Genotoxic Potential Research Articles

Multiparametric in vitro genotoxicity assessment of different variants of amorphous silica nanomaterials in rat alveolar epithelial cells

The hazard posed to human health by inhaled amorphous silica nanomaterials (aSiO2 NM) remains uncertain. Herein, we assessed the cyto- and genotoxicity of aSiO2 NM variants covering different sizes (7, 15, and 40 nm) and surface modifications (unmodified, phosphonate-, amino- and trimethylsilyl-modified) on rat alveolar epithelial (RLE-6TN) cells. Cytotoxicity was evaluated at 24 h after exposure to the aSiO2 NM variants by the lactate dehydrogenase (LDH) release and WST-1 reduction assays, while genotoxicity was assessed using different endpoints: DNA damage (single- and double-strand breaks [SSB and DSB]) by the comet assay for all aSiO2 NM variants; cell cycle progression and γ-H2AX levels (DSB) by flow cytometry for those variants that presented higher cytotoxic and DNA damaging potential. The variants with higher surface area demonstrated a higher cytotoxic potential (SiO2_7, SiO2_15_Unmod, SiO2_15_Amino, and SiO2_15_Phospho). SiO2_40 was the only variant that induced significant DNA damage on RLE-6TN cells. On the other hand, all tested variants (SiO2_7, SiO2_15_Unmod, SiO2_15_Amino, and SiO2_40) significantly increased total γ-H2AX levels. At high concentrations (28 µg/cm2), a decrease in G0/G1 subpopulation was accompanied by a significant increase in S and G2/M sub-populations after exposure to all tested materials except for SiO2_40 which did not affect cell cycle progression. Based on the obtained data, the tested variants can be ranked for its genotoxic DNA damage potential as follows: SiO2_7 = SiO2_40 = SiO2_15_Unmod > SiO2_15_Amino. Our study supports the usefulness of multiparametric approaches to improve the understanding on NM mechanisms of action and hazard prediction.

Characterization of hepatotoxic effects induced by pyraclostrobin in human HepG2 cells and zebrafish larvae

Pyraclostrobin is a highly effective and broad-spectrum strobilurin fungicide. With the widespread use of pyraclostrobin to prevent and control crop diseases, its environmental pressure and potential safety risks to humans have attracted much attention. Herein, the toxicological risks of pyraclostrobin toward HepG2 cells and the mechanisms of intoxication in vitro were investigated. The liver toxicity of pyraclostrobin in zebrafish larvae was also evaluated. It was found that pyraclostrobin induced DNA damage and reactive oxygen species generation in HepG2 cells, indicating the potential genotoxicity of pyraclostrobin. The results of fluorescent staining experiments and the expression of cytochrome c, Bcl-2 and Bax demonstrated that pyraclostrobin induced mitochondrial dysfunction, resulting in cell apoptosis. Monodansylcadaverine staining and autophagy marker-related proteins LC3, p62, Beclin-1 protein expression showed that pyraclostrobin promoted cell autophagy. Furthermore, immunoblotting analysis suggested that pyraclostrobin induced autophagy accompanied with activation of adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK)/mTOR signaling pathway. Visualization of zebrafish liver and oil red staining indicated that pyraclostrobin could induce liver degeneration and liver steatosis in zebrafish. Collectively, these results help to better understand the hepatotoxicity of pyraclostrobin and provide a scientific basis for its safe applications and risk control.

Evaluation of the cytotoxicity and genotoxicity potential of synthetic diacetyl food flavoring in silico, in vivo, and in vitro.

Diacetyl is a common ingredient that creates a buttery flavor in baked goods and other food products. The cytotoxic impact of diacetyl on a normal human liver cell line (THLE2) indicated an IC50 value of 41.29mg/ml through MTT assay and a cell cycle arrest in the G0/G1 phase relative to the control. Administration of diacetyl at two-time points (acute-chronic) led to a significant increase in DNA damage indicated by the increase in tail length, tail DNA%, and tail moment. The mRNA and protein expression levels of genes in the rats' livers were then measured using real-time PCR and western blotting. The results showed an activation of the apoptotic and necrosis mechanism, with an upregulation of p53, Caspase 3, and RIP1 and a downregulation of Bcl-2 at the mRNA level. The ingestion of diacetyl disrupted the liver's oxidant/antioxidant balance, as evidenced by alterations in levels of GSH, SOD, CAT, GPx, GR, MDA, NO, and peroxynitrite. Additionally, heightened levels of inflammatory cytokines were shown. Histopathological examinations revealed necrotic foci and congested portal areas in the rats' liver cells after treatment with diacetyl. Diacetyl may interact moderately with Caspase, RIP1, and p53 core domain through In-silico, possibly resulting in upregulated gene expression.

Oxidative, Genotoxic and Cytotoxic Damage Potential of Novel Borenium and Borinium Compounds

In this study, the biological properties of novel borenium and borinium compounds in terms of their oxidative, genotoxic, and cytotoxic effects were assessed on cultured human peripheral blood cells, as well as several types of cancer cells. Our results revealed that the borinium compounds yielded the best results in terms of supporting total antioxidant capacity (TAC). In fact, borenium 1, borenium 2, borenium 3, borinium 4, and borinium 5 compounds elevated TAC levels of cultured human blood cells at rates of 42.8%, 101.5%, 69.8%, 33.3%, and 49.2%, respectively. There were no statistically significant differences (p > 0.05) between the negative control and the groups treated with all borinium and borenium concentrations from the micronucleus (MN) and chromosome aberration (CA) assays, demonstrating the non-genotoxic effects. Moreover, borenium 1 (60.7% and 50.7%), borenium 2 (70.4% and 57.2%), borenium 3 (53.1% and 45.2%), borinium 4 (55.1% and 48.1%), and borinium 5 (51.0% and 36.1%) minimized the mitomycin C(MMC)-induced genotoxic damages at different rates as determined using CA and MN assays, respectively. Again, it was found that the borinium compounds exhibited higher cytotoxic activity on cancer cells when compared to borenium compounds. Consequently, in light of our in vitro findings, it was suggested that the novel borinium and borenium compounds could be used safely in pharmacology, cosmetics, and various medical fields due to their antioxidant and non-genotoxic features, as well as their cytotoxicity potential on cancer cells.

Cytotoxicity and genotoxicity evaluation of chloroform using Vicia faba roots.

Chloroform is a widely used industrial chemical that can also pollute the environment. The aims of this study were to examine the potential cytotoxicity and genotoxicity of chloroform on plant cells, using the Vicia faba bioassay. Chloroform was evaluated at concentrations of 0.1, 0.5, 1, 2, and 5mg·L-1. The following parameters were analyzed: the mitotic index (MI), micronucleus (MN) frequency, chromosomal aberration (CA) frequency, and malondialdehyde (MDA) content. The results showed that exposure to increasing concentrations of chloroform caused a decrease in MI and an increase in the frequency of MN in Vicia faba root tip cells, relative to their controls. Moreover, various types of CA, including C-mitosis, fragments, bridges, laggard chromosomes, and multipolar mitosis, were observed in the treated cells. The frequency of MN was positively correlated with the frequency of CA in exposure to 0.1-1mg·L-1 chloroform. Furthermore, chloroform exposure induced membrane lipid peroxidation damage in the Vicia faba radicle, and a linear correlation was observed between the MDA content and the frequency of MN or CA. These findings indicated that chloroform exposure can result in oxidative stress, cytotoxicity, and genotoxicity in plant cells.

Evaluation of the cytogenetic activity of the food dye Azorubine in a micronucleus test in mice

Introduction. Azorubine E122 (Carmoisine, Food Red 3) monoazo dye is used in the manufacture of desserts, caramels, sweets, marmalades, ice cream, alcoholic and non-alcoholic drinks, etc. The safety assessment of food additives includes the study of genotoxic potential. At the same time, for substances with a high or a small but long-term exposure (including food additives), in vivo tests are required.
 Materials and methods. The genotoxic activity of aqueous solutions of synthetic food azo dyes E122 Azorubin was studied by the micronuclear method on bone marrow cells in mice (males, hybrids F1 CBA × C57Bl6/j). The studied substances were injected into the stomach of mice at doses of 250–2000 mg/kg twice with an interval of 24 hours, with the preparation of bone marrow preparations 24 hours after the last administration. The frequency of micronucleated (MN) polychromatophilic erythrocytes (PCEs) was estimated on the base of the results of the analysis of 4000 PСЕ. The proportion of PСE among all red blood cells was determined by analyzing 500 cells per animal. 
 Results. There was no statistically significant increase in the frequency of PCE with MN over the current control with a double administration of Azorubine in all studied doses. After exposure at doses of 1000 and 2000 mg/kg, the incidence of MN PCEs slightly exceeded the upper limit of the 95% CI of the accumulated negative control and the effect was dose-dependent and statistically significant, which does not allow recognizing the answer as clearly negative.
 Limitations of the study are due to the methodology of the test: only cytogenetic disorders in a single tissue were analyzed under conditions of double enteral administration of the studied sample.
 Conclusion. An analysis of the frequency of MN PCEs in the bone marrow of mice after a double injection of Azorubine at doses of 250–2000 mg/kg made it possible to qualify the result of the experiment as uncertain.

An evaluation of the genotoxicity and 90-day repeated-dose toxicity of a CBD-rich hemp oil.

Currently, there is much interest in the sales and study of consumable Cannabis sativa L. products that contain relatively high levels of cannabidiol (CBD) and low levels of Δ-9-tetrahydrocannabinol. While there are published safety evaluations for extracts containing low concentrations of CBD, toxicological assessments for those with higher concentrations are still scant in the public domain. In this paper, genotoxicity tests and a 90-day repeated-dose toxicity study of an ethanolic extract of C. sativa containing ~85% CBD were performed following relevant OECD guidelines. No increased gene mutations were observed in a bacterial reverse mutation assay compared to controls up to the maximum recommended concentration of the guideline. An in vitro chromosomal aberration assay showed no positive findings in the short-term (3 h) treatment assays. Long-term treatment (20 h) showed an increased number of cells containing aberrations at the highest dose of 2 μg/mL, which was outside of historical control levels, but not statistically significantly different from the controls. An in vivo micronucleus study showed no genotoxic potential of the test item in mice. A 90-day repeated-dose gavage study using 0, 75, 125, and 175 mg/kg bw/day showed several slight findings that were considered likely to be related to an adaptive response to consumption of the extract by the animals but were not considered toxicologically relevant. These included increases in liver and adrenal weights compared to controls. The NOAEL was determined as 175 mg/kg bw/day, the highest dose tested (equivalent to approximately 150 mg/kg bw/day of CBD).

Evaluation of In Vitro Genotoxicity of Polystyrene Nanoparticles in Human Peripheral Blood Mononuclear Cells.

According to the trade association PlasticEurope, global plastics production increased to 390.7 million tons in 2021. Unfortunately, the majority of produced plastics eventually end up as waste in the ocean or on land. Since synthetic plastics are not fully biodegradable, they tend to persist in natural environments and transform into micro- and nanoplastic particles due to fragmentation. The presence of nanoplastics in air, water, and food causes ecotoxicological issues and leads to human exposure. One of the main concerns is their genotoxic potential. Therefore, this study aimed to evaluate the internalization rates, cytotoxicity, and genotoxicity of polystyrene nanoparticles (PS-NPs) in human peripheral blood mononuclear cells (PBMCs) in vitro. The uptake of PS-NPs was confirmed with flow cytometry light scattering analysis. None of the tested nanoparticle concentrations had a cytotoxic effect on human PBMCs, as evaluated by a dual ethidium bromide/acridine orange staining technique. However, an alkaline comet assay results revealed a significant increase in the levels of primary DNA damage after 24 h of exposure to PS-NPs in a dose-dependent manner. Moreover, all tested PS-NPs concentrations induced a significant amount of micronucleated cells, as well. The results of this study revealed the genotoxic potential of commercially manufactured polystyrene nanoparticles and highlighted the need for more studies with naturally occurring plastic NPs.

In Vitro Mutagenic and Genotoxic Assessment of Anatoxin-a Alone and in Combination with Cylindrospermopsin.

Anatoxin-a (ATX-a) is a cyanobacterial toxin whose occurrence has been reported worldwide and has attracted increasing scientific interest due to its toxicity. Moreover, in nature, ATX-a usually appears together with other cyanotoxins, such as cylindrospermopsin (CYN), so possible interaction phenomena could happen and should be considered for risk assessment purposes. For this reason, the aim of this work was to explore the potential mutagenicity and genotoxicity of pure ATX-a and an ATX-a/CYN mixture using a battery of in vitro assays, including the bacterial reverse-mutation assay in Salmonella typhimurium (OECD 471) and the micronucleus test (MN) (OECD 487) on L5178Y Tk+/- cells. The results showed that ATX-a was not mutagenic either alone or in combination with CYN under the conditions tested. Nevertheless, genotoxic effects were observed for both ATX-a and its mixture with CYN following the in vitro MN assay. The genotoxicity exhibited by ATX-a was only observed in the absence of S9 mix, whereas in the cyanotoxin mixture the concentration-dependent genotoxicity of ATX-a/CYN in vitro was observed only in the presence of S9. Thus, the toxicity induced by cyanotoxin mixtures may vary from that produced by toxins alone, and consequently more studies are necessary in order to perform more realistic risk assessments.

MIC accident: lesson may guide for evaluation of genotoxic potential of the industrial chemicals for prevention of industrial accidents.

Most of the individual and/or amalgamated compounds present in the atmospheric air are not known for their toxicologic potential and impact on human health. The toxicologic strength of methyl isocyanate (MIC) gas was unknown till its accidental leakagethat instantly claimed thousands of lives. Cytogenetic study showed increased chromosome aberrations (CA) and sister chromatid exchanges (SCEs) and delayed cell replication index (RI) in a multicentre genetic screeningprogram on gas victimsimmediate post-disaster. A surveillance study after 30 years displayed reduction in CA compared to the initial status in survivors of the severely and moderately exposed strata. Altogether, cytogenetic damage was significantly predominant in the severely exposed population. Stable and replicable aberrations and chromatid exchanges were detected in both studies, which collectively indicate genetic instability. The variation in individual cytogenetic spectrum from similar exposure status could be the result of inter-individual response to the external factors over 30 years post-disaster. The spectrum of CA detected after 30 years might be the cumulative effect of occupational, environmental and life-style factors at a background of one episode of acute MIC exposure. Had MIC's toxicologic potential was known before, fatality and health effects could have been averted. In vitro assessment of toxicity of tin showed a positive correlation with dose and age of exposure, which was aggravated by smoking. Age has shown a significant effect on CA in the general population. The present report recommends evaluation of toxicity prior to use, and reduction of pollution at source for a maintaining a sustainable environmental context.

Heavy Metals in the Striped Snakehead Murrel Channa striata and Sediments of Lake Mainit Philippines with Notes on Piscine Micronuclei Occurrence

Lake Mainit is the deepest lake in the Philippines, with sporadic documentation of various types of aquatic pollution. This paper reports the heavy metal content in the muscles of the striped snakehead murell Channa striata and bottom sediments from five stations across Lake Mainit using quadrupole-inductively coupled plasma-mass spectrometry (Q-ICP-MS). The micronuclei (MN) formation in erythrocytes of C. striata was also assessed for potential genotoxicity. The relative order of the average concentrations of heavy metals in C. striata samples across all stations is Zn>Cu>Cr>Ni>As>Pb>Cd. As per international safety standards, fish muscle samples across all stations have exceeded the permissible limits of Cu, Ni, and Zn, which has implications for health risks in humans consuming this important fish commodity. The relative order of the concentrations of heavy metals in sediments is Cr>Ni>Cu>Zn>As>Pb>Cd, where Cr, Ni, Cu, Pb, and Zn exceeded safety standards. The MN assay revealed that 98% of the erythrocytes assessed (N=147,000 RBCs) have normal cell morphology. Of the 2% of erythrocytic nuclear alterations (ENAs), 55% are found to be fragmented-apoptotic cells, while 31% are elongated or reshaped. The MNs in C. striata are relatively minimal at 2%. While the MN assay implies that C. striata from Lake Mainit are not at risk for potential genotoxic injury, the present study calls for seasonal monitoring for heavy metals in sediments and other fishery resources in Lake Mainit.

Tradescantia-Based Test Systems Can Be Used for the Evaluation of the Toxic Potential of Harmful Algal Blooms

Harmful algal blooms (HABs) are overgrowths of toxic strains of algae (diatoms, green) and cyanobacteria (blue-green algae). While occurring naturally, human-induced environmental changes have resulted in more frequent occurrences of such blooms worldwide. Meantime, the ecotoxicological risk of HABs is rarely evaluated by means of standard test methods. For the first time, the genotoxic potential of the HAB event 2020 was assessed using two different Tradescantia-based test systems (Trad-SHM and Trad-MN, 24-h exposure). An integrated analysis of biological (algal abundance) and ecotoxicological (testing) data revealed linkages among algal proliferation, changes in Tradescantia stamen hairs (mutations and suppressed growth) and chromosomal aberrations during microsporogenesis (appearance of micronuclei) that were likely to be caused by toxic algal groups. Green alga Botryococcus braunii and the cyanobacterial species Anabaena and Oscillatoria could suppress stamen hair growth; Cyanobacteria Phormidium and Aphanothece sp. could trigger mutations in stamen hairs (appearance of pink and colorless cells); and Oscillatoria sp. could be responsible for the occurrence of chromosomal damage. Diatom proliferation in the spring was not related to the genotoxic response in Tradescantia. Both tests, the Trad-SHM and Trad-MN, are suitable for the evaluation of the toxic potential of HABs.

Desempenho zootécnico, grau de esteatose e potencial genotóxico de lambaris Astyanax lacustris alimentados com diferentes níveis de L-carnitina

ABSTRACT L-carnitine perform a major role in transporting long-chain fatty acids into the mitochondria, where they are oxidized. It has been used in animal diets to decrease fat and increase muscle protein. The aim of this study was to evaluate the zootechnical performance, degree of steatosis in the liver, and genotoxic potential in Astyanax lacustris fed with different levels of L-carnitine (LC). Yellowtail tetra juveniles (n = 140) were distributed in 20 tanks of 70 L, with seven fish in each, in a water recirculation system with controlled temperature (27±0.1⁰C). The treatments with different levels of L-carnitine supplementation were: 0 (control), 250, 500, 750, and 1000 mg of LC per kg of food. The diets were provided twice a day for 60 days. The results showed that the different levels of LC did not affect (P>0.05) weight gain, survival, viscerosomatic index, and the liver hepatocytes showed a normal appearance. However, the use of LC supplementation showed genotoxic potential with a significant difference (P<0.05) for cell alterations when compared to the control at concentrations above 500mg kg-1.

Exposure to Cadmium Telluride Quantum Dots and Gene Expression Profile of Huh-7 Hepatocellular Carcinoma Cell Line.

Nanoparticles have shown promising potential for efficient drug delivery, circumventing biological interferences like immunological and renal clearance and mechanical and enzymatic destruction. However, a handful of research papers have questioned the biomedical use of metal-based nanoparticles like cadmium telluride quantum dots (CdTe-QDs) for their cytotoxic, genotoxic, and carcinogenic potential. Herein, we examined the effects of CdTe-QD NPs on gene expression profile of hepatocellular carcinoma (Huh-7) cell line. Huh-7 cells were treated with CdTe-QD NPs (10μg/ml for 6, 12, and 24hours, and 25μg/ml for 6 and 12hours), and transcriptomic analysis was performed using microarray to evaluate the global gene expression profile. Differential expressed genes (DEGs) were observed for both the doses (10 and 25μg/ml) of CdTe-QD NPs at different time points. Gene ontology (GO) analysis revealed that genes involved in molecular function of cell cycle, organizational injury and abnormalities, cell death and survival, gene expression, cancer, organismal survival, and cellular development were differentially expressed. Overall, we have demonstrated differential expression of several genes, involved in maintaining cell survival, metabolism, and genome integrity. These findings were confirmed by RT-qPCR study for some canonical pathway genes signifying possible implication in NP toxicity-mediated cell survival and inhibition of cell death.

The Safety Assessment of Cosmetic Perfumes by Using In Chemico and In Vitro Methods in Combination with GC-MS/MS Analysis.

Animal testing has been prohibited for the safety assessment of cosmetic ingredients or finished products. Thus, alternative non-animal methods, followed by confirmatory clinical studies on human volunteers, should be used as the sole legally acceptable approach within the EU. The safety assessment of cosmetic products requires the involvement of multiple scientific disciplines, including analytical chemistry and biomedicine, as well as in chemico, in vitro and in silico toxicology. Recent data suggest that fragrance components may exert multiple adverse biological effects, e.g. cytotoxicity, skin sensitisation, (photo)genotoxicity, mutagenicity, reprotoxicity and endocrine disruption. Therefore, a pilot study was conducted with selected samples of fragrance-based products, such as deodorant, eau de toilette and eau de parfum, with the aim of integrating results from a number of alternative non-animal methods suitable for the detection of the following toxicological endpoints: cytotoxicity (with 3T3 Balb/c fibroblasts); skin sensitisation potential (in chemico method, DPRA); skin sensitisation potential (LuSens in vitro method, based on human keratinocytes); genotoxicity potential (in vitro Comet assay with 3T3 Balb/c cells); and endocrine disruption (in vitro YES/YAS assay). The presence of twenty-four specific known allergens in the products was determined by using GC-MS/MS. The strategies for estimation of the NOAEL of a mixture of allergens, which were proposed by the Scientific Committee on Consumer Products in their 'Opinion on Tea tree oil' document and by the Norwegian Food Safety Authority in their 'Risk Profile of Tea tree oil' report, were used as models for the NOAEL estimation of the mixtures of allergens that were identified in the individual samples tested in this study.

In vitro Assessment of Antibacterial Activity and Potential Genotoxic Effect of Fruit Extracts of Capparis spinosa L. Plant

This research was carried out to assess the minimum inhibitory concentration (MIC) and the genotoxic potential of ethanolic and aqueous (cold and hot) fruit extracts of Capparis spinosa L. (C. spinosa) plant against different types of bacterial strains. The antimicrobial effect of these extracts against the tested bacteria was investigated using broth microdilution method. The potential genotoxic effect was evaluated by ERIC-PCR technique. Results of the current study revealed that the MIC values of ethanolic fruit extract against the tested bacterial had a range of 12.5 mg/ml to 25 mg/ml. However, aqueous fruit extracts had an MIC with a range of 50 mg/ml to 100 mg/mL. The potential genotoxic activity of cold aqueous extract was determined according to the changes in ERIC-PCR profile of E. coli strain treated with extract in comparison to that untreated (negative control). Results of this study suggest the genotoxic effect of aqueous fruit extract on E. coli. Further research is required to assess and identify the biological molecules and their mechanisms in the context of the genotoxicity. In vivo genotoxicity assessment or with the presence of liver extract is recommended to evaluate the safety of using fruits for therapeutic purposes and a valuable nutrient source.

Genotoxicity Evaluation of The Novel Psychoactive Substance MTTA.

MTTA, also known as mephtetramine, is a stimulant novel psychoactive substance characterized by a simil-cathinonic structure. To date, little has been studied on its pharmaco-toxicological profile, and its genotoxic potential has never been assessed. In order to fill this gap, the aim of the present work was to evaluate its genotoxicity on TK6 cells in terms of its ability to induce structural and numerical chromosomal aberrations by means of a cytofluorimetric protocol of the "In Vitro Mammalian Cell Micronucleus (MN) test". To consider the in vitro effects of both the parental compound and the related metabolites, TK6 cells were treated with MTTA in the absence or presence of an exogenous metabolic activation system (S9 mix) for a short-term time (3 h) followed by a recovery period (23 h). No statistically significant increase in the MNi frequency was detected. Specifically, in the presence of S9 mix, only a slight increasing trend was observable at all tested concentrations, whereas, without S9 mix, at 75 µM, almost a doubling of the negative control was reached. For the purposes of comprehensive evaluation, a long-term treatment (26 h) was also included. In this case, a statistically significant enhancement in the MNi frequency was observed at 50 µM.

Anti-Oxidant, Anti-Mutagenic Activity and Safety Evaluation of Antrocin

Antrocin is a novel compound isolated from Antrodia cinnamomea, and is classified as a sesquiterpene lactone. The therapeutic efficacy of antrocin has been studied, and it has shown an antiproliferative effect on various cancers. The aim of this study was to evaluate the anti-oxidant activity, potential genotoxicity, and oral toxicity of antrocin. Ames tests with five different strains of Salmonella typhimurium, chromosomal aberration tests in CHO-K1 cells, and micronucleus tests in ICR mice were conducted. The results of anti-oxidant capacity assays showed that antrocin has great anti-oxidant activity and is a moderately strong antimutagenic agent. In the results of the genotoxicity assays, antrocin did not show any mutagenic potential. In the 28-day oral toxicity test, Sprague Dawley rats were gavaged with 7.5 or 37.5 mg/kg of antrocin for 28 consecutive days. In addition, 7.5 mg/kg sorafenib, an anti-cancer drug, was used as a positive control for toxicity comparison. At the end of the study, antrocin did not produce any toxic effects according to hematology, serum chemistry, urine analysis, or histopathological examinations. According to the results of the genotoxicity and 28-day oral toxicity study, antrocin, at a dose of 37.5 mg/kg, did not cause adverse effects and can be a reference dose for therapeutic agents in humans.

Comparative evaluation of natural and artificial sweeteners from DNA damage, oxidative stress, apoptosis, to development using Drosophila melanogaster

The overconsumption of added sugars makes people vulnerable to a myriad of diseases. Several biochemical and developmental assays were performed in the current study to assess the effect of fructose on Drosophila melanogaster and to find substitutes for fructose by comparing it to well-known sweeteners. Drosophila was exposed separately to the same ratio of sugar 9.21% (w/v) of several types of sweeteners (sucrose, fructose, glucose syrup, high-fructose corn syrup and stevia). Results revealed that fructose might induce recombination, whereas stevia lacks genotoxic potential. No developmental delay, growth defects, or neurotoxic effects were recorded for any of the sweeteners. We also observed no striking differences in reactive oxygen species levels. Thus, stevia seems to be an alternative sweetener to fructose that can be consumed to reduce fructose-induced anomalies.

Genotoxicity Assessment of Hydroalcoholic Extract of Bark of <i>Terminalia arjuna</i> (Combretaceae)

The mutagenic potential of Terminalia arjuna hydroalcoholic bark extract was evaluated using Ames test with strains TA98, TA100, TA102, TA1535, and TA1537. The genotoxic potential was assessed by performing Chromosomal Aberration test with cell lines of CHO. In the assessment of mutagenic potential by the Ames test, hydroalcoholic extract of Terminalia arjuna induced a negative response and in Chromosomal Aberration test, it did not show any significant genotoxic activity in the CHO cell lines.