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Burning vegetative debris is a worldwide long-standing practice. The current study was designed to examine the cytotoxicity and mutagenicity of particulate matter with an aerodynamic diameter below 10 µm (PM10) released from the burning of pruning residues common in Portugal and other countries of the Mediterranean region. Field measurements were conducted to collect PM10 samples from open burning of vines, olive, willow and acacia pruning branches. To assess the cytotoxicity of the PM10 total organic extract, the A549 cell line, representative of the alveolar type II pneumocytes of the human lung, was used. The cytotoxicity was checked using two complementary methods: water-soluble tetrazolium (WST-8) test to evaluate the cell metabolic activity and lactate dehydrogenase (LDH) activity assay to assess the loss of cell membrane integrity. The mutagenicity of the PM10-bound polycyclic aromatic hydrocarbons (PAHs) was screened through the Ames test. PM10 organic extracts induced LDH release in a dose-dependent manner. Regarding the cellular metabolic activity, dose-dependency was lacking for the majority of the samples. Combined WST-8 and LDH data indicate that PM10 exposure induce a necrotic cell death pathway in which the cell membrane integrity is lost. Direct and indirect mutagenicity towards the TA98 Salmonella strain has been recorded for the PAH extracts of PM10 collected from combustion of vine and willow branches during the ignition/flaming combustion stage. Significant correlations were found between the cytotoxic responses (WST-8 and LDH) and the PM10 organic component.

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 mu L/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.

In the present study, the antimicrobial and antibiofilm activities of two 4,5-dihydro-1H-pyrazole-1-carboximidamide hydrochloride, (trifluoromethyl) phenyl-substituted (compound 1) and bromophenyl-substituted (compound 2), were evaluated against four Salmonella spp. serotypes through broth microdilution and biofilm-forming activity. Further, the cytotoxicity of the compounds was evaluated by cell viability assays using cultures of HeLa and Vero cell lines, and the mutagenic potential was assessed by the Ames test. In the broth microdilution test, compound 1 inhibited 90% of the strains tested at the minimum inhibitory concentration of 62.5 μg mL−1. Furthermore, both compounds prevented biofilm formation, with a reduction of up to 5.2 log10. HeLa and Vero cells exhibited 100% viability in the presence of compound 1. In contrast, low cell viability was observed in the presence of 15 µg mL−1 of compound 2. Furthermore, no mutagenic potential was detected at any of the tested concentrations of compound 1.

Environmental mutagens are chemical and physical substances in the environment that has a potential to induce a wide range of mutations and generate multiple physiological, biochemical, and genetic modifications in humans. Most mutagens are having genotoxic effects on the following generation through germ cells. The influence of germinal mutations on health will be determined by their frequency, nature, and the mechanisms that keep a specific mutation in the population. Early prenatal lethal mutations have less public health consequences than genetic illnesses linked with long-term medical and social difficulties. Physical and chemical mutagens are common mutagens found in the environment. These two environmental mutagens have been associated with multiple neurological disorders and carcinogenesis in humans. Thus in this study, we aim to unravel the molecular mechanism of physical mutagens (UV rays, X-rays, gamma rays), chemical mutagens (dimethyl sulfate (DMS), bisphenol A (BPA), polycyclic aromatic hydrocarbons (PAHs), 5-chlorocytosine (5ClC)), and several heavy metals (Ar, Pb, Al, Hg, Cd, Cr) implicated in DNA damage, carcinogenesis, chromosomal abnormalities, and oxidative stress which leads to multiple disorders and impacting human health. Biological tests for mutagen detection are crucial; therefore, we also discuss several approaches (Ames test and Mutatox test) to estimate mutagenic factors in the environment. The potential risks of environmental mutagens impacting humans require a deeper basic knowledge of human genetics as well as ongoing research on humans, animals, and their tissues and fluids.

Introduction. The bacterial reverse gene mutations test (the Ames test) is widely used to assess chemicals’ mutagenic activity. The spontaneous mutation level of test strains is a mandatory characteristic that has to be monitored in a laboratory performing mutagenicity studies using the Ames test. In this regard, it is important to assess the factors affecting the spontaneous mutation level in the experiment and, therefore, on the general conclusion on the test item mutagenicity. Material and methods. A plate incorporation test version was used both in the presence and absence of a metabolic activation system. Results. We summarized the historical control data obtained in the laboratory in 2016-2020, determine the fluctuation limits in the number of revertant colonies for each strain, and identify the factors affecting the negative control variability. No significant differences were found in the spontaneous background of test strains when using DMSO or water as solvents, polypropylene or polystyrene tubes, as well as Petri dishes of different types. In the case of the TA1535, TA102 and TA100 cultures, no influence of the presence of the S9 mixture on the spontaneous reversion range was revealed (p≤0.05). Statistically significant differences in the number of spontaneous revertants (at + S9 or -S9) were found for the strains that allow detecting frameshift mutations, TA97 and TA98. It has been shown that the volume of the selective medium and the brand of gelling agent in its composition are important factors leading to the variability of the historical negative control. Conclusion. To ensure the quality of experiments according to the principles of good laboratory practice and the reliability of the data obtained using the bacterial reverse mutation method, it is necessary to standardize the operations in advance of experiments. Contribution: Egorova O.V. — the concept and design of the study, collection and processing of material, statistical analysis, writing a text; Demidova Yu.V. — collection of material; Ilyushina N.A. — processing of material, writing a text. All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version Conflict of interest. The authors declare no conflict of interest. Acknowledgment. The study had no sponsorship.

Short-term in vitro genotoxicity assays are useful tools to assess whether new and emerging tobacco products potentially have reduced toxicity. We previously demonstrated that potency ranking by benchmark dose (BMD) analysis quantitatively identifies differences among several known carcinogens and toxic chemicals representing different chemical classes found in cigarette smoke. In this study, six whole smoke solution (WSS) samples containing both the particulate and gas phases of tobacco smoke were generated from two commercial cigarette brands under different smoking-machine regimens. Sixty test cigarettes of each brand were machine-smoked according to the International Organization for Standardization (ISO) puffing protocol. In addition, either 60 or 20 test cigarettes of each brand were machine-smoked with the Canadian Intense (CI) puffing protocol. All six WSSs were evaluated in the bacterial reverse mutation (Ames) test using Salmonella typhimurium strains, in the presence or absence of S9 metabolic activation. The resulting S9-mediated mutagenic concentration–responses for the four WSSs from 60 cigarettes were then compared using BMD modelling analysis and the mutagenic potency expressed as number of revertants per μl of the WSS. The quantitative approaches resulted in a similar rank order of mutagenic potency for the Ames test in both TA98 and TA100. Under the conditions of this study, these results indicate that quantitative analysis of the Ames test data can discriminate between the mutagenic potencies of WSSs on the basis of smoking-machine regimen (ISO vs. CI), and cigarette product (differences in smoke chemistry).

Nicotiana tabacum is the most cultivated tobacco species in the state of Rio Grande do Sul, Brazil. Workers who handle the plant are exposed to the leaf components during the harvesting process and when separating and classifying the dried leaves. In addition to nicotine, after the drying process, other components may be found including tobacco-specific nitrosamines, polycyclic aromatic hydrocarbons, as well as pesticides residues. The objective of this study was to examine the genotoxicity attributed to the aqueous extract of dried tobacco leaves obtained from tobacco barns using Chinese hamster lung fibroblast cells (V79) as a model system by employing alkaline comet assay, micronucleus (MN) and Ames test. MTT assay was used to assess cytotoxicity and establish concentrations for this study. Data demonstrated cell viability > 85% for concentrations of 0.625-5 mg/ml while the comet assay indicated a significant increase in DNA damage at all concentrations tested. A significant elevation of MN and nuclear buds (NBUD) was found for 5 mg/ml compared to control and other dry tobacco leaves concentrations (0.625-2.5 mg/ml). Mutagenicity was not found using the Salmonella/Microsome test (TA98, TA100, and TA102 strains) with and without metabolic activation. The concentration of inorganic elements was determined employing the PIXE technique, and 13 inorganic elements were detected. Using CG/MS nicotine amounts present were 1.56 mg/g dry tobacco leaf powder. Due to the observed genotoxicity in V79 cells, more investigations are needed to protect the health of tobacco workers exposed daily to this complex mixture of toxic substances present in dry tobacco leaves.

Exposure to solar ultraviolet (UV) radiation is a major contributing factor to the increasing number of skin cancer cases. Interest has grown to use plant extracts as natural ingredients in cosmetic formulations due to their photoprotective effect, antioxidant and anti-inflammatory activity, as well as other biological activities. The aim of this study was to evaluate the biological activity of two South African plant extracts, Helichrysum odoratissimum (L.) Sweet. and Buddleja saligna Willd., and to successfully incorporate these extracts into sunscreen formulations (o/w emulsions) due to their reported biological activity. Ethanolic extracts were prepared from the leaves and stems of H. odoratissimum and B. saligna and evaluated for their antioxidant activity, mutagenic potential and antiproliferative activity against human dermal fibroblasts (MRHF). The extracts were further characterized using gas chromatography-mass spectrometry (GC-MS). Thereafter, the extracts were incorporated into separate sunscreen formulations to evaluate the in vivo dermal irritancy potential, in vivo sun protection factor, in vitro UVA protection, photostability and long term stability of the formulation, to confirm that by incorporating the extracts, the stability or photoprotective effect of the sunscreen formulation was not reduced and that these formulation were considered safe for topical application. Three separate sunscreen formulations were prepared; the base sunscreen formulation (formulation A), the base sunscreen formulation containing B. saligna (formulation B) and H. odoratissimum (formulation C) respectively. Both extracts showed significant radical scavenging activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay with a fifty percent inhibitory concentration (IC50) of 5.13 ± 0.07 and 8.16 ± 0.34 µg/mL for H. odoratissimum and B. saligna respectively. No mutagenic activity was observed when the extracts were tested in the Ames assay using Salmonella typhimurium (TA98 and TA100). The PrestoBlue® cell viability assay was used to determine the antiproliferative activity of the extracts against MRHF cells, both extracts showed an IC50 value >90 µg/mL. Photoprotective activity was measured using in vivo sun protection factor (SPF) test method according to South African (SANS 1557) and International (ISO 24444) standards as well as the in vitro UVA SPF testing procedure (ISO 24443). The SPF results showed that the formulations had broad-spectrum UV protection with SPF values of 15.8±0.41, 16.1±0.66 and 16.0±0.49 and UVAPF values of 6.47±0.06, 6.45±0.06 and 6.47±0.07 for formulation A, B and C respectively. Furthermore, the formulations remained stable under normal and extreme conditions and the plant extracts did not affect the photoprotective effect of the sunscreen formulations and contributed towards the formulations stability. Additionally, each of the formulations were photostable, whereas the formulations with the addition of the extracts showed an incremental increase in photostability when compared to the base formulation. Both these extracts have been previously reported to display antiproliferative activity against skin cancer cell lines (previously published data), with an IC50 value of 31.80 ± 0.35 µg/mL (human malignant melanoma, UCT-MEL-1) for B. saligna and IC50 values of 15.50 ± 0.20 (human epidermoid carcinoma, A431) and 55.50 ± 6.60 µg/mL (human malignant melanoma, A375) for H. odoratissimum, contributing towards the medicinal benefit of using these extracts as ingredients into sunscreen formulations. Therefore, Helichrysum odoratissimum and Buddleja saligna could be considered as useful and viable additives to sunscreen formulations due to their reported biological activity.

Background Conflicting results between bacterial mutagenicity tests (the Ames test) and mammalian carcinogenicity tests might be due to species differences in metabolism, genome structure, and DNA repair systems. Mutagenicity assays using human cells are thought to be an advantage as follow-up studies for positive results in Ames tests. In this collaborative study, a thymidine kinase gene mutation study (TK6 assay) using human lymphoblastoid TK6 cells, established in OECD TG490, was used to examine 10 chemicals that have conflicting results in mutagenicity studies (a positive Ames test and a negative result in rodent carcinogenicity studies). Results Two of 10 test substances were negative in the overall judgment (20% effective as a follow-up test). Three of these eight positive substances were negative after the short-term treatment and positive after the 24 h treatment, despite identical treatment conditions without S9. A toxicoproteomic analysis of TK6 cells treated with 4-nitroanthranilic acid was thus used to aid the interpretation of the test results. This analysis using differentially expressed proteins after the 24 h treatment indicated that in vitro specific oxidative stress is involved in false positive response in the TK6 assay. Conclusions The usefulness of the TK6 assay, by current methods that have not been combined with new technologies such as proteomics, was found to be limited as a follow-up test, although it still may help to reduce some false positive results (20%) in Ames tests. Thus, the combination analysis with toxicoproteomics may be useful for interpreting false positive results raised by 24 h specific reactions in the assay, resulting in the more reduction (> 20%) of false positives in Ames test.

The presented study aims to extend the knowledge of toxicological profile of rare earth elements salts (REEs). The basal toxicity test performed comprised assessment of cytotoxicity (3T3 Balb/c Neutral Red Uptake Test) that allows for calculation of LD50 (rats) on the basis of the concentration which leads to a 50% reduction in cell growth (IC50). Environmental toxicity was addressed by the Tubifex tubifex (T.t.) express test. The in vitro skin irritation (OECD TG 439) and skin corrosion tests (OECD TG 431) utilizing the 3D in vitro reconstructed human epidermal model EpiDerm (MatTek IVSL, SK) were used for assessment of skin irritation and corrosion potential hazard of REEs. Mutagenic effects were determined using the bacterial reverse mutation assay (Ames Test) on 5 Salmonella typhimurium strains with and without metabolic activation (OECD TG 471). Endocrine disruption was evaluated by means of a yeast-based assay YES/YAS (Xenometrix, CH). Skin sensitization was assessed using the LuSens assay, based on a genetically modified human keratinocyte cell line (OECD TG 442D). The tested REEs have no potential of mutagenicity or skin sensitization, exhibit very weak endocrine disruption potential and only exceptional local irritation/corrosion effects for thulium (III) chloride anhydrous, but have acute and chronic toxic effects on the aquatic environment.

Made available in DSpace on 2021-06-25T10:50:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Myrcia bella is a medicinal plant used for the treatment of diabetes, hemorrhages, and hypertension in Brazilian folk medicine. Considering that plant extracts are attractive sources of new drugs, the aim of the present study was to verify the influence of incorporating 70% hydroalcoholic of M. bella leaves in nanostructured lipid systems on the mutagenic and antifungal activities of the extract. In this work, we evaluated the antifungal potential of M. bella loaded on the microemulsion against Candida sp for minimum inhibitory concentration, using the microdilution technique. The system was composed of polyoxyethylene 20 cetyl ether and soybean phosphatidylcholine (10%), grape seed oil, cholesterol (10%: proportion 5/1), and purified water (80%). To investigate the mutagenic activity, the Ames test was used with the Salmonella Typhimurium tester strains. M. bella, either incorporated or free, showed an important antifungal effect against all tested strains. Moreover, the incorporation surprisingly inhibited the mutagenicity presented by the extract. The present study attests the antimicrobial properties of M. bella extract, contributing to the search for new natural products with biological activities and suggesting caution in its use for medicinal purposes. In addition, the results emphasize the importance of the use of nanotechnology associated with natural products as a strategy for the control of infections caused mainly by the genus Candida sp. University of Araraquara (UNIARA) Department of Biological Sciences and Health São Paulo State University (UNESP) Department of Biological Sciences Faculty of Science São Paulo State University (UNESP) Department of Biological Sciences School of Pharmaceutical Sciences, Campus Araraquara São Paulo State University (UNESP) Department of Drugs and Medicines School of Pharmaceutical Sciences, Campus Araraquara University of Brasilia Nanobiotechnology Laboratory Department of Genetics and Morphology University of Araraquara Department of Biological Sciences and Health UNIARA Carlos Gomes 1338, Rua Carlos Gomes, 1217 São Paulo State University (UNESP) Department of Biological Sciences Faculty of Science São Paulo State University (UNESP) Department of Biological Sciences School of Pharmaceutical Sciences, Campus Araraquara São Paulo State University (UNESP) Department of Drugs and Medicines School of Pharmaceutical Sciences, Campus Araraquara

This study investigated the microbial synthesis of Ag/AgCl nanoparticles, their mutagenicity, antimutagenicity and antibacterial effects. For this purpose, bacteria isolates were taken from soil samples of Zarshouran gold mine. One silver resistant bacterial strain was selected for the production of silver nanoparticles. Results of the phylogenetic analysis showed that the strain ZAR1 was 99% similar to Staphylococcus pasteuri. The synthesis of the nanoparticles was confirmed using UV–vis spectroscopy, FTIR, XRD, DLS, and SEM-EDX. The analysis of the nanoparticles synthesized with extracellular extract of bacteria showed that the nanoparticles were spherical with a size of approximately 20–85 nm. Also, the ability of TSB media for the synthesis of silver nanoparticles was evaluated. The formation of Ag/AgCl nanoparticle composites was detected cubic form in the range of 20–50 nm. Antibacterial activity of the nanoparticles was determined against sixty-six pathogenic bacteria. Our results indicated that the nanoparticles had high antimicrobial activity against pathogenic bacteria. Moreover, the nanoparticles stabilized using sodium alginate showed high antimicrobial effects. Ames test revealed that Ag NPs did not display a mutagenic effect. Moreover, the results showed that the nanoparticles were antimutagenic. The percent inhibition of mutagenicity was approximate 97–99%.

Dioscorea Rhizome is commonly used in traditional herbal medicines for the treatment of diabetes, hyperthyroidism, liver damage, neuropathy, and asthma. Here, we investigated the genotoxicity potential of D. Rhizome water extract (DRWE) using three standard battery systems in accordance with the test guidelines of the Organisation for Economic Cooperation and Development and Ministry of Food and Drug Safety as well as the principles of Good Laboratory Practice. A bacterial reverse mutation test (Ames test) was performed using the direct plate incorporation method in the presence or absence of a metabolic activation system (S9 mixture). The tester strains used included four histidine auxotrophic strains of Salmonella typhimurium, TA100, TA1535, TA98, and TA1537, along with a tryptophan auxotrophic strain of Escherichia coli, WP2 uvrA. An in vitro chromosome aberration test was performed using CHL/IU cells originally derived from the lung of a female Chinese hamster in the presence or absence of the S9 mixture. An in vivo mouse bone marrow micronucleus test was performed using male ICR mice. The micronucleus was confirmed after observation of the micro-nucleated polychromatic. The Ames test showed that DRWE did not induce gene mutations at any dose level in any of the tested strains. Additionally, DRWE did not result in any chromosomal aberrations specified in the in vitro chromosomal aberration and in vivo micronucleus tests. These results showed that DRWE exhibited neither mutagenic nor clastogenic potential in either the in vitro or in vivo test systems.

Swapping the substituents in positions 2 and 4 of the previously synthesized but yet undisclosed 5-cyano-4-(methylthio)-2-arylpyrimidin-6-ones 4, ring closure, and further optimization led to the identification of the potent antitubercular 2-thio-substituted quinazolinone 26. Structure-activity relationship (SAR) studies indicated a crucial role for both meta-nitro substituents for antitubercular activity, while the introduction of polar substituents on the quinazolinone core allowed reduction of bovine serum albumin (BSA) binding (63c, 63d). While most of the tested quinazolinones exhibited no cytotoxicity against MRC-5, the most potent compound 26 was found to be mutagenic via the Ames test. This analogue exhibited moderate inhibitory potency against Mycobacterium tuberculosis thymidylate kinase, the target of the 3-cyanopyridones that lies at the basis of the current analogues, indicating that the whole-cell antimycobacterial activity of the present S-substituted thioquinazolinones is likely due to modulation of alternative or additional targets. Diminished antimycobacterial activity was observed against mutants affected in cofactor F-420 biosynthesis (fbiC), cofactor reduction (fgd), or deazaflavin-dependent nitroreductase activity rv3547 , indicating that reductive activation of the 3,5-dinitrobenzyl analogues is key to antimycobacterial activity.

Introduction: Edible and medicinal plants contain active principles that can act as antimutagens, and hence their intake may be useful for human cancer prevention. Green leafy vegetables are important sources of carotenoids which possess antioxidant, anti-inflammatory and antimutagenic properties. Objective: To study the antimutagenic activity of oxycarotenoid-rich extracts isolated from Coriandrum sativum (coriander leaves) and Murraya koenigii (curry leaves). Methods: Oxycarotenoid-rich extracts isolated from Coriandrum sativum (coriander leaves) and Murraya koenigii (curry leaves) were investigated for antimutagenic activity in vitro by Ames test using Salmonella typhimurium strains TA 98 and TA 1535. Mutagens used were, nitro-o- phenylenediamine (NPD) (20μg/plate) and N-methyl- N-nitro-N-nitrosoguanidine (MNNG) (1μg/ plate). Results: The results revealed that oxycarotenoid-rich extracts isolated from Coriandrum sativum (coriander leaves) and Murraya koenigii (curry leaves) administered at doses of 1.0 mg, 2.5 mg and 5 mg per plate significantly inhibited mutagenicity induced by NPD and MNNG.Conclusion: These findings suggest that oxycarotenoid- rich extracts isolated from Coriandrum sativum (coriander leaves) and Murraya koenigii (curry leaves) have antimutagenic properties.

Graphical abstract, Highlights • The P2Et extract was assessed for genotoxicity and mutagenicity activity. • The P2Et extract showed no genotoxicity in Micronucleus assay. • The P2Et extract showed no mutagenicity in Ames test., The standardized P2Et extract obtained from Caesalpinia spinosa has shown antioxidant, and direct antitumor activity, but also activation of specific immune response through the induction of tumor immunogenic cell death in breast and melanoma cancer models. The present work evaluated the mutagenicity and genotoxicity profile of P2Et to continue the development of the P2Et. Genotoxicity was evaluated by OECD 1997 a guideline and mutagenicity by OECD 2016. At P2Et’s doses of 500, 1000, and 2000 mg/kg body weight in mice (Mus musculus), the difference between the number of micronuclei in PCE of the groups were not statistically significant (17 (negative control), 15 (500 mg/kg), 15 (1000 mg/kg), 19 (2000 mg/kg) and 271 (positive control). Similarly, P2Et did not induce gene mutations by base pair changes or frameshifts in the genome of Salmonella Typhimurium strains TA98, TA100, TA102, TA1535 and TA1537 at the tested range of concentrations up to 5000 μg/plate in the absence and presence of metabolic activation. Therefore, the P2Et was considered as non- mutagenic and non-genotoxic at the conditions of the tests.

Background: A series of new six thiazolyl-2-amine-based Schiff base derivatives (4a-4f) were synthesized by a sequential multistep reaction starting with Salicylaldehyde. Methods: All the Schiff base derivatives were screened in-vitro for their antibacterial activity against Mycobacterium tuberculosis (H37RV strain) ATCC No-27294. The synthesized compounds were characterized by FTIR, 1H-NMR, 13C-NMR and Mass spectrometry. Results: Among the compounds tested, 4c and 4f derivatives exhibited potent antitubercular activity against M. tuberculosis at MIC 6.25 μg/mL. Conclusion: We extended our study to explore the inhibition mechanism by conducting molecular docking analysis by using Schrodinger’s molecular modeling software. All the newly synthesized compounds were found to be in-silico AMES test non-toxic and non-carcinogens. The good Qikprop’s Absorption, Distribution, Metabolism and Excretion (ADMET) would definitely help the researchers in order to make more potent Anti-TB agents.

—The use of novel carriers for gene delivery has been rapidly growing; thus, investigation of potential phytotoxic and mutagenic action of gene delivery carriers is important for preventing their negative side effects. We found that poly-DMAEMA carriers used in a 0.0025% dose exhibited a weak cytotoxic effect towards Allium cepa plants. In a higher dose (0.025%), they slightly (by 26–55%) increased the level of catalase activity but did not affect the level of superoxide dismutase activity and malondialdehyde content in roots of A. cepa. Results of the ana-telophase test in A. cepa demonstrated no genotoxic activity of the polymeric carriers used in a 0.0025% concentration and the higher dose (0.025%). Slight genotoxic activity was detected only for BGP24 and BGP26, PEG-containing poly-DMAEMA carriers, used in 0.025% concentration. The Ames test (–S9 and +S9) revealed no mutagenic potential of the DMAEMA-based polymers. Thus, low phytotoxicity and no mutagenicity of novel polymeric carriers suggest their potential as promising nanocarriers for gene delivery into plant cells.

14-O-[(4,6-Diaminopyrimidine-2-yl)thioacetyl] mutilin (DPTM) is a promising drug candidate with excellent antibacterial activity against Gram-positive bacteria. The present study was designed to characterize its Cytochrome P450 (CYP) enzymes inhibition activities and the genotoxicity with the standard Ames test. We determined the inhibitory effects of DPTM on CYP1A2, CYP2D1/6, CYP2E1, CYP2C11/9 and CYP3A/4 in rat liver microsomes (RLMs) and in human liver microsomes (HLMs). The mRNA expressions of the above CYP isoforms and their transcriptional regulators were also evaluated using the Hep G2 cell model. The results showed DPTM exhibited a moderate inhibitory potential against CYP3A/4 (IC50 values of 10 ± 2 and 8 ± 2 μM, respectively) and weak against the other CYP enzymes based on their IC50 values. Compared to the control, CYP isoforms and their transcriptional regulators mRNA expressions significantly increased when the Hep G2 cells were treated with DPTM for a certain period of time. In the Ames test, Salmonella strains TA97, TA98, TA100, TA102 and TA1535 were treated with or without the metabolic activation (S9). Analysis showed the average number of revertant colonies per plate was less in double in the groups treated with DPTM than that in the negative control plate and showed no dose-related increase.

Background Taraxacum species (commonly known as dandelion) used as herbal medicine have been reported to exhibit an antiproliferative effect on hepatoma cells and antitumor activity in non-small-cell lung cancer cells. Although several investigations have demonstrated the safety of Taraxacum officinale, the safety of tissue-cultured plants of T. formosanum has not been assessed so far. Therefore, the present study examines the safety of the water extract of the entire plant of tissue cultured T. formosanum based on acute and subacute toxicity tests in rats, as well as the Ames tests. Results No death or toxicity symptoms were observed in the acute and subacute tests. The results of the acute test revealed that the LD50 (50% of lethal dose) value of the T. formosanum water extract for rats exceeded 5 g/kg bw. No abnormal changes in the body weight, weekly food consumption, organ weight, or hematological, biochemical, and morphological parameters were observed in the subacute toxicity test. Thus, the no observed adverse effect level (NOAEL) of T. formosanum water extract was estimated to be higher than 2.0 g/kg. Finally, the results of the Ames test revealed that T. formosanum water extract was not genotoxic at any tested concentration to any of five Salmonella strains. Conclusions The water extract of tissue-cultured T. formosanum was non-toxic to rats in acute and subacute tests and exhibited no genotoxicity to five Salmonella strains. How to cite: Tsai WC, Chang HC, Tseng YH, et al. Toxicity evaluation of water extract of tissue-cultured Taraxacum formosanum by acute, subacute administration, and Ames test. Electron J Biotechnol 2020;45. https://doi.org/10.1016/j.ejbt.2020.04.001

Abstract Indole alkaloids extract (IAAS) was prepared from leaves of Alstonia scholaris (L.) R. Br., an evergreen tropical plant widely distributed throughout the world. This plant has been used historically by the Dai ethnic people of China to treat respiratory diseases. This study evaluated the genotoxicity and safety pharmacology of IAAS to support clinical use. The bacterial reverse mutation (Ames) test, in vitro mammalian chromosomal aberration test, and in vivo mammalian erythrocyte micronucleus (MN) test were performed to evaluate genotoxicity. Mice were administered IAAS (240, 480, or 960 mg/kg bw) once orally to observe adverse central nervous system effects. Furthermore, beagle dogs were administered IAAS (10, 30, 60 mg/kg bw) once via the duodenum to evaluate its effects on the cardiovascular and respiratory systems. IAAS with or without S9-induced metabolic activation showed no genotoxicity in the Ames test up to 500 μg/plate, in the mammalian chromosomal aberration test up to 710 μg/mL, or in the MN test up to 800 mg/kg bw. No abnormal neurobehavioral effects were observed in mice following treatment with up to 960 mg/kg bw of IAAS. Moreover, blood pressure, heart rate, electrocardiogram parameters, and depth and rate of breathing in anesthetized beagle dogs did not differ among the IAAS doses or from the vehicle group. These data indicated that IAAS did not induce mutagenicity, clastogenicity, or genotoxicity, and no pharmaco-toxicological effects were observed in the respiratory, cardiovascular, or central nervous systems. Our results increased understanding of safety considerations associated with IAAS, and may indicate that IAAS is a possible drug candidate. Graphic Abstract

The current study aims to assess the aqueous extract of Pinus wallichiana stem for the synthesis of small spherical-shaped (10–30 nm) silver nanoparticles (AgNPs) and their in-vitro and in-vivo biomedical applications. The biosynthesized AgNPs were nonmutagenic and safe at all test doses as per Ames and acute toxicity assay (20, 40, 60, and 80 mg/kg). The percent writhing inhibitory effect generated by AgNPs was 42.51, 50.84, and 59.06 at test doses of 10, 20, and 30 mg/kg, respectively. The percent decreased in gastrointestinal tract motility observed was 41.34%, 32.69%, and 28.48% at 10, 20, and 30 mg/kg, respectively. They also showed a significant antipyretic effect after 1, 2, and 3 h in comparison to normal saline. The AgNPs of P. wallichiana showed good antibacterial activity against Acinetobacter baumannii (60% with MIC50 = 2.36 mg/ml and MBC = 5.0 mg/ml). These nanoparticles also possessed good antioxidant activity of 61.77 ± 0.828% and 70.25 ± 0.56% at 400 and 500 µg/ml, respectively and lack phytoagglutinin potential. Because of their high potency as biomedical agents, these nanoparticles can be a good alternative to the currently available drugs and approaches.

Probiotic Escherichia coli strain Nissle 1917 (EcN) has a long history of safe use. However, the recently discovered presence of a pks locus in its genome presumably producing colibactin has questioned its safety, as colibactin has been implicated in genotoxicity. Here, we assess the genotoxic potential of EcN. Metabolic products were tested in vitro by the Ames test, a mutagenicity assay developed to detect point mutation-inducing activity. Live EcN were tested by an adapted Ames test. Neither the standard nor the adapted Ames test resulted in increased numbers of revertant colonies, indicating that EcN metabolites or viable cells lacked mutagenic activity. The in vivo Mammalian Alkaline Comet Assay (the gold standard for detecting DNA-strand breaks) was used to determine potentially induced DNA-strand breaks in cells of the gastro-intestinal tract of rats orally administered with viable EcN. Bacteria were given at 109–1011 colony forming units (CFU) per animal by oral gavage on 2 consecutive days and daily for a period of 28 days to 5 rats per group. No significant differences compared to negative controls were found. These results demonstrate that EcN does not induce DNA-strand breaks and does not have any detectable genotoxic potential in the test animals.

The various species that comprise the genus Glycyrrhiza (Licorice) have long been used as oriental herbal medicines in Asian countries. Wongam (WG), which is a new variety of Glycyrrhiza, was developed in Korea to overcome the limitations of low productivity, environmental restrictions, and an insufficient presence of glycyrrhizic acid and liquiritigenin. In this study, we evaluated WG extract’s genotoxicity through an in vitro bacterial reverse mutation (AMES) test, an in vitro chromosome aberration test, and an in vivo mouse bone marrow micronucleus test. In the AMES test, WG extract at concentrations of up to 5000 µg/plate showed no genotoxicity regardless of S9 mix. No chromosome aberrations appeared after 6 h in 1400 µg/mL WG extract regardless of S9 mix or in 1100 µg/mL WG extract after 24 h without S9 mix. Nor was there a significant increase in the number of micronucleated polychromatic erythrocytes to total erythrocytes up to 5000 mg/kg/day for 2 days detected in the micronucleus test. These results confirm that WG extract is safe for use as an herbal medicine, as it precipitates no detectable genotoxic effects.

Thailand is located in the tropics and a wide variety of fruits are grown commercially. However, studies regarding the phytonutrients, anti-mutagenic and chemopreventive effects of these fruits are limited. Thus, phytochemical profiles and inhibition of key enzymes involved in obesity and diabetes, together with anti-mutagenic and chemopreventive properties of eight tropical fruit extracts cultivated in Thailand, including Psidium guajava ‘Kimju’, Psidium guajava ‘Keenok’, Ananas comosus ‘Pattavia’, Ananas comosus ‘Phulae’, Durio zibethinus ‘Chanee’, Durio zibethinus ‘Monthong’, Carica papaya ‘Khaekdum’ and Mangifera indica ‘Namdokmai’ were investigated. Different cultivars were also compared. Results showed that M. indica ‘Namdokmai’ was the most antioxidant-rich extract containing abundant 4-hydroxybenzoic acid and its derivative, gallic acid, as the main phenolics. M. indica ‘Namdokmai’ also exhibited high inhibitory capacities (&gt, 60% inhibition under studied conditions) against lipase, α-amylase and α-glucosidase, key enzymes as drug targets for controlling obesity and type 2 diabetes. Interestingly, all fruit extracts suppressed food mutagen-induced DNA mutations assayed by the Ames test, especially M. indica ‘Namdokmai’ and C. papaya ‘Khaekdum’ (&gt, 50% inhibition at 200 µg/plate). The M. indica ‘Namdokmai’ was also the most potent extract for suppression of cancer promotion (&gt, 90% inhibition at 200 µg/mL) followed by P. guajava ‘Kimju’, P. guajava ‘Keenok’ and C. papaya ‘Khaekdum’. Results potentially indicated that fruit intake after overcooked meat consumption might supplement nutrients and fiber and also reduce DNA mutation sources.

In this research, mutagenic properties of blueberry polyphenol extract were studied in gene mutation induction test (Ames test) on four strains of Salmonella typhimurium TA98, TA100, TA1535, TA1537. None of the strains of Salmonella typhimurium showed statistically reliable dose-dependent increase in number of revertant colonies in the presence of investigated drug in the studied dose range from 4,0 to 40,0 mg/ml relative to baseline of spontaneous mutations. The blueberry extract does not have any mutagenic activity in the researched dose range in relation to TA98, TA100, TA1535, TA1537 strains of Salmonella typhimurium.

Preparation and comprehensive characterization of three Schiff base ligands; with trimethoxy substitution (1E,1'E)-N,N'-(naphthalene-1,5-diyl)bis(1-(3,4,5-trimethoxyphenyl)methanimine), 1, with ortho-hydroxy substitution 6,6'-((1E,1'E)-(naphthalene-1,5-diylbis(azaneylylidene))bis(methaneylyl- idene))bis(2-methoxyphenol), 2 and 3,4-bis(((E)-2-hydroxy-3-methoxybenzylidene)amino)benzoic- acid, 3 and their Ni(II), Cu(II), Co(II), Zn(II), Fe(II), Mn(II) complexes have been reported. Their spectral properties were studied in solution and solid-state by a combination of different analytical techniques; FT-IR spectroscopy, 1H-NMR and 13C-NMR spectroscopy, elemental analysis and thermal analysis. Diamagnetic and paramagnetic natures of the complexes were also determined by magnetic susceptibility measurements in solid-state. Promising photophysical properties were observed as; Amax. were recorded at 226 nm for 2; at 795 nm for 2-Ni, at 782 nm for 2-Cu, at 784 nm for 2-Co, at 702 nm for 2-Zn, at 784 nm for 2-Fe, at 702 nm for 2-Mn and at 289 nm for 3, at 786 nm for 3-Ni, at 797 nm for 3-Cu, at 746 nm for 3-Co, at 794 nm for 3-Zn, at 699 nm for 3-Fe, at 781 nm for 3-Mn ; and Imax were also recorded at; 380, 490, 725 nm for 2 and 2-Metal; 375 nm, 510 nm, 725 nm for 3 and 3-Metal when excitated at 220 nm. Antibacterial activities against different microorganisms; Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC 70603, Staphylococcus aureus ATCC 43300 (MRSA), Salmonella enteritidis ATTC 13076, Sarcina lutea ATCC 9341, Bacillus cereus ATTC 11778, and antifungal activities against Candida albicans NRRL Y-417 of the compounds 1, 2, 3, 2-Cu, 2-Fe, 3-Zn, 3-Fe were determined. Mutagenic properties of the compounds were also studied and according to the results 2-Cu and 3 have been found non-mutagenic in Ames test but also they have strong antimicrobial potential against pathogen microorganisms. For 2-Cu MIC values were ranging between 0.39-0.024 mg/ml and the lowest minimum inhibitory concentration (0.024 mg/ml) was determined against E. coli. The 3 numbered compound revealed strong antimicrobial activity at doses of ranging between 0.39-0.097 mg/ml and E. coli was the most sensitive bacterium against this chemical.

Rheum emodi Wall. Ex. Meissen is an important food and medicinal plant growing wild in western Himalayas. The current endeavor was undertaken to carry out bioassay guided fractionation and isolation of antimutagenic principle(s) present in the rhizomes of Rheum emodi. The antimutagenicity assay was performed to check the recuperating effect of 80% aqueous methanol extract of rhizomes of R. emodi and its fractions i.e. n-hexane, chloroform, ethyl acetate, n-butanol and water fractions, against 4-nitro-o-phenylenediamine (NPD), sodium azide, and 2AF-induced mutagenicity in TA98 and TA100 strains using the Ames Salmonella his+ reversion assay. The crude 80% aqueous methanol extract and ethyl acetate fraction exhibited strong antimutagenicity. The antimutagenic potential of the extracts and fractions was markedly enhanced when indirect acting mutagen was used in both the tester strains of Salmonella typhimurium. In an effort to identify the molecules responsible for the antimutagenic effect displayed by R. emodi, four anthraquinones i.e. aloe-emodin, chrysophanol, emodin and rhein detected in the ethyl acetate fraction by HPLC analysis were also tested for their antimutagenic potential in both the tester strains. It was found that the anthraquinones in their pure form showed lower antimutagenicity than the mother fraction from which they were isolated. The results point towards a synergistic effect of the principles present in the mother fraction.

A comparative analysis was performed of the impact on living organisms, in terms of genotoxicity, of two factors, namely, radiation (residual 137Cs) and a chemical factor (pesticides) . The experiment was conducted both on individual pesticide samples and the aqueous extracts of soil samples. Soil samples were collected in the territory of the Orel region (Russia) from May 12 to May 15, 2010, i.e., years after the Chernobyl accident. Changes in the mutagenic index derived from Ames assay (Salmonella/microsomes) and the data obtained from mammalian cell culture in the Cricetulus griseus test (Chinese hamster) of the soil samples arising from the cesium radioactivity and the content of pesticides in these samples were considered. No direct correlation was found between dose level and genotoxicity at low radiation doses (20 μR/h), i.e., correlation with DNA mutations produced, both in terms of frameshifting and the base replacement type. On the contrary, a clear tendency was observed for increase in genotoxicity with increase in the concentration of pesticides and the entire amount of anthropogenic toxicants in the studied soil samples.

To solve the problem of utilizing massive harmful algal blooms (HABs) biomass, we developed a technique involving physical purification methods, namely, microfiltration (2 μm, 0.45 μm), ultrafiltration (100 kDa cutoff), and low-temperature precipitation (4 °C, 48 h), as the core methodology and used the toxic Microcystis biomass in Dianchi Lake for batch preparations of microcystins (MCs) and phycocyanin (PC). The results were as follows: (1) From 1.0 kg of Dianchi Lake cyanobacteria, 57 g of PC powder with a purity (A620/A280) of 1.78 was prepared. An acute oral toxicity test in mice showed that the LD50 of the prepared PC was >5.25 g kg−1, practically non-toxic. The LD50 of PC administered by intraperitoneal injection was >4.71 g kg−1. The Ames test showed that the mutagenic effect was negative independent of the addition of S9. The overall results of the toxicity tests suggested that the prepared PC was not potentially toxic. (2) From 666.7 g of cyanobacteria, 2.262 g of MC extract powder (containing 192.7 mg of MC-RR and 54.3 mg of MC-LR) was prepared. MC-RR and MC-LR accounted for 8.52% and 2.40%, respectively, and the extraction ratios for MC-RR and MC-LR were 40.1% and 83.1%, respectively. A further purification by preparative HPLC was carried out, obtaining 2.338 mg of pure MC-RR (chromatographic purity >85%) from 70 mg of MC extract powder, which was used as an HPLC chromatographic standard and in routine toxicology experiments. The efficiency of the ultrafiltration treatment of cyanobacterial powder reached 49.3 kg d−1, and the cost of producing food-grade PC powder in this study was 17.6 US$ g−1, which is only 13.6% of the current price of food-grade PC on the international market. Thus, the cost advantage was significant. Therefore, this study provides an approach for the safe disposal of HABs and the large-scale commercial utilization of HAB biomass.

The high diversity of species in the marine environment gives rise to compounds with unique structural patterns not found as natural products in other systems and with great potential for pharmacological, cosmetic and nutritional use. The genus Tubastraea (Class Anthozoa, Order Scleractinia, Family Dendrophylliidae) is characterized as a hard coral without the presence of zooxanthellae. In species of this genus alkaloids derived from the compound aplysinopsin with pharmacological activity are known. In Brazil T. coccinea and T. tagusensis are characterized as non-indigenous and invasive and are currently found along the Brazilian coast, from Santa Catarina to Bahia states. This study aims to analyze the mutagenic, cytotoxic and genotoxic potential of methanolic and ethanolic extracts from T. coccinea and T. tagusensis collected in Ilha Grande Bay, Rio de Janeiro state, Brazil. Bacterial reverse mutation assay on the standard strains TA97, TA98, TA100, TA102 and TA104, in vitro micronucleus formation test and colorimetric assays for cytotoxic signals on the cell lines HepG2 and RAW264.7 were used. We also synthesized an oxoaplysinopsin derivate alkaloid (APL01) for comparative purposes. No mutagenic (250; 312.5; 375; 437.5 and 500 μg/plate) or genotoxic (0.05; 0.5; 5.0; 50 and 500 μg/mL) effects were observed in any sample tested for all measured concentrations. Cytotoxic responses were observed for eukaryotic cells in all tested samples at 500 and 5000 μg/mL concentrations. Cytotoxicity found in the WST-1 assay was independent of the metabolism of substances present in samples compositions. The cytotoxicity observed in the LDH release assay depended on metabolism.

—Limonium plants have commonly used as antibacterial, styptical, anti-swelling agent, digestive system regulator, anticarcinogenic, and antiviral in alternative medicine. Beside this, different extracts of Limonium plants have mutagenic, cytotoxic and genotoxic effects. Different parts of Limonium effusum (Boiss.) Kuntze ethanol extracts were used to determine the mutagenic and cytotoxic effects. Ames test was carried out with Salmonella typhimurium TA98, and TA100 strains to determine the mutagenicity. Ames test results showed that root 1000 μg/plate of plant has mutagenic effect and root extracts have weak mutagenicity with TA98 strain. MTT test was carried out with MDBK (Madin-Darby Bovine Kidney) cells to determine the cytotoxicity. Cytotoxicity results showed that root 50, 25, 12.5 and 6.25 μg/mL extracts were increased the proliferation rates in 24 h treatment. All doses of root extracts decreased the rates in 96 h. All concentrations of stem decreased the proliferation rates in 96 h. Leaf extracts increased the rates in 24 h but decreased in 48, 72, and 96 h.

Nar (Punica granatum L.) günlük diyet içerisinde sıklıkla tüketilen bir meyve olup, ayrıca nar ekşisi formunda da yemek ve salatalarda çeşni olarak sıklıkla kullanılmaktadır. Bu çalışmada ise Hatay ve Şanlıurfa yörelerinde geleneksel olarak üretilen iki farklı nar ekşisi örneğinin potansiyel antibakteriyel ve antimutajenik etkisinin tespiti amaçlanmıştır. Antibakteriyel aktivite analizi agar kuyu difüzyon metoduna göre yapılmış olup, test bakterisi olarak Bacillus subtilis subsp. spizizenii ATCC 6633, Staphylococcus aureus ATCC 6538, Bacillus licheniformis ATCC 14580 ve Enterobacter aerogenes ATCC 13048 kullanılmıştır. Antimutajen etki analizlerinde ise Ames Testi kullanılmış ve Salmonella Typhimurium TA 98 ve TA 100 suşları tercih edilmiştir. Analiz sonuçlarına göre Hatay nar ekşisinin test bakterilerine karşı 18.09-25.63 mm inhibisyon zonu meydana getirdiği tespit edilirken, Şanlıurfa nar ekşisinde bu değer 14.84-22.32 mm olarak gözlemlenmiştir. Ayrıca; Ames testi sonucunda TA 98’e karşı Hatay nar ekşisinin 10 μg plak-1 ve 20 μg plak-1 dozlarında, Şanlıurfa nar ekşisinin ise denenen tüm dozlarında antimutajenik etki tespit edilirken; TA 100’e karşı ise Hatay ve Şanlıurfa nar ekşisinin denenen hiçbir dozunun antimutajenik özellik göstermediği belirlenmiştir. Sonuç olarak, nar ekşisinin sahip olduğu antibakteriyel ve antimutajenik etkilerden dolayı sağlık için faydalı olduğu düşünülmekle beraber; bu sonuçların farklı bakterilerle ve nar ekşileriyle yapılan çalışmalarla desteklenmesi faydalı olacaktır.

As di(2-ethylhexyl) phthalate (DEHP), one of phthalates, is classified as probable human carcinogens in EPA, acetyltriethyl citrate(ATEC), one of aliphatic esters, could be applied to DEHP substitute. ATEC is used as plasticizers in cosmetics and nail products. Here, we studied whether ATEC might have genotoxic potential and induce glucose tolerance as compared to DEHP. Genotoxicity was determined by Ames test with histidine-requiring Salmonella typhimurium (TA98, TA100, TA1535 and TA1537) and tryptophan-requiring Escherichia coli (WP2uvrA(pKM101)) strains, chromosomal aberration assay with Chinese hamster lung(CHL/IU) cells, and micronucleus test with bone marrow cells of CD-1 mice. The number of revertants was not significantly changed in Ames test. The frequency of cells with chromosome aberrations was less than 5% in ATEC- or DEHP-treated cells for 6 or 24 h. In addition, no statistically significant increase was observed for the incidence of micronucleated polychromatic erythrocytes (MNPCE) in polychromatic erythrocytes (PCE) and for the ratio of PCE among total erythrocytes at 24 or 48 h after the treatment of mice with ATEC or DEHP. In the meanwhile, blood glucose level (BGL) was increased by the treatment of mice with DEHP or ATEC for 5 consecutive days. Additional 7 days later, BGL by DEHP was recovered to normal level, but not that by ATEC. Then, taken together, our results suggest that ATEC could disrupt glucose metabolism under our experimental conditions. Therefore, although DEHP and ATEC may not be genotoxic, our data should be helpful for persons with the problem in glucose metabolism to choose products containing DEHP or ATEC.

En los últimos quince años se ha registrado un alto grado de deterioro en el río Balsas, México, debido a los asentamientos humanos e industriales. El objetivo de este estudio fue evaluar la condición actual del río y establecer la relación entre los parámetros de la legislación vigente y las pruebas de toxicidad y mutagenicidad. Se determinaron parámetros de campo (color, olor, temperatura, CE, pH, OD), fisicoquímicos (SST, SDT, SSe, DBO5, DQO y GyA, Ntotal, Norg, Ptotal, Porg, ortofosfatos), metales pesados (As, Cd, Cu, Cr, Cr+6, Hg, Ni, Pb, Zn, Al, Fe, Mn), microbiológicos (coliformes fecales) con base en los límites máximos permisibles (LMP) en la NOM-001-ECOL-1996 y en los CE-CCA-001/89, análisis de toxicidad (Vibrio fischeri, Daphnia magna) y mutagenicidad (prueba de Ames con microsomas/Salmonella typhimurium), en época de sequía en nueve estaciones a lo largo de 85km. Los valores para OD, DBO5 y DQO fueron de 2-6, 11-270 y 22-1841mg·l-1, respectivamente. Los metales fueron detectados por debajo de los LMP en todas las estaciones, excepto en la estación 5 (0,002mg·l-1 de Hg) y la 8 (0,13mg·l-1 de Cr+6). Los coliformes fecales rebasaron los LMP en todas las estaciones. Los valores para Vibrio fischeri, Daphnia magna se encuentran en 2-28 y 1-4UT, y para la prueba de Ames entre 5 y 63RM, indicando toxicidad y mutagenicidad desde la estación 3 a la 9. Se registró contaminación alta según la legislación vigente y un riesgo para la salud pública de acuerdo a los parámetros toxicológicos y mutagénicos.

Peltogyne mexicana heartwood might be a novel purple pigment source. The results of the present study demonstrate that the purple pigment is an important source of phenolic compounds (698.22 ± 2.99 mg GAE/g) and flavonoids (48.01 ± 0.51 mg EPE/g). UV–Vis spectrum and color parameters (L* a* b*) showed that purple pigment has different shades of purple–red (H° value 19.32 ± 0.02 in methanol and 22.85 ± 0.01 in ethanol) depending on the solvent and the pH. Also, the purple pigment did not exhibit acute oral toxicity at a single dose (2000 mg/kg body weight). No mutagenicity was observed in the Ames test with three Salmonella typhimurium strains. The purple pigment exhibited considerable coloring properties with a wider range of citric acid-dependent color hues in gelatin (H° from 280.3 to 319.9 and from 68.0 to 88.1), and higher color intensity than commercial anthocyanin. Minor variations in the hue were found in yogurt, for purple pigment with H° values from 317.5 to 315.0, and commercial anthocyanin from 82.6 to 88.7 and 276.9 to 295.5. However, purple pigment required lower concentrations to achieve superior effects. For gelatin and yogurt samples, similar variations in the color parameters L*, a*, b*, and pigment degradation were observed for purple pigment and commercial anthocyanin in the stability assay.

From the methanolic extract of the leaves of Lansium domesticum, three new onoceranoid-type triterpenoids, lansium acids X-XII and a new cycloartane-type triterpene, lansium acid XIII, were isolated. The chemical structures of the isolated new compounds were elucidated on the basis of chemical/physicochemical evidence. For new onoceranoid-type triterpenoids, the absolute configurations were established by comparison of experimental and predicted electronic circular dichroism (ECD) data. The isolated onoceranoid-type triterpenoids showed antimutagenic effects in the Ames assay against 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1).

PubMed ID: 31049867 Nanoparticles are very effective compounds to transform and detoxicate common environmental contaminants. For this reason, crude urban liquid wastewater sludges were treated by silver nanoparticles (Ag-NPs, 100 nm) for 24 h. Both Ag-NPs’ treated and untreated sludges were examined for the evaluation if there are possible mutagenic/anti-mutagenic, cytotoxic, and genotoxic/anti-genotoxic effects by Ames and Allium cepa tests. The results were then subjected to statistical analyses by using SPSS software and p < 0.05 was accepted as a significant value. The data obtained from the Ames test showed that while untreated crude liquid sludge had a significant mutagenic effect, Ag-NP-treated one decreased its mutagenicity. Similar effects were also observed in the chromosome aberration-Allium cepa tests. Significant chromosome aberrations observed were C-metaphase, sticky metaphase, sticky anaphase, anaphase bridge, vagrant chromosome, and multipolar anaphases. Both tests demonstrated that silver nanoparticle treatment decreased the major mutagenicity and genotoxicity detected in the liquid wastewater sludges. © 2019, Springer-Verlag GmbH Germany, part of Springer Nature.

Ethnopharmacological relevance Traditionally, Verbena officinalis L. has been used for reproductive and gynaecological purposes. However, the mutagenicity and genotoxicity of V. officinalis have not been extensively investigated. Aim of the study To assess the in vitro mutagenicity and in vivo genotoxicity of aqueous extract of V. officinalis leaves using a modified Ames test and rat bone marrow micronucleus assay according to OECD guidelines. Materials and methods In vitro Ames test was carried out using different strains of Salmonella (TA97a, TA98, TA100, and TA1535) and Escherichia coli WP2 uvrA (pKM101) in the presence or absence of metabolic activation (S9 mixture). For micronucleus experiment, male and female Sprague-Dawley rats (n = 6/group) were received a single oral daily dose of 500, 1000, and 2000 mg/kg of V. officinalis extract for three days. Negative and positive control rats were received distilled water or a single intraperitoneal injection of 50 mg/kg of cyclophosphamide, respectively. Following dissection, femurs were collected and bone marrow cells were stained with May-Grunwald-Giemsa solution for micronucleus assessment. Results Ames test results demonstrated that 5, 2.5, 1.25 and 0.625 mg/ml of V. officinalis extract induced a significant mutagenic effect against TA100 and TA98 strains (with and without metabolic activation). Findings of the animal study showed there were no significant increase in the micronucleated polychromatic erythrocytes (MNPE) and no significant alterations in the polychromatic erythrocytes (PCE) to normochromatic erythrocytes (NCE) ratio of treated rats as compared with their negative control. Meanwhile, significantly increased in the MNPEs was seen in the cyclophosphamide-treated group only. Conclusion Aqueous extract of V. officinalis has mutagenic effect against TA98 and TA100 strains as demonstrated by Ames test, however, there is no in vivo clastogenic and myelotoxic effect on bone marrow micronucleus of rats indicating that the benefits of using V. officinalis in traditional practice should outweigh risks.

The bacterial strains and mutagenicity test procedure developed by Bruce Ames, and published in 1973, greatly enhanced the ability of laboratories to test chemicals for mutagenicity. The test that became known as the "Ames Test" was simple to perform, took only two days, was relatively inexpensive, and was easily transferrable to other laboratories. Their demonstration that the test was effective at identifying potentially carcinogenic chemicals led its immediate adoption, and requirement, by regulatory authorities world-wide. Despite the development of other microbial and mammalian cell tests to measure mutation or other genetic damage, the Ames test still retains a primary role in the testing of chemicals for commercial use.

UV-C was investigated to inactivate microorganisms in grape must. The aim was to evaluate the effect of UV-C on wine-related yeasts under winery conditions, to investigate the toxicological potential and to analyze the effects on chemical and sensory wine properties. Metschnikowia pulcherrima showed the highest UV-C tolerance, Saccharomyces cerevisiae the lowest. With an initial cell count of 106 cfu/mL, M. pulcherrima required a UV-C dose of >1.2 kJ/L, S. cerevisiae Industrial relevance UV-C treatment is a non-thermal processing technique for the preservation of food. In this study, the applicability of UV-C treatment for the inactivation of spoilage yeast in grape must was demonstrated in pilot scale. The treatment provided sufficient microbial inactivation without having toxicological impact as examined by the Ames test and without causing off-flavor formation in the wines using relevant doses.

5-Hydroxy-L-tryptophan (5-HTP) is a serotonin pathway metabolite of L-tryptophan in the brain. In the knowledge that the biological properties of some compounds can be modified upon metal complexation, a new solid metal complex, [Cu(5-hydroxytryptophan) 2 ].H 2 O (Cu5HTP), has been synthesized and characterized to analyze the modification of some biological properties. The conformational investigations (optimized in gas phase at B3LYP/6-311G** theory level) suggest the coexistence of two conformers of Cu5HTP with cis- and trans- arrangements of the amino acids in the equatorial plane. The trans- Cu5HTP1 complex is the most stable conformer. The complexation led to an enhancement of the antioxidant properties of the ligand. The metal complex also improved the anticancer behavior of the ligand (tested in cancer cell lines derived from human lung (A549), cervix (HeLa) and colon (HCT-116)). It did not show toxicity against either the non-malignant human lung fibroblast (MRC-5) cell line or Artemia salina and did not behave as mutagenic agent (Ames test). Cellular reactive oxygen species production may be one of the possible mechanisms of action. Besides, the metal complex exerted neuroprotective action on cortical neurons from embryonic 18 days rats exposed to glutamate. Fil: Martínez Medina, Juan José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Chaco Austral; Argentina Fil: Naso, Luciana Gissella. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Química Inorgánica "Dr. Pedro J. Aymonino". Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Química Inorgánica "Dr. Pedro J. Aymonino"; Argentina Fil: Pérez, Ana Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rizzi, Alberto Claudio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Okulik, Nora Beatriz. Universidad Nacional del Chaco Austral; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Valcarcel, María. Innoprot Sl; España Fil: Salado, Clarisa. Innoprot Sl; España Fil: Ferrer, Evelina Gloria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Química Inorgánica "Dr. Pedro J. Aymonino". Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Química Inorgánica "Dr. Pedro J. Aymonino"; Argentina Fil: Williams, Patricia Ana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Química Inorgánica "Dr. Pedro J. Aymonino". Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Química Inorgánica "Dr. Pedro J. Aymonino"; Argentina

The objective of this study was to explore the mutagenic and cytotoxic effects of Napoleon 4EC pesticide used in Turkey to control insect pest by using two standard assays. The Allium cepa test was used for determined the cytotoxic effects of this pesticide. For this test, onion seeds were exposed to Napoleon 4EC (100, 200, and 400 ppm) for 24, 48, and 72 hours. For each test group root tip cells were stained with Feulgen and five slides were prepared for each concentration and counted microscopically. The concentrations Napoleon 4EC was compared with the value for the negative control using Dunnet-t test, 2 sided. The results indicated that mitotic index was clearly decreased with increasing the concentration of Napoleon 4EC in each treatment group as compared to the controls. The percentage of mitotic phases has been markedly impacted. Five different doses of the pesticide (50, 100, 200, 400, 800 μg/plate) were examined with Ames test using Salmonella typhimurium strains TA98 and TA100 with and without S9 metabolic activation for mutagenic activity. Ames test results showed a dose dependent effect, but not twice the negative control for S. typhimurium TA98 and TA100, with or without S9 mix except 800 μg/plate doses. In 800 μg/plate doses, colony numbers are two-fold increase according to colony number of control group. So, this places the this compound as a weak mutagen according to the parameters.

Kirkland et al. [Mutation Research/Genetic Toxicology and Environmental Mutagenesis 847 (2019) 403035, https://doi.org/10.1016/j.mrgentox.2019.03.008 ; Mutation Research/Genetic Toxicology and Environmental Mutagenesis 839 (2019): 21–35, https://doi.org/10.1016/j.mrgentox.2019.01.007 ] made recommendations on the use of the in vivo comet and transgenic rodent (TGR) gene mutation assays to screen for in vivo mutagenicity. Although it is not directly stated in either of these publications, we are concerned that the reports could potentially be used to support assertions that it is equally acceptable to follow up a positive bacterial reverse mutation (Ames) finding for an investigational drug with either the in vivo TGR mutation assay or an in vivo comet assay. For regulatory genotoxicity assessment, the in vivo follow-up for an in vitro bacterial mutation-positive drug, drug-related metabolite, or impurity should be based upon evaluating a similar endpoint (i.e., mutagenicity) as the intent is to determine if the findings of in vitro gene mutation correlate with findings of in vivo gene mutation (i.e., biologically relevant to the in vitro results). Thus, the most scientifically appropriate in vivo assays would be the TGR mutation assay or, in some circumstances, the in vivo Pig-a assay. An in vivo rodent comet assay or combination of the in vivo micronucleus and in vivo rodent comet assays would generally not be an appropriate follow-up test.

Ficus deltoidea var. deltoidea is used as traditional medicine for diabetes, inflammation, and nociception. However, the antimutagenic potential and cytoprotective effects of this plant remain unknown. In this study, the mutagenic and antimutagenic activities of F. deltoidea aqueous extract (FDD) on both Salmonella typhimurium TA 98 and TA 100 strains were assessed using Salmonella mutagenicity assay (Ames test). Then, the cytoprotective potential of FDD on menadione-induced oxidative stress was determined in a V79 mouse lung fibroblast cell line. The ferric-reducing antioxidant power (FRAP) assay was conducted to evaluate FDD antioxidant capacity. Results showed that FDD (up to 50 mg/mL) did not exhibit a mutagenic effect on either TA 98 or TA 100 strains. Notably, FDD decreased the revertant colony count induced by 2-aminoanthracene in both strains in the presence of metabolic activation (p &lt, 0.05). Additionally, pretreatment of FDD (50 and 100 µg/mL) demonstrated remarkable protection against menadione-induced oxidative stress in V79 cells significantly by decreasing superoxide anion level (p &lt, 0.05). FDD at all concentrations tested (12.5–100 µg/mL) exhibited antioxidant power, suggesting the cytoprotective effect of FDD could be partly attributed to its antioxidant properties. This report highlights that F. deltoidea may provide a chemopreventive effect on mutagenic and oxidative stress inducers.

Made available in DSpace on 2021-06-26T04:30:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-17 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Copaifera langsdorffii Desf. is a plant found in South America, especially in Brazil. Oleoresin and the leaves of this plant is used as a popular medicinal agent. However, few studies on the chemical composition of aerial parts and related biological activities are known. This study aimed to examine the cytotoxic, genotoxic, and antigenotoxic potential of C. langsdorffii aerial parts hydroalcoholic extract (CLE) and two of its major compounds afzelin and quercitrin. The cytotoxic and antigenotoxic potential of CLE was determined as follows: 1) against genotoxicity induced by doxorubicin (DXR) or methyl methanesulfonate (MMS) in V79 cells; 2) by direct and indirect-acting mutagens in Salmonella typhimurium strains; and 3) by MMS in male Swiss mice. The protective effects of afzelin and quercitrin against DXR or MMS were also evaluated in V79 and HepG2 cells. CLE was cytotoxic as evidenced by clonogenic efficiency assay. Further, CLE did not induce a significant change in frequencies of chromosomal aberrations and micronuclei; as well as number of revertants in the Ames test demonstrating absence of genotoxicity. In contrast, CLE was found to be antigenotoxic in mammalian cells. The results also showed that CLE exerted inhibitory effect against indirect-acting mutagens in the Ames test. Afzelin and quercitrin did not reduce genotoxicity induced by DXR or MMS in V79 cells. However, treatments using afzelin and quercitrin decreased MMS-induced genotoxicity in HepG2 cells. The antigenotoxic effect of CLE observed in this study may be partially attributed to the antioxidant activity of the combination of major components afzelin and quercitrin. Univ Franca, Lab Mutagenese, Franca, SP, Brazil Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Sao Paulo, Brazil Univ Estadual Paulista, Fac Ciencias Farmaceut Araraquara, Araraquara, SP, Brazil Univ Estadual Paulista, Fac Ciencias Farmaceut Araraquara, Araraquara, SP, Brazil FAPESP: 2011/1360-7

Neuropathic pain is described as the “most terrible of all tortures that a nerve wound may inflict.” The aim of the present study was to demonstrate the antinociceptive effect of Symplocos chinensis f. pilosa Ohwi water extract (SCW) and synthesized derivatives of the isolated compound. The antinociceptive effect was tested using the acetic acid-induced writhing and 5% formalin tests. Antinociceptive effects on neuropathic pain were evaluated using the von Frey test with chronic constriction injury (CCI) and surgical nerve injury (SNI) models and tail-flick test with a vincristine-induced pain model. An Ames test was also conducted. 5-hydroxymethylfurfural (5-HMF) was isolated and derivatives were synthesized with various acid groups. Among the plant water extracts, SCW showed significantly effective activity. Additionally, SCW presented antinociceptive effects in the neuropathic pain models. The SCW water fraction resulted in fewer writhes than the other fractions, and isolated 5-HMF was identified as an effective compound. Because 5-HMF revealed a positive response in the Ames test, derivatives were synthesized. Among the synthesized derivations, 5-succinoxymethylfurfural (5-SMF) showed the best effect in the neuropathic pain model. Our data suggest that SCW and the synthesized compound, 5-SMF, possess effective antinociceptive activity against neuropathic pain.

Three specimens of H. officinalis subsp. aristatus were collected in three areas of the Abruzzo region (Italy) and subjected to macroscopic and microscopic observation to support their botanical identification. The essential oils (EOs) obtained from the aerial parts of the samples were characterized with the object to define their phytochemical and pharmaceutical biology profile. They highlight three different chemotypes, including one never seen in previous literature (CIV17-EO, distilled from sample harvested in 2017 at Civitaretenga), that showed a fingerprinting with the predominance of (-)-limonen-10-yl-acetate (67.9%). In 2017 European Food Safety Authority (EFSA) reported the genotoxicity of similar compounds, therefore, to dismiss any safety concern for the CIV17-EO use as flavouring substance, the Ames test was performed with no evidence of mutagenic activity. Safety of use coupled with chemical characterization of this new chemotype set the stage for a better standardization of H. officinalis EOs. The ethanolic extracts, on the other hand, with qualitatively similar chemical profiles in which caftaric, chlorogenic and rosmarinic acid were the main molecules, showed interesting antioxidant activity and a slight cytotoxicity towards the A549 cell line that could indicate a starting point for the evaluation of an additional preventive tool for maintaining health status.