA (pDNA), modulation of rat hemodynamic parameters and tension of isolated arteries induced by products of interaction of selenite with thiols. We found that the products of selenite interaction with thiols had significant reducing properties that could be attributed mainly to the selenide and that selenite had catalytic properties in the access of thiols. The potency of thiols to reduce •cPTIO in the interaction with selenite was cysteine > homocysteine > glutathione reduced > N-acetylcysteine. Thiol/selenite products cleaved pDNA, with superoxide dismutase enhancing these effects suggesting a positive involvement of superoxide anion in the process. The observed •cPTIO reduction and pDNA cleavage were significantly lower when selenomethionine was used instead of selenite. The products of glutathione/selenite interaction affected several hemodynamic parameters including rat blood pressure decrease. Notably, the products relaxed isolated mesenteric artery, which may explain the observed decrease in rat blood pressure. In conclusion, we found that the thiol/selenite interaction products exhibited significant reducing properties which can be used in further studies of the treatment of pathological conditions caused by oxidative stress. The results of decreased rat blood pressure and the tension of mesenteric artery may be perspective in studies focused on cardiovascular disease and their prevention.

Synergistic Antimicrobial Effect of Cold Atmospheric Plasma and Redox-Active Nanoparticles.

Cold argon plasma (CAP) and metal oxide nanoparticles are well known antimicrobial agents. In the current study, on an example of Escherichia coli, a series of analyses was performed to assess the antibacterial action of the combination of these agents and to evaluate the possibility of using cerium oxide and cerium fluoride nanoparticles for a combined treatment of bacterial diseases. The joint effect of the combination of cold argon plasma and several metal oxide and fluoride nanoparticles (CeO2, CeF3, WO3) was investigated on a model of E. coli colony growth on agar plates. The mutagenic effect of different CAP and nanoparticle combinations on bacterial DNA was investigated, by means of a blue-white colony assay and RAPD-PCR. The effect on cell wall damage, using atomic force microscopy, was also studied. The results obtained demonstrate that the combination of CAP and redox-active metal oxide nanoparticles (RAMON) effectively inhibits bacterial growth, providing a synergistic antimicrobial effect exceeding that of any of the agents alone. The combination of CAP and CeF3 was shown to be the most effective mutagen against plasmid DNA, and the combination of CAP and WO3 was the most effective against bacterial genomic DNA. The analysis of direct cell wall damage by atomic force microscopy showed the combination of CAP and CeF3 to be the most effective antimicrobial agent. The combination of CAP and redox-active metal oxide or metal fluoride nanoparticles has a strong synergistic antimicrobial effect on bacterial growth, resulting in plasmid and genomic DNA damage and cell wall damage. For the first time, a strong antimicrobial and DNA-damaging effect of CeF3 nanoparticles has been demonstrated.

Ratio-dependent effects of photoactivated hypericin and manumycin A on their genotoxic and mutagenic potential

Natural compounds originated from plants and microorganisms and their combinations are currently being investigated as a possible treatment for several diseases including cancer. Hypericin (photodynamically-active pigment from Hypericum perforatum L.) and manumycin A (inhibitor of farnesyltransferase from Streptomyces parvulus) belong to the chemicals potentially applicable in clinical practice. In this study we evaluated potential cytotoxic (via trypan blue exclusion test), genotoxic (via DNA-topology and comet assays), and mutagenic effects (via bacterial reverse mutation test) of these compounds and their combinations considering the molecular mechanism of their action in cell-free and cellular systems. Our results did not reveal neither cytotoxic nor mutagenic activities of tested compounds and their combinations. Regarding the genotoxic potential, no damage of plasmid DNA in cell-free system was detected. On the other hand, photoactivated hypericin and manumycin A were able to induce primary DNA damage in human lymphocytes analyzed by comet assay. The possible antagonistic interactions between these two metabolites were estimated using SynergyFinder software analysis and experimental data obtained from comet assay. Our findings indicate that not only the presence of substances, but also their ratio plays an important role in resulting effects of the combined treatment in cellular system.

Induction of DNA strand breaks and oxidative base damages in plasmid DNA by ultra-high dose rate proton irradiation

Purpose Radiation cancer therapy with ultra-high dose rate (UHDR) exposure, so-called FLASH radiotherapy, appears to reduce normal tissue damage without compromising tumor response to therapy. The aim of this study was to clarify whether a 59.5 MeV proton beam at an UHDR of 48.6 Gy/s could effectively reduce the DNA damage of pBR322 plasmid DNA in solution compared to the conventional dose rate (CONV) of 0.057 Gy/s. Materials and Methods A simple system, consisting of pBR322 plasmid DNA in 1× Tris-EDTA buffer, was initially employed for proton beam exposure. We then used formamidopyrimidine-DNA glycosylase (Fpg) enzymes. which convert oxidative base damages of oxidized purines to DNA strand breaks, to quantify DNA single strand breaks (SSBs) and double strand breaks (DSBs) by agarose gel electrophoresis. Results Our findings showed that the SSB induction rate (SSB per plasmid DNA/Gy) at UHDR and the induction of Fpg enzyme sensitive sites (ESS) were significantly reduced in UHDR compared to CONV. However, there was no significant difference in DSB induction and non-DSB cluster damages. Conclusions UHDR of a 59.5 MeV proton beam could reduce non-clustered, non-DSB damages, such as SSB and sparsely distributed ESS. However, this effect may not be significant in reducing lethal DNA damage that becomes apparent only in acute radiation effects of mammalian cells and in vivo studies.

Cobalt-mediated oxidative DNA damage and its prevention by polyphenol antioxidants

Although cobalt is a required nutrient, it is toxic due to its ability to generate reactive oxygen species (ROS) and damage DNA. ROS generation by Co2+ often has been compared to that of Fe2+ or Cu+, disregarding the reduction potential differences among these metal ions. In plasmid DNA damage studies, a maximum of 15% DNA damage is observed with Co2+/H2O2 treatment (up to 50 μM and 400 μM, respectively) significantly lower than the 90% damage observed for Fe2+/H2O2 or Cu+/H2O2 treatment. However, when ascorbate is added to the Co2+/H2O2 system, a synergistic effect results in 90% DNA damage. DNA damage by Fe2+/H2O2 can be prevented by polyphenol antioxidants, but polyphenols both prevent and promote DNA damage by Cu+/H2O2. When tested for cobalt-mediated DNA damage affects, eight of ten polyphenols (epicatechin gallate, epigallocatechin gallate, propyl gallate, gallic acid, methyl-3,4,5-trihydroxybenzoate, methyl-4,5-dihydroxybenzoate, protocatechuic acid, and epicatechin) prevent cobalt-mediated DNA damage with IC50 values of 1.3 to 27 μM and two (epigallocatechin and vanillic acid) prevent little to no DNA damage. EPR studies demonstrate cobalt-mediated formation of •OH, O2•ˉ, and •OOH, but not 1O2 in the presence of H2O2 and ascorbate. Epigallocatechin gallate and methyl-4,5-dihydroxybenzoate significantly reduce ROS generated by Co2+/H2O2/ascorbate, consistent with their prevention of cobalt-mediated DNA damage. Thus, while cobalt, iron, and copper are all d-block metal ions, cobalt ROS generation and its prevention is significantly different from that of iron and copper.

Phytochemical screening and in vitro evaluation of antioxidant and DNA inhibition activity of Caralluma bhupenderiana Sarkaria

Introduction and Aim: Phytochemicals isolated from medicinal plants have been receiving greater attention owing to their potential therapeutic efficiency. Despite rapid developments in the field of modern medicine various herbal medications are being used to treat different types of health disorders. Caralluma bhupenderiana Sarkaria is an endemic species of the genus Caralluma. Although selected species of Caralluma have been investigated for their medicinal properties, no studies have addressed the composition of bioactive phytochemicals and its therapeutic properties. The objective was to screen the endogenous composition of C. bhupenderiana extract and evaluate its antioxidant and DNA damaging inhibitory activity. Materials and Methods: Antioxidant potential of C. bhupenderiana was determined by measuring the ability of the extracts to scavenge free radicals. Chemical bonding and structural aspects of the phytochemicals were determined by FT-IR and GC-MS. Plasmid DNA (pBR322) damage inhibition was determined by photolyzing H2O2 and exposing to ultra-violet radiation. Results: The extract was rich in flavonoids, phenols, quinones, glycosides and saponins. FT-IR analysis revealed the presence of functional groups to be ‘polymeric’ OH stretch, /Methylene C-H asymmetric, esters with double bond, cyclic alkene/organic nitrates, nitro compound, ammonium ion, sulfonates. C. bhupenderiana exhibited significant antioxidant activity as evidenced by its free radical scavenging activity. The extract also exhibited complete protection against DNA damage caused by UV irradiation. Conclusion: C. bhupenderiana contains various endogenous phytochemicals that offer high antioxidant potential along with significant ability to attenuate DNA damage. C. bhupenderiana is highly therapeutic in nature and can be a good candidate for developing novel pharmacological and therapeutic molecules.

Green synthesis, characterizations of silver nanoparticles using sumac (Rhus coriaria L.) plant extract and their antimicrobial and DNA damage protective effects.

Today, antimicrobial resistance against bacteria has become an important global public health problem. In this sense, the development of new biomedical solutions is becoming increasingly important. Especially plant-based nanoparticles produced by green synthesis are used in many fields. AgNPs have an important place in nanoscience and nanotechnology, especially in nanomedicine. Therefore, the present study was conducted to synthesize AgNPs using the medicinal plant extract sumac and to characterize them using advanced techniques and to determine the antibacterial activity of some bacteria that cause disease. Nanoparticles produced by green synthesis are used in a wide area around the world due to their many advantages such as environmentally friendly, economically and non-toxically. In this study, AgNPs were biosynthesized using sumac extract and evaluated for their antibacterial potency against Bacillus cereus, Bacillus subtilis, Enterococcus faecalis, Pseudomonas aeruginos, and Candida albicans. UV-Vis spectroscopy of the prepared sumac-mediated silver solution showed the absorption maximum at about 400 nm. According to the TEM results obtained, it was observed that the particles were spherical, approximately 4 nm in size, and showed a homogeneous distribution. The sizes of nanoparticles formed by XRD pattern were supported and silver nanoparticles were obtained. According to the obtained XRD results, the crystal nature of nanoparticles in face-centered cubic structure was confirmed by the peaks in the XRD model corresponding to the planes (111) (200) (220) and (311). It was observed that the synthesized AgNPs provided a strong protection against plasmid DNA damage. It was determined that the inhibition zone diameters of biosynthesized nanoparticles measured in terms of antibacterial activity were between 10 and 14 mm. As a result, the study revealed significant antibacterial activity of the synthesized AgNPs due to extensive membrane damage.

Anti-tumor and antioxidant activity of kaempferol-3-O-alpha-L-rhamnoside (Afzelin) isolated from Pithecellobium dulce leaves

BackgroundPithecellobium dulce (Roxb.), an evergreen medium-sized, spiny tree which have vast nutritional values and widely used in ayurvedic medicines and home remedies. The plant has also been a rich source of biologically active compounds. The present study was designed to isolate pure compound from ethyl acetate fraction of methanol extract of leaves and to know the efficacy as antioxidant as well as its anti-tumor activity on Ehrlich ascites carcinoma cell (EAC). MethodsThe leaves were extracted with methanol and fractionated with different solvents. The isolation of the compound was carried out by column chromatography from ethyl acetate fraction (EAF) and structure was revealed by 1H-NMR and 13C NMR. The antioxidant activity was investigated by the scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals as well as the inhibition of oxidative damage of pUC19 plasmid DNA, hemolysis and lipid peroxidation induced by a water-soluble free radical initiator 2,2’-azo (2-asmidinopropane) dihydrochloride (AAPH) in human erythrocytes. In vivo anti-tumor activity of the compound was also evaluated by determining the viable tumor cell count, hematological profiles of experimental mice along with observing morphological changes of EAC cells by fluorescence microscope.ResultsThe isolated compound kaempferol-3-O-alpha-L-rhamnoside effectively inhibited AAPH induced oxidation in DNA and human erythrocyte model and lipid per oxidation as well as a stronger DPPH radical scavenging activity. In anti-tumor assay, at a dose of 50 mg/kg body weight exhibit about 70.89 ± 6.62% EAC cell growth inhibition, whereas standard anticancer drug vincristine showed 77.84 ± 6.69% growth inhibition.ConclusionThe compound may have a great importance as a therapeutic agent in preventing oxidative damage of biomolecules and therapeutic use in chemotherapy.

Radiosensitization Effect of Gold Nanoparticles on Plasmid DNA Damage Induced by Therapeutic MV X-rays.

Gold nanoparticles (AuNPs) can be used with megavolt (MV) X-rays to exert radiosensitization effects, as demonstrated in cell survival assays and mouse experiments. However, the detailed mechanisms are not clear; besides physical dose enhancement, several chemical and biological processes have been proposed. Reducing the AuNP concentration while achieving sufficient enhancement is necessary for the clinical application of AuNPs. Here, we used positively charged (+) AuNPs to determine the radiosensitization effects of AuNPs combined with MV X-rays on DNA damage in vitro. We examined the effect of low concentrations of AuNPs on DNA damage and reactive oxygen species (ROS) generation. DNA damage was promoted by 1.4 nm +AuNP with dose enhancement factors of 1.4 ± 0.2 for single-strand breaks and 1.2 ± 0.1 for double-strand breaks. +AuNPs combined with MV X-rays induced radiosensitization at the DNA level, indicating that the effects were physical and/or chemical. Although −AuNPs induced similar ROS levels, they did not cause considerable DNA damage. Thus, dose enhancement by low concentrations of +AuNPs may have occurred with the increase in the local +AuNP concentration around DNA or via DNA binding. +AuNPs showed stronger radiosensitization effects than −AuNPs. Combining +AuNPs with MV X-rays in radiation therapy may improve clinical outcomes.

Quantifying the DNA-damaging Effects of FLASH Irradiation With Plasmid DNA

To investigate a plasmid DNA nicking assay approach for isolating and quantifying the DNA-damaging effects of ultrahigh-dose-rate (ie, FLASH) irradiation relative to conventional dose-rate irradiation. We constructed and irradiated phantoms containing plasmid DNA to nominal doses of 20 Gy and 30 Gy using 16 MeV electrons at conventional (0.167 Gy/s) and FLASH (46.6 Gy/s and 93.2 Gy/s) dose rates. We delivered conventional dose rates using a standard clinical Varian iX linear accelerator and FLASH dose rates (FDRs) using a modified Varian 21EX C-series linear accelerator. We ran the irradiated DNA and controls (0 Gy) through an agarose gel electrophoresis procedure that sorted and localized the DNA into bands associated with single strand breaks (SSBs), double strand breaks (DSBs), and undamaged DNA. We quantitatively analyzed the gel images to compute the relative yields of SSBs and DSBs and applied a mathematical model of plasmid DNA damage as a function of dose to compute the relative biological effectiveness (RBE) of SSB and DSB (RBESSB and RBEDSB) damage for a given endpoint and FDR. Both RBESSB and RBEDSB were less than unity with the FDR irradiations, indicating FLASH sparing. With regard to the more deleterious DNA DSB damage, the DSB RBEs of FLASH beams at dose rates of 46.6 Gy/s and 93.2 Gy/s relative to the conventional 16 MeV beam dose rate were 0.54 ± 0.15 and 0.55 ± 0.17, respectively. This study demonstrated the feasibility of using a DNA-based phantom to isolate and assess the FLASH sparing effect on DNA. We also found that FLASH irradiation causes less damage to DNA compared with a conventional dose rate. This result supports the notion that the protective effect of FLASH irradiation occurs at least partially via fundamental biochemical processes.

In vitro cytotoxicity, macromolecular interaction and antioxidant potential of dual coated selenium nanoparticles.

The present research focuses on synthesizing selenium nanoparticles (Se NPs) coated with Methionine and Folic acid. selenium-methionine-folate nanoparticles (Se-Met-Fa NPs) were prepared by the chemical precipitation method. These were characterized by UV-Vis spectroscopy, FTIR, X-ray diffraction, Zeta potential, ICP-AES, TEM, and Raman spectroscopy. The average diameter of nanoparticles was determined by TEM was 50 nm. In Vitro viability of cells exposed to Se-Met-Fa NPs were studied using MTT and AO/EB assay. Approximately 80% of cells were viable at 100 μg/ml concentration after 24 h of incubation, suggestive of the safety of nanoparticles. Macromolecular interaction studies were carried out with plasmid DNA and Bovine serum albumin (BSA) protein. UV-Vis Spectroscopy showed ground state complex formation of Se-Met-Fa NPs with BSA. Intrinsic fluorescence of BSA was quenched by Se-met-Fa NPs via Static quenching and was observed under Spectro-fluorimetry. Conformational structural change in α-Helices of BSA was observed to be 4.4% after the interaction with Se-Met-Fa NPs, and it was studied using CD spectroscopy. At 250 μg/ml Se-Met-Fa NPs prevented oxidative damage of Plasmid DNA. The total antioxidant property of Se-Met-Fa NPs expressed in terms of scavenging of free DPPH radicals. Ten micrograms per milliliter could inhibit 41% of DPPH, proving its scavenger role at the lowest concentration. Nanoparticles comprising antioxidant semi-conducting cores and encapsulated by biomaterials that are highly bioavailable can be promising therapeutic agents for inflammation and oxidative stress disorders studies.

Synthesis and characterization of Sophora alopecuroides L. green synthesized of Ag nanoparticles for the antioxidant, antimicrobial and DNA damage prevention activity

In this study, it was aimed to investigate the amount of antioxidant, protective properties against DNA damage and antibacterial properties against various pathogens after the interaction of Ag metal (Ag NPs/Sa) of Sophora alopecuroides L. (S. alopecuroides L) plant seed, which is grown in Iğdır and used in the treatment of many diseases. The DPPH radical quenching activity of Ag NPs/Sa was performed by using Blois method, DNA damage prevention activity by gel electrophoresis and antibacterial property by disk diffusion method. With the green synthesis method, AgNPs obtained as a result of the reaction of the plant and Ag metal are UV visible spectrophotometer (UV-vis), fourier-transformed infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and scanning electron microscope (SEM). DPPH radical quenching activity of Ag NPs/Sa was investigated in the concentration range of 25-250 μg/ml. The radical quenching activity at a concentration of 250 μg/ml was 85,215 ± 0,101%, while this value was 93,018% for the positive control BHA. It has been observed that the protective property of pBR322 plasmid DNA damage against OH radicals originating from H2O2 increases with concentration. It has been observed that Ag NPs/Sa has significant antimicrobial properties against some pathogens (B. subtilis ATCC 6633 E. coli ATCC 25952, B. cereus ATCC 10876, P. aeruginosa ATCC 27853, E. faecalis ATCC 29212, S. aureus ATTC 29213 and C. albicans ATTC 90028) that cause disease and even some pathogens are more effective than antibiotics.

Evaluation of Leea rubra Leaf Extract for Oxidative Damage Protection and Antitumor and Antimicrobial Potential.

Background The leaves of Leea rubra contain an abundance of phenolic constituents and have medicinal uses as antipyretic and diaphoretic agents and are also used in the treatment of stomach ache, rheumatism, arthritis etc. In spite of the traditional uses, data on the scientific evaluation of the plant are not sufficient. So, the present study was designed to evaluate the protective role of the extract against oxidative damage to DNA and human erythrocytes as well as antitumor and antibacterial activities against some resistant bacteria. Methods The protective activity of the ethyl acetate fraction (EAF) of the extract was investigated by evaluating the inhibition of oxidative damage of pUC19 plasmid DNA as well as hemolysis and lipid peroxidation damage to human erythrocytes induced by 2,2′-azobis-2-amidinopropane (AAPH). Antitumor activity was assessed by evaluating the percentage inhibition of cell growth, morphological changes of Ehrlich's ascites carcinoma (EAC) cells, and hematological parameters. Antimicrobial activity was determined by the disc diffusion method against different resistant microorganisms. Results EAF effectively inhibited AAPH-induced oxidative damage to DNA because it can inhibit the transformation of the supercoiled form of plasmid DNA to open circular and further linear form. The oxidative hemolysis caused by AAPH in human erythrocytes was inhibited by EAF extract in a time-dependent manner, and the production of malondialdehyde (MDA) was significantly reduced, which indicates the prevention of lipid peroxidation. In antitumor assay, 76% growth of inhibition of EAC was observed compared with the control mice (p < 0.05) at a dose of 100 mg/kg body weight. Antimicrobial activity was evaluated against two pathogenic resistant microorganisms (Escherichia coli and Pseudomonas aeruginosa), and the highest antimicrobial activity was observed against Pseudomonas spp. Conclusion EAF may have great importance in preventing oxidative damage to DNA, erythrocytes, and other cellular components as well as can be a good candidate in cancer chemotherapy and treating infectious diseases caused by antibiotic-resistant bacteria.

In vitro cytogenotoxic and mutagenic effects of Commiphora myrrha essential oil

Commiphora myrrha, located in the tropical zone, is a widely used tree for medicinal purposes in the Arabian Peninsula and a large part of Africa. In this research, cytogenotoxic effects of the commercially available Commiphora myrrha essential oil (myrrh) were studied using micronucleus (MN), comet, and total oxidant (TOS), and total antioxidant (TAS) assays on human peripheral lymphocytes under in vitro conditions. In addition, pure pBR322 plasmid DNA was used to investigate DNA damaging/protecting activity of the essential oil. Finally, a bacterial reversion (Ames) test was performed using Salmonella typhimurium mutant strains TA98 and TA100 to determine the potential effect of the agent in the induction of gene mutations. The high concentration of Commiphora myrrha (0.125 µL/mL) induced MN formation significantly compared to the untreated control in both treatment times (24 or 48 h). Only at the highest concentration, nuclear division index (NDI) values were found lower than the controls. In the Comet test performed on healthy lymphocytes, only the highest concentration of myrrh caused significant increases in the percentage of damaged cells and genetic damage index (GDI) values. Myrrh oil showed no significant mutagenic effect on mutant Salmonella strains. In addition, the substance did not directly damage plasmid DNA but also protected DNA against damaging factors such as H2O2 and UV. Finally, in the TAS and TOS assays, no significant differences on the oxidative stress parameters were found in cell culture compared to the control. The results of this study showed that myrrh oil exerts cytogenotoxic risk only at higher concentrations.

Variation of Particle-Induced Oxidative Potential of PM2.5 in Xinjiang, NW-China

In order to evaluate the toxicity of PM2.5 in the Dushanzi area, PM2.5 samples were collected from December 2015 to July 2016, and a plasmid DNA damage assessment method was used to analyze the variation in the oxidative damage ability and its relationship with sampling conditions and toxic components (polycyclic aromatic hydrocarbons, and heavy metals) loaded on the surface of PM2.5. The results showed that the TD30 values (toxic dosage of PM2.5 causing 30% of plasmid DNA damage) of both the whole samples and the water-soluble fractions were lower during the heating period (369 μg/mL and 536 μg/mL, respectively), but higher in the dust period and non-heating period (681 μg/mL and 498 μg/mL, respectively; and 804 μg/mL and 847 μg/mL, respectively). Studies on the effect of meteorological parameters showed an increasing trend in TD30 values for the whole samples and the water-soluble fractions as relative humidity, temperature and wind speed decrease. TD30 values for the whole samples and the water-soluble fractions were negatively correlated with Flu (r = −0.690,r = −0.668; p &lt; 0.05), Flt (r =−0.671, r = −0.760; p &lt; 0.05), BaP (r = −0.672, r = −0.725; p &lt; 0.05), IcdP (r = −0.694, r = −0.740; p &lt; 0.05), Pyr (r = −0.727, r = −0.768; p &lt; 0.01) and BghiP (r = −0.874, r = −0.845; p &lt; 0.01) during the heating period, while As (r = 0.792, r = 0.749; p &lt; 0.05) and Sr (r = 0.776, r = 0.754; p &lt; 0.05) during the dust period showed significant positive correlation. In addition, the TD30 values of PM2.5 collected during sand blowing weather was the highest (1458 μg/mL and 1750 μg/mL), while the average TD30 value of PM2.5 collected on hazy days were the lowest (419.8 μg/mL and 488.6 μg/mL). Particles collected on the first day after snowfall showed a lower oxidizing capacity (676 μg/mL and 1330 μg/mL). The characteristic TD30 values combined with back trajectory analysis indicated that hazy days were heavily influenced by air masses originating from the southern continent and local emissions, whereas the sand blowing weather came from the north of the Taklimakan Desert.

Protective Effects of Amino Acids on Plasmid DNA Damage Induced by Therapeutic Carbon Ions.

Radioprotectors with few side effects are useful for carbon-ion therapy, which directly induces clustering damage in DNA. With the aim of finding the most effective radioprotector, we investigated the effects of selected amino acids which might have chemical DNA-repair functions against therapeutic carbon ions. In the current study, we employed five amino acids: tryptophan (Trp), cysteine (Cys), methionine (Met), valine (Val) and alanine (Ala). Samples of supercoiled pBR322 plasmid DNA with a 17 mM amino acid were prepared in TE buffer (10 mM Tris, 1 mM ethylenediaminetetraacetic acid, pH 7.5). Phosphate buffered saline (PBS) was also used in assays of the 0.17 mM amino acid. The samples were irradiated with carbon-ion beams (290 MeV/u) on 6 cm spread-out Bragg peak at the National Institute of Radiological Sciences and Heavy Ion Medical Accelerator in Chiba, Japan. Breaks in the DNA were detected as changes in the plasmids and quantified by subsequent electrophoresis on agarose gels. DNA damage yields and protection factors for each amino acid were calculated as ratios relative to reagent-free controls. Trp and Cys showed radioprotective effects against plasmid DNA damage induced by carbon-ion beam, both in PBS and TE buffer, comparable to those of Met. The double-strand break (DSB) yields and protective effects of Trp were comparable to those of Cys. The yields of both single-strand breaks and DSBs correlated with the scavenging capacity of hydroxyl radicals (rate constant for scavenging hydroxyl radicals multiplied by the amino acid concentration) in bulk solution. These data indicate that the radioprotective effects of amino acids against plasmid DNA damage induced by carbon ions could be explained primarily by the scavenging capacity of hydroxyl radicals. These findings suggest that some amino acids, such as Trp, Cys and Met, have good potential as radioprotectors for preventing DNA damage in normal tissues in carbon-ion therapy.

Caffeic Acid Phenyl Ester (CAPE) Protects against Iron-Mediated Cellular DNA Damage through Its Strong Iron-Binding Ability and High Lipophilicity.

Caffeic acid phenethyl ester (CAPE) and its structurally-related caffeic acid (CA), ferulic acid (FA) and ethyl ferulate (EF) are constituents of honeybee propolis that have important pharmacological activities. This study found that CAPE—but not CA, FA, and EF—could effectively prevent cellular DNA damage induced by overloaded iron through decreasing the labile iron pool (LIP) levels in HeLa cells. Interestingly, CAPE was found to be more effective than CA in protecting against plasmid DNA damage induced by Fe(II)–H2O2 or Fe(III)–citrate–ascorbate-H2O2 via the inhibition of hydroxyl radical (•OH) production. We further provided more direct and unequivocal experimental evidences for the formation of inactive CAPE/CA–iron complexes. CAPE was found to have a stronger iron-binding ability and a much higher lipophilicity than CA. Taken together, we propose that the esterification of the carboxylic moiety with phenethyl significantly enhanced the iron-binding ability and lipophilicity of CAPE, which is also responsible for its potent protection against iron-mediated cellular DNA damage. A study on the iron coordination mechanism of such natural polyphenol antioxidants will help to design more effective antioxidants for the treatment and prevention of diseases caused by metal-induced oxidative stress, as well as help to understand the structure–activity relationships of these compounds.