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AbstractSince the safety of new-generation polyester-based internal coatings regarding the migration of non-intentionally added substances (NIAS) is poorly documented, studies are needed to identify NIAS originating from these food-contact materials (FCM). The aim of this study was to identify volatile and semi-volatile NIAS from polyester-based coatings in order to assess their hazard and ensure consumers’ safety with regard to exposure from canned food. Extraction and migration tests were carried out on a single polyester-coated tin plate (5 batches) using two solvents: acetonitrile and ethanol 95%, then FCM’s extracts and migrates were analysed by GC-MS. An antioxidant degradation (hydrolysis) product, 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionic acid or fenozan (CAS RN: 20170-32-5), was identified and confirmed by reference standard in all migrates. To assess fenozan’s toxicity, several in vitro bioassays, such as the Ames test (to assess point mutation), the micronucleus assay (to detect chromosomal aberrations), and the iodide uptake assay (to study one mode of action for thyroid disruption) were conducted. Fenozan was negative in the Ames test on three strains of S. typhimurium (TA98, TA100, and TA1535) and on one strain of E.coli (WP2), with and without metabolic activation system (S9 mix) using direct incorporation and pre-incubation methods. The in vitro micronucleus assay conducted on HepG2 cells also exhibited a negative response following a 4-hour treatment with the S9 mix, and a 48-hour treatment without the S9 mix. A weak inhibitory effect was obtained when testing fenozan in the iodide uptake assay using rat thyroid FRTL-5 cells. Significant inhibition started from 800 µM of fenozan, with a maximal inhibition of almost 47% at 1000 µM. The findings indicate that fenozan exhibits an anti-thyroid activity in vitro. [ABSTRACT FROM AUTHOR]

AbstractThe effects of XQ528 tartrate on the embryonic and fetal development of fertile Sprague-Dawley (SD) rats, along with their embryos and littermates, were evaluated using an embryo-fetus developmental toxicity assay. fertile SD rats exhibited no significant general toxic effects when administered doses of 0.25, 1.25, and 5.0 mg/kg intranasally from days 6 to 15 of gestation. The genotoxicity of the compound was evaluated through an amalgam of tests that included the Ames test, the Chinese hamster ovary (CHO) cell chromosome aberration test, and the micronucleus test in ICR mice. The results from the Ames test indicated non-mutagenicity at concentrations of 5000, 500, 50.0, 5.0, and 0.5 μg/dish across strains TA97, TA98, TA100, TA102, and TA1535. Additionally, the chromosomal aberration rates in CHO cells were not significantly altered at concentrations of 50.5, 101.0, and 202.0 μg/mL. No micronuclei induction was observed in ICR mice at dosage levels of 11.25, 22.50, and 45.00 mg/kg post intranasal administration. In conclusion, the no observed adverse effect level (NOAEL) for developmental toxicity of XQ528 tartrate in fertile SD rats, embryos, and littermates under the test conditions in this study was established at 5.0 mg/kg/day. Under these test conditions, XQ528 tartrate did not exhibit any significant genotoxic or carcinogenic potential. [ABSTRACT FROM AUTHOR]

AbstractCanagliflozin, Dapagliflozin, and Empagliflozin, glucagon-like peptide-1 receptor agonists, are indicated for managing type II diabetes. Although the genotoxicity profiles of these drugs are well-explored, limited information exists regarding the genotoxic potential of their impurities. In this investigation, the dimer impurities of Canagliflozin, Dapagliflozin, and Empagliflozin underwent both in silico and in vitro assessments for mutagenic potential. Tester strains of Salmonella typhimurium and Escherichia coli were subjected to the Ames test, utilizing concentrations of up to 1 µg per plate, with and without the presence of metabolic activation. Evaluation of micronucleus induction in TK6 cells was conducted through a micronucleus test, exploring concentrations up to 500 µg/mL, with or without the presence of exogenous metabolic activation. Under the specific test conditions, the dimer impurities of Canagliflozin, Dapagliflozin, and Empagliflozin showed no evidence of mutagenicity or clastrogenicity, establishing their in vitro classification as nonmutagenic. These findings align with negative in silico predictions from quantitative structure–activity relationship (QSAR) analyses for mutagenicity and genotoxicity of the dimer impurities. Collectively, these studies contribute clinically relevant safety information by confirming that the dimer impurities of Canagliflozin, Dapagliflozin, and Empagliflozin are nonmutagenic and nongenotoxic, emphasizing the consistency between in silico and in vitro data. [ABSTRACT FROM AUTHOR]

7-Methylxanthine (7-MX, CAS No. 552-62-5, purity 99.46%) is the first orally administered drug candidate, which showed anti-myopic activity in different pre-clinical studies. In the present study, we investigated the in-vivo genotoxic and mutagenic toxicity of 7-MX in Wistar rats using comet/single-cell gel electrophoresis, chromosomal aberration and micronucleus assays after oral administration. For the single-dose study (72 h), two doses of 7-MX 300 and 2000 mg/kg body weight were selected. For a repeated dose 28 d study, three doses (250, 500, and 1000 mg/kg) of 7-MX were selected. The doses were administered via oral gavage in the suspension form. Blood and major vital organs such as bone marrow, lung and liver were used to perform comet/single cell gel electrophoresis, chromosomal aberration, and micronucleus assays. The in-vitro Ames test was performed on TA98 and TA100 strains. In the chromosomal aberration study, a non-significant increase in deformities such as stickiness, ring chromosome, and endoreduplication was observed in bone marrow cells of 7-MX treated groups. These chromosomal alterations were observed upon treatment with doses of 2000 mg/kg single dose for 72 h and 1000 mg/kg repeated dose for 28 d. At a dose of 500 mg/kg, DNA damage in terms of tail length, tail moment, % tail DNA and the olive tail moment was also found to be non-significant in 7-MX treated groups. The Ames test showed the non-mutagenic nature of 7-MX in both strains of TA98 and TA100 of Salmonella typhimurium with or without metabolic activation. Thus, the present work is interesting in view of the non- genotoxicity and non-mutagenicity of repeated doses of 7-MX. [ABSTRACT FROM AUTHOR]

Abstract\nHighlightsDoxorubicin (Dox), an effective anticancer agent, is known for its genotoxic effects on normal cells. Phenolic compounds, renowned for their antitumor, antioxidant, and antigenotoxic properties, have gained prominence in recent years. This study investigates the individual and combined protective effects of rosmarinic acid (RA) and epigallocatechin gallate (EGCG) against Dox-induced genotoxicity using various in vitro test systems. The synergistic/antagonistic interaction of these combinations on Dox’s chemotherapeutic effect is explored in breast cancer cell lines. Both RA and EGCG significantly mitigate Dox-induced genotoxicity in comet, micronucleus, and Ames assays. While Dox exhibits higher selectivity against MCF-7 cells, EGCG and RA show greater selectivity against MDA-MB-231 cells. The coefficient of drug interaction reveals a synergistic effect when RA or EGCG is combined with Dox in breast cancer cells. In conclusion, both EGCG and RA effectively reduce Dox-induced genetic damage and enhance Dox’s cell viability-reducing effect in breast cancer cells.Rosmarinic acid (RA) showed protective effect against doxorubicin-induced genotoxicity.Epigallocatechin gallate (EGCG) demonstrated pro-oxidant properties at high concentrations.EGCG and RA selectively targeted MDA-MB-231 cells.Synergistic effect was observed when EGCG or RA was administered together with Dox on breast cancer cells. [ABSTRACT FROM AUTHOR]

Tellimagrandin-I (TL) and camptothin A (CA) are ellagitannins widely found in diverse plant species. Numerous studies demonstrated their significant biological activities, which include antitumor, antioxidant, and hepatoprotective properties. Despite this protective profile, the effects of TL and CA on DNA have not been comprehensively investigated. Thus, the aim of this study was to determine the mutagenic and antimutagenic effects attributed to TL and CA exposure on Salmonella enterica serovar Typhimurium strains using the Ames test. In addition, the cytotoxic and genotoxic effects were examined on human lymphocytes, employing both trypan blue exclusion and CometChip assay. The antigenotoxic effect was determined following TL and CA exposure in the presence of co-treatment with doxorubicin (DXR). Our results from the Ames test indicated that TL or CA did not display marked mutagenic activity. However, TL or CA demonstrated an ability to protect DNA against the damaging effects of the mutagens 4-nitroquinoline-1-oxide and sodium azide, thereby exhibiting antimutagenic properties. In relation to human lymphocytes, TL or CA did not induce significant cytotoxic or genotoxic actions on these cells. Further, these ellagitannins exhibited an ability to protect DNA from damage induced by DOX during co-treatment, indicating their potential beneficial usefulness as antigenotoxic agents. In conclusion, the protective effects of TL or CA against mutagens, coupled with their absence of genotoxic and cytotoxic effects on human lymphocytes, emphasize their potential therapeutic value in chemopreventive strategies. [ABSTRACT FROM AUTHOR]

Oenothein B (OeB) is a dimeric ellagitannin with potent antioxidative, antitumor, immunomodulatory, and anti-inflammatory properties. Despite the promising activities of OeB, studies examining the genotoxic or protective effects of this ellagitannin on DNA are scarce. Therefore, to further comprehensively elucidate the chemopreventive profile of OeB, the aim of this study was to evaluate the mutagenic and antimutagenic actions of OeB using Salmonella typhimurium strains with the Ames test. The micronucleus (MN) test and comet assay were used to assess the anticytotoxic and antigenotoxic effects of OeB on mouse bone marrow cells following differing treatments (pre-, co-, and post-treatment) in response to cyclophosphamide (CPA)-induced DNA damage. In addition, histopathological analyses were performed to assess liver and kidney tissues of Swiss Webster treated mice. Our results did not detect mutagenic or antimutagenic activity attributed to OeB at any concentration in the Ames test. Regarding the MN test, data showed that this ellagitannin exerted antigenotoxic and anticytotoxic effects against CPA-induced DNA damage under all treatment conditions. However, no anticytotoxic action was observed in MN test after pre-treatment with the highest doses of OeB. In addition, OeB demonstrated antigenotoxic effects in the comet assay for all treatments. Histopathological analyses indicated that OeB attenuated the toxic effects of CPA in mouse liver and kidneys. These findings suggest that OeB exerted a chemoprotective effect following pre- and co-treatments and a DNA repair action in post-treatment experiments. Our findings indicate that OeB protects DNA against CPA-induced damaging agents and induces post-damage DNA repair. [ABSTRACT FROM AUTHOR]

Cervical cancer ranks fourth in incidence among women worldwide. Cisplatin is currently the first-line drug of treatment for cervical cancer; however, it causes serious adverse effects. Therefore, it is crucial to explore natural products for cervical cancer treatment. Prosopis laevigata is a medicinal plant frequently used for ophthalmological and gastrointestinal infections. In this study, we used the MTT cell viability assay to evaluate the cytotoxic effect of a wood extract from Prosopis laevigata (Extract T7) in SiHa, HeLa, Ca Ski, and C-33 A cancer cell lines. Phosphatidylserine translocation and cell cycle evaluations were performed to determine the mechanism of cellular death. The extract's safety was evaluated using the Ames test with Salmonella typhimurium strains, in vivo acute toxicity assay, and repeated dose toxicity assay in mice. We also identified phenolic compounds of Extract T7 through liquid chromatography/mass spectrometry. Naringin, catechin, and eriodictyol demonstrated a higher concentration in Extract T7. Additionally, Extract T7 exhibited a cytotoxic effect against cervical cancer cells, where C-33 A was the most sensitive (IC50= 22.58 ± 1.10 µg/mL and 14.26 ± 1.11 µg/mL at 24 h and 48 h respectively). Extract T7 induced death by apoptosis and cell cycle arrest in the G2 phase in C-33 A. Extract T7 was not mutagenic. No toxicological effects were observed during acute toxicity and repeated dose toxicity for 28 days. Therefore, further evaluations of Extract T7 should be conducted to identify the complete mechanism of action for potential anti-tumoral activity and safety before conducting studies in animal models. [ABSTRACT FROM AUTHOR]

Sevelamer hydrochloride (SH) and calcium carbonate (CaCO3) are two agents included in the phosphate-binding group which are frequently prescribed in the treatment of patients with hyperphosphatemia. However, there are no satisfactory studies on the genotoxic effects of SH in vitro. This study was conducted to reveal the genotoxic and/or cytotoxic potential of these two drugs in cultured human peripheral lymphocytes. Human peripheral lymphocytes were treated with SH and CaCO3 at sublethal concentrations for 24 or 48 h for micronucleus assay and 1 h in the comet assay. CaCO3 and SH stimulated a slight increase in micronucleus formation however this increase was not significant compared to the control group. According to the findings of the comet test, only one concentration of the SH caused significant DNA damage (2 mg/ml, 48 h) whereas CaCO3 did not cause important DNA breakage. No significant oxidative damage or anti-radical effect caused by test substances was observed on the pure pBR322 plasmid DNA in a cell-free medium. Also, it was found that the drugs were devoid of mutagenic activity in the Ames test, but had a weak cytotoxic effect. Both test substances, particularly SH, significantly reduced the nuclear division index compared to the control group. In conclusion, the cytotoxic effect of SH was evident on the basis of in vitro tests and slightly higher than CaCO3. [ABSTRACT FROM AUTHOR]

The opioid agonist hydromorphone is indicated for the management of severe acute and chronic pain given that alternate treatments are insufficient. While the genotoxicity profile of hydromorphone is well investigated, little is known about the genotoxic potential of its impurities. In this study, 2,2-bishydromorphone was tested in silico and in vitro for both its mutagenic potential in an Ames test performed with Salmonella typhimurium and Escherichia coli tester strains up to a maximum concentration of 5 mg per plate in the absence and presence of metabolic activation. Furthermore, it was tested for its ability to induce micronuclei in TK6 cells in a micronucleus test up to a maximum concentration of 500 µg/mL with or without an exogenous metabolic activation system. 2,2-Bishydromorphone did not reveal any potential for inducing mutagenicity or clastogenicity under the conditions of the respective tests and is therefore considered non-mutagenic and non-clastogenic/aneugenic in vitro. These results are in line with negative in silico quantitative structure-activity relationship (QSAR) prediction for 2,2-bishydromorphone mutagenicity and clastogenicity and provide evidence of good correlation of in silico and in vitro data. Conclusively, these studies add important new clinically relevant information on the safety of hydromorphone as the impurity of 2,2-bishydromorphone is proven to be non-mutagenic and non-clastogenic. [ABSTRACT FROM AUTHOR]

Brazilian brown propolis (BBP) is a natural product derived predominantly from the south region of Brazil, where Araucaria forests are dominant. Despite its potential as a source of bioactive compounds with leishmanicidal, anti-inflammatory, nociceptive, and antimicrobial properties, BBP has not been comprehensively studied compared to green propolis. Therefore, this study aimed to determine the safety and chemopreventive potential of BBP. The cytotoxicity attributed to BBP was assessed using two different assays, while the Salmonella/microsome assay was employed to evaluate mutagenicity. The acute toxicity attributed to BBP was determined using a zebrafish model, while the chemopreventive potential was investigated utilizing Chinese hamster lung (V79) cell lines. Data demonstrated that BBP exerted cytotoxic effects at concentrations greater than or equal to 10 µg/ml and did not exhibit mutagenicity in Salmonella typhimurium strains TA98 and TA100. However, at the highest concentration tested (4000 µg/plate), BBP induced a significant increase in revertant colonies in S. typhimurium TA102 strain. The LC50 equivalent to 8.83 mg/L was obtained in the acute toxicity evaluation in zebrafish. BBP also showed antigenotoxic effect by significantly reducing chromosomal damage induced by the mutagen doxorubicin in V79 cell cultures at a concentration of 2.5 μg/ml. Compared to Brazilian green and red propolis, BBP exhibited greater toxicity. On the other hand, at lower concentrations, BBP displayed chemopreventive potential, which may be associated with the antioxidant capacity of the extract. These findings contribute to a better understanding of the biological properties and potential applications of BBP in treating various diseases. [ABSTRACT FROM AUTHOR]

Hymenaea genus has been used in folk medicine in Brazil, but few studies investigated its toxicity profile. Thus, the aim of this study was to determine toxicological parameters of Hymenaea courbaril stem bark hydroalcoholic extract by utilizing three cell lines including murine macrophages (RAW 264.7), mouse fibroblast cells (L929) and human lung fibroblast (MRC-5), as well as Salmonella/microsome assay, and in vivo Caenorhabditis elegans model. The predominant detected phytoconstituents in the extract were coumarins, flavonoids, phenolics, tannins and saponins and by HPLC analysis, astilbin (AST) was found to be the main component. The DPPH assay demonstrated that H. courbaril hydroalcoholic extract exhibited potent antioxidant activity, with an IC50 of 3.12 μg/ml. The extract at concentrations of 400 and 800 μg/ml decreased cell viability 48 hr after treatment in L929 and MRC-5 cell lines. In the Raw 264.7 strain, just the highest concentration (800 μg/ml) lowered cell viability within 48 hr following exposure. The concentration of 100 μg/ml did not markedly affect cell viability in the trypan blue assay. In the alkaline comet assay the extract was found to be non-genotoxic. In the Ames test, the extract exhibited low mutagenic potential without metabolic activation, since only the highest concentrations produced an effect. H. courbaril extract only affected the survival of C. elegans at concentrations of 800 and 1600 μl/ml. These findings demonstrate that H. courbaril extract appears to exert low toxicity as evidenced in vitro and mutagenicity assays; however, the biological relevance of the response of C. elegans survival to safety assessments needs further studies. [ABSTRACT FROM AUTHOR]

The current study aimed to investigate the phenolic and flavonoid contents, cytotoxicity, genotoxicity, and mutagenicity of ethanolic and aqueous extracts of A. roseum (Camb.). The MTT assay was used to determine cytotoxicity, Comet assay was used to determine genotoxicity and mutagenicity of extracts was assessed through Ames test. The results showed that total phenolic contents were higher in aqueous extract (538.26 mg GAE/100 g); however, total flavonoid contents were significantly higher in ethanolic extract (6553.29 mg Eq. of Quercetin/100 g). The cells survival percentage was significantly (P <.05) decreased as concentrations of ethanolic extract increased. However, no significant toxicity was observed in groups treated with 0.031–0.25 mg/ml of ethanolic extract of A. roseum compared to negative control (P >.05). IC50 of ethanolic and aqueous extract of A. roseum was 5.54 mg/ml and 11.39 mg/ml, respectively. The results indicated that all concentration groups of ethanolic and aqueous extract of A. roseum significantly (P <.05) lowered DNA damage than positive control group. The lowest DNA damage was observed at 0.031 and 0.062 mg/ml in both aqueous and ethanolic extracts. Moreover, at higher concentrations of 4, 8, and 16 mg/ml of both ethanolic and aqueous extract observed 100% DNA damage cells (comet cells). Moreover, Ames mutagenicity showed that no significant (P >.05) changes were observed in revertant colonies of both bacterial strains (T 98; T 100) at 0.25, 0.5, and 1 mg/ml concentrations of both aqueous and ethanolic extract of A. roseum as MI index was less than 2. The research concluded that A. roseum extracts exhibited no cytotoxicity, genotoxicity, and mutagenicity at lower concentrations (≤2 mg/ml). [ABSTRACT FROM AUTHOR]

Two series of nicotinonitrile and/or arene-linked bis(thiazoles) possessing chromene units are efficiently prepared in this study. A one-pot protocol was designed to prepare the target hybrids involved in the reaction of the respective salicylaldehydes, 2-cyanoethanethioamide, and the appropriate bis(α-haloketone). The reaction was irradiated by microwaves at 100 °C for 40–90 min in dioxane and mediated by piperazine to produce the desired products in 84–95% yields. Nicotinonitrile-linked hybrids 2b, and 2c attached to 6-(4-chlorophenyl) or 6-(4-nitrophenyl) units, respectively, had the best antibacterial activity, particularly against Staphylococcus aureus, and Enterococcus faecalis strains. They showed equipotent efficacy to ciprofloxacin with MIC/MBC values in the ranges from 5.9 to 12.0 µM. Additionally, 2c demonstrated comparable inhibitory activity to linezolid against MRSA and VRE strains with MIC/MBC values ranging from 2.9 to 11.8 µM. The Ames mutagenicity test was used to conclude that 2c was not mutagenic to the Salmonella strains. [ABSTRACT FROM AUTHOR]

Direct interference with Kelch-like ECH-associated protein 1 (Keap1)-Nrf2 protein-protein interaction (PPI) has recently been introduced as an attractive approach to control life-threatening diseases like myocarditis. The present study aimed to investigate the potential application in myocarditis of a series of novel non-naphthalene derivatives as potential Keap1-Nrf2 PPI inhibitors. Our results indicated that the optimal compound K22 displayed the highest metabolic stability and showed notable Keap1-Nrf2 PPI inhibitory activities in vitro. K22 effectively triggered Nrf2 activation and increased the protein and mRNA expression of Nrf2-regulated genes in H9c2 cells. Moreover, pre-treatment with K22 was shown to mitigate LPS-induced damage to H9c2 cells, causing a marked decrease in the levels of inflammatory factors as well as reactive oxygen species (ROS). Furthermore, K22 was also shown to be non-mutagenic in the Ames test. Overall, our findings suggest that K22 may be a promising drug lead as a Keap1-Nrf2 PPI inhibitor for myocarditis treatment. [ABSTRACT FROM AUTHOR]

Coumarins and chalcones are compounds widely found in plants or obtained by synthetic methods which possess several biological properties including antioxidant, anti-inflammatory, and antitumor effects. A series of coumarin-chalcone hybrids were synthesized to improve their biological actions and reduce potential adverse effects. Considering the applications of these molecules, a coumarin-chalcone hybrid [7-methoxy-3-(E)-3-(3,4,5-trimethoxyphenyl) acryloyl-2 H-chromen-2-one] (4-MET) was synthesized and the genotoxic, cytotoxic, and protective effects assessed against damage induced by different mutagens. First, in silico tools were used to predict biological activity of 4-MET which indicated a chemopreventive potential. Subsequently, the genotoxic/antigenotoxic activities of 4-MET were determined both in vitro (Ames test) and in vivo (micronucleus (MN) test and comet assay). In addition, molecular docking simulations were performed between 4-MET and glutathione reductase, an important cellular detoxifying enzyme. Our results indicated that 4-MET was not mutagenic in the Ames test; however, when co-treated with sodium azide or 4-nitroquinoline 1-oxide (4-NQO), 4-MET significantly reduced the harmful actions of these mutagens. Except for a cytotoxic effect after 120 hr treatment, 4-MET alone did not produce cytotoxicity or genotoxicity in the MN test and comet assay. Nonetheless, all treatments of 4-MET with cyclophosphamide (CPA) showed a chemoprotective effect against DNA damage induced by CPA. Further, molecular docking analysis indicated a strong interaction between 4-MET and the catalytic site of glutathione reductase. These effects may be related to (1) damage prevention, (2) interaction with detoxifying enzymes, and (3) DNA-repair induction. Therefore, data demonstrated that 4-MET presents a favorable profile to be used in chemopreventive therapies. [ABSTRACT FROM AUTHOR]

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

Rabeprazole sodium is a widely used drug for gastrointestinal disorders. Several analytical methods for identifying rabeprazole sodium and its impurities have been reported. However, the genotoxicity of rabeprazole sodium and its impurities is still unclear. Thus, it is necessary to develop analytical methods that can identify the structures of its impurities and evaluate their genotoxicity. Here, we used high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry for identifying the impurities in rabeprazole sodium enteric-coated tablets. Impurities in the samples were matched with synthesized impurities based on the exact mass and secondary mass spectrometry characteristics and then subjected to in silico analysis using the Derek and Sarah software, as well as in vitro genotoxicity evaluations. Our method successfully identified the impurities as 2-[[4-(3-methoxy propane)-3-methyl-N-oxido-2-pyridyl] methyl sulfonyl]-1H-benzimidazole (impurity I), 2-[[4-(3-methoxy propane)-3-methyl-2-pyridyl]methyl sulfonyl]-benzimidazole (impurity II), 2-[[4-(3-methoxy propane)-3-methyl-2-pyridyl] methionyl]-1H-benzimidazole (impurity III), and 2-mercapto benzimidazole (impurity IV). In silico analysis predicted that impurity III demonstrated a structural alert; thus, this impurity was evaluated for in vitro genotoxicity using the Ames test and chromosomal aberration assay. Impurity III at concentrations of 7.5–30 μg/mL had an aberration rate of over 5% with or without S-9 mix. Furthermore, impurity III at concentrations of 40–1000 μg/plate significantly increased the number of mutagenic colonies with or without S-9 mix. These results indicated that impurity III should be regulated to the limit of 0.01%. [ABSTRACT FROM AUTHOR]

The self-assembled-micelle inhibitory RNA-targeting amphiregulin (SAMiRNA-AREG) is a novel small-interfering RNA (siRNA) nanoparticle that is used for treatment of pulmonary fibrosis. We investigated the potential genotoxicity of SAMiRNA-AREG based on the guidelines published by the Organization for Economic Cooperation and Development. In the bacterial reverse mutation assay (Ames test), SAMiRNA-AREG did not induce mutations in Salmonella typhimurium TA100, TA1535, TA98, and TA1537 and Escherichia coli WP2uvrA at concentrations of up to 3000 μg/plate with or without metabolic activation. The SAMiRNA-AREG (concentrations up to 500 μg/mL) did not induce chromosomal aberrations in cultured Chinese hamster lung cells with or without metabolic activation. In the in vivo mouse bone marrow micronucleus assay, the SAMiRNA-AREG (concentrations up to 300 mg/kg body weight) did not affect the proportions of polychromatic erythrocytes and total erythrocytes, nor did it increase the number of micronucleated polychromatic erythrocytes in ICR mice. Collectively, these results suggest that SAMiRNA-AREG is safe with regard to genotoxicity such as mutagenesis or clastogenesis under the present experimental conditions. These results might support the safety of SAMiRNA-AREG as a potential therapeutic agent for pharmaceutical development. [ABSTRACT FROM AUTHOR]

Pistacia lentiscus L. is one of the most popular medicinal plants attributed to its beneficial properties on human health. However, few toxicogenetic studies have been carried out. Therefore, the aim of this study was to examine the potential genotoxic/antigenotoxic and mutagenic/antimutagenic properties of oil, ethyl acetate and ethanolic extracts of P. lentiscus L. fruits using in vitro the Ames and Umu assays, as well as in vivo micronucleus (MN) test. Extracts did not exert any significant mutagenic/genotoxic effects but provided protection against standard mutagenic and genotoxic agents including 2 nitrofluorene (2-NF) at 2.5 and 5 µg/ml; sodium azide at 5 and 10 µg/ml; 3-methylcholanthrene (3-MC) at 25 and 50 μg/ml; cyclophosphamide (CP) at 50 and 100 μg/ml; 4-nitroquinoline 1-oxide (4-NQO) at 0.05 µg/ml and 2-amino-anthracene (AA) at 0.2 µg/ml. Further, cytotoxicity and selectivity were examined on human hepatocarcinoma (HepG2), and MCF-7 breast cancer cell lines as well as a human normal-like fibroblast cell line (TelCOFS02MA) using MTT assay. Among all extracts, PF1 (ethanolic) showed the most significant selectivity index (SI) (HepG2:11.98; MCF7:4.83), which led to further investigations using an animal model. Oral administration of PF1 (125–1000 mg/kg b.w.) significantly decreased the number of micronucleated cells in CP -initiated (50 mg/kg b.w.) mice, while the number of micronucleated reticulocytes (MNRET), micronucleated polychromatic erythrocytes (MNPCE) or mitotic index (MI) were not markedly affected. Further, PF1 significantly enhanced catalase (CAT) and superoxide dismutase (SOD) activities in the livers and kidneys of these animals. The obtained results indicated the beneficial properties of P. lentiscus L. fruits for use in therapy against harmful effects of genotoxic and mutagenic agents. However, while promising it should be noted that the obtained results are preliminary and need to be confirmed prior to therapeutic use. [ABSTRACT FROM AUTHOR]

Forsythia suspensa leaves (FSL), rich in phillyrin, forsythiaside A, phillygenin, rutin, and other compounds, is a known traditional Chinese medicine (TCM). It has been effective in heat retreat and detoxification. In this study, we performed the mutagenic and teratogenic toxicity evaluation of FSL aqueous extract (FSLAE) using the bacterial reverse mutation assay (Ames test), mouse bone marrow micronucleus assay, spermatocyte chromosomal aberration assay in mice. Kunming mice and SD rats were used were for the mutagenic and the teratogenic studies, respectively. We found that FSLAE was not mutagenic and did not induce unfavorable chromosomal events. Additionally, the Ames test revealed FSLAE was not genotoxic and showed no mutagenic activity in histidine dependent strains of Salmonella typhimurium at concentrations up to 5000 μg/plate. Likewise, in vivo test revealed no induced micronucleus of mouse bone marrow or chromosome aberration in spermatocytes up to the dose of 10.00 g/kg BW. For the teratogenic evaluations, pregnant rats were treated with 1.04, 2.08, and 4.17 g/kg FSL, and fetuses were examined on the 6–15 day of pregnancy. We observed no maternal toxicity and embryotoxicity related to the treatment. Based on these in vitro and in vivo studies, we concluded the genotoxic and teratogenic safety of FSL. [ABSTRACT FROM AUTHOR]

Patent Blue V (PBV) is a water-soluble synthetic dyestuff that is used as a coloring agent in the food industry and for medical imaging in health monitoring. The aim of this study was to investigate the in vitro clastogenic, aneugenic and cytotoxic effects of PBV in human peripheral lymphocytes using micronucleus assay, comet assay, as well as plasmid DNA interaction and bacterial AMES tests. In addition to in vitro tests, the affinity of PBV against DNA was determined by molecular docking analysis in silico. PBV produced significant MN formation only at high doses and longer treatment time, however, it did not significantly affect the nuclear division index (NDI). Furthermore, PBV was unable to cause DNA single-strand breaks and significant oxidative damage on the pBR322 plasmid DNA and it didn't reverse the harmful effects caused by the clastogenic treatment of UV + H2O2 on plasmid DNA. In the Ames test, no significant increase was detected in the number of revertant colonies of mutant strains, TA98 and TA100, following PBV treatment. No significant molecular interaction between B-DNA and PBV occured in molecular docking simulations. In conclusion, PBV had no significant genotoxic and cytotoxic effects in this study. However, considering that the information intensity related to the genotoxic effects of PBV in the literature is still insufficient, reports of further studies with different genotoxicity endpoints will be needed to elucidate the exact genotoxic feature. [ABSTRACT FROM AUTHOR]

Due to its calcium-rich and diverse multimineral profile, Aquamin (derived from the red seaweed Lithothamnion sp.) is used globally as a dietary food supplement. Published reports on the genetic and prenatal developmental toxicity of Lithothamnion sp. do not exist. In accordance with the standardized protocols set by the Ministry of Health of the People's Republic of China (GB-15193), the following studies were performed: the Ames test, the mammalian erythrocyte micronucleus test, the mammalian spermatocyte chromosome test, and prenatal developmental toxicity testing. The results showed that Lithothamnion sp. did not induce a significant increase in the following: revertant colony numbers for Salmonella typhimurium strains TA 97, 98, 100, 102 and 1535; frequency of micronucleated polychromatic erythrocytes (MNPCE); spermatocyte chromosomal aberration rate. In the prenatal developmental toxicity study, no mortality, no abnormal changes in behavior and activities, and the absence of toxic symptoms and abnormalities in macroscopic autopsy were observed in each dam/all pups. Compared to the negative control group, Lithothamnion sp. at all tested doses had no effects on body weight gain, number of corpora lutea and implantations, fetal body weight and length, external, visceral and skeletal malformations. In conclusion, Lithothamnion sp. did not cause genetic toxicity. Furthermore, the prenatal developmental toxicity no observed adverse effect level (NOAEL) was determined to be greater than 2000 mg/kg.bw. [ABSTRACT FROM AUTHOR]

Pedunculagin (PD), an ellagitannin found in different plant species, possesses several pharmaceutical properties, including antitumor, antioxidant, gastroprotective, hepatoprotective, and anti-inflammatory properties. However, the effects of PD alone on DNA remain to be determined. The aim of this study was to investigate the potential cytotoxic, genotoxic, and antigenotoxic activities of PD isolated from Plinia cauliflora seeds using in silico and in vitro assays. To elucidate the biological activities of PD, in silico tools indicative of antioxidant, antineoplastic, and chemopreventive activities of PD were used. Subsequently, the mutagenic/antimutagenic effects of PD were later assessed using bacteria with the Ames test, and the cytotoxic, genotoxic, and antigenotoxic effects utilizing human lymphocytes as evidenced by trypan blue exclusion test and CometChip assay. In silico analysis indicated potential antioxidant, chemopreventive, free radical scavenger, and cytostatic activities of PD. In the Ames test, PD was found to be not mutagenic; however, this plant component protected DNA against damage-mediated by mutagens 4-nitroquinoline-1-oxide and sodium azide. Regarding human lymphocytes, PD alone was cytotoxic and genotoxic; however, it also reduced DNA damage induced by doxorubicin at co- and post-treatment. In conclusion, PD showed genotoxic, antigenotoxic and cytotoxic effects in human lymphocytes and antimutagenic effects in bacteria. [ABSTRACT FROM AUTHOR]

Recycling residual sludge in drinking water treatment plants (DWTPs) may release excessive heavy metals and organic matter, which are substances of concern because of their toxic and carcinogenic potential. The aim of this study was to investigate potential genotoxic, cytotoxic, and mutagenic effects of recycled residual sludge in terms of quality of water in potable water works. Genotoxic effects of reusing residual sludge were evaluated using: the Ames test, sperm abnormality test in mice, micronucleus assay, comet assay, and single-cell gel electrophoresis assay. The results of the Ames assay show that the disinfected water sample displays bacteriostasis at a dose of 7 L/dish regardless of treatment styles, but mutagenicity ratio (MR) < 2 can still be judged as negative. The micronucleus rates of conventional treatment were slightly genotoxic but only at 4 and 40 L/kg·bw, whereas micronucleus rates of filtered water and disinfectant from the recycling process were negative in all of the dose groups. The levels of DNA damage that are caused by different treatment processes were equivalent. Reusing residual sludge for DWTPs did not contribute to the release of genotoxic or mutagenic compounds, but it did have a remarkable effect on saving the drug dose and increasing drinking water yield. Thus, reusing residual sludge for DWTPs should be widely recommended. [ABSTRACT FROM AUTHOR]

Ocotea minarum (Nees & Mart.) Mez., a native species of Brazil, is used in the treatment of infections and oxidative stress; however, there is no scientific evidence of its toxicological characteristics. We assessed the cytotoxicity, mutagenic activity, and acute oral toxicity of the aqueous extract isolated from O. minarum leaves. The cytotoxicity of this extract was evaluated in tumour and non-tumour cell lines, while the Ames test with a Salmonella Typhimurium was used to determine the mutagenic activity. Wistar rats received a single 2 g/kg dose as part of an LD50 toxicity assessment. Our results showed that the aqueous extract of O. minarum leaves did not present cytotoxic and mutagenic properties and was not toxic, with an LD50 greater than 2 g/kg. Therefore, the O. minarum extracts are pharmacologically safe and can continue to be investigated for the development of new drugs and herbal medicines. [ABSTRACT FROM AUTHOR]

Xylan is the second most abundant polysaccharide group in plants and has a wide variety of food and pharmaceutical applications. However, little information on the safety assessment of extracted xylan as dietary supplement is available. As part of a comprehensive toxicological assessment, this study examined the potential toxicity of xylan extracted from sugarcane bagasse by three genotoxicity studies (Ames test, in vivo mice bone marrow micronucleus test, and mice sperm abnormality test) and a teratogenicity study in rats. In the Ames test, xylan showed no mutagenic activity on histidine dependent strains of Salmonella typhimurium at concentrations up to 5000 μg/plate; results of the in vivo mice bone marrow micronucleus test and mice sperm abnormality test indicated no significant effect on sperm morphology and micronucleus rate of polychromatic erythrocytes in mice at doses up to 5 g/kg body weight. In the teratogenicity study, a total of 60 pregnant rats were exposed to 10, 5, and 2.5% xylan in diet, from gestation days 7 to 16, and the no-observed-adverse-effect levels (NOAEL) of xylan was determined to be 9.8 g/kg body weight. The safe dose of xylan for human was estimated to be 98 mg/kg/day (i.e., 6.86 g/day for a 70-kg person), using a 100-fold safety factor. Taken together, results of this study indicated that xylan is practically nontoxic in terms of potential dietary consumption by humans in food or as a dietary supplement. [ABSTRACT FROM AUTHOR]

The screening of aqueous extract of Clerodendrum serratum revealed its broad-spectrum antimicrobial potential against Gram positive, Gram negative bacteria, and yeast. Optimizing the extraction strategies, revealed 15% concentration of aqueous extract prepared at 40 °C by extracting for 40 min, as optimum parameters and its statistical optimization by Box–Behnken design led to 1.16–1.35 folds enhancement in activity. Organic solvent extraction further improved the activity where methanol proved to be the best organic extractant which was effective against all the 13 pathogens tested with inhibition zone ranging from 14 to 32 mm. Minimum Inhibitory Concentration (MIC) study endorsed the methanolic extract to be the best organic extractant, as it showed the lowest MIC (0.5–10 mg/ml) in comparison to aqueous extract (1–10 mg/ml) as well as Partially Purified Phytoconstituents i.e., flavonoids (1–5 mg/ml), diterpenes (5–10 mg/ml) and cardiac glycosides (5–10 mg/ml). All these were found to be biosafe in both In-vitro (Ames and MTT assay) and In-vivo toxicity studies. Acute oral toxicity testing of flavonoids (2000 mg/ml) on Wistar rats did not reveal any significant change in relative organ weight, biochemical, hematological parameters and organs' architecture in comparison to control. Antiproliferative potential of flavonoids against human cancerous cell lines i.e., HeLa, HCT-15, and U87-MG, further increase the importance of this plant as a promising candidate for drug development. The overall study justified the medicinal importance of this plant. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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

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. [ABSTRACT FROM AUTHOR]

The aim of this interdisciplinary research project in North Rhine-Westphalia (NRW), Germany, entitled "Elimination of pharmaceuticals and organic micropollutants from waste water" involved the conception of cost-effective and innovative waste-water cleaning methods. In this project in vitro assays, in vivo assays and chemical analyses were performed on three municipal waste-water treatment plants (WWTP). This publication focuses on the study of the in vitro bioassays. Cytotoxic, estrogenic, genotoxic and mutagenic effects of the original as well as enriched water samples were monitored before and after wastewater treatment steps using MTT and PAN I, ER Calux and A-YES, micronucleus and Comet assays as well as AMES test. In most cases, the measured effects were reduced after ozonation, but in general, the biological response depended upon the water composition of the WWTP, in particular on the formed by-products and concentration of micropollutants. In order to be able to assess the genotoxic and/or mutagenic potential of waste-water samples using bioassays like Ames test, Comet assay or micronucleus test an enrichment of the water sample via solid-phase extraction is recommended. This is in agreement with previous studies such as the "ToxBox"-Project of the Environmental Agency in Germany. [ABSTRACT FROM AUTHOR]

The increasing medical and food applications of silver nanoparticles (AgNPs) raise concerns about their safety, including the potential health consequences of human exposure. Previous studies found that AgNPs were negative in the Ames test due to both their microbicidal activity and the inability of nanoparticles to penetrate bacterial cell walls. Thus, the mutagenicity of AgNPs is still not completely clear, though they do induce chromosome damage, as suggested by many previous genotoxicity studies. In this study, whole-genome sequencing (WGS) was used to analyze the mutagenicity of AgNPs in mouse lymphoma cells expanded from single-cell clones. The cells were treated with AgNPs, 4-nitroquinolone-1-oxide (4-NQO) as the positive control, and vehicle controls. Both AgNPs and 4-NQO significantly increased mutation frequencies over their concurrent controls by 1.12-fold and 4.89-fold with mutation rates at 4-fold and 130-fold, respectively. AgNP-induced mutations mainly occurred at G:C sites with G:C > T:A transversions, G:C > A:T transitions, and deletions as the most commonly observed mutations. AgNPs also induced higher fold changes in tandem mutations. The results suggest that the WGS mutation assay conducted here can detect the low-level mutagenicity of AgNPs, providing substantial support for the use of the WGS method as a possible alternative assay with respect to the mutagenic assessment of nanomaterials. [ABSTRACT FROM AUTHOR]

The influence of the UV–Vis radiation on the toxicity of agomelatine, loxapine, clozapine and tiapride was studied. The phototransformation procedure was conducted with the use of simulated solar radiation. In the case of each compound irradiation time necessary to decompose half of the initial concentration was chosen. The embryotoxicity and acute toxicity were evaluated using zebrafish (Danio rerio) embryos and larvae. The mutagenicity assay was done with the use of a micro-plate Ames test. Generally, the embryotoxicity decreased after the irradiation procedure. The obtained results showed that tiapride is the least toxic compound to zebrafish, however, its toxicity toward larvae increases after the irradiation. Similarly, the mutagenic potential of the mixture of tiapride photoproducts is higher than in the case of parent compound. The phototransformation of loxapine resulted in the change of the acute toxicity profile and increased the rate of reverse mutations in the Ames test. Oppositely, the irradiation of agomelatine caused the decrease of mutagenic potential and acute toxicity was also lower in the postirradiated mixture. The phototransformation of clozapine did not alter the mutagenicity and decreased the acute toxicity to the zebrafish larvae. In silico calculations showed a satisfactory prediction ability in some instances, especially in the case of mutagenic potential of the tiapride phototransformation products. [ABSTRACT FROM AUTHOR]

Strychnos pseudoquina is a plant species whose stem bark is used as bitter tonic beverage. The phytochemical analysis, as well as quantification of phenolic constituents and antioxidant activity of ethanolic extracts from S. pseudoquina stem bark, and leaves were conducted. The extracts were tested for mutagenicity (Ames test) and DNA-damaging activity (Plasmid Cleavage test). Leaves recorded the largest amount of flavonoids. The performed high-performance liquid chromatography (HPLC) showed flavonoids such as isorhamnetin and strychnobiflavone (phytochemical markers of the investigated species) in stem barks, but not in leaves. The proanthocyanidin content and antioxidant activity were significantly higher in stem barks than in leaves. Stem bark and leaf extracts presented mutagenic activity against TA98 and TA100 strains with, and without, metabolic activation (S9). The Plasmid Cleavage test did not indicate DNA-damaging activity. Our results suggest that extracts deriving from S. pseudoquina should be used with extreme caution, mainly the stem bark extract, which is widely used in folk medicine. [ABSTRACT FROM AUTHOR]

Commercially processed meat and fish products are common sources of human exposure to chemical food mutagens. In this study, we investigated the mutagenic potential of 20 different commercially processed meat and fish products (7 product types with 2–3 lots of each), along with the presence of four principal polycyclic aromatic hydrocarbons (PAHs) (benzo[a]pyrene [BaP], benzo[b]fluoranthene [BbF], benzo[a]anthracene [BaA] and chrysene [CHR]) in them. Sample extraction was carried out by an accelerated solvent extraction method, while the concentrations of the 4 PAHs were determined by gas chromatography-tandem mass spectrometry (GC-MS/MS). The mutagenic potential of food extracts was assessed by the standard plate incorporation assay (Ames test) using two strains of Salmonella typhimurium (TA 100 and TA 98) both in the presence and the absence of metabolic activation (S9-mix). The results show that in the majority of food items investigated, PAH levels were below the limit of quantification (0.78 μg/kg), except for smoked fish (0.8–15 μg/kg for the 3 lots), one batch of which even exceeded the maximum limits for both the sum of the 4 PAHs (44 μg/kg vs. 30 μg/kg) and BaP (8.2 μg/kg vs. 5.0 μg/kg). Furthermore, all three batches of smoked fish were also found to be mutagenic on both strains of Salmonella, both in the presence and the absence of metabolic activation. Overall, the data from both assays were in a fairly good agreement with one another, suggesting that PAHs are major contributors to mutagenicity of processed food products and the set maximum levels for PAHs are usually protective against food mutagenicity, although food samples harboring PAHs at levels approaching the maximum limits may exhibit mutagenic potential. Since the number of samples investigated was relatively small, further studies are warranted to verify the conclusions. [ABSTRACT FROM AUTHOR]

Chalcones are chemically defined as α,β–unsaturated ketones with a 1,3-diphenyl-2-propen-1-one nucleus. These compounds occur naturally in plants and are considered precursors of flavonoids. Given that evaluating genetic toxicology tests is essential in investigating the safe use and chemopreventive potential of different natural and synthetic compounds, this study aimed to assess the genotoxic, cytotoxic, antigenotoxic, and anticytotoxic activity of the chalcone 1E,4E-1-(4-chlorophenyl)-5-(2,6,6-trimethylcyclohexen-1-yl)penta-1,4-dien-3-one (CAB7β). The CAB7β was synthesized via Claisen–Schmidt reaction. The Ames test was applied using the co-treatment model as well as a micronucleus assay of mouse bone marrow with co-, pre- and post-treatment models. Our results indicate no genotoxic effect for CAB7β in any of the tests applied. At all the concentrations used, CAB7β showed a significant DNA protective effect against the mutagenic action of 4-nitroquinoline-1-oxide and sodium azide according to the Ames test, and against doxorubicin in the co-, pre- and post-treatment models of the micronucleus assay. CAB7β alone displayed cytotoxic activity in the micronucleus test. At concentrations of 12,5 and 50 µg/plate, CAB7β showed a moderate cytotoxic profile only in Salmonella typhimurium strain TA98. However, an anticytotoxic effect was observed against S. typhimurium strain TA100 for all the concentrations tested and during co-, pre- and post-treatment in the micronucleus assay. It was concluded that CAB7β exhibited a slightly cytotoxic effect in S. typhimurium strain TA98 and significant antigenotoxic and anticytotoxic effects in cells of mouse, making it a promising candidate in chemoprevention and possibly in the development of new cancer treatments. [ABSTRACT FROM AUTHOR]

Pinus wallichiana, silver nanoparticles, Ames assay, acute toxicity assay, antibacterial, antifungal. [Extracted from the article]

A method for combining statistical-based QSAR predictions of two or more binary classification models is presented. It was assumed that all models were independent. This facilitated the combination of positive and negative predictions using a quantitative weight of evidence (qWoE) procedure based on Bayesian statistics and the additivity of the logarithms of the likelihood ratios. Previous studies combined more than one prediction but used arbitrary strengths for positive and negative predictions. In our approach, the combined models were validated by determining the sensitivity and specificity values, which are performance metrics that are a point of departure for obtaining values that measure the weight of evidence of positive and negative predictions. The developed method was experimentally applied in the prediction of Ames mutagenicity. The method achieved a similar accuracy to that of the experimental Ames test for this endpoint when the overall prediction was determined using a combination of the individual predictions of more than one model. Calculating the qWoE value would reduce the requirement for expert knowledge and decrease the subjectivity of the prediction. This method could be applied to other endpoints such as developmental toxicity and skin sensitisation with binary classification models. [ABSTRACT FROM AUTHOR]

The U.S. Air Force (USAF) has pursued development of alternative fuels to augment or replace petroleum-based jet fuels. Hydroprocessed esters and fatty acids (HEFA) renewable jet fuel is certified for use in commercial and USAF aircraft. HEFA feedstocks include camelina seed oil (Camelina sativa, HEFA-C); rendered animal fat (tallow, HEFA-T); and mixed fats and oils (HEFA-F). The aim of this study was to examine potential toxic effects associated with HEFA fuels exposures. All 3 HEFA fuels were less dermally irritating to rabbits than petroleum-derived JP-8 currently in use. Inhalation studies using male and female Fischer-344 rats included acute (1 day, with and without an 11-day recovery), 5-, 10- or 90-day durations. Rats were exposed to 0, 200, 700 or 2000 mg/m3 HEFA-F (6 hr/day, 5 days/week). Acute, 5 – and 10-day responses included minor urinalysis effects. Kidney weight increases might be attributed to male rat specific hyaline droplet formation. Nasal cavity changes included olfactory epithelial degeneration at 2000 mg/m3. Alveolar inflammation was observed at ≥700 mg/m3. For the 90-day study using HEFA-C, no significant neurobehavioral effects were detected. Minimal histopathological effects at 2000 mg/m3 included nasal epithelium goblet cell hyperplasia and olfactory epithelium degeneration. A concurrent micronucleus test was negative for evidence of genotoxicity. All HEFA fuels were negative for mutagenicity (Ames test). Sensory irritation (RD50) values were determined to be 9578 mg/m3 for HEFA-C and greater than 10,000 mg/m3 for HEFA-T and HEFA-F in male Swiss-Webster mice. Overall, HEFA jet fuel was less toxic than JP-8. Occupational exposure levels of 200 mg/m3 for vapor and 5 mg/m3 for aerosol are recommended for HEFA-based jet fuels. [ABSTRACT FROM AUTHOR]

Over the past years, the European Food Safety Authority (EFSA) released to the public domain several databases, with the main objectives of collecting and storing hazard data on the substances considered in EFSA's risk assessment and secondly to serve as a basis for further development of in silico tools such as quantitative structure–activity relationship (QSAR) models. In this work, we evaluated the ability of freely available QSAR models to estimate genotoxicity and carcinogenicity properties and their possible use for screening purposes on three different EFSA's databases. With an accuracy close to 90%, the results showed good capabilities of QSAR models to predict genotoxicity in terms of bacterial reverse mutation test, while statistics for in vivo micronucleus test are not satisfactory (accuracy in the predictions close to 50%). Interestingly, results on the carcinogenicity assessment showed an accuracy in prediction close to 70% for the best models. In addition, an example of the potential application of in silico models is presented in order to provide a preliminary screening of genotoxicity properties of botanicals intended for use as food supplements. [ABSTRACT FROM AUTHOR]

Some of the chemicals in materials used for packaging food may leak into the food, resulting in human exposure. These include so-called Non-intentionally Added Substances (NIAS), many of them being unidentified and toxicologically uncharacterized. This raises the question of how to address their safety. An approach consisting of identification and toxicologically testing all of them appears neither feasible nor necessary. Instead, it has been proposed to use the threshold of toxicological concern (TTC) Cramer class III to prioritise unknown NIAS on which further safety investigations should focus. Use of the Cramer class III TTC for this purpose would be appropriate if amongst others sufficient evidence were available that the unknown chemicals were not acetylcholinesterase inhibitors or direct DNA-reactive mutagens. While knowledge of the material and analytical chemistry may efficiently address the first concern, the second could not be addressed in this way. An alternative would be use of a bioassay capable of detecting DNA-reactive mutagens at very low levels. No fully satisfactory bioassay was identified. The Ames test appeared the most suitable since it specifically detects DNA-reactive mutagens and the limit of biological detection of highly potent genotoxic carcinogens is low. It is proposed that for a specific migrate, the evidence for absence of mutagenicity based on the Ames test, together with analytical chemistry and information on packaging manufacture could allow application of the Cramer class III TTC to prioritise unknown NIAS. Recommendations, as well as research proposals, have been developed on sample preparation and bioassay improvement with the ultimate aim of improving limits of biological detection of mutagens. Although research is still necessary, the proposed approach should bring significant benefits over the current practices used for safety evaluation of food contact materials. [ABSTRACT FROM AUTHOR]

Assessing chemical toxicity is a multidisciplinary process, traditionally involving in vivo, in vitro and in silico tests. Currently, toxicological goal is to reduce new tests on chemicals, exploiting all information yet available. Recent advancements in machine learning and deep neural networks allow computers to automatically mine patterns and learn from data. This technology, applied to (Q)SAR model development, leads to discover by learning the structural-chemical-biological relationships and the emergent properties. Starting from Toxception, a deep neural network predicting activity from the chemical graph image, we designed SmilesNet, a recurrent neural network taking SMILES as the only input. We then integrated the two networks into C-Tox network to make the final classification. Results of our networks, trained on a ~20K molecule dataset with Ames test experimental values, match or even outperform the current state of the art. We also extract knowledge from the networks and compare it with the available mutagenic structural alerts. The advantage over traditional QSAR modelling is that our models automatically extract the features without using descriptors. Nevertheless, the model is successful if large numbers of examples are provided and computation is more complex than in classical methods. [ABSTRACT FROM AUTHOR]

Myrciaria dubia is a native plant from the Amazon region which produces red-purplish fruit rich in antioxidant compounds such as ascorbic acid, carotenoids, and phenolic. M. dubia fruit is used to prepare juices considered to possess high nutritional content providing health benefits. The aim of this study was to examine the ability of M. dubia juice to protect DNA against genomic instability induced by sub-acute ethanol consumption attributed to oxidative stress. Mice were treated for 28 days with juice at 25% and 50% diluted in distilled water or with the diluted combination juice plus ethanol (5 g/kg). The genotoxic/antigenotoxic and mutagenic/antimutagenic effects were assessed using comet assay in blood, liver, and kidney and micronucleus (MN) test with bone marrow. In addition, the mutagenicity was also evaluated using Salmonella/microsome assay. Phytochemical compounds were determined using HPLC/PDA/MS/MS. The juice did not induce genotoxic effects in blood, kidney, and liver cells at both doses. In combination with ethanol, the juice reduced the alcohol-mediated DNA damage in all tissues analyzed. Further, the juice did not produce mutagenic effects and decreased mutagenicity induced by ethanol in the bone marrow. The anthocyanins were major compounds detected by HPLC/PDA/MS/MS, which modulated genotoxic and mutagenic effects initiated by ethanol and at least in part appeared responsible for the observed antigenotoxic and antimutagenic effects of M. dubia juice. [ABSTRACT FROM AUTHOR]

A major challenge in the safety assessment of food contact materials (FCM) is the evaluation of unknown non-intentionally added substances (NIAS). Even though consumer exposure levels may be quantitatively low, these substances are considered to be of high toxicological concern if they act as DNA reactive mutagens. From a safety assessment perspective, it is therefore important to detect their presence in FCM migrates. The present study applied the Ames MPF assay to assess the mutagenicity of migrates obtained from 30 food contact material samples out of 3 categories: plastics, composite materials and coatings. As a food simulant, 95% ethanol (EtOH) had a superior performance to less volatile simulants when evaluating recovery rates of representative model substances in different volatility categories. To monitor possible interference of the FCM matrix with Ames MPF results, migrates were spiked with reference substances and recovery rates were established. Out of 30 samples tested, two caused significant inhibition of revertant formation in the presence of the spiking control. Overall detection limits of the applied test method were estimated by determination of the lowest effective concentrations (LEC) for 10 Ames-positive substances. Even though the current limits of detection are not sufficient to entirely fulfil regulatory and safety requirements, three out of 30 FCMs showed evidence of dose-dependent effects in the Ames MPF assay. Overall, the data obtained supported the relevance of testing FCM migrates for DNA reactive contaminants and showed the value of the Ames MPF assay for the safety assessment of FCMs. [ABSTRACT FROM AUTHOR]

In this study, the potential genotoxic and mutagenic effects of cocoa industry wastewater was assessed using SOS Chromo and Ames fluctuation tests, respectively. The physico-chemical analyses of the wastewater were also carried out. The SOS Chromo and Ames fluctuation tests results showed that cocoa industry wastewater can induce different levels of DNA damage and mutagenic potential at the different concentrations used in this study. Salmonella typhimurium TA98 was more sensitive than TA100 strain, indicating frameshift mutation as a possible mechanism of DNA damage. High concentrations of Pb, Cd, Cr, Ni and Cu in the tested sample were possibly responsible for the observed mutagenicity and genotoxicity. [ABSTRACT FROM AUTHOR]

Triclosan has been used in a large number of consumer products and concerns have been raised over regarding potential genotoxicity. However, the genotoxicity of triclosan has not been assessed in normal human cells. The aim of this study was to examine the potential genotoxicity using the comet assay and micronucleus (MN) test to detect DNA damage and chromosomal breakage attributed to triclosan in human keratinocyte HaCaT and hepatic L02 cells. The concentrations of triclosan selected for the comet assay and MN test were based upon preliminary results from cytotoxicity testing in order to reduce cytotoxic effects. The mutagenicity of triclosan was assessed in Salmonella reverse mutation assay (Ames test). Results of comet assay showed that 5, 7.5 or 10 μM triclosan did not markedly affect olive tail moment (OTM) in HaCaT and L02 cells. In addition, no significant alterations in MN frequency were found in cells treated with triclosan. Further, treatment with 10 μg/plate triclosan produced inhibitory effects in bacterium using Ames test, while 1 and 0.1 μg/plate triclosan did not markedly affect the number of colonies or mutant frequencies of Salmonella strains. Taken together, triclosan did not cause DNA and chromosomal damage in HaCaT and L02 cells and did not induce gene mutations in Salmonella strains under our experimental conditions. [ABSTRACT FROM AUTHOR]

Pinocembrin (1) and cardamonin (2) from Sozuku showed a suppressive effect on umu gene expression of SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen furylfuramide. Compounds 1 and 2 suppressed 52% and 36% of SOS-inducing activity at a concentration of 0.20 μmol/mL. The ID50 value of 1 was 0.18 μmol/mL. These compounds showed the suppression of 2-amino-3,4-dimethylimidazo-[4,5-f]quinolone (MeIQ) and UV irradiation-induced SOS response. Pinostrobin (3) and 5,7-dimethoxyflavanone (4), methyl ethers of 1, showed similar activity to 1 against MeIQ-induced SOS response, but that of furylfuramide and UV irradiation were decreased. On the other hand, compounds 1–4 did not show the suppression of activated MeIQ-induced SOS response. Furthermore, compounds 1–4 showed potent antimutagenic activity against MeIQ mutagenesis in Ames test using the S. typhimurium TA100 and TA98 strains. [ABSTRACT FROM AUTHOR]

Context: Influenza is a severe, life-threatening viral disease that can be prevented by vaccination. However, the anti-influenza human vaccine failed to show the required efficacy both in infants under 5 years old and in the elder population, who are among those with the highest risk of developing severe complications after influenza infection. Therefore, it is of high importance to improve the vaccine efficacy and ensure its safety in these susceptible populations. GK-1, a novel 18-aa peptide adjuvant, has been proved to increase the immunogenicity of the human influenza vaccine in both young and aged mice. Objective: A preclinical study of the toxicity profile of GK-1 following the World Health Organization guidelines to support its use was herein conducted. Material and methods: GK-1 was synthetically produced following Good Manufacturing Practices. The toxicological evaluation of GK-1 peptide was performed in rats after repeated dose-ranging trials by the subcutaneous route. The mutagenic potential of GK-1 was assessed by the micronucleus, chromosomal aberration, and Ames tests, in accordance with OECD Guidelines. Results: GK-1 did not show toxic effects at doses up to 12.5mg/kg, corresponding to 25 times the dose intended for human use. No indications of mutagenic potential were observed. GK-1 after dermal administration was well tolerated locally. Conclusion: The efficacy of GK-1 to improve influenza vaccine protection, along with the absence of toxicity and mutagenicity, as reported herein, support the evaluation of this peptide in a clinical trial as a novel adjuvant for human use. [ABSTRACT FROM AUTHOR]

Non-intentionally added substances (NIAS) are chemical impurities which can migrate from packaging materials (FCM) into food. Safety assessment of NIAS is required by European law, but currently there is no comprehensive testing strategy available. In this context, one key element is to get insight on the potential presence of genotoxic NIAS in FCM migrates. This raises questions about the limit at which genotoxins can be detected in complex mixtures such as FCM migrates, and if such limits of detection (LOD) would be compatible with safety. In this context, the present review assesses the suitability of the Ames assay to address genotoxicity of FCM migrates. Lowest effective concentrations of packaging-related and other chemicals in test media were retrieved from scientific literature and used as surrogates of LODs to be benchmarked against a value of 0.01 mg kg−1 (10 ppb) in migrates. This is a pragmatic threshold used in FCM safety evaluation to prioritise substances requiring proper identification and risk assessment. The analysis of the data shows that only potent genotoxins can theoretically be detectable at a level of 0.01 mg kg−1 in migrates or food. Only a minority (10%) of genotoxic chemicals reported to be associated with FCMs could be picked up at a level of 0.01 mg kg−1 or lower. Overall, this review shows that the Ames test in its present form cannot be used as standalone method for evaluating the genotoxic potential of FCM migrates, but must be used together with other information from analytical chemistry and FCM manufacturing. [ABSTRACT FROM AUTHOR]