Evaluation Of Genotoxicity Research Articles

Modern alternative research methods in genetic toxicology (literature review)

The review represents the current principles of assessment of chemicals genotoxicity. The main attention is paid to alternative research methods. The international experience of the application of alternative approaches and prospects of their use for regulatory purposes are discussed. The data for this review were collected from the Russian and foreign literature, as well as Internet resources, concerning the development of the new alternative methods for testing chemicals for genotoxicity. The OECD database, Scopus, Medline, Google Scholar, RISC, CyberLeninka were used for the information retrieval. Although the evaluation of genotoxicity of chemical substances is the well-established and based on the battery of validated methods, the studies for improving the existing tests and developing new technologies, including the alternative approaches, continue unabated up to now. In general, three trends of development of genetic toxicology can be outlined, including creating of new methods based on the whole-genome sequencing and the application of genome editing technologies; implementation of quantitative system of effects assessment in addition to the existing qualitative approach (mutagenic/non-mutagenic) and testing of various combinations of genotoxicity evaluation methods to identify a battery of tests with a greater predictive activity regarding carcinogenic effects. To use the developed alternative models for regulatory purposes, it is necessary to provide convincing evidence that the data obtained are good predictors of the organism’s actual response to the effects of toxicants/genotoxicants, validation of methods, standardization, and harmonization of research protocols, and changes to the existing regulatory framework are required.

Acute toxicity of trypsin inhibitor from tamarind seeds in embryo and adult zebrafish (Danio rerio)

The trypsin inhibitor isolated from tamarind seeds (TTI) is being investigated for potential applications in the treatment of noncommunicable diseases (NCD), such as hypertension, obesity, and diabetes. This study aimed to assess TTI embryotoxicity and acute toxicity in adult zebrafish (Danio rerio). TTI was extracted and isolated from tamarind seeds. Embryonic and adult zebrafish were exposed for 96 hours to three concentrations of TTI (12.5, 25, and 50 mg/L). Zebrafish embryos (n=60 per group) were evaluated for survival, hatching, malformations, and potential developmental marker alterations, in addition to cardiotoxicity and neurotoxicity tests. For acute toxicity assessment in adults (n=20 per group), survival and locomotor and anxiety-like behaviors were assessed, along with genotoxicity (micronucleus) evaluation. Embryos exposed to TTI showed no significant adverse effects, presented normal heart rates and positive reflex response in the neurotoxicity tests. In adult fish, TTI did not cause mortality or significant behavioral changes, suggesting no neurotoxicity and no genotoxicity. Histopathological analyses of the whole body showed only changes in the liver and spinal cord, similar to those observed in the control group not exposed to TTI. These findings indicate TTI's biosafety and therapeutic potential in complex organisms. Further research is required to evaluate its long-term effects and efficacy in treating non-communicable diseases.

Scoping review on the genotoxicity of silver nanoparticles in endodontics: therapeutic saviors or genetic saboteurs?

Silver nanoparticles (AgNPs) have gained prominence in endodontics due to their exceptional antimicrobial properties. However, concerns regarding their genotoxic potential have prompted investigations into their safety profiles. This scoping review aims to consolidate current knowledge on the genotoxic effects of AgNPs in the field of endodontics. A thorough literature search across seven electronic databases was conducted using specific keywords. Inclusion criteria included experimental studies published in English from January 1960 to March 2024, addressing the genotoxicity of AgNPs in endodontic applications. Study selection and data extraction were conducted independently. The Quality Assessment Tool For In Vitro Studies (QUIN) tool was employed to evaluate the risk of bias in each study. 5 articles were selected, of which 3 were in vitro experimental designs, while the remaining were ex vivo studies. All were published between 2009 and 2021. AgNPs have been used as root canal irrigating solutions, pulp-capping materials, and root canal sealers. Most studies employed the comet assay for genotoxic evaluation. One study was found to have a low risk of bias, while others were categorized as having a medium risk. Mixed findings were noted on the genotoxic effects of AgNPs using various assays. The genotoxic potential of AgNPs somewhat poses concerns for endodontic practices. This review highlights the need for further research to develop safer alternatives and optimize their concentrations and exposure durations.

Mutagenicity and genotoxicity evaluation of 15 nitrosamine drug substance-related impurities in human TK6 cells

Nitrosamine drug substance-related impurities (NDSRIs) are a sub-category of N-nitrosamine drug impurities that share structural similarity to the corresponding active pharmaceutical ingredient. The mutagenicity of NDSRIs is poorly understood. We previously tested a series of NDSRIs using the Enhanced Ames Test (EAT). In this follow-up study, we further examined the genotoxicity and mutagenicity of 15 of these NDSRIs in human TK6 cells. Seven EAT-positive NDSRIs, including N-nitroso-nortriptyline, N-nitroso-fluoxetine, N-nitroso-desmethyl-diphenhydramine, N-nitroso-duloxetine, N-nitroso-lorcaserin, N-nitroso-varenicline, and N-nitroso-sertraline, induced concentration-dependent increases in micronuclei after bioactivation with hamster liver S9. These NDSRIs were also mutagenic in the TK and HPRT gene mutation assays, consistent with their positive EAT results. In the presence of hamster liver S9, the eight EAT-negative NDSRIs were negative in the micronucleus assay and negative for mutation induction. Using TK6 cells endogenously expressing a single human cytochrome P450 (CYP), we found that CYP2C19, CYP2B6, CYP2A6, and CYP3A4 are key enzymes activating the genotoxicity and mutagenicity of these NDSRIs. Overall, the hamster S9-mediated TK6 cell mutagenicity results agreed with those observed in the EAT, indicating consistency in the mutagenic responses produced by NDSRIs across different testing systems. These data support the use of EAT for hazard identification and safety assessment of NDSRIs.

P21-32 Genotoxicity and phytotoxicity evaluation of Norfloxacin (NOR) and intermediate of NOR after ZnO treatment

P21-32 Genotoxicity and phytotoxicity evaluation of Norfloxacin (NOR) and intermediate of NOR after ZnO treatment

P19-52 In silico and in vitro genotoxicity evaluation of vildagliptin impurities: vildagliptin cyclic amidine, vildagliptin diketopiperazine and vildagliptine amide

P19-52 In silico and in vitro genotoxicity evaluation of vildagliptin impurities: vildagliptin cyclic amidine, vildagliptin diketopiperazine and vildagliptine amide

P20-34 Genotoxicity evaluation of Propyl gallate: dissecting ambiguous in vitro findings and validating in vivo assays

P20-34 Genotoxicity evaluation of Propyl gallate: dissecting ambiguous in vitro findings and validating in vivo assays

Evaluation of genotoxicity and teratogenicity of phillyrin

Phillyrin is extracted from Forsythia suspensa (Thunb.) Vahl, is significantly higher in (unripe Forsythiae Fructus) Qing qiao than in (ripe Forsythiae Fructus) Lao qiao fruits of the plant. However, the toxicity of phillyrin has not been adequately investigated. The study investigates the genetic and teratogenic effects of phillyrin to determine its safety profile. Assessing the genotoxicity and teratogenicity of phillyrin involved various tests, such as the bacterial reverse mutation assay, mammalian erythrocyte micronucleus assay, spermatocyte chromosome aberration assay, and teratogenicity assay. The results demonstrated that phillyrin exhibited no discernible impact on the following: number of colonies that spontaneously revert for Salmonella typhimurium TA 97, TA98, TA100, TA102, and TA1535, frequency of bone marrow polychromatic erythrocytes, and the rate of chromosomal aberrations. In the teratogenicity test, the pregnant rats exhibited no signs of toxicity or abnormal changes, and the growth, embryonic development, and visual anatomy of each pup were normal. In comparison with the negative control group, there were no significant differences in fetal body weight, mortality, deformity rate, malformed nest rate, gravid uterus weight, average number of fetuses per litter, fetal body length, or visceral and skeletal development in each dose group. In conclusion, these findings provide evidence that phillyrin does not exhibit genotoxic or teratogenic effects, supporting its potential safety for pharmacological applications.

Cyto- and genotoxicity evaluation of water samples collected from two rivers in the Kosovo.

River water in Kosovo is exposed to various discharges from industrial and agricultural activities as well as to urban wastewater. Rivers Sitnica and Drenica are among the most affected ones and water samples drawn from these rivers show the presence of various toxic substances. Genotoxic effects are seen in fish living in these rivers indicating a cytotoxic and mutagenic potential of the river water. Aiming at substantiating these observations, we assessed cyto- and genotoxic effects of water samples collected at different locations from Drenica and Sitnica river. Samples drawn from Lake Badovc served for comparison. To address seasonal effects, samples were collected at different seasons/time points during the period summer 2016 - spring 2018. The water samples were analyzed employing primary rat hepatocytes as reliable in vitro cell model for the assessment of cytotoxic effects (mitotic arrest and cell death) and DNA damage/genotoxicity (micronucleus assay, Comet assay). The results do not account for significant effects associated with specific locations but demonstrate seasonal differences of the genotoxic potential of the water samples collected along both rivers, which are accompanied by a limited cytotoxic potential. Our data provide substantial support to earlier observations and strongly warrant the need for continuous chemical as well as biological monitoring of the river water in Kosovo, focusing on an improved toxicant profiling of the river water and investigations addressing the observed seasonal variations.

Genotoxicity Evaluation of a Hyperosmotic Solution Proposed as Endodontic Irrigant

Genotoxicity Evaluation of a Hyperosmotic Solution Proposed as Endodontic Irrigant

Assessment of cytotoxic and genotoxic effects of glyphosate-based herbicide on glioblastoma cell lines: Role of p53 in cellular response and network analysis

Glyphosate, the world's most widely used herbicide, has a low toxicity rating despite substantial evidence of adverse health effects. Furthermore, glyphosate-based formulations (GBFs) contain several other chemicals, some of which are known to be harmful. Additionally, chronic, and acute exposure to GBFs among rural workers may lead to health impairments, such as neurodegenerative diseases and cancer. P53 is known as a tumor suppressor protein, acting as a key regulator of the cellular response to stress and DNA damage. Therefore, mutations in the TP53 gene, which encodes p53, are common genetic alterations found in various types of cancer. Therefore, this study aimed to evaluate the cytotoxicity and genotoxicity of GBF in two glioblastoma cell lines: U87MG (TP53-proficient) and U251MG (TP53-mutant). Additionally, the study aimed to identify the main proteins involved in the response to GBF exposure using Systems Biology in a network containing p53 and another network without p53. The MTT assay was used to study the toxicity of GBF in the cell lines, the clonogenic assay was used to investigate cell survival, and the Comet Assay was used for genotoxicity evaluation. For data analysis, bioinformatics tools such as String 12.0 and Stitch 5.0 were applied, serving as a basis for designing binary networks in the Cytoscape 3.10.1 program. From the in vitro test analyses, it was observed a decrease in cell viability at doses starting from 10 ppm. Comet Assay at concentrations of 10 ppm and 30 ppm for the U251MG and U87MG cell lines, respectively observed DNA damage. The network generated with systems biology showed that the presence of p53 is important for the regulation of biological processes involved in genetic stability and neurotoxicity, processes that did not appear in the TP53-mutant network.

Elderberry Hydrolate: Exploring Chemical Profile, Antioxidant Potency and Antigenotoxicity for Cosmetic Applications

Elderberry (Sambucus nigra L.) hydrolate, derived from domestic steam distillation, holds promise as a multifunctional ingredient for skincare and cosmetic applications. This study investigates the chemical composition and biological activities of elderberry hydrolate obtained through steam distillation. Despite the growing interest in elderberry hydrolate, there has been a lack of comprehensive studies elucidating its chemical composition and potential bioactive constituents. To address this gap, we conducted a detailed analysis of elderberry hydrolate’s composition, antioxidant activity, and antigenotoxicity. Genotoxic evaluation and antioxidant assays (ABTS, DPPH) were conducted to assess its biological properties. We obtained elderberry hydrolate with a notable transfer of aromatic compounds through the steam distillation process, highlighting its efficacy and sustainability. The chemical characterization identified vital compounds, including phenylacetaldehyde, 2-acetyl-pyrrole, and an unidentified major component, collectively contributing to the hydrolate’s aromatic and biological properties. The genotoxic evaluation using the Comet assay demonstrated the hydrolate’s protective effects against DNA damage induced by hydrogen peroxide and streptonigrin. The optimal DNA protection was observed at 10% (w/v), attributed to the antioxidant activity of the identified compounds. The hydrolate exhibited significant antioxidant potential, demonstrating concentration-dependent responses and correlating higher concentrations with increased antioxidant activity. These findings underscore the multifaceted attributes of elderberry hydrolate, positioning it as a promising natural ingredient for skincare. This study supports elderberry hydrolate as a valuable natural and sustainable product development resource.

Evaluation of Genotoxicity by Comet Assay in Tissues of Clarias gariepinus Exposed to Cassava Effluent

Untreated wastewater discharge into waterbodies poses a number of risks, including the potential for aquatic organisms' DNA to be damaged. Using the comet assay, this study assessed the genotoxic impact of cassava wastewater on the gonads, liver, and gills of post-juvenile Clarias gariepinus. The post-juvenile C. gariepinus fish were acquired from a fish farm in Edo State, Nigeria, and were subjected to different concentrations of cassava wastewater (0.2%, 0.3%, 0.5%, and 0.7%) for a duration of 96 hours. In contrast to the typical actions seen in the control groups, the fish exposed to the effluent exhibited restless, erratic movements, and gasping for air. The first set of repetitions showed that catfish mortality increased with concentration; after 96 hours, exposure to the lowest concentration (0.2%) and the highest concentration (0.7%) of cassava wastewater caused 10% and 40% of the fish to die, respectively. The second set of repetitions showed that catfish mortality increased with increasing concentration; after 96 hours, exposure to the lowest concentration (0.2%) and the highest concentration (0.7%) of cassava wastewater resulted in 20% and 50% catfish mortality, respectively. Following the exposure time, the fish were brought to the lab where their gonads, liver, and gills were removed in order to use the comet assay for genotoxic assessment. The genotoxins present in the wastewater can be the reason for the DNA impairment found in the genotoxic assessment result at different effluent concentrations. The investigation also showed that DNA damage increased with increasing concentration, indicating a dose-dependent genotoxic effect of cassava wastewater on post-juveniles of C. gariepinus. This research demonstrates the critical importance of treating wastewater before to discharge.

Toxicity Evaluation of a Non-Pain Pharmacopuncture Extract Using a Bacterial Reverse Mutation Test.

The objective of this study was to assess the genotoxicity of a no-pain pharmacopuncture (NPP) extract developed in 2022 using a bacterial reverse mutation assay, aiming to further substantiate the safety profile of NPP. The genotoxicity evaluation involved a bacterial reverse mutation assay to assess the mutagenic potential of NPP extracts with and without metabolic activation. Histidine-requiring Salmonella typhimurium strains (TA98, TA100, TA1535, and TA1537) and tryptophan-requiring Escherichia coli strains (WP2uvrA) were used in the assay. The NPP extract did not induce a revertant colony count exceeding two times that of the negative control at any dose level in any of the tested strains, both with and without metabolic activation. Additionally, no growth inhibition or precipitation was observed in the presence of NPP. Based on the findings, it can be concluded that the NPP extract exhibited no mutagenic potential in the in vitro genotoxicity tests conducted.

Evaluation of infliximab-induced genotoxicity and possible action on BCL-2 and P53 genes

ABSTRACT Although the last pandemic created an urgency for development of vaccines, there was a continuous and concerted effort to search for therapeutic medications among existing drugs with different indications. One of the medications of interest that underwent this change was infliximab (IFM). This drug is used as an anti-inflammatory, predominantly in patients with Crohn ‘s disease, colitis ulcerative, and rheumatoid arthritis. In addition to these patients, individuals infected with Coronavirus Disease (COVID-19) were administered this chimeric monoclonal antibody (IMF) to act as an immunomodulator for patients in the absence of comprehensive research. Consequently, the present study aimed to examine the genotoxic effects attributed to IFM treatment employing different assays in vivo using mouse Mus musculus. Therefore, IFM was found to induce genotoxic effects as evidenced by the comet assay but did not demonstrate genotoxic potential utilizing mouse bone marrow MN test. The results of evaluating the expression of the P53 and BCL-2 genes using RT-qPCR showed stimulation of expression of these genes at 24 hr followed by a decline at 48 hr. Although the comet assay provided positive results, it is noteworthy that based upon negative findings in the micronucleus test, the data did not demonstrate significant changes in the genetic material that might affect the therapeutic use of IFM. The stimulation of expression of P53 and BCL-2 genes at 24 hr followed by a decline at 48 hr suggest a transient, if any, effect on genetic material. However, there is still a need for more research to more comprehensively understand the genotoxic profile of this medication.

Photoprotective efficacy of Sunset Yellow via inhibition of type-I and type-II pathway under exposure of sunlight.

Exposure to phototoxicants and photosensitizers can result in the generation of reactive oxygen species (ROS), leading to oxidative stress, DNA damage, and various skin-related issues such as aging, allergies, and cancer. While several photo-protectants offer defense against ultraviolet radiation (UV-R), their effectiveness is often limited by photo-instability. Sunset Yellow (SY), an FDA-approved food dye, possesses significant UV-R and visible light absorption properties. However, its photoprotective potential has remained unexplored. Our investigation reveals that SY exhibits remarkable photostability for up to 8 h under both UV-R and sunlight. Notably, SY demonstrates the ability to quench ROS, including singlet oxygen (1O2), superoxide radicals ( ), and hydroxyl radicals (·OH) induced by rose bengal, riboflavin and levofloxacin, respectively. Moreover, SY proves effective in protecting against the apoptotic and necrotic cell death induced by the phototoxicant chlorpromazine (CPZ) in HaCaT cells. Further, it was observed that SY imparts photoprotection by inhibiting intracellular ROS generation and calcium release. Genotoxicity evaluation provides additional evidence supporting SY's photoprotective effects against CPZ-induced DNA damage. In conclusion, these findings underscore the potential of SY as a promising photoprotective agent against the toxic hazards induced by phototoxicants, suggesting its prospective application in the formulation of broad-spectrum sunscreens.

Antioxidant and Antileishmanial Activities, and Evaluation of Acute Oral Toxicity and Genotoxicity of Bixa orellana Leaf Extracts

Antioxidant and Antileishmanial Activities, and Evaluation of Acute Oral Toxicity and Genotoxicity of Bixa orellana Leaf Extracts

Evaluation of oxidative damage and genotoxicity in populations exposed to arsenic in drinking water from Santa Fe province, Argentina

The presence of arsenic in the environment is a public health problem. Groundwater of certain regions of Argentina contains arsenic of natural origin in concentrations that exceed the guide level recommended by World Health Organization (WHO, 10 µg/L). Pathologies derived from chronic arsenic consumption justify the planning of human biomonitoring. Hence, the aim of this study was to evaluate oxidative damage and genotoxicity and its relationship with nutritional variables in populations exposed to arsenic through drinking water in Santa Fe province, Argentina. A total of 322 participants were analyzed for arsenic in urine together with biomarkers of genotoxicity (Comet assay in blood and frequency of Micronuclei and other Nuclear Abnormalities in exfoliated buccal cells) and oxidative stress (modified Comet assay with Endonuclease III, Lipid peroxidation and antioxidant enzyme activity), as well as nutritional and biochemical variables. Results showed that 45 % of participants excreted arsenic in the urine. Consumption of water with arsenic, whether currently or previously, was associated with statistically significant increase of oxidative DNA damage and lipid peroxidation. MN in exfoliated buccal cells serve as an early biomarker of genotoxicity and showed significant differences in the current exposed group. Biochemical results indicate dyslipidemias potentially linked to dietary choices, and insufficient intake of fruits and vegetables rich in antioxidants, was also noted. This study advocates risk communication to the population, educators, and health authorities, emphasizing the need for preventive health strategies and improved food education.

Genotoxicity and cytotoxicity evaluation of a heat-not-burn product

‘Heat-not-burn’ products (HnBP) contain lower levels of harmful substances than traditional cigarettes, but the use of these products warrants further toxicological evaluation. We have compared the cytotoxicity and genotoxicity of a heat-not burn product with conventional cigarettes, in vivo and in vitro. Male Sprague Dawley rats were exposed to mainstream smoke from conventional cigarettes or a HnBP, for 4 or 28 days, followed by isolation of bone marrow polychromatic erythrocytes (PCE) and histological examination of the testes. Chinese hamster lung fibroblast cells were exposed in vitro to total particulate matter from cigarette smoke obtained through Cambridge filters. The cytotoxicity and genotoxicity of total particulate matter were assessed by the neutral red uptake assay, chromosome aberration assay, in vitro micronucleus test, comet assay, and Ames assay. In the short-term exposure rat models, only the conventional-cigarettes group showed a significant increase in the ratio of micronuclei to total PCE. There was no significant difference in rat testis histology in the long-term exposure models. In vitro, in the neutral red uptake assay, the HnBP product showed lower cytotoxicity than conventional cigarettes. Conventional cigarettes showed greater genotoxicity in the chromosome aberration assay, high-dose Ames tests with exogenous metabolic activation, and micronucleus tests. In summary, our results suggest that HnBP have lower cytotoxicity and genotoxicity than conventional cigarettes.

Non-clinical safety assessment of Annona atemoya leaf extract: evaluation of genotoxicity.

The online version contains supplementary material available at 10.1007/s43188-024-00241-4.