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Heated tobacco products (HTPs) have emerged as an alternative to cigarettes in the last 10 years. HTPs heat rather than burn tobacco to produce a nicotine-containing aerosol. Avoiding combustion reduces the amount of harmful and potentially harmful constituents (HPHCs) in HTP aerosol compared to cigarette smoke. As heating technologies evolve, it is important to thoroughly assess new iterations to confirm that the aerosol they produce contains fewer and lower levels of HPHCs than cigarette smoke. Tobacco Heating System (THS) 3.0 is an induction heating device used with specially designed tobacco sticks. This paper describes the chemical composition, physical properties, cytotoxicity, genotoxicity, and mutagenicity of the aerosols produced by THS 3.0 using the International Organization for Standardization (ISO) intense puffing regimen for regular and menthol tobacco sticks. Targeted chemical testing and physics evaluations confirmed that >95% of the aerosol droplets are respirable with substantial HPHC yield reductions compared to cigarette smoke. A standard battery of in vitro toxicology tests was performed to assess the aerosols' cellular effects. Results from the neutral red uptake, micronucleus, and Ames assays showed that THS 3.0 aerosol fractions induced substantially less cytotoxicity and have substantially lower genotoxic and mutagenic potentials than cigarette smoke fractions. Collectively, the evidence demonstrates that inductively heated tobacco sticks used with THS 3.0 produce aerosol emissions and substantially lower levels of HPHCs similar to those with THS 2.2—a previous version of HTP with resistive heating—leading to reduced in vitro cytotoxicity and genotoxicity compared to cigarette smoke. [ABSTRACT FROM AUTHOR]

Rickettsia felis , responsible for flea-borne spotted fever, is a rising zoonotic pathogen posing an increasing global threat due to its expanding geographical distribution. The rise in antibiotic-resistant strains of this pathogen underscores the urgent need for new therapeutic interventions. This study employed a comprehensive subtractive proteomics analysis of the R. felis proteome, aiming to identify essential, non-host homologous, and pathogen-specific proteins, which were subsequently evaluated as potential new drug targets. These findings offer valuable insights into the development of therapeutic strategies against rickettsiosis. The analysis revealed 343 proteins that are non-homologous to the host, including 108 essential proteins, 25 unique metabolic pathways, and 11 distinct proteins. Out of these, 10 proteins were druggable in which two associated with virulence, and one related to resistance (succinate dehydrogenase). Through a rigorous screening process and extensive literature review, succinate dehydrogenase emerged as a promising drug target. Protein interaction partners for succinate dehydrogenase were identified using the STRING database. To further assess the functionality of succinate dehydrogenase, structure-based studies were conducted. Approximately 18,000 ZINC compounds were screened, leading to the finding of six potential inhibitors: ZINC67847806, ZINC67982856, ZINC67974679, ZINC67895371, ZINC05668040, and ZINC05670149. Absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiling confirmed that most compounds met the preferred pharmacokinetic properties, except for ZINC67895371 and ZINC67847806, which exhibited positive ames test results, and ZINC05670149, ZINC67895371, and ZINC67847806, showed hepatotoxicity. All compounds were found to be non-sensitizing to the skin. Based on these findings, further experimental validation of ZINC67974679, ZINC67982856, and ZINC05668040 is recommended. [ABSTRACT FROM AUTHOR]

Introduction: Modern oral nicotine pouch products (ONPs) are a category of oral nicotine products which contain pharmaceutical-grade nicotine, flavors, and other food-grade ingredients but no tobacco leaf. Recent reports indicate that ONPs in general do not contain (or only at minimal levels) the harmful and potentially harmful constituents (HPHCs) identified in cigarette smoke, suggesting their potential as alternative products for reducing harm from cigarette smoking. Methods: We assessed in vitro toxicological effects of eight ONPs, designated as modern oral (MO) 1 to 8 along with an ONP, an oral tobacco (snus), and a combustible cigarette market comparator using established regulatory toxicological assays including Ames, in vitro micronucleus (ivMN), and neutral red uptake (NRU) assays. Results: The MO test products 1-7 ZYN wintergreen, and General Snus white mint were negative for mutagenicity (Ames assay), genotoxicity (ivMN), and cytotoxicity (NRU). The combustible cigarette was positive in all three assays. The MO-8 test product was negative for mutagenicity; however, it was positive in the ivMN and NRU assays at concentrations either 42 to 135-fold based on the ivMN i to iv treatment schedule or 60-fold higher, respectively, when compared to combustible cigarettes. Discussion: Thus, the MO test products are likely to be less harmful than combustible cigarettes and are alternatives to cigarettes. However, understanding of long-term effects of ONPs in general requires additional research. [ABSTRACT FROM AUTHOR]

Mutations are the result of effects produced by mutagenic agents, which may be responsible for the generation of chronic-degenerative diseases such as cancer. The aim of this work was to detect and characterize compounds with antimutagenic activity in white shrimp (Litopenaeus vannamei) cephalothorax. A serial extraction was carried out where different organic solvents, and its biological activity was evaluated. The antioxidant capacity was evaluated by DPPH and ABTS assay, where the methanol and water-soluble extracts had the highest antioxidant capacity. Antimutagenicity was evaluated by the Ames test, where the methanol-soluble extract showed the highest antimutagenic activity. The methanolic extract was fractionated by open column chromatography, the fractions obtained were determined antioxidant and antimutagenic activity and the ability to protect DNA from oxidative damage. Fraction H27 provided DNA the highest protection against the damage caused by H2O2. Partial chemical-characterization of fraction H27 suggests the presence of aromatic, hydroxylated, unsaturated compounds. [ABSTRACT FROM AUTHOR]

In recent years, nitrosamine impurities in pharmaceuticals have been subject to intense regulatory scrutiny, with nitrosamine drug substance-related impurities (NDSRIs) treated as cohort of concern impurities, regardless of predicted mutagenic potential. Here, we describe a case study of the NDSRI N-nitroso-hydrochlorothiazide (NO-HCTZ), which was positive in the bacterial reverse mutation (Ames) test but is unstable under the test conditions, generating formaldehyde among other products. The mutagenic profile of NO-HCTZ was inconsistent with that expected of a mutagenic nitrosamine, exhibiting mutagenicity in the absence of metabolic activation, and instead aligned well with that of formaldehyde. To assess further, a modified Ames system including glutathione (3.3 mg/plate) to remove formaldehyde was developed. Strains used were S. typhimurium TA98, TA100, TA1535, and TA1537, and E. coli WP2 uvrA/pKM101. In this system, formaldehyde levels were considerably lower, with a concomitant increase in levels of S-(hydroxymethyl)glutathione (the adduct formed between glutathione and formaldehyde). Upon retesting NO-HCTZ in the modified system (1.6–5000 µg/plate), a clear decrease in the mutagenic response was observed in the strains in which NO-HCTZ was mutagenic in the original system (TA98, TA100, and WP2 uvrA/pKM101), indicating that formaldehyde drives the response, not NO-HCTZ. In strain TA1535, an increase in revertant colonies was observed in the modified system, likely due to a thiatriazine degradation product formed from NO-HCTZ under Ames test conditions. Overall, these data support a non-mutagenic designation for NO-HCTZ and demonstrate the value of further investigation when a positive Ames result does not align with the expected profile. [ABSTRACT FROM AUTHOR]

Since 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]

The contamination of ground and surface waters with per- and polyfluoroalkyl substances (PFASs) is of major concern due to their potential adverse effects on human health. The carbon–fluorine bond makes these compounds extremely stable and hardly degradable by natural processes. Therefore, methods for PFAS removal from water are desperately needed. In this context, plasma treatment of water has been proposed as an effective method with reported removal rates exceeding 90%. However, the high reactivity of plasma discharge results in the formation of many reactive species, like radicals, ozone, or even solvated electrons, which lead to a complex reaction cascade and, consequently, to the generation of a wide variety of different chemical products. The toxicological properties of these PFAS breakdown products are largely unknown. The present study focuses on a toxicological assessment of PFAS-containing plasma-treated water samples. Aqueous solutions of long-chain perfluorooctanesulfonate (PFOS) were treated with various plasma-atmospheric regimes. Subsequently, plasma-treated water samples were subjected to in vitro bioassays. Cytotoxicity and genotoxicity were assessed with the MTS assay using human liver cells (HepG2) and the Ames MPFTM assay using Salmonella Typhimurium strains. Our results demonstrate varying cyto- and genotoxic properties of water containing PFAS breakdown products depending on the atmosphere present during plasma treatment. Based on the results of this study, the atmosphere used during plasma treatment affects the toxicological properties of the treated sample. Further studies are therefore needed to uncover the toxicological implications of the different treatment parameters, including the PFAS starting compound, the atmosphere during treatment, as well as the quantity of plasma energy applied. [ABSTRACT FROM AUTHOR]

Abstract Background Although the in silico predictive ability of the Ames test results has recently made remarkable progress, there are still some chemical classes for which the predictive ability is not yet sufficient due to a lack of Ames test data. These classes include simple heterocyclic compounds. This study aimed to investigate the mutagenicity and structure-mutagenicity relationships for some heterocycles in the Ames test. In the present study, we selected 12 quinoline analogues containing one or two nitrogen atoms in the naphthalene ring and 12 indole analogues containing one to three nitrogen atoms in the indole ring, without any side moiety. Results The Ames test was performed with five standard bacterial strains (TA100, TA1535, TA98, TA1537, and WP2uvrA) using the pre-incubation method with and without rat liver S9. Five quinoline and two indole analogues were mutagenic. Among the five quinoline analogues, four were mutagenic in the presence of S9 mix with TA100, whereas cinnoline was mutagenic in the absence of S9 mix with TA1537. Among the two indole analogues, indazole was mutagenic in the presence and absence of S9 mix with WP2uvrA and 4-azaindole was mutagenic in the absence of S9 mix with TA1537. The mechanisms underlying the induction of mutagenesis appear to differ between quinoline and indole analogues. In addition, we performed in silico analysis of the mutagenicity of all these analogues using DEREK Nexus 6.1.1 (Lhasa Limited) and GT_EXPERT from CASE Ultra 1.8.0.5 (MultiCASE Inc.) as knowledge-based models and GT1_BMUT from CASE Ultra 1.8.0.5 (MultiCASE Inc.) as a statistical-based model. The knowledge-based model showed low sensitivity for both the quinoline and indole analogues (DEREK Nexus and GT_EXPERT: 20% for quinolines and 0% for indoles). Conversely, the statistical model showed high sensitivity (100% for both quinolines and indoles) and low specificity (43% for quinolines and 10% for indoles). Conclusion Based on the Ames test results, we proposed structural alerts noting that quinoline analogues were mutagenic when they had nitrogens in any of the positions 2, 5, 7, or 8 in addition to 1, and indole analogues were mutagenic when they had nitrogens at positions 2 or 4 in addition to 1.

Abstract The Ames test is used worldwide to initially screen the mutagenic potential of new chemicals. In the standard Ames test, S. typhimurium strains (TA100, TA98, TA1535, and TA1537) and Escherichia coli (WP2uvrA) are treated with substances with/without cytochrome P450s (CYPs)-induced rat S9 fractions for identifying mutagens and pro-mutagens. However, many substances show completely different toxicity patterns depending on whether the liver S9 fraction belongs to rats or humans. The natural product Polygoni Multiflori Radix (PMR) can also show bacterial reverse mutation, followed by the rat or human liver S9 fraction. While PMR elicits reverse mutations in the TA1537 strain in rat liver S9 but not in human liver S9, this mechanism has not been verified yet. To explain this, the differences in metabolic enzymes compositions commonly observed between rats and humans have been implicated. This study aimed to explore the key factors that cause differences in the genotoxicity of PMR between rat and human liver S9 metabolic enzymes. The results of next-generation sequencing (NGS) analysis showed that both rat and human metabolic enzymes caused similar mutations in TA1537. However, when the metabolic enzymes in each S9 fraction were analyzed using ion mobility tandem mass spectrometry (IM-MS), rat- and human-specific enzymes were identified among the cytochrome (CYP) family, especially aryl hydrocarbon receptor (AHR)-related CYPs. These findings suggest that CYP1A1 isoforms contribute to the mechanism of PMR in the Ames test. Therefore, an in vitro Ames test might be more reliable in predicting genotoxicity for both rodents and humans. This will also help overcome the limitations of laboratory animal-based toxicity evaluations, which provide unreliable results due to interspecies differences between humans and rodents.

Background: The rose ketone β-damascone (β-Dam) elicits positive results in the in vitro micronucleus (MN) assay using human lymphocytes, but shows negative outcomes in the Ames test and combined in vivo MN and comet assays. This has led to the interpretation that the in vitro MN result is a misleading positive result. Oxidative stress has been suggested as an indirect mode of action (MoA) for in vitro MN formation, with the α, β-unsaturated carbonyl moiety of the β-Dam chemical structure expected to cause misleading positive results through this MoA. In this study, we investigated the role of oxidative stress in β-Dam-induced in vitro MN formation by co-treatment with the antioxidant N-acetyl-l-cysteine (NAC), thereby highlighting a possible link between mitochondrial dysfunction and aneugenicity. Results: β-Dam induced MN formation in both CHL/IU and BEAS-2B cells, with the response completely inhibited by co-treatment with NAC. Moreover, β-Dam induced oxidative stress-related reporter activity in the ToxTracker assay and increased reactive oxygen species levels, while decreasing glutathione levels, in BEAS-2B cells in the high-content analysis. All of these effects were suppressed by NAC co-treatment. These findings indicate that β-Dam elicits oxidative stress, which causes DNA damage and ultimately leads to MN induction. However, no significant DNA damage-related reporter activities were observed in the ToxTracker assay, nor was there an increased number of γH2AX foci in the high-content analysis. These data suggest that MN formation is not a DNA-reactive MoA. Considering recent reports of aneuploidy resulting from chromosome segregation defects caused by mitochondrial dysfunction, we investigated if β-Dam could cause such dysfunction. We observed that the mitochondrial membrane potential was dose-dependently impaired in BEAS-2B cells exposed to β-Dam. Conclusions: These findings suggest that the oxidative stress induced by β-Dam exposure may be explained through an aneugenic MoA via mitochondrial dysfunction, thereby contributing to MN formation in mammalian cells. [ABSTRACT FROM AUTHOR]

This study performed advanced toxicological assessments of the new substance AP-238 using nine 'green' in silico methods, focusing on acute toxicity, organ-specific effects, skin and eye irritation, genetic toxicity, and cardiotoxicity. A quantitative assessment of AP-238's acute toxicity (AT) was performed by predicting theoretical LD50 values for both rats and mice across different administration routes using various in silico methods. Results indicated the highest toxicity via intravenous administration in mice, with a t-LD50 of 53 mg/kg, while oral administration in rats exhibited a lower toxicity range, with t-LD50 values between 666.43 and 1838.77 mg/kg, depending on the predictive model used. The identification of toxicophores (the fragment connecting the benzene ring to the piperazine ring, including the α, β, and γ carbon atoms near the nitrogen atom) in AP-238 suggests a high likelihood of lung toxicity (61%), with additional risks to the cardiovascular (58%) and renal systems (56%), emphasizing specific molecular fragments associated with these adverse effects. Genotoxic evaluations presented a mixed view, with low to moderate probabilities of a positive Ames test, suggesting some uncertainty but generally indicating a reduced risk of genetic toxicity. Eye and skin irritation risks were deemed minimal, supported by several models with high confidence. Cardiotoxicity assessments revealed varied information on the potential effects of AP-238 on the hERG channel, with some studies suggesting a nonsignificant impact, while others indicated moderate risk, although with low reliability in the predictions. This highlights the nuanced challenges in assessing the safety of novel substances through 'green' in silico methods. [ABSTRACT FROM AUTHOR]

Background: Primary aromatic amines (PAAs) present significant challenges in the prediction of mutagenicity using current standard quantitative structure activity relationship (QSAR) systems, which are knowledge-based and statistics-based, because of their low positive prediction values (PPVs). Previous studies have suggested that PAAs are metabolized into genotoxic nitrenium ions. Moreover, ddE, a relative-energy based index derived from quantum chemistry calculations that measures the stability nitrenium ions, has been correlated with mutagenicity. This study aims to further examine the ability of the ddE-based approach in improving QSAR mutagenicity predictions for PAAs and to develop a refined method to decrease false positive predictions. Results: Information on 1,177 PAAs was collected, of which 420 were from public databases and 757 were from in-house databases across 16 laboratories. The total dataset included 465 Ames test-positive and 712 test-negative chemicals. For internal PAAs, detailed Ames test data were scrutinized and final decisions were made using common evaluation criteria. In this study, ddE calculations were performed using a convenient and consistent protocol. An optimal ddE cutoff value of -5 kcal/mol, combined with a molecular weight ≤ 500 and ortho substitution groups yielded well-balanced prediction scores: sensitivity of 72.0%, specificity of 75.9%, PPV of 65.6%, negative predictive value of 80.9% and a balanced accuracy of 74.0%. The PPV of the ddE-based approach was greatly reduced by the presence of two ortho substituent groups of ethyl or larger, as because almost all of them were negative in the Ames test regardless of their ddE values, probably due to steric hindrance affecting interactions between the PAA and metabolic enzymes. The great majority of the PAAs whose molecular weights were greater than 500 were also negative in Ames test, despite ddE predictions indicating positive mutagenicity. Conclusions: This study proposes a refined approach to enhance the accuracy of QSAR mutagenicity predictions for PAAs by minimizing false positives. This integrative approach incorporating molecular weight, ortho substitution patterns, and ddE values, substantially can provide a more reliable basis for evaluating the genotoxic potential of PAAs. [ABSTRACT FROM AUTHOR]

Background: Although the in silico predictive ability of the Ames test results has recently made remarkable progress, there are still some chemical classes for which the predictive ability is not yet sufficient due to a lack of Ames test data. These classes include simple heterocyclic compounds. This study aimed to investigate the mutagenicity and structure-mutagenicity relationships for some heterocycles in the Ames test. In the present study, we selected 12 quinoline analogues containing one or two nitrogen atoms in the naphthalene ring and 12 indole analogues containing one to three nitrogen atoms in the indole ring, without any side moiety. Results: The Ames test was performed with five standard bacterial strains (TA100, TA1535, TA98, TA1537, and WP2uvrA) using the pre-incubation method with and without rat liver S9. Five quinoline and two indole analogues were mutagenic. Among the five quinoline analogues, four were mutagenic in the presence of S9 mix with TA100, whereas cinnoline was mutagenic in the absence of S9 mix with TA1537. Among the two indole analogues, indazole was mutagenic in the presence and absence of S9 mix with WP2uvrA and 4-azaindole was mutagenic in the absence of S9 mix with TA1537. The mechanisms underlying the induction of mutagenesis appear to differ between quinoline and indole analogues. In addition, we performed in silico analysis of the mutagenicity of all these analogues using DEREK Nexus 6.1.1 (Lhasa Limited) and GT_EXPERT from CASE Ultra 1.8.0.5 (MultiCASE Inc.) as knowledge-based models and GT1_BMUT from CASE Ultra 1.8.0.5 (MultiCASE Inc.) as a statistical-based model. The knowledge-based model showed low sensitivity for both the quinoline and indole analogues (DEREK Nexus and GT_EXPERT: 20% for quinolines and 0% for indoles). Conversely, the statistical model showed high sensitivity (100% for both quinolines and indoles) and low specificity (43% for quinolines and 10% for indoles). Conclusion: Based on the Ames test results, we proposed structural alerts noting that quinoline analogues were mutagenic when they had nitrogens in any of the positions 2, 5, 7, or 8 in addition to 1, and indole analogues were mutagenic when they had nitrogens at positions 2 or 4 in addition to 1. [ABSTRACT FROM AUTHOR]

An ethanol extract of Piper auritum leaves (PAEE) inhibits the mutagenic effect of three food-borne aromatic amines (2-amino-1-methyl-6-phenylimidazo[4,5- b ]pyridine (PhIP); 2-amino-3,8-dimethylimidazo[4,5- f ]quinoxaline (MeIQx); 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx)) in the TA98 Salmonella typhimurium strain. Preincubation with MeIQx demonstrated in mutagenesis experiments that inhibition of Cytochrome P450 (CYP), as well as direct interaction between component(s) of the plant extract with mutagens, might account for the antimutagenic observed effect. Gas chromatography/mass spectrometry analysis revealed that safrole (50.7%), α-copaene (7.7%), caryophyllene (7.2%), β-pinene (4.2%), γ-terpinene (4.1%), and pentadecane (4.1%) as the main components (PAEE). Piper extract and safrole were able to inhibit the rat liver microsomal CYP1A1 activity that participates in the amines metabolism, leading to the formation of the ultimate mutagenic/ molecules. According to this, safrole and PAEE-inhibited MeIQx mutagenicity but not that of the direct mutagen 2-nitrofluorene. No mutagenicity of plant extract or safrole was detected. This study shows that PAEE and its main component safrole are associated with the inhibition of heterocyclic amines activation due in part to the inhibition of CYP1A subfamily activity. [ABSTRACT FROM AUTHOR]

Doxorubicin (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. HIGHLIGHTS: 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]

Gamisoyo-san is an herbal formula widely used to treat psychological issues, menopausal symptoms, and dysmenorrhea. However, there is insufficient information on its safety profile. This study aimed to confirm the genotoxic and acute toxic potential of Gamisoyo-san. We performed a battery of tests, which included a bacterial reverse mutation test (Ames test) using five bacterial strains, an in vitro chromosomal aberration test using Chinese hamster lung (CHL) cells, an in vivo micronucleus test in mice, and human Cytochrome P450 (CYP450) and UDP-glucuronosyltransferase (UGT) assays. In the acute toxicity study, male and female rats were orally administered Gamisoyo-san 1000, 2000, or 5000 mg/kg and observed for 14 days. The activities of human CYP450s and UGTs were evaluated using recombinant baculosomes. Gamisoyo-san showed no signs of genotoxicity in the five bacterial strains, CHL cells, or mouse bone marrow cells. The acute toxicity test showed that the median lethal dose (LD50) of Gamisoyo-san was greater than 5000 mg/kg in rats. Gamisoyo-san inhibited the activities of CYP1A2, CYP2C19, and UGT1A1. In conclusion, Gamisoyo-san may not exert severe toxicological events or genotoxic effects at doses up to 5000 mg/kg in rats. [ABSTRACT FROM AUTHOR]

Introduction: Phosphoinositide-3-kinase (PI3K) is overexpressed in many tumors and is, thus, an ideal target for cancer treatments. Accordingly, there is an urgent need for the development of PI3K inhibitors with high potency and low toxicity. Methods: In this study, we designed and synthesized a series of 2,4-dimorpholinopyrimidine-5-carbonitrile derivatives, which were evaluated for their PI3K inhibitory potency. Results and discussion: Compound 17p demonstrated comparable PI3Kα inhibitory activity (IC50: 31.8 ± 4.1 nM) to the positive control, BKM-120 (IC50: 44.6 ± 3.6 nM). In addition, 17p showed significant inhibitory activity against PI3Kδ (IC50: 15.4 ± 1.9 nM) and significant isoform selectivity against PI3Kβ, PI3Kγ, and mTOR. Furthermore, 17p exhibited good antiproliferative activities against cancer cell activity and good safety in the Ames and hERG tests while having outstanding liver microsomal stability in vitro , with half-lives of 38.5 min in rats and 127.9 min in humans. In addition, in an apoptosis assay, 17p could induce dose-dependent cytotoxicity in the ovarian cancer cell line A2780. In a pharmacokinetic study, 17p was stable (T ½: 2.03 h) and showed high bioavailability (46.2%). Collectively, these results indicate that 17p could be a promising PI3K agent for cancer treatment. [ABSTRACT FROM AUTHOR]

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Objective: To evaluate the immunomodulatory function and safety of compound preparation containing Ganoderma lucidum polysaccharides. Methods: According to the Technical Standards for Testing and Assessment of Health Food (2003 Edition), the immunomodulatory function of compound preparation containing Ganoderma lucidum polysaccharides was evaluated comprehensively from cellular immunity, humoral immunity and non-specific immunity by measuring the ability of lymphocyte proliferation, degree of ear swelling, number of hemolytic plaques, phagocytic percentage, phagocytic index and natural killer (NK) cells activity in normal mice. Its safety was evaluated comprehensively by acute oral toxicity test in mice, Ames test and 30 d feeding test in rats. Results: In cellular immunity test, the tested substance had no significant effect on lymphocyte proliferation induced by concanavalin A and degree of ear swelling (P>0.05). In the humoral immunity test, the low dose group (0.17 g/kg), the middle dose group (0.33 g/kg) and the high dose group (1.00 g/kg) could increase the level of antibody-producing cells by 10.98%, 12.08%, 17.63% (P<0.05). In the non-specific immunity test, the middle dose group and the high dose group could significantly increase the phagocytosis ability of mononuclear macrophages (P<0.05) and the activity of NK cells (P<0.01). The maximum tolerance dose (MTD) of the tested substance to the acute oral dose of both male and female mice was more than 42.56 g/kg, which belonged to non-toxic substances. The results of Ames test were negative and no mutagenicity was found. In the 30 d feeding test, each dose group had no significant effect on body weight, weekly food utilization rate, blood routine index, blood biochemical index and main organ coefficient (P>0.05), and no obvious pathological changes were found by histological examination. Conclusion: Compound preparation containing Ganoderma lucidum polysaccharides had a positive effect on immunomodulatory function in mice, and it had a good safety as a health food. [ABSTRACT FROM AUTHOR]

ABSTRACT In pre‐clinical and medical quality control, it is of interest to assess the stability of the process under monitoring or to validate a current observation using historical control data. Classically, this is done by the application of historical control limits (HCL) graphically displayed in control charts. In many applications, HCL are applied to count data, for example, the number of revertant colonies (Ames assay) or the number of relapses per multiple sclerosis patient. Count data may be overdispersed, can be heavily right‐skewed and clusters may differ in cluster size or other baseline quantities (e.g., number of petri dishes per control group or different length of monitoring times per patient). Based on the quasi‐Poisson assumption or the negative‐binomial distribution, we propose prediction intervals for overdispersed count data to be used as HCL. Variable baseline quantities are accounted for by offsets. Furthermore, we provide a bootstrap calibration algorithm that accounts for the skewed distribution and achieves equal tail probabilities. Comprehensive Monte‐Carlo simulations assessing the coverage probabilities of eight different methods for HCL calculation reveal, that the bootstrap calibrated prediction intervals control the type‐1‐error best. Heuristics traditionally used in control charts (e.g., the limits in Shewhart c‐ or u‐charts or the mean ± 2 SD) fail to control a pre‐specified coverage probability. The application of HCL is demonstrated based on data from the Ames assay and for numbers of relapses of multiple sclerosis patients. The proposed prediction intervals and the algorithm for bootstrap calibration are publicly available via the R package predint. [ABSTRACT FROM AUTHOR]

Polymers derived from renewable sources, particularly those based on furan, have attracted significant attention within the scientific community due to their potential as an ecologically viable alternative, aiming to minimize the negative impacts arising from the accumulation of plastics derived from fossil hydrocarbons. However, few toxicogenetic studies have been conducted. Therefore, the objective of this study was to assess the cytotoxicity and mutagenicity of starting materials (hexamethylenediamine, maleic anhydride, furan and furan maleic anhydride adduct), as well as a new polymer derived from the furan maleic anhydride adduct. Hexamethylenediamine, maleic anhydride, and furan maleic anhydride adduct exhibited comparable cytotoxic profiles, whereas furan demonstrated lower cytotoxic potential (IC50 of 72.94 ± 2.49 mg/mL), yet it was the only reagent capable of inducing mutagenicity. Furan's mutagenic activity by Ames test was observed in the TA100 strain, both with and without metabolization. The polymer, in turn, exhibited low cytotoxic effect (cell viability exceeding 70%) and demonstrated non-mutagenicity under the experimental conditions, emphasizing its safety concerning toxicological risks. In conclusion, this study contributes to the genotoxicological safety assessment of starting materials, and a new polymer sourced from renewable origins. The absence of mutagenicity and the low cytotoxic potential of the assessed samples contribute to the battery of tests required to indicate safe applications, thereby fostering acceptance, and minimizing associated risks. Among the evaluated reagents, special caution is warranted in handling furan due to its potential to induce gene mutations. [ABSTRACT FROM AUTHOR]

The dye-contaminated industrial effluent causes serious health issues when it gets mixed with underground water without primary treatment. The current project was designed to treat reactive blue-19 dye aqueous solutions in the presence of hydrogen peroxide (H2O2) and titanium dioxide (TiO2) under UV-LED light. The characterization of the photocatalyst was carried out via X-ray diffraction (XRD) and Scanning Electron Microscopy (SEM) for structure, purity, and surface study. The effect of various factors such as pH, TiO2 dose, UV-LED light exposure time, H2O2, and dye concentration, on the degradation rate and cytotoxicity reduction was evaluated and optimized through the Response Surface Methodology (RSM). The maximum degradation of dye solution and chemical oxygen demand (COD) reduction was achieved at 98.81 and 86.22 %, respectively for 50 ppm solution, using UV-LED/H2O2(3 %)/TiO2(6 g/L) hybrid process. The toxicity evaluation through the Allium cepa test demonstrated a 62.40, 65.2, and 56.97 % increase in root length (RL), root count (RC), and mitotic index (MI), respectively, following treatment with the UV-LED/H₂O₂/TiO₂ combined process for 150 min. The hemolytic and brine shrimp tests revealed a reduction in toxicity up to 92.18 and 84.08 %, respectively, after applying the same treatment. Additionally, the Ames test indicated up to 80.94 % reduction in mutagenicity for TA98 and an 84.04 % reduction for TA100 strain when dye samples were treated with UV-LED light in the presence of H₂O₂ and TiO₂ for 150 min. The findings suggested that UV-LED light in conjunction with H2O2 and TiO2 can be a useful tool for the degradation and detoxification of toxic pollutants found in textile wastewater. [ABSTRACT FROM AUTHOR]

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The Ames test is used worldwide to initially screen the mutagenic potential of new chemicals. In the standard Ames test, S. typhimurium strains (TA100, TA98, TA1535, and TA1537) and Escherichia coli (WP2uvrA) are treated with substances with/without cytochrome P450s (CYPs)-induced rat S9 fractions for identifying mutagens and pro-mutagens. However, many substances show completely different toxicity patterns depending on whether the liver S9 fraction belongs to rats or humans. The natural product Polygoni Multiflori Radix (PMR) can also show bacterial reverse mutation, followed by the rat or human liver S9 fraction. While PMR elicits reverse mutations in the TA1537 strain in rat liver S9 but not in human liver S9, this mechanism has not been verified yet. To explain this, the differences in metabolic enzymes compositions commonly observed between rats and humans have been implicated. This study aimed to explore the key factors that cause differences in the genotoxicity of PMR between rat and human liver S9 metabolic enzymes. The results of next-generation sequencing (NGS) analysis showed that both rat and human metabolic enzymes caused similar mutations in TA1537. However, when the metabolic enzymes in each S9 fraction were analyzed using ion mobility tandem mass spectrometry (IM-MS), rat- and human-specific enzymes were identified among the cytochrome (CYP) family, especially aryl hydrocarbon receptor (AHR)-related CYPs. These findings suggest that CYP1A1 isoforms contribute to the mechanism of PMR in the Ames test. Therefore, an in vitro Ames test might be more reliable in predicting genotoxicity for both rodents and humans. This will also help overcome the limitations of laboratory animal-based toxicity evaluations, which provide unreliable results due to interspecies differences between humans and rodents. [ABSTRACT FROM AUTHOR]

• Obliquumol had an LD 50 >2000 mg/kg since there were no mortalities after 14 days. • At the highest dose of obliquumol, the mass, behaviour and food intake of the mice were not affected. • Gross necropsy and histopathological analysis on organs indicated hardly any effects of obliquumol. • P. obliquum leaf extracts, fractions (hexane, chloroform and 30 % H 2 O in MeOH) and isolated compounds (obliquumol and a mixture of lupeol and β-amyrin) had no genotoxic activity against the salmonella typhimurium strains (TA 100, TA 100, and TA 102). Obliquumol (12- O -acetylptaeroxylinol) isolated from Ptaeroxylon obliquum leaves has excellent antifungal activity and low cellular toxicity. As a next step in the potential development of a framework antifungal product, the present work investigated the acute animal toxicity of obliquumol according to OECD 423 guidelines. Furthermore, the genotoxicity of P. obliquum acetone leaf extracts, fractions (hexane, chloroform and 30 % H 2 O in MeOH) and isolated compounds (obliquumol and a mixture of lupeol and β-amyrin) was determined using Ames test.. A single dose of obliquumol was orally administered to mice at levels of 50, 300 and 2000 mg/kg and observed for 14 days. The three S. typhimurium tester strains TA 98, TA 100 and TA 102 were used without metabolic activation to determine the genotoxicity. Even at the highest dose of obliquumol, the mass, behaviour and food intake of the animals were not affected. Gross necropsy and histopathological analysis on organs indicated hardly any effects. No samples had genotoxic activity against the S. typhimurium strains tested. Obliquumol had an LD 50 >2000 mg/kg since there were no mortalities after 14 days. This encourages the possible development of a new class of antifungal compounds from the obliquumol framework. [Display omitted] [ABSTRACT FROM AUTHOR]

The Janus kinase 3 (JAK3) family, particularly JAK3, is pivotal in initiating autoimmune diseases such as rheumatoid arthritis. Recent advancements have focused on developing antirheumatic drugs targeting JAK3, leading to the discovery of novel pyrazolopyrimidine-based compounds as potential inhibitors. This research employed covalent docking, ADMET (Absorption, Distribution, Metabolism, Excretion, Toxicity) analysis, molecular dynamics modeling, and MM/GBSA (Molecular Mechanics Generalized Born Surface Area) binding free energy techniques to screen 41 in silico-designed pyrazolopyrimidine derivatives. Initially, 3D structures of the JAK3 enzyme were generated using SWISS-MODEL, followed by virtual screening and covalent docking via AutoDock4 (AD4). The selection process involved the AMES test, binding affinity assessment, and ADMET analysis, narrowing down the candidates to 27 compounds that passed the toxicity test. Further covalent docking identified compounds 21 and 41 as the most promising due to their high affinity and favourable ADMET profiles. Subsequent development led to the creation of nine potent molecules, with derivatives 43 and 46 showing exceptional affinity upon evaluation through molecular dynamics simulation and MM/GBSA calculations over 300 nanoseconds, comparable to tofacitinib, an approved RA drug. However, compounds L21 and L46 demonstrated stable performance, suggesting their effectiveness in treating rheumatoid arthritis and other autoimmune conditions associated with JAK3 inhibition. [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]

Green medicine, the drugs derived from plants attained a wide spread interest, as believed to be safe and dependable, compared with costly synthetic drugs that have adverse effects. In this study, Annona Squamosa (A. squamosa) have been investigated for their phytochemical, antioxidant, antimutagenic activities. Qualitative analysis of various phytochemical constituents and quantitative analysis of total phenol were determined by the well-known test protocol available in the literature. The extraction yield was found in the order: methanol>water>ethyl acetate>hexane. The extracts found to possess saponin glycosides, cardiac glycosides, phenolics, terpenoids, sterols, and flavonoids. The total phenolic content of hexane, ethyl acetate, methanolic and aqueous extract of A. squamosa were 3.04±0.37, 5.83±0.29, 31.42±2.35, and 27.21±1.06 GAE mg/g, respectively. The methanolic extracts exhibited highest amount of total polyphenol content compared to all other solvent extracts. The ethyl acetate, methanolic and aqueous extracts were evaluated for their antioxidant potential in terms of DPPH radical scavenging capabilities. The scavenging was found to dose dependent with IC50 value of 185.24, 103.09 and 160.17 µg/ml for the ethyl acetate, methanol and aqueous extracts respectively. The antimutagenic potential of the methanolic extract was studied using Ames test using T98 strain of Salmonella typhimurium applying NPD as the mutagen. The number of reverant colonies were counted using a digital colony counter and the reduction in colonies was calculated as the activity (%) of the extract at four doses. The reverent colonies were visible as lawn like structures on the surface of the plate and were significantly lower compared to positive control in the extract plate and the negative control. The extract was able to control the growth of reverant colonies upto 79.30% in comparison to the mutagenic control. The results show that the methanolic extract of A. squamosa leaf was able to exhibit a dose regulated antimutagenic activity by inhibiting the growth of His+ reverant colonies of Salmonella typhimurium. [ABSTRACT FROM AUTHOR]

To fulfil the European Green Deal targets and implement a circular economy, there is an urgent need to increase recycling rates of packaging materials. However, before recycled materials can be used in food contact applications, they must meet high safety standards. According to the European Food Safety Authority (EFSA), a worst-case scenario must be applied and unknown substances must be evaluated as being potentially genotoxic. The Ames test, which detects direct DNA-reactive effects, together with chromatographic analysis is very promising to complement risk assessment. This study aims to evaluate the effectiveness of functional barriers in ten different samples, including virgin and recycled LDPE foils. FT-IR analysis did not show major differences between virgin and recycled films. Light microscopy revealed differences in quality and an increased number of particles. GC-MS analysis detected and quantified 35 substances, including eight unknowns. Using a miniaturized version of the Ames test, four of ten samples tested positive in two individual migrates up to a dilution of 12.5%. All virgin LDPE materials tested negative; however, recycled material F showed an increased mutagenic activity, with an n-fold induction up to 28. Samples with functional barriers lowered migration and reduced mutagenicity. Nonetheless, further investigations are needed to identify possible sources of contamination. [ABSTRACT FROM AUTHOR]

N-nitrosamines and nitrosamine drug substance related impurities (NDSRIs) became a critical topic for the development and safety of small molecule medicines following the withdrawal of various pharmaceutical products from the market. To assess the mutagenic and carcinogenic potential of different N-nitrosamines lacking robust carcinogenicity data, several approaches are in use including the published carcinogenic potency categorization approach (CPCA), the Enhanced Ames Test (EAT), in vivo mutagenicity studies as well as read-across to analogue molecules with robust carcinogenicity data. We employ quantum chemical calculations as a pivotal tool providing insights into the likelihood of reactive ion formation and subsequent DNA alkylation for a selection of molecules including e.g., carcinogenic N-nitrosopiperazine (NPZ), N-nitrosopiperidine (NPIP), together with N-nitrosodimethylamine (NDMA) as well as non-carcinogenic N-nitrosomethyl-tert-butylamine (NTBA) and bis (butan-2-yl) (nitros)amine (BBNA). In addition, a series of nitrosomethylaminopyridines is compared side-by-side. We draw comparisons between calculated reaction profiles for structures representing motifs common to NDSRIs and those of confirmed carcinogenic and noncarcinogenic molecules with in vivo data from cancer bioassays. Furthermore, our approach enables insights into reactivity and relative stability of intermediate species that can be formed upon activation of several nitrosamines. Most notably, we reveal consistent differences between the free energy profiles of carcinogenic and non-carcinogenic molecules. For the former, the intermediate diazonium ions mostly react, kinetically controlled, to the more stable DNA adducts and less to the water adducts via transition-states of similar heights. Non-carcinogenic molecules yield stable carbocations as intermediates that, thermodynamically controlled, more likely form the statistically preferred water adducts. In conclusion, our data confirm that quantum chemical calculations can contribute to a weight of evidence approach for the risk assessment of nitrosamines. [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]

Purpose: Histone deacetylase 6 (HDAC6) has emerged as a therapeutic target for neurodegenerative diseases such as Alzheimer's disease. Noninvasive imaging of HDAC6 in the brain by positron emission tomography (PET) would accelerate research into its roles in these diseases. We recently discovered an 18F-labeled derivative of the selective HDAC6 inhibitor SW-100 ([18F]FSW-100) as a potential candidate for brain HDAC6 radioligand. As a mandatory step prior to clinical translation, we performed preclinical validation of [18F]FSW-100. Methods: Process validation of [18F]FSW-100 radiosynthesis for clinical use and assessment of preclinical toxicity and radiation dosimetry estimated from mouse distribution data were performed. In vitro selectivity of FSW-100 for 28 common receptors in the brain and HDAC isoforms was characterized. [18F]FSW-100 PET imaging was performed in non-human primates in a conscious state to estimate the feasibility of HDAC6 imaging in humans. Results: Three consecutive validation runs of the automated radiosynthesis gave [18F]FSW-100 injections with radiochemical yields of 12%, and the injections conformed to specified quality control criteria for batch release. No acute toxicity was observed for non-radiolabeled FSW-100 or radioactivity decayed [18F]FSW-100 injection, and the former was negative in the Ames test. The whole-body effective dose estimated from biodistribution in mice was within the range of that of previously reported 18F-radioligands in humans. In vitro selectivity against common receptors and other HDAC isoforms was confirmed. [18F]FSW-100 demonstrated good penetration in monkey brain, and in vivo blocking studies suggested that the uptake was specific. Conclusion: These results support the clinical utility of [18F]FSW-100 for in vivo imaging of HDAC6 in the brain. [ABSTRACT FROM AUTHOR]

Platinum(II) and palladium(II) complexes have been investigated as potential anticancer drugs since the serendipitous discovery of the antineoplastic activities of cisplatin in the 1960s. Skin cancer is considered the most common malignant neoplasm that affects humans, and melanoma is the most lethal type of skin cancer. Surgical excision is the main form of treatment, which also may include radiotherapy, systemic chemotherapy, and immunotherapy. In this work, new insights concerning the structural characterization and in vitro anti-proliferative activity of the palladium(II) complex with the amino acid deoxyalliin (Pd-sac) against a panel of thirteen human tumor cells, with emphasis on skin cancer cell lines, are presented. The composition of the complex was confirmed by elemental analysis as [Pd(C6H10NO2S)2]. The structure of the complex was elucidated for the first time by a single-crystal X-ray diffraction technique. Each deoxyalliin molecule coordinates in a bidentate N,S-mode to palladium(II) in a trans-configuration analogous to the platinum(II) deoxyalliin complex early reported. As the main result, the Pd-sac complex showed a selective anti-proliferative activity against melanoma (UACC-62, TGI = 63.5 µM), while both deoxyalliin and K2PdCl4 were inactive against all cell lines. Moreover, Pd-sac did not affect the proliferation of non-tumorigenic keratinocytes (HaCaT, TGI > 586 µM) and was non-mutagenic in the Ames assay. The results open new perspectives for in vivo studies concerning the application of the Pd-sac complex in the treatment of melanoma. [ABSTRACT FROM AUTHOR]

Litsea martabanica (Kurz) Hook.f. has traditionally been used as an anti-insecticidal agent and as a medication due to its hepatoprotective properties by highland communities in Thailand. This study examined the mutagenicity, as well as the acute and chronic toxicity, of the L. martabanica water leaf extract in Sprague-Dawley rats. The pharmacognostic evaluation of L. martabanica was performed in this study to ensure its authenticity and purity. Then, the sample was extracted using decoction with water to obtain the crude water extract. The assessment of acute toxicity involved a single oral administration of 5000 mg/kg, whereas the chronic toxicity assessment comprised daily oral doses of 250, 750, and 2250 mg/kg over 270 days. Various physiological and behavioral parameters, as well as body and organ weights, were systematically monitored. The endpoint assessments involved hematological and biochemical analyses plus gross and histopathological assessments of the internal organs. Our results exhibited no mutagenic activation by the L. martabanica water leaf extract in the Ames test, and no acute toxicity was observed. In the chronic toxicity tests, no abnormalities were found in rats receiving the L. martabanica water leaf extract across multiple measures, comprising behavioral, physiological, and hematological indices. Crucially, the histopathological assessment corroborated previous studies, reporting an absence of any tissue abnormalities. The results revealed that the L. martabanica water leaf extract had no adverse effects on rats over 270 days of oral administration. This demonstrates its safety and crucial scientific evidence for informing public policy and enabling its potential future commercial use in both highland and lowland communities. [ABSTRACT FROM AUTHOR]

Human adenoviruses pose a threat to ophthalmic clinics and are responsible for sporadic, localized respiratory illness across the world. According to recent research, differences in the interferon-mediated regulation of viral replication could contribute to the emergence of persistent infections and reactivation. In addition to being contagious, adenoviral genomes include potent oncogenes promoting the growth of tumors, albeit precision is yet unclear. Adenoviruses infect a wide variety of cells thus used as vectors, allows for the development of innovative treatments for diseases including cancer and heart issues. Intriguingly, Withania somnifera (WS) has shown promising antiviral effects against numerous viral infections, including adenovirus, which are contributed by its phytochemicals. W. somnifera, often known as Indian ginseng, contain large amounts of a class of pharmacologically active chemicals called withanolides in its roots and leaves. Withanolides, which are C-28 phytochemicals, highly oxygenated steroidal lactones shown to have anticancer, immunomodulatory, and other properties. Four phytochemicals from Withania somnifera were examined for their molecular characteristics in this research. The objective was to evaluate their capacity to bind and potentially block the human adenovirus 2 protease protein, which is essential for the reproduction of the adenoviruses, through the use of in-silico work. Using computational methodologies, the drug-likeness characteristics of the phytochemicals were ascertained; followed by molecular docking investigations. These results suggested that out of selected phytochemicals withanolide B is the most promising drug candidate (with binding energy of -8.3kcal/mol) for inhibiting the catalytic activity of human adenovirus 2 protease protein resulting thus preventing infection cycle of adenovirus. The ADME analysis also showed no violation of Lipinski rule of five, no permeation of blood brain barrier, and ames test gave negative value indicating no mutagenic activity by any of the candidate phytocompounds. [ABSTRACT FROM AUTHOR]

Chrysin is hypothesized to possess the ability to prevent different illnesses, such as diabetes, cancer, and neurodegenerative disorders. Nonetheless, chrysin has a low solubility under physiological conditions, resulting in limited bioavailability. In a previous study, we utilized an oil-in-water emulsion system (chrysin-ES or chrysin-NE) to encapsulate chrysin, thereby increasing its bioaccessibility and preserving its antioxidant and anti-Alzheimer's properties. To promote the chrysin-ES as a supplementary and functional food, it was obligatory to carry out a safety assessment. Cytotoxicity testing showed that chrysin-ES was harmless, with no killing effect on 3T3-L1 (adipocytes), RAW 264.7 (macrophages), HEK293 (kidney cells), and LX-2 (hepatic stellate cells). The acute toxicity evaluation demonstrated that the 50% lethal dose (LD50) for chrysin-ES was greater than 2000 mg/kg BW. Genotoxicity assessments found that chrysin-ES did not induce DNA mutations in vitro or in vivo. Furthermore, chrysin and chrysin-ES exhibited anti-mutagenic properties against PhIP-induced and IQ-induced mutagenesis in the Ames test, while they inhibited urethane-, ethyl methanesulfonate-, mitomycin C-, and N-nitrosomethylurea-mediated mutations in Drosophila. The present study illustrates the safety and anti-genotoxicity properties of chrysin-ES, allowing for the further development of chrysin-based food supplements and nutraceuticals. [ABSTRACT FROM AUTHOR]

Atmospheric nonthermal plasma (ANTP) has rapidly evolved as an innovative tool in biomedicine with various applications, especially in treating skin diseases. In particular, the formation of reactive oxygen species (ROS) and nitrogen species (RNS), which are generated by ANTP, plays an important role in the biological signaling pathways of human cells. Unfortunately, excessive amounts of these reactive species significantly result in cellular damage and cell death induction. To ensure the safe application of ANTP, preclinical in vitro studies must be conducted before proceeding to in vivo or clinical trials involving humans. Our study aimed to investigate adverse effects on genetic substances in murine fibroblast cells exposed to ANTP. Cell viability and proliferation were markedly reduced after exposing the cells with plasma. Both extracellular and intracellular reactive species, especially RNS, were significantly increased upon plasma exposure in the culture medium and the cells. Notably, significant DNA damage in the cells was observed in the cells exposed to plasma. However, plasma was not classified as a mutagen in the Ames test. This suggested that plasma led to the generation of both extracellular and intracellular reactive species, particularly nitrogen species, which affect cell proliferation and are also known to induce genetic damage in fibroblast cells. These results highlight the genotoxic and mutagenic effects of ANTP, emphasizing the need for the cautious selection of plasma intensity in specific applications to avoid adverse side effects resulting from reactive species production. [ABSTRACT FROM AUTHOR]

Tomatidine is an aglycone of α-tomatine, a glycoalkaloid present in tomato plants, and has muscle atrophy inhibitory effect and anticancer activity. Tomatidine-Rich Tomato Leaf Extract powder (TRTLE) contains 60% tomatidine, which is converted to tomatidine by acid hydrolysis after extracting α-tomatine from tomato leaves. The purpose of this study was to evaluate the safety of TRTLE by conducting a series of toxicity studies in mice and bacteria to support its safe food use. Single-dose and 90-day repeated-dose toxicity study were conducted in 6-week-old ICR male and female mice to calculate the LD50 and non-toxic dose (NOAEL) of TRTLE. In the single-dose toxicity study, a single oral dose of 667, 2,000, and 5,000 mg TRTLE/kg body weight (bw) was administered, and in the 90-day repeated-dose toxicity study, 133 mg TRTLE/kg bw was administered orally daily. In addition, the Ames test was performed with Salmonella Typhimurium and Escherichia coli to determine the genotoxic activity. The single-dose toxicity study indicated the LD50 was 833 mg TRTLE/kg bw (tomatidine equivalent: 500 mg/kg bw). In the 90-day repeated-dose toxicity study, no abnormalities due to TRTLE were observed in each laboratory test, including general symptoms, body weight changes, hematology, urinalysis, and histopathological examination. In the Ames test, TRTLE was confirmed not to be mutagenic with or without metabolic activation. Based on these data, the NOAEL in mice was determined to be 133 mg TRTLE/kg bw (tomatidine equivalent: 80 mg/kg bw). [ABSTRACT FROM AUTHOR]

Halicin, the first antibacterial agent discovered by artificial intelligence, exerts broad-spectrum antibacterial effects and has a unique structure. Our study found that halicin had a good inhibitory effect on clinical isolates of drug-resistant strains and Clostridium perfringens (C. perfringens). The safety of halicin was evaluated by acute oral toxicity, genotoxicity and subchronic toxicity studies. The results of acute toxicity test indicated that halicin, as a low-toxicity compound, had an LD50 of 2018.3 mg/kg. The results of sperm malformation, bone marrow chromosome aberration and cell micronucleus tests showed that halicin had no obvious genotoxicity. However, the results of the 90-day subchronic toxicity test indicated that the test rats exhibited weight loss and slight renal inflammation at a high dose of 201.8 mg/kg. Teratogenicity of zebrafish embryos showed that halicin had no significant teratogenicity. Analysis of intestinal microbiota showed that halicin had a significant effect on the intestinal microbial composition, but caused a faster recovery. Furthermore, drug metabolism experiments showed that halicin was poorly absorbed and quickly eliminated in vivo. Our study found that halicin had a good therapeutic effect on intestinal infection model of C. perfringens. These results show the feasibility of developing oral halicin as a clinical candidate drug for treating intestinal infections. [ABSTRACT FROM AUTHOR]

Background: Colombia is joining global initiatives to mitigate climate change through bioethanol production, as it has large sugar cane plantations and sugar mills, particularly in the Valle del Cauca region. One of the main by-products of the bioethanol industry is vinasse, which consists mainly of water, organic solids and heavy metals. Some of the compounds present in vinasses, such as melanoidins and phthalates, show genotoxic, mutagenic and carcinogenic activity in onion cells, tilapia and aquatic organisms. Various methods, such as bio-oxidation and Fenton reaction, have been used to reduce the organic load of vinasses. Among the most commonly used assays to study genotoxicity and mutagenicity are single cell gel electrophoresis (comet assay) and the Ames test. Objective: In this study, the genotoxicity in human lymphocytes and the mutagenicity in Salmonella typhimurium induced by different dilutions of vinasse produced at the bioethanol production plant in Frontino, Antioquia, before and after being subjected to biooxidation and Fenton processes, were evaluated. Methods: Genotoxicity was evaluated by the comet assay in human lymphocytes, and mutagenic activity was evaluated by the Ames test using Salmonella typhimurium strains TA98 and TA100, with and without the addition of microsomal enzymes (S9). Both tests were applied to each type of vinasse considered in this study, including raw vinasse (RV), bio-oxidised vinasse (BV) and Fenton oxidised vinasse (FV). Results: The results showed that at RV doses above 3%, viability decreased to values between 70% and 88%, whereas for BV and FV, viability remained above 93% and 94%, respectively. Vinasse was also found to have a dose-dependent effect on genotoxicity. However, no mutagenic activity was observed in any of the Salmonella strains evaluated, indicating that vinasse does not induce mutations. Conclusion: The importance of addressing vinasse pollution and treatment methods to reduce its toxicity is emphasised. However, further research is needed to fully understand the risks associated with vinasse exposure and to develop effective mitigation strategies. [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]

• Thirty-three plant species were analysed for mutagenicity and toxicity. • A. villicaulis, G. occidentalis and G. senegalensis were not toxic nor mutagenic. • Three plant combinations were found to be non-toxic in the Brine Shrimp Lethality Assay. • Toxicity was more frequently observed than expected. Medicinal plants play an important role in the primary healthcare of lay people around the world, including the rural community of Maputaland, South Africa. According to an ethnobotanical survey conducted in 2014, the lay people in northern Maputaland use plant species independently and in combination to treat gynaecology and obstetric ailments. These plant species were generally regarded as safe by the lay people except for one plant species, Trichilia dregeana Harv. & Sond. The aim of this study was to investigate the safety of 16 plant combinations using the brine shrimp lethality assay (BSLA) for toxicity and Ames test using S. typhimurium TA98 and TA100 strains for mutagenicity. Toxicity studies on 33 medicinal plant species independently needed to be analysed first in order to make a comparison of combined with single use. The aqueous and organic (1:1 methanol-dichloromethane) extracts were prepared from 51 plant samples (including leaf samples collected as a substitution for the roots). There were three plant species (Acalypha villicaulis Horchst. Ex A.Rich. root , Grewia occidentalis L. root and Gymnosporia senegalensis Loes. leaves) indicated neither toxicity nor mutagenicity when tested at 1 and 5 mg/ml using the BSLA and Ames test, respectively. Hermannia boraginiflora Hook. , Sapium integerrimum (Hochst. ex Krauss) , Scadoxus puniceus (L.) Friis Nordal and Tabernaemontana elegans Stapf demonstrated toxicity even after dilution to the lowest concentration of 0.031 mg/ml. The three plant combinations which were found to be non-toxic in the BSLA (both aqueous and organic extracts) were Euphobia tirucalli L. (root) with Ozoroa engleri R.Fern & A.Fern. (bark), S. puniceus (bulb) and Senecio serratuloides DC. (whole plant); Bridelia cathartica G.Bertol. (root) with Opuntia stricta Haw. (stem) with Searsia nebulosa (Schoenland) Moffett (bark); and B. cathartica (root) with Erythrina humeana Spreng (root). In the Ames test, the plant samples which appeared to be non-mutagenic against both S. typhimurium TA98 and TA100 strains were A. villicaulis root , Cyperus natalensis Hochst. ex Krauss root, Euclea natalensis A.DC. leaves, G. occidentalis root , Ochna natalitia Walp. leaves , S. integerrimum leaves and S. puniceus bulb. This study indicated that medicinal plant species (independently and in combination) may have toxic and/ or mutagenic effects, even without any obvious signs after consumption. More importantly, it was determined that toxicity can be reduced by carefully managing the dose. [ABSTRACT FROM AUTHOR]

In this article, we present polymer materials consisting of polylactide (PLA) and nisaplin (N), as well as PLA and natamax (X). These materials were obtained using the solvent method and tested by various test methods, i.e., functional properties—water vapor permeability, light transmission, gloss, and bactericidal activity against strains E. coli (ATCC 8739P), S. aureus (ATCC 65388), and P. aeruginosa (ATCC 8739). Furthermore, analyses were conducted to evaluate their efficacy against pathogenic fungi, including A. niger, A. flavus, A. glaucus, and A. versicolor. Mutagenicity analyses were performed using the standard Ames Test with Salmonella typhimurium. The main test methods used were ISO 22196, ISO 846. The results obtained confirm the potential suitability of the films of PLA with nisaplin and natamax for applications in the food packaging industry. [ABSTRACT FROM AUTHOR]