Base-pair Substitution Mutations Research Articles

Contribution of the SOS response and the DNA repair systems to norfloxacin induced mutations in E. coli

Antibiotic-resistant bacteria severely threaten human health. Besides spontaneous mutations generated by endogenous factors, the resistance might also originate from mutations induced by certain antibiotics, such as the fluoroquinolones. Such antibiotics increase the genome-wide mutation rate by introducing replication errors from the SOS response pathway or decreasing the efficiency of the DNA repair systems. However, the relative contributions of these molecular mechanisms remain unclear, hindering understanding of the generation of resistant pathogens. Here, using newly-accumulated mutations of wild-type and SOS-uninducible Escherichia coli strains, as well as those of the strains deficient for the mismatch repair (MMR) and the oxidative damage repair pathways, we find that the SOS response is the major mutagenesis contributor in mutation elevation, responsible for ~ 30–50% of the total base-pair substitution (BPS) mutation-rate elevation upon treatment with sublethal levels of norfloxacin (0 ~ 50 ng/mL). We further estimate the significance of the effects on other mutational features of these mechanisms (i.e., transversions, structural variations, and mutation spectrum) in E. coli using linear models. The SOS response plays a positive role in all three mutational features (mutation rates of BPSs, transversions, structural variations) and affects the mutational spectrum. The repair systems significantly reduce the BPS mutation rate and the transversion rate, regardless of whether antibiotics are present, while significantly increasing the structural variation rate in E. coli. Our results quantitatively disentangle the contributions of the SOS response and DNA repair systems in antibiotic-induced mutagenesis.

Implications of reactive oxygen species in lung cancer and exploiting it for therapeutic interventions.

Lung cancer is the second (11.4%) most commonly diagnosed cancer and the first (18%) to cause cancer-related deaths worldwide. The incidence of lung cancer varies significantly among men, women, and high and low-middle-income countries. Air pollution, inhalable agents, and tobacco smoking are a few of the critical factors that determine lung cancer incidence and mortality worldwide. Reactive oxygen species are known factors of lung carcinogenesis resulting from the xenobiotics and their mechanistic paths are under critical investigation. Reactive oxygen species exhibit dual roles in cells, as a tumorigenic and anti-proliferative factor, depending on spatiotemporal context. During the precancerous state, ROS promotes cancer origination through oxidative stress and base-pair substitution mutations in pro-oncogenes and tumor suppressor genes. At later stages of tumor progression, they help the cancer cells in invasion, and metastases by activating the NF-kB and MAPK pathways. However, at advanced stages, when ROS exceeds the threshold, it promotes cell cycle arrest and induces apoptosis in cancer cells. ROS activates extrinsic apoptosis through death receptors and intrinsic apoptosis through mitochondrial pathways. Moreover, ROS upregulates the expression of beclin-1 which is a critical component to initiate autophagy, another form of programmed cell death. ROS is additionally involved in an intermediatory step in necroptosis, which catalyzes and accelerates this form of cell death. Various therapeutic interventions have been attempted to exploit this cytotoxic potential of ROS to treat different cancers. Growing body of evidence suggests that ROS is also associated with chemoresistance and cancer cell immunity. Considering the multiple roles of ROS, this review highlights the exploitation of ROS for various therapeutic interventions. However, there are still gaps in the literature on the dual roles of ROS and the involvement of ROS in cancer cell immunity and therapy resistance.

Correcting a PLCζ Mutation in the Human Germ Line to Overcome Hereditary Infertility

Human germline gene correction by CRISPR/Cas9 holds great promise to eliminate transmission of genetic mutations. Recent studies have however made troublesome observations of mosaicism (the occurrence of multiple genetic events in one embryo) and loss-of-heterozygosity (LOH, loss of the allele of one of the parents), the latter being either caused by inter-homologue homologous recombination (IH-HR) or chromosome loss events. As a proof-of-concept we targeted a heterozygous base pair substitution mutation in PLCZ1, leading to subfertility. After CRISPR/Cas9 editing, we detected in 26% (7/27) of the targeted embryos that originated from mutant sperm only, wild-type alleles upon analysis, which were not obtained by exogenous repair template use. In these seven embryos, we observed a significant amount of LOH events (14%) that spanned regions beyond the PLCZ1 gene, but did not stretch the complete analysed region, pointing to IH-HR, next to a large proportion of samples (57%) showing potential but non-detectable (i.e. below the resolution of our assay) LOH. In addition, a minority of embryos (29%) showed LOH across the whole analysed region. Remarkably, also 10% of the embryos originating from wild-type sperm showed LOH events, underscoring the subpar specificity of the CRIPSR/Cas9 system. Single-cell analysis demonstrated mosaicism in 80% (4/5) of the targeted embryos, warranting caution for trophectoderm biopsy interpretation in future human embryo editing attempts. Lastly, a correction rate of 75% was observed in our single-cell data which illustrates that the correction rate was possibly underestimated when looking at the whole-embryo level.

In vitro screening of genotoxicity and mutagenicity of pyriproxyfen in human lymphocytes and Salmonella typhimurium TA98 and TA100 strains

Pyriproxyfen (PPX) is a pesticide/larvicide used to increase productivity in agriculture against insects by inhibiting development of insects’ larvae. In this study, cytotoxic, genotoxic, and mutagenic effects of PPX were investigated in human peripheral lymphocytes and Salmonella typhimurium strains by performing chromosomal aberration, micronucleus (MN) tests, and Ames test, respectively. For the chromosome aberration (CA) and MN methods, blood from four healthy donors (two men and two women, nonsmokers) were used. Two hundred microliters of blood was inoculated into PbMax medium and prepared according to International Guidelines. For the Ames test, S. typhimurium TA98 and TA100 strains were used to detect frameshift and base pair substitution mutagens, respectively. PPX induced both the CA percentage and MN frequency in human peripheral lymphocytes and exhibited cytotoxic effects. In addition, it showed a mutagenic effect at all doses in TA98 and TA100 strains in the presence of S9mix; however, no such effect was observed in the absence of S9mix. According to the obtained results, it can be said that PPX has genotoxic and mutagenic potentials.

Mutagenicity of silver nanoparticles synthesized with curcumin (Cur-AgNPs)

IntroductionSilver nanoparticles (AgNPs) are of particular interest for their antibacterial properties and are produced by the action of reducing agents on silver ions. Curcumin from Curcuma longa (Zingiberaceae) has been used as a precursor for obtaining biogenic AgNPs, to act as a potential drug. ObjectivesThis study aimed to evaluate the toxicity of AgNPs synthesized with curcumin (Cur-AgNPs 0.081 mg/mL, ~130 nm) through the Salmonella/microsome (Ames test), one of the first required assays for evaluating toxicity. MethodsThe study design was experimental and in vitro. After defining the preliminary toxicity, the mutagenicity was assessed in a concentration range of 0.0010–0.0081 mg/plate Cur-AgNPs using histidine negative (His−) Salmonella Typhimurium strains TA97a, TA98, TA100, and TA102, with (+S9) and without metabolic activation (−S9), in triplicate. Assays were monitored by positive and negative controls. The results were statistically analyzed by Salanal software with p < 0.05 values considered significant. ResultsThe data obtained in the absence of metabolic activation showed that Cur-AgNPs is not mutagenic, but when exposed to the presence of S9, Cur-AgNPs became mutagenic to TA98 and TA100 strains, showing the significance of metabolizer enzymes to activate Cur-AgNPs on these bacteria, which recovered their abilities in synthesizing histidine (His+). ConclusionCur-AgNPs is mutagenic in the presence (+S9), but not in the absence (−S9) of metabolic activation, being able to act as indirect mutagens potentially to organisms that share the same genotype vulnerabilities found in TA98 and TA100 strains to cause a frameshift and base-pair substitution mutations, respectively.

Ecotoxicological evaluation of fruit extracts from yerba mate progenies (Ilex paraguariensis a St-Hil.): a natural biopesticide

This study evaluated the biocidal activity of aqueous fruit extracts from Ilex paraguariensis progenies at different ripening stages in two ecotoxicity assays using Pomacea canaliculata juveniles and Danio rerio larvae. Mutagenicity was verified by the Salmonella/microsome assay (TA 98 and TA 100) in the presence and absence of hepatic metabolism in vitro. The results showed that the snails were more sensitive than the larvae to the P.1 progeny extracts in all three evaluated ripening stages. P.3 progeny extracts were the most toxic to Danio rerio larvae after 96 hours of exposure. The toxicity results indicate that the progenies' selection influenced the metabolic contents present in the fruits in all ripening stages over time, indicating high molluscicidal activity for the P.1 progeny. In the Salmonella/microsome assays, the semi-ripe and ripe extracts from progenies P.1 and P.3 showed mainly base pair substitution mutation in assays with metabolism, but at higher doses than the toxic concentrations detected for Danio rerio or Pomacea canaliculata. Therefore, the toxicological investigation of the progenies' extracts can be interesting, given the selection of plant materials influenced the response of the bioassays.

Unexpected Discovery of Hypermutator Phenotype Sounds the Alarm for Quality Control Strains.

Microbial strains with high genomic stability are particularly sought after for testing the quality of commercial microbiological products, such as biological media and antibiotics. Yet, using mutation–accumulation experiments and de novo assembled complete genomes based on Nanopore long-read sequencing, we find that the widely used quality-control strain Shewanella putrefaciens ATCC-8071, also a facultative pathogen, is a hypermutator, with a base-pair substitution mutation rate of 2.42 × 10−8 per nucleotide site per cell division, ∼146-fold greater than that of the wild-type strain CGMCC-1.6515. Using complementation experiments, we confirm that mutL dysfunction, which was a recent evolutionary event, is the cause for the high mutation rate of ATCC-8071. Further analyses also give insight into possible relationships between mutation and genome evolution in this important bacterium. This discovery of a well-known strain being a hypermutator necessitates screening the mutation rate of bacterial strains before any quality control or experiments.

MutVis: Automated framework for analysis and visualization of mutational signatures in pathogenic bacterial strains.

In recent years, mutational signature analysis has become a routine practice in cancer genomics for classification and diagnosis. Characterizing mutational signatures across species or within genomes of a bacteria helps in understanding their evolution and adaptation. However, an integrated framework for analysis and visualization of mutational signatures in bacterial genome is lacking. Hence, we aim to develop an integrated, automated, open-source and user-friendly framework called MutVis to analyze mutational signatures from bacterial whole genome next generation sequencing data. The current framework integrates various publicly available packages using Snakemake workflow management software, Python and R scripting. MutVis supports variant calling, transition (Ti) and transversion (Tv) graphical representation, generation of mutational count matrix, graphical visualization of base-pair substitution spectrum (BPSs) and mutation signatures extraction. TvTi plots provide the 6 base substitution classification for both genome and gene level. Further resolution of base pair substitution classification is provided as 96-profile BPSs plot. Mutation signatures is derived based on the characteristic pattern observed in BPSs using non-negative matrix factorization. Relative contribution of signatures is given as hierarchically clustered heatmap. This provides information on active signatures in the individual given sample and classify samples according to signature contributions. We demonstrated the MutVis framework using geographically different strains of Mycobacterium tuberculosis, downloaded from PATRIC TB-ARC Antibiotic Resistance Catalog (n=963). The current framework can be used to study mutation biases and characteristic mutational signatures in bacterial genomes and is freely available at https://github.com/AkshathaPrasanna/MutVis.

ACB-PCR Quantification of Low-Frequency Hotspot Cancer-Driver Mutations.

Allele-specific competitive blocker PCR (ACB-PCR) is a sensitive and quantitative approach for the selective amplification of a specific base substitution. Using the ACB-PCR technique, hotspot cancer-driver mutations (tumor-relevant mutations in oncogenes and tumor suppressor genes, which confer a selective growth advantage) are being developed as quantitative biomarkers of cancer risk. ACB-PCR employs a mutant-specific primer (with a 3'-penultimate mismatch relative to the mutant DNA sequence, but a double 3'-terminal mismatch relative to the wild-type DNA sequence) to selectively amplify rare mutant DNA molecules. A blocker primer having a non-extendable 3'-end and a 3'-penultimate mismatch relative to the wild-type DNA sequence, but a double 3'-terminal mismatch relative to the mutant DNA sequence is included in ACB-PCR to selectively repress amplification from abundant wild-type molecules. Consequently, ACB-PCR can quantify the level of a single base pair substitution mutation in a DNA population when present at a mutant:wild-type ratio of 1×10-5 or greater. Quantification of rare mutant alleles is achieved by parallel analysis of unknown samples and mutant fraction (MF) standards (defined mixtures of mutant and wild-type DNA sequences). The ability to quantify specific mutations with known association to cancer has several important applications in evaluating the carcinogenic potential of chemical exposures in rodent models. Further, the measurement of cancer-driver mutant subpopulations is important for precision cancer treatment (selecting the most appropriate targeted therapy and predicting the development of therapeutic resistance). This chapter provides a step-by-step description of the ACB-PCR methodology as it has been used to measure human PIK3CA codon 1047, CAT→CGT (H1047R) mutation.

Nonclinical evaluation of IQG-607, an anti-tuberculosis candidate with potential use in combination drug therapy

New effective compounds to treat tuberculosis are urgently needed. IQG-607 is an orally active anti-tuberculosis drug candidate, with promising preliminary safety profile and anti-mycobacterial activity in both in vitro and in vivo models of tuberculosis infection. Here, we evaluated the mutagenic and genotoxic effects of IQG-607, and its interactions with CYP450 isoforms. Moreover, we describe for the first time a combination study of IQG-607 in Mycobacterium tuberculosis-infected mice. Importantly, IQG-607 had additive effects when combined with the first-line anti-tuberculosis drugs rifampin and pyrazinamide in mice. IQG-607 presented weak to moderate inhibitory potential against CYP450 isoforms 3A4, 1A2, 2C9, 2C19, 2D6, and 2E1. The Salmonella mutagenicity test revealed that IQG-607 induced base pair substitution mutations in the strains TA100 and TA1535. However, in the presence of human metabolic S9 fraction, no mutagenic effect was detected in any strain. Additionally, IQG-607 did not increase micronucleus frequencies in mice, at any dose tested, 25, 100, or 250 mg/kg. The favorable activity in combination with first-line drugs and mild to moderate toxic events described in this study suggest that IQG-607 represents a candidate for clinical development.

Induction of Micronuclei, Base-pair Substitution Mutation and Excision-repair Deficient by Polluted Water from Asa River in Nigeria

Abstract Asa river is a major river designated to supply millions of people of Ilorin, Kwara State, Nigeria potable water for drinking but its managements is of grave concern due to anthropogenic activities. Thus, evaluation of genotoxicity of this river was carried out by subjecting the water samples and fish therein to three bioassays (Micronucleus (MN) assay, Ames test and SOS-chromo test). Physicochemical parameters and heavy metals were analysed at three different stations (Aliara (SI), Unity (SII) and Tuyil (SIII)) of the river. In SII, most of the heavy metals analysed were above the acceptable limits compare to SI and SIII. The peripheral erythrocyte of the fishes (Oreochromis niloticus, Synodontis batensoda, Synodontis eupterus, Clarias gariepinus and Clarias angullaris) at SI and SII stations showed a significant (p&lt;0.05) induction of MN and different nuclear abnormalities (NA). Water samples from the three stations subjected to Ames test (Salmonella typhimurium TA100) and SOS chromotests (Escherichia coli PQ37) at 25%, 50% and 100% concentrations showed statistically significant (p&lt;0.05) induction of DNA damage at all concentrations in the two tester strains, thus indicating base-pair substitution mutation and excision-repairdeficient, respectively, by the water samples. Therefore, drinking of this water and/or consumption of fish from this river should be taken with caution to avoid a carcinogenic risk.

STUDY OF MUTAGENIC ACTIVITY NANO- AND MICROPARTICLES IN THE AMES TEST (SALMONELLA / MICROSOME)

Introduction. One of the important steps in assessing the nanoparticles (NP) safety is the analysis of mutagenic activity, including the evaluation of gene, chromosomal, and genomic mutations. Material and methods. The purpose of this investigation is to study the ability of different NP aqueus suspensions and the same compounds in microforms to unduce gene mutations in Salmonella/microsome test (Ames test). Anatase titanium dioxide NP coated with simethicone (33.16 ± 16.7 nm, 5-50000 μg/ml), magnetite NP coated with silicate (10 nm, 0.92-575 μg/ml), silver NP coated with аrabian gum (14 ± 0.2 nm, 5-50000 μg/ml), aluminum hydroxide nanofibres (50-70 nm, 24-3000 μg/ml) and multi-walled carbon nanotubes (Taunit MWСNTs, outer diameter 15-40 nm, inner diameter 3-8 nm, length 2 and more microns, 5-50000 μg/ml). In parallel, the mutagenic activity of equivalent microparticles was evaluated in experiments. Ames test (Salmonella/microsomes) registers gene mutations induced by a different mechanism of action, in the variant with preincubation. A set of Salmonella typhimurium indicator strains: TA 100 (base pair substitution mutations), TA 98 and TA 97 (mutations of the frameshift type of the genetic code) were used. Using addition the S9 microsomal activating mixture during the experiment makes it is possible to determine the effect not only of the substances themselves, but also of their metabolites. Conclusion. The investigated nanomaterials as well as their micro analogs in the studied dose range did not induce gene mutations in the Ames test both in presence and absence microsomal activating mixture.

Influence of chicken growth hormone (cGH) SNP genotypes on morphometric and growth traits of three chicken breeds in Nigeria.

This study was conducted to identify the diversity of chicken growth hormone (cGH) single nucleotide polymorphs’ (SNP) and their association on morphometric and growth traits of three Nigerian chicken breeds namely Funaab Alpha (FA), Shikabrown (SB) and Nigerian local chicken (NLC). Morphometric traits measured include shank length (cm), breast girth (cm) and breast width (cm) while growth traits measured were final body weight, av. weight gain, feed intake and feed conversion ratio all at laying stages respectively. Morphometric and growth traits were significantly influenced (P less than 0.05) between the different breeds. The SNP panel showed two base pair substitution mutation (GT and CA) on locus 5 chromosome for the FA and locus 6 chromosome for the SB and NLC respectively. The G – T alignment produced genotypes GG and GT, with TT not observed. Also C – A alignment produced genotypes CC and CA, with AA not observed. The NLC breed had the highest number of haplotypes of 6 while FA and SB had 4 haplotypes each implying that the local breed had more allelic variation than the other two breeds. Also FA had heterozygosity of 0.48 while SB and NLC had a value of 0.44, indicating higher genetic variability of FA breed than the rest with negative theta value (-0.0843) of F-statistics indicating high level of outbreeding. The conducted association of SNP genotypes showed no significant association effects (P&gt;0.05) on the morphometric traits, except for SB breed where CA genotypes was significantly associated with CC for bodyweight, and in NLC breed where CC genotype was also significantly associated with CA genotypes for shank length.

Genotoxicity and mutagenicity of nickel(II) and iron(III) and interactions between these microelements

Abstract. Background: Nickel and iron are essential and necessary elements required for correct functioning of the human body when found in trace amounts, but their excess can be toxic for cells. They are widely used in numerous industrial applications as components in orthopedic implants and in dietary supplementation. The aim of this study was to examine the effect of nickel(II) and iron(III) and their combinations on genotoxicity and mutagenicity in BALB/3T3 and HepG2 cells. Materials and methods: Genotoxicity of nickel and iron and their mixture was assessed by analyzing induction of ­micronucleus formation and DNA damage by comet assay in BALB/3T3 and HepG2 cells. Mutagenicity of iron(III) and nickel(II) and their mixtures was assessed by Ames assay. Results: A statistically significant increase of DNA damage through use of both microelements in both cell lines was observed. The micronucleus assay performed with the use of all concentrations showed a statistically significant induction of chromosomal aberrations in both tested microelements in both cell lines. The results obtained from both cell lines in both tests show that BALB/3T3 cells are more sensitive when compared to the HepG2 cells after incubation with both microelements. Additions of iron(III) at 200 µM plus nickel(II) at 1,000 µM showed antagonistic effects in a decrease of genotoxicity as assessed by comet and micronuclei assays in both cell lines. In the case of nickel(II) at 200 µM plus iron(III) at 1,000 µM, a synergistic effect was observed. The results of Ames assay show that both tested microelements caused an increased number of reverse mutations. Conclusion: Nickel(II) and iron(III) are genotoxic. Nickel chloride evokes frameshift mutation more often than base-pair substitution mutation. However, in the case of iron chloride, the frequency in frameshift and base-pair substitution mutation was similar. Iron(III) at concentrations of 200 µM plays a protective role against nickel(II) genotoxicity and mutagenicity.

Nutritional composition of Eragrostis teff and its association with the observed antimutagenic effects.

Eragrostis teff is an Ethiopian native grass plant (Poaceae or Gramineae family) whose importance as a crop grain has increased in recent years. The aim of this study is to analyze the nutritional composition of its seeds and the mutagenic/antimutagenic activity of the hydroalcoholic extract of the seed flour. Chemical elements (colloquially known as minerals) were determined using Particle-Induced X-ray Emission (PIXE) and Flame Atomic Absorption Spectroscopy (FAAS), while the content of amino acids (aminogram) and fatty acids (profile of fatty acids) were quantified by HPLC. Mutagenic activities were tested using Salmonella/microsome assay. Mutagens doxorubicin, 4-nitroquinolin N-oxide, methylmethanosulphonate, and aflatoxin B-1 were used in Salmonella typhimurium TA98 and TA100 strains to assess antimutagenic activities. The major elements observed were K, P, S, Mg, and Ca. Almost all essential amino acids were observed and the predominance of unsaturated fatty acids in the total oil content of 2.72% (w/w) is also noted, including the two essential fatty acids alpha-linolenic acid (an omega-3 fatty acid) and linoleic acid (an omega-6 fatty acid). Hydroalcoholic extract of E. teff seed flour showed antimutagenic activity, protecting against frameshift and base pair substitution mutations. These findings provide valuable information for further development of healthier foods that can be produced with increasing yields and minimal environmental impact.

Mutation Landscape of Base Substitutions, Duplications, and Deletions in the Representative Current Cholera Pandemic Strain.

Pandemic cholera is a major concern for public health because of its high mortality and morbidity. Mutation accumulation (MA) experiments were performed on a representative strain of the current cholera pandemic. Although the base-pair substitution mutation rates in Vibrio cholerae (1.24 × 10−10 per site per generation for wild-type lines and 3.29 × 10−8 for mismatch repair deficient lines) are lower than that previously reported in other bacteria using MA analysis, we discovered specific high rates (8.31 × 10−8 site/generation for wild-type lines and 1.82 × 10−6 for mismatch repair deficient lines) of base duplication or deletion driven by large-scale copy number variations (CNVs). These duplication–deletions are located in two pathogenic islands, IMEX and the large integron island. Each element of these islands has discrepant rate in rapid integration and excision, which provides clues to the pandemicity evolution of V. cholerae. These results also suggest that large-scale structural variants such as CNVs can accumulate rapidly during short-term evolution. Mismatch repair deficient lines exhibit a significantly increased mutation rate in the larger chromosome (Chr1) at specific regions, and this pattern is not observed in wild-type lines. We propose that the high frequency of GATC sites in Chr1 improves the efficiency of MMR, resulting in similar rates of mutation in the wild-type condition. In addition, different mutation rates and spectra were observed in the MA lines under distinct growth conditions, including minimal media, rich media and antibiotic treatments.

Gamma-Decanolactone Improves Biochemical Parameters Associated with Pilocarpine-Induced Seizures in Male Mice.

Gamma-decanolactone (GD) is a monoterpene effective against seizures induced by pentylenetetrazole. The mechanism of action of GD is likely to be via glutamate antagonism. GD also inhibits intracellular reactive oxygen species (ROS) generation and the lipopolysaccharide-induced expression of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-α) in vitro. Considering the neuropharmacological profile of GD studied so far, we investigated the effect of intraperitoneal administration of GD 100 and 300 mg/kg on pilocarpine (PIL)-induced status epilepticus (SE) in mice. GD was administered 30 min before PIL. Behavioral (latency to first seizure and the percentage of clonic forelimb seizures), biochemical, and oxidative stress parameters were evaluated. DNA damage in the cerebral cortex of mice was assessed using the comet assay and mutagenic activity of GD was evaluated using Salmonella/microsome assay in TA100, TA98, TA97a, TA102, and TA1535 strains, with and without metabolic activation (S9 mix). The behavioral results showed that only the latency to the first clonic seizure increased in the groups treated with GD 300 mg/kg, but not when the animals received GD 100 mg/kg. Both GD doses were able to increase superoxide dismutase and catalase activities, inducing a decrease in ROS and nitrite production and in DNA damage in the cerebral cortex. GD was not able to induce base pair substitution and frameshift mutations in the absence or in the presence of metabolic activation. These findings demonstrate that GD does not improve behavioral parameters in the PIL model, but it was able to protect seizure-related oxidative stress and DNA damage in mice, without inducing gene mutations.

The Mutagenicity and Antimutagenicity of Canarium odontophyllum (Dabai) Acetone Leaves Extracts

Canarium odontophyllum or locally known as Dabai in Sarawak is a fruit largely consumed by the locals. Based on previous studies, the plant possessed various biological activities, such as antimicrobial, antioxidant, antifungal and anticancer. Our aim was to investigate the mutagenicity and antimutagenicity of C. odontophyllum acetone leaves extracts by using the Ames test (Salmonella reverse mutagenicity assay).The Ames test also involved the pre-incubation method against Salmonella typhimurium TA98 and TA100 bacterial strains in the absence and presence of metabolic activator S9 system. C. odontophyllum crude acetone extracts were diluted with 10% DMSO to obtain three different concentrations of 3.125, 12.5 and 50 mg/ml. To determine the mutagenicity effects of the extracts, each concentration of the extract was evaluated based on the two-fold value of the number of revertant’s colony in negative control plate as the cut-of point. No mutagenic activity was observed for the frameshift mutation (TA98) and base-pair substitution mutation (TA100) in all concentrations of C. odontophyllum in the presence and absence of metabolic activator S9 system. Antimutagenicity test was carried out to determine the potential of C. odontophyllum extracts to inhibit the mutation induced by specific mutagens. The highest antimutagenic activity was seen in the presence of metabolic activator S9 system with inhibition percentage greater than 50% in both bacteria strains TA98 (62.38%) and TA100 (58.24%). In conclusion, C. odontophyllum acetone leaves extract was not mutagenic and had significant inhibitory effects on mutagenicity in both bacterial strains with and without the metabolic activator S9 system. Our results could contribute to the safe use of C. odontophyllum. In addition, based on the significant antimutagenic activity demonstrated by the C. odontophyllum acetone leaves extracts, the extract could also be developed as a chemopreventive agent.

The differential role of reactive oxygen species in early and late stages of cancer.

The large doses of vitamins C and E and β-carotene used to reduce reactive oxygen species (ROS) production and oxidative damages in cancerous tissue have produced disappointing and contradictory results. This therapeutic conundrum was attributed to the double-faced role of ROS, notably, their ability to induce either proliferation or apoptosis of cancer cells. However, for a ROS-inhibitory approach to be effective, it must target ROS when they induce proliferation rather than apoptosis. On the basis of recent advances in redox biology, this review underlined a differential regulation of prooxidant and antioxidant system, respective to the stage of cancer. At early precancerous and neoplastic stages, antioxidant activity decreases and ROS appear to promote cancer initiation via inducing oxidative damage and base pair substitution mutations in prooncogenes and tumor suppressor genes, such as RAS and TP53, respectively. Whereas in late stages of cancer progression, tumor cells escape apoptosis by producing high levels of intracellular antioxidants, like NADPH and GSH, via the pentose phosphate pathway to buffer the excessive production of ROS and related intratumor oxidative injuries. Therefore, antioxidants should be prohibited in patients with advanced stages of cancer and/or undergoing anticancer therapies. Interestingly, the biochemical and biophysical properties of some polyphenols allow them to selectively recognize tumor cells. This characteristic was exploited to design and deliver nanoparticles coated with low doses of polyphenols and containing chemotherapeutic drugs into tumor-bearing animals. First results are encouraging, which may revolutionize the conventional use of antioxidants in cancer.

Genetic damage induced by electronic waste leachates and contaminated underground water in two prokaryotic systems

The inappropriate and unsafe management practices related to disposal and recycling of electronic wastes in Nigeria has led to environmental and underground water contamination. Reports on the level and type of contamination as well as the possible DNA damage effects of this contamination are insufficient. This study evaluated the DNA damaging potential of e-waste simulated and raw leachates, and its contaminated underground water using the SOS chromotest on Escherichia coli PQ37 and the Ames Salmonella fluctuation test on Salmonella typhimurium strains TA98 and TA100, without and with metabolic activation. Physico-chemical parameters of the samples were also analyzed. The result of the Ames test showed induction of base pair substitution and frameshift mutation by the test samples. However, the TA100 was the more responsive strain for the three samples in terms of mutagenic index in the absence and presence of metabolic activation. The SOS chromotest results were in agreement with those of the Ames Salmonella fluctuation test. Nevertheless, the E. coli PQ37 system was slightly more sensitive than the Salmonella assay for detecting genotoxins in the tested samples. Lead, cadmium, manganese, copper, nickel, chromium, arsenic, and zinc contents analyzed in the samples were believed to play a significant role in the observed DNA damage in the microbial assays. The results of this study showed that e-waste simulated and raw leachates, and its contaminated underground water are of potential mutagenic and genotoxic risks to the exposed human populace.