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Mutagenesis Mar 2024The two-test in vitro battery for genotoxicity testing (Ames and micronucleus) has in the majority of cases replaced the three-test battery (as two-test plus mammalian...
The two-test in vitro battery for genotoxicity testing (Ames and micronucleus) has in the majority of cases replaced the three-test battery (as two-test plus mammalian cell gene mutation assay) for the routine testing of chemicals, pharmaceuticals, cosmetics, and agrochemical metabolites originating from food and feed as well as from water treatment. The guidance for testing agrochemical groundwater metabolites, however, still relies on the three-test battery. Data collated in this study from 18 plant protection and related materials highlights the disparity between the often negative Ames and in vitro chromosome aberration data and frequently positive in vitro mammalian cell gene mutation assays. Sixteen of the 18 collated materials with complete datasets were Ames negative, and overall had negative outcomes in in vitro chromosome damage tests (weight of evidence from multiple tests). Mammalian cell gene mutation assays (HPRT and/or mouse lymphoma assay (MLA)) were positive in at least one test for every material with this data. Where both MLA and HPRT tests were performed on the same material, the HPRT seemed to give fewer positive responses. In vivo follow-up tests included combinations of comet assays, unscheduled DNA synthesis, and transgenic rodent gene mutation assays, all gave negative outcomes. The inclusion of mammalian cell gene mutation assays in a three-test battery for groundwater metabolites is therefore not justified and leads to unnecessary in vivo follow-up testing.
Topics: Mice; Animals; Hypoxanthine Phosphoribosyltransferase; Mutagenicity Tests; Comet Assay; Rodentia; Lymphoma; Agrochemicals; Micronucleus Tests; DNA Damage
PubMed: 38183270
DOI: 10.1093/mutage/gead037 -
Reports of Practical Oncology and... 2023Angiotensin-converting enzyme inhibitors (ACE-I) and their pharmacologically related sartans have been associated with an increased cancer incidence in several clinical...
BACKGROUND
Angiotensin-converting enzyme inhibitors (ACE-I) and their pharmacologically related sartans have been associated with an increased cancer incidence in several clinical observations. In 2018, sartans were revealed as being significantly contaminated with nitrosamines. Nitrosamines are potent human mutagens that can be formed ex vivo and, more concerningly, also in vivo from nitrosatable drug precursors. Their formation in sartans may justify the reported cancer risk and, by analogy, this may also apply to ACE-Is.
MATERIALS AND METHODS
We investigated a commonly used ACE-I, ramipril (RAM). We checked its susceptibility to in vivo interaction with nitrite, potentially resulting in the generation of mutagenic N-nitrosamines. To that end, in silico simulation of mutagenicity of RAM nitroso-derivatives was performed using VEGA-GUI software. Then, the Nitrosation Assay Procedure was conducted which served as a model of endogenous reaction. The resulting post-nitrosation mixtures were subjected to a bacterial reverse mutation test employing Salmonella typhimurium strains TA98 and TA100 with and without metabolic activation.
RESULTS
Our results showed that studied samples did not induce point mutations in the test bacteria, regardless of the catalytic cytochrome activity.
CONCLUSION
We concluded that RAM endogenous nitrosation is not the reason for increased cancer incidence. However, other ACE-Is must be verified in a similar manner.
PubMed: 38179284
DOI: 10.5603/rpor.97433 -
PloS One 2024Deficient water, sanitation, and hygiene (WASH) significantly account for a high burden of disease across the globe. Lebanon, an Eastern Mediterranean...
Deficient water, sanitation, and hygiene (WASH) significantly account for a high burden of disease across the globe. Lebanon, an Eastern Mediterranean lower-middle-income country with a polluted environment, a fragmented healthcare system, and an ongoing severe economic crisis, faces serious challenges in sustaining safe water supplies, especially in vulnerable communities, while also hosting the world highest refugee population per capita. This study aimed to examine the mutagenicity, and the estrogenic and androgenic activities of water supplies, across both a Palestinian refugee camp and a Syrian informal settlement. Water samples were collected from two targeted camps in Dbayeh and Choueifat, North and South of the Capital City Beirut, respectively, between the months of September and October 2022. Microbial and physicochemical properties of samples were determined, including fecal contamination, total dissolved solids, and various minerals and salts. Organic pollutants were extracted using pre-packed solid phase extraction (SPE) columns, and then mutagenicity of extracts was examined using the Ames test in two Salmonella typhi bacterial strains. The estrogenic and androgenic activities of extracts were assessed using the yeast estrogen and androgen screen tests assays (YES/YAS). Results show excessive levels of total coliforms and total dissolved solids (TDS) in samples from both sites. In addition, the water supply from the Dbayeh Palestinian refugee camp is mutagenic, while the water supply from the Choueifat Syrian informal settlement shows anti-androgen activity. Our findings provide valuable WASH baseline data in two major vulnerable communities in Lebanon, and highlight the importance of a water toxicity testing approach concomitant with a water safety plan, based on a holistic strategy that covers all stages of the water supply chain.
Topics: Humans; Lebanon; Refugee Camps; Syria; Arabs; Water Supply; Refugees
PubMed: 38165866
DOI: 10.1371/journal.pone.0294679 -
Food and Chemical Toxicology : An... Feb 2024Spermidine is a polyamine consumed in the diet, endogenously biosynthesized in most cells, and produced by the intestinal microbiome. A variety of foods contribute to...
Spermidine is a polyamine consumed in the diet, endogenously biosynthesized in most cells, and produced by the intestinal microbiome. A variety of foods contribute to intake of spermidine along with other polyamines. Spermidine trihydrochloride (spermidine-3HCl) of high purity can be produced using an engineered strain of Saccharomyces cerevisiae. Spermidine has a demonstrated history of safe use in the diet; however, limited information is available in the public literature to assess the potential toxicity of spermidine-3HCl. To support a safety assessment for this spermidine-3HCl as a dietary source of spermidine, authoritative guideline and good laboratory practice (GLP) compliant in vitro genotoxicity assays (bacterial reverse mutation and mammalian micronucleus assays) and a 90-day oral (dietary) toxicity study in rats were conducted with spermidine-3HCl. Spermidine-3HCl was non-genotoxic in the in vitro assays, and no adverse effects were reported in the 90-day oral toxicity study up to the highest dose tested, 12500 ppm, equivalent to 728 mg/kg bw/day for males and 829 mg/kg bw/day for females. The subchronic no observed adverse effect level (NOAEL) is 728 mg/kg bw/day.
Topics: Male; Female; Rats; Animals; Spermidine; Saccharomyces cerevisiae; No-Observed-Adverse-Effect Level; Micronucleus Tests; Mammals; Mutagenicity Tests
PubMed: 38163454
DOI: 10.1016/j.fct.2023.114428 -
Free Radical Biology & Medicine Feb 2024Accumulation of DNA damage is a critical feature of genomic instability, which is a hallmark of various cancers. The enzyme-modified comet assay is a recognized method...
Accumulation of DNA damage is a critical feature of genomic instability, which is a hallmark of various cancers. The enzyme-modified comet assay is a recognized method to detect specific DNA lesions at the level of individual cells. In this cross-sectional investigation, we explore possible links between clinicopathological and treatment related factors, nutritional status, physical activity and function, and DNA damage in a cohort of colorectal cancer (CRC) patients with non-metastatic disease. Levels of DNA damage in peripheral mononuclear blood cells (PBMCs) assessed 2-9 months post-surgery, were compared across tumour stage (localized (stage I-II) vs. regional (stage III) disease), localization (colon vs. rectosigmoid/rectum cancer), and adjuvant chemotherapy usage, with the last dosage administrated 2-191 days prior to sampling. Associations between DNA damage and indicators of nutritional status, physical activity and function were also explored. In PBMCs, DNA base oxidation was higher in patients diagnosed with regional compared with localized tumours (P = 0.03), but no difference was seen for DNA strand breaks (P > 0.05). Number of days since last chemotherapy dosage was negatively associated with DNA base oxidation (P < 0.01), and patients recently receiving chemotherapy (<15 days before blood collection) had higher levels of DNA base oxidation than those not receiving chemotherapy (P = 0.03). In the chemotherapy group, higher fat mass (in kg and %) as well as lower physical activity were associated with greater DNA base oxidation (P < 0.05). In conclusion, DNA base oxidation measured with the enzyme-modified comet assay varies according to tumour and lifestyle related factors in CRC patients treated for non-metastatic disease.
Topics: Humans; Cross-Sectional Studies; DNA Damage; Comet Assay; DNA; Colorectal Neoplasms
PubMed: 38141887
DOI: 10.1016/j.freeradbiomed.2023.12.016 -
Pharmaceutics Nov 2023The growing problem of bacterial resistance to antimicrobials actualizes the development of new approaches to solve this challenge. Supramolecular chemistry tools can...
The growing problem of bacterial resistance to antimicrobials actualizes the development of new approaches to solve this challenge. Supramolecular chemistry tools can overcome the limited bacterial resistance and side effects of classical sulfonamides that hinder their use in therapy. Here, we synthesized a number of pillar[5]arenes functionalized with different substituents, determined their ability to self-association using DLS, and characterized antimicrobial properties against , , , , via a resazurin test. Biofilm prevention concentration was calculated for an agent with established antimicrobial activity by the crystal-violet staining method. We evaluated the mutagenicity of the macrocycle using the Ames test and its ability to affect the viability of A549 and LEK cells in the MTT-test. It was shown that macrocycle functionalized with sulfonamide residues exhibited antimicrobial activity an order higher than pure streptocide and also revealed the ability to prevent biofilm formation of and . The compound did not show mutagenic activity and exhibited low toxicity to eukaryotic cells. The obtained results allow considering modification of the macrocyclic platforms with classic antimicrobials as an opportunity to give them a "second life" and return to practice with improved properties.
PubMed: 38140001
DOI: 10.3390/pharmaceutics15122660 -
Toxins Nov 2023Humans are constantly exposed to mixtures of different xenobiotics through their diet. One emerging concern is the mycotoxin alternariol (AOH), which can occur in foods...
Humans are constantly exposed to mixtures of different xenobiotics through their diet. One emerging concern is the mycotoxin alternariol (AOH), which can occur in foods typically contaminated by the process contaminant acrylamide (AA). AA is a byproduct of the Maillard reaction produced in carbohydrate-rich foods during thermal processing. Given the genotoxic properties of AOH and AA as single compounds, as well as their potential co-occurrence in food, this study aimed to assess the cytotoxic, genotoxic, and mutagenic effects of these compounds in combination. Genotoxicity was assessed in HepG2 cells by quantifying the phosphorylation of the histone γ-H2AX, induced as a response to DNA double-strand breaks (DSBs). Mutagenicity was tested in strains TA98 and TA100 by applying the Ames microplate format test. Our results showed the ability of AOH and AA to induce DSBs and increase revertant numbers in TA100, with AOH being more potent than AA. However, no synergistic effects were observed during the combined treatments. Notably, the results of the study suggest that the compounds exert mutagenic effects primarily through base pair substitutions. In summary, the data indicate no immediate cause for concern regarding synergistic health risks associated with the consumption of foods co-contaminated with AOH and AA.
Topics: Humans; Mycotoxins; Mutagens; Alternaria; DNA Damage; Lactones; Acrylamides
PubMed: 38133174
DOI: 10.3390/toxins15120670 -
Toxicology Jan 2024The T-2 toxin is a mycotoxin produced by molds belonging to Fusarium. Among the Fusarium mycotoxins, trichothecenes are frequently reported in food and feed, being the...
The T-2 toxin is a mycotoxin produced by molds belonging to Fusarium. Among the Fusarium mycotoxins, trichothecenes are frequently reported in food and feed, being the T-2 toxin (T-2) the mycotoxin which possesses the highest toxicity. According to EFSA, T-2 is found in various cereal grains used in food and feed products, mainly in oats, and it has a high environmental impact due to its mechanisms of toxicity. However, recent information on its genotoxic and mutagenic effects is lacking. This work aimed to evaluate the genotoxic and mutagenic potential of T-2 in vitro. For this purpose, HepG2 cells were exposed to 15, 30, and 60 nM T-2 for 24 h, then the DNA damage was evaluated by the micronucleus and the comet assays. In addition, point mutation analysis was performed by the bacterial reverse mutation test using 0.15-60 nM of T-2 concentrations. The results showed chromosomal damage at 60 nM T-2 since significantly more MN appeared at this concentration than in the control samples. Regarding the comet assay, DNA double helix breaks appeared at all concentrations tested and, in a concentration-dependent manner. However, no mutagenic effects were observed at any of the concentrations tested for the Salmonella typhimurium (S. Typhimurium) strains TA98, TA100, TA1535, TA1537, or the Escherichia coli (E. Coli) WP2 strain in the absence or presence of a metabolic activation system. Therefore, these results showed that T-2 mycotoxin produced genotoxic effects by MN and comet assay, while no mutagenicity was observed. However, further research simulating different metabolic activation pathways and the combined exposure of this mycotoxin with other mutagenic chemicals that could be present in the diet is necessary to discard the mutagenic potential of T-2 fully. These results highlight the carcinogenic potential and danger associated with T-2 exposure and should be considered to prevent associated food risks for the human population.
Topics: Humans; Mutagens; Mutagenicity Tests; Hep G2 Cells; Escherichia coli; T-2 Toxin; DNA Damage; Micronucleus Tests
PubMed: 38128774
DOI: 10.1016/j.tox.2023.153712 -
Brazilian Journal of Biology = Revista... 2023Chicken (Gallus gallus domesticus) is one of the primary sources of animal protein for the Brazilian population. Thus, the safety of this food is highly relevant. This...
Chicken (Gallus gallus domesticus) is one of the primary sources of animal protein for the Brazilian population. Thus, the safety of this food is highly relevant. This study was based on the evidence of severe contamination of these animals by metals such as lead in Santo Amaro, Bahia. This exploratory study aimed to evaluate associations between lead levels in blood of chicken exposed to a contaminated area with the occurrence of chromosomal alterations, evidencing genotoxic effects. Serum lead analysis was performed by GF-AAS after dilution with a matrix modifier solution (Triton X-100 0.2% v/v and HNO3 0.1% v/v), while chromosomal damage was evaluated using the comet assay. The results showed genotoxic effects (positive comet assay) only for the specimen sample with higher serum lead concentrations (33.9 µg dL-1), suggesting the occurrence of toxic effects at this level of exposure. This work evaluated a relationship between the reduction of serum lead levels in chicken and increased distance from the primary polluting source - a lead processing plant (COBRAC). It also showed that lead is bioavailable in this territory, contaminating chicken and causing genotoxic effects in these animals, further expanding the concern with the local biota and the health of the residents of Santo Amaro.
Topics: Animals; Chickens; Lead; Brazil; Comet Assay; Chromosomes
PubMed: 38126633
DOI: 10.1590/1519-6984.274806 -
Mutagenesis Mar 2024The robust control of genotoxic N-nitrosamine (NA) impurities is an important safety consideration for the pharmaceutical industry, especially considering recent drug...
The robust control of genotoxic N-nitrosamine (NA) impurities is an important safety consideration for the pharmaceutical industry, especially considering recent drug product withdrawals. NAs belong to the 'cohort of concern' list of genotoxic impurities (ICH M7) because of the mutagenic and carcinogenic potency of this chemical class. In addition, regulatory concerns exist regarding the capacity of the Ames test to predict the carcinogenic potential of NAs because of historically discordant results. The reasons postulated to explain these discordant data generally point to aspects of Ames test study design. These include vehicle solvent choice, liver S9 species, bacterial strain, compound concentration, and use of pre-incubation versus plate incorporation methods. Many of these concerns have their roots in historical data generated prior to the harmonization of Ames test guidelines. Therefore, we investigated various Ames test assay parameters and used qualitative analysis and quantitative benchmark dose modelling to identify which combinations provided the most sensitive conditions in terms of mutagenic potency. Two alkyl-nitrosamines, N-nitrosodimethylamine (NDMA) and N-nitrosodiethylamine (NDEA) were studied. NDMA and NDEA mutagenicity was readily detected in the Ames test and key assay parameters were identified that contributed to assay sensitivity rankings. The pre-incubation method (30-min incubation), appropriate vehicle (water or methanol), and hamster-induced liver S9, alongside Salmonella typhimurium strains TA100 and TA1535 and Escherichia coli strain WP2uvrA(pKM101) provide the most sensitive combination of assay parameters in terms of NDMA and NDEA mutagenic potency in the Ames test. Using these parameters and further quantitative benchmark dose modelling, we show that N-nitrosomethylethylamine (NMEA) is positive in Ames test and therefore should no longer be considered a historically discordant NA. The results presented herein define a sensitive Ames test design that can be deployed for the assessment of NAs to support robust impurity qualifications.
Topics: Humans; Animals; Cricetinae; Nitrosamines; Mutagens; Diethylnitrosamine; Mutagenesis; Mutagenicity Tests; Carcinogens
PubMed: 38112628
DOI: 10.1093/mutage/gead033