Synthesis of l-indospicine, [5,5,6-(2)H3]-l-indospicine and l-norindospicine.

Indospicine is a non-proteogenic amino acid that accumulates as the free amino acid in livestock grazing Indigofera plant species and causes both reproductive losses and hepatotoxic effects. An efficient synthetic route to l-indospicine from l-homoserine lactone is described. The methodology is applicable for the synthesis of both deuterium labelled isotopomers and structural analogues for utilisation in biological studies. The key steps are a zinc mediated Barbier reaction with acrylonitrile and a Pinner reaction that together introduce the target amidine moiety.

Continued Studies on the Effects of Simazine on the Liver Histological Structure and Metamorphosis in the Developing Xenopus laevis.

This study continued our previous work (Sai et al. in Bull Environ Contam Toxicol 95:157-163, 2015a) by analysing the effects of simazine on the liver histological structure and metamorphosis in the developing Xenopus laevis. Tadpoles (Nieuwkoop-Faber stage 46) were exposed to simazine at 0.1, 1.2, 11.0 and 100.9 µg/L for 100 days. When tadpoles were exposed to simazine at 11.0 and 100.9 µg/L, an increased mortality and damaged liver tissues were observed together with significant inhibition of percent of X. laevis completing metamorphosis on days 80 and 90 and prolonged time of completing metamorphosis. On the other hand, we found that simazine has no significant effects on liver weight and altered hepatosomatic index. Results of this study may be considered to inform risk assessment of the effects of simazine on the development of X. laevis.

Micronucleus formation by single and mixed heavy metals/loids and PAH compounds in HepG2 cells.

Humans and other organisms are exposed to multi-chemical mixtures including commonly found carcinogens such as polycyclic aromatic hydrocarbons (PAHs) and heavy metal/loids. The joint effects of these chemicals as beyond the binary mixtures have not been well characterised. In this study, we evaluated the combined genotoxicity of mixtures of PAHs and heavy metal/loids containing benzo(a)pyrene (B[a]P), naphthalene (Nap), phenanthrene (Phe), pyrene (Pyr), arsenic (As), cadmium (Cd) and chromium (Cr) using in vitro micronucleus (MN) test in HepG2 cells. The induction of aryl hydrocarbon receptor (AhR) by single and mixed PAHs was also measured. The results indicated that individual and mixed Nap, Phe and Pyr did not induce significant MN frequencies. PAHs mixture containing B[a]P and B[a]P alone caused significant but similar level of MN frequencies. The same pattern was found in their AhR induction. Individual metal/loids induced significant cytostasis and MN formation of which Cd was found the most potent inducer. Mixture of metal/loids caused higher frequency of MN suggesting a possible additive effect among metal/loids. In addition, binary mixture of metal/loids and B[a]P, namely As/B[a]P, Cd/B[a]P and Cr/B[a]P, increased MN formation. Mixture of Cd and B[a]P induced the highest level of MN. Exposure of cells to the mixture containing B[a]P and Cd/Cr/As at lower concentration (0.25 µM) resulted in significant MN frequency, the level of which was equal to that by Cd/B[a]P at 1.0 µM. The results of the study suggested that an additive effect may exist between PAHs and heavy metal/loids in a compound- and concentration-dependent manner. The compounds with highest potencies of genotoxicity in the mixture seem dominant as driving sources in the final combined genotoxicity of PAHs and heavy metal/loids.

Assessing atrazine-induced toxicities in Bufo bufo gargarizans Cantor.

Atrazine (AZ), a widely used herbicide has drawn attentions for its potential impacts on amphibians. This study aims to investigate the toxicity of AZ in Bufo bufo gargarizans Cantor (B. bufo gargarizans), a species of toad commonly found in China and countries in East Asia. We treated tadpoles with 0.1, 1, 10 and 100 µg/L AZ for 85 days and examined related parameters. The results showed that the mortality of the toads in the treatment group increased dramatically in a U-shaped dose-response relationship. The hindlimb extension and metamorphosis rate of the toads were significantly inhibited by AZ at 10 and 100 µg/L. Under the same condition, there were significant progressive changes in the testicular structures. Moreover, we found that AZ has no significant effects on growth, sex ratios, gonadal morphology, forelimb emergence and histology in the ovaries. Our results support the idea that environmental contaminants including AZ may be relevant to global amphibian decline.

The Effects of Simazine, a Chlorotriazine Herbicide, on the Expression of Genes in Developing Male Xenopus laevis.

Simazine was investigated for gene expression concurrent with simazine-induced phenotype changes during development of male Xenopus laevis. X. laevis tadpoles (Nieuwkoop-Faber stage 46) were exposed to 0.1, 1.2, 11.0 and 100.9 µg/L simazine for 100 days. The results showed that an increased mortality of X. laevis, decreased gonad weight and altered gonadosomatic index of males significantly (p<0.05) when exposed to simazine at 11.0 and 100.9 µg/L. Significant degeneration in testicular tissues was observed when tadpoles were exposed to simazine at 100.9 µg/L. To investigate the molecular mechanisms behind the testicular degeneration by simazine, we evaluated gene expression in animals treated with 100.9 µg/L simazine and found that 1,315 genes were significantly altered (454 upregulated, 861 downregulated). Genes involved in the cell cycle control, and amino acid metabolism pathways were significantly downregulated. These results indicate that simazine affects the related gene expressions which may be helpful for the understanding of the reason for the reproductive toxicity of simazine on male X. laevis.

Mixture effects of benzene, toluene, ethylbenzene, and xylenes (BTEX) on lung carcinoma cells via a hanging drop air exposure system.

A recently developed hanging drop air exposure system for toxicity studies of volatile chemicals was applied to evaluate the cell viability of lung carcinoma A549 cells after 1 and 24 h of exposure to benzene, toluene, ethylbenzene, and xylenes (BTEX) as individual compounds and as mixtures of four or six components. The cellular chemical concentrations causing 50% reduction of cell viability (EC50) were calculated using a mass balance model and came to 17, 12, 11, 9, 4, and 4 mmol/kg cell dry weight for benzene, toluene, ethylbenzene, m-xylene, o-xylene, and p-xylene, respectively, after 1 h of exposure. The EC50 decreased by a factor of 4 after 24 h of exposure. All mixture effects were best described by the mixture toxicity model of concentration addition, which is valid for chemicals with the same mode of action. Good agreement with the model predictions was found for benzene, toluene, ethylbenzene, and m-xylene at four different representative fixed concentration ratios after 1 h of exposure, but lower agreement with mixture prediction was obtained after 24 h of exposure. A recreated car exhaust mixture, which involved the contribution of the more toxic p-xylene and o-xylene, yielded an acceptable, but lower quality, prediction as well.

Uncovering the impact of mega-scale shipbreaking yards on soil and crop quality in Bangladesh: A spatiotemporal dynamics and associated health risks of metal/loid contamination.

The uncontrolled release of harmful metal/loids from mega-scale shipbreaking activities in Bangladesh is a significant concern. This study investigated the impact of shipbreaking activities on soil and crop quality and human health in relation to metal/loid contamination. This work covered an area of 1221 km2 surrounding the shipbreaking yards in Chittagong during the wet and dry seasons between 2019 and 2020. Amongst the sixteen elements measured, the concentrations of Pb, Cd, As, V, Cr, Mn, Cu, Zn, Fe, Co, Ni, and Sn in the soil, rice, and vegetables from the four exposure sites were significantly higher compared to the control site in both seasons. Soil pollution indices indicated moderate to higher contamination levels of Pb, Zn, Cd, As, and Se in 30-50% of soil, supporting their accumulation in food crops. Source apportionment analysis identified uncontrolled shipwrecking operations as the primary anthropogenic activity mainly contributing to metal/loid pollution. Health risk analysis showed inorganic arsenic (estimated), Cd, and Pb in food crops could pose potential health threats to the general population. Spinach leaf and gourd were identified as the highest-risk contributing vegetables in the dry and wet seasons. These findings help to inform management strategies to protect agroecosystems and public health.

Genotoxicity of hydroquinone in A549 cells.

Hydroquinone (HQ) is found in natural and anthropogenic sources including food, cosmetics, cigarette smoke, and industrial products. In addition to ingestion and dermal absorption, human exposure to HQ may also occur by inhaling cigarette smoke or polluted air. The adverse effects of HQ on respiratory systems have been studied, but genotoxicity HQ on human lung cells is unclear. The aim of this study was to investigate the cytotoxicity and genotoxicity of HQ in human lung alveolar epithelial cells (A549). We found that HQ induced a dose response in cell growth inhibition and DNA damage which was associated with an increase in oxidative stress. Cytotoxicity results demonstrated that HQ was most toxic after 24 h (LC50 = 33 µM) and less toxic after 1 h exposure (LC50 = 59 µM). Genotoxicity of HQ was measured using the Comet assay, H2AX phosphorylation, and chromosome aberration formation. Results from the comet assay revealed that DNA damage was highest during the earlier hours of exposure (1 and 6 h) and thereafter was reduced. A similar pattern was observed for H2AX phosphorylation suggesting that damage DNA may be repaired in later exposure hours. An increase in chromosomal aberration corresponded with maximal DNA damage which further confirmed the genotoxic effects of HQ. To investigate whether oxidative stress was involved in the cytotoxic and genotoxic effects of HQ, cellular glutathione and 8-Oxo-deoguanisone (8-Oxo-dG) formation were measured. A decrease in the reduced glutathione (GSH) and an increase oxidized glutathione (GSSG) was observed during the early hours of exposure which corresponded with elevated 8-Oxo-dG adducts. Together these results demonstrate that HQ exerts its cytotoxic and genotoxic effects in A549 lung cells, probably through DNA damage via oxidative stress.

Interaction effects of lead on bioavailability and pharmacokinetics of arsenic in the rat.

Arsenic (As) and lead (Pb) are common contaminants found in mine waste materials. For an evidence-based risk assessment, it is important to better understand the potential interaction of mixed contaminants; and this interaction study was investigated in an in vivo rat model. Following co-administration of a fixed dose of As(V) as in sodium arsenate and different doses of Pb as lead acetate to Sprague-Dawley rats, blood arsenic concentration and bioavailability decreased. A decrease in As blood concentration when lead was co-administered was observed with increasing lead doses. Pharmacokinetic parameters for As in the blood showed faster absorption and elimination of this metalloid in the presence of Pb. The elimination half-life of As decreased from 67 days in As solo group to 27-30 with doses of Pb. Bioavailability of As was also decreased by 30-43 % in the presence of Pb. Decreased urinary excretion of Pb and tissue accumulation were also observed. It indicates lower absorption of As when co-administered with Pb. A probable explanation for these findings is that As co-administration with Pb could have resulted in the formation of less soluble lead arsenate. However, such an interaction between As and Pb could only explain about one-third of the variation when real mine waste materials containing both of these elements were administered to rats. This suggests that other effects from physical and chemical parameters could contribute to the bioavailability of arsenic in complex real environmental samples.

Effect of basal metabolic rate on lifespan: a sex-specific Mendelian randomization study.

Observationally, the association of basal metabolic rate (BMR) with mortality is mixed, although some ageing theories suggest that higher BMR should reduce lifespan. It remains unclear whether a causal association exists. In this one-sample Mendelian randomization study, we aimed to estimate the casual effect of BMR on parental attained age, a proxy for lifespan, using two-sample Mendelian randomization methods. We obtained genetic variants strongly (p-value < 5 × 10-8) and independently (r2 < 0.001) predicting BMR from the UK Biobank and applied them to a genome-wide association study of parental attained age based on the UK Biobank. We meta-analyzed genetic variant-specific Wald ratios using inverse-variance weighting with multiplicative random effects by sex, supplemented by sensitivity analysis. A total of 178 and 180 genetic variants predicting BMR in men and women were available for father's and mother's attained age, respectively. Genetically predicted BMR was inversely associated with father's and mother's attained age (years of life lost per unit increase in effect size of genetically predicted BMR, 0.46 and 1.36; 95% confidence interval 0.07-0.85 and 0.89-1.82), with a stronger association in women than men. In conclusion, higher BMR might reduce lifespan. The underlying pathways linking to major causes of death and relevant interventions warrant further investigation.

Clinical outcome of a quadrifocal (trifocal) intraocular lens in Chinese patients: prospective, observational case series.

PURPOSE: To report the visual outcomes and quality of vision and life after bilateral implantation of a single-piece trifocal intraocular lens (IOL) in Chinese patients. SETTING: Hong Kong Sanatorium & Hospital, Hong Kong, China. DESIGN: Prospective, observational case series. METHODS: Patients with bilateral implantation of AcrySof IQ PanOptix multifocal IOL were included. Distance, intermediate (60 cm), and near (40 cm) visual acuities (VAs) and contrast sensitivity (CS), defocus curve, preoperative higher-order aberration (HOA), dysphotopsia (0 to 5), satisfaction (1 to 5), spectacle independence, and quality of life were evaluated. The association between preoperative HOA and postoperative halos was also assessed. RESULTS: 54 eyes of 27 patients were included. The mean binocular distance, intermediate, and near uncorrected VA was -0.05 ± 0.06 (20/18), 0.06 ± 0.10 (20/23), and 0.04 ± 0.05 (20/22), respectively. No eyes lost more than 1 line of vision. Binocular CS was comparable with the monocular population norm of older adults. The defocus curve demonstrated that the binocular VA of 20/25 or better was achieved at a power of -3.00 to +0.50 diopters. The mean scores for halos, glare, and starbursts were 2.4 ± 1.4, 0.2 ± 0.8, and 1.4 ± 1.4 (of 5), respectively. The mean satisfaction score was 4.3 ± 0.7 (of 5). All the patients (100%) reported total spectacle independence. The mean vision-targeted composite score of the vision-related quality-of-life questionnaire was 97.2 ± 9.7 (of 100). Preoperative HOA was not associated with postoperative halos. CONCLUSIONS: Implantation of the trifocal IOL provided satisfactory visual outcomes and quality of vision and life, which resulted in a high rate of spectacle independence.

Relationships between growth indicators, liver and kidney function markers, and blood concentrations of essential and potentially toxic elements in environmentally exposed young children.

Environmental exposure to multiple metals and metalloids is widespread, leading to a global concern relating to the adverse health effects of mixed-metals exposure, especially in young children living around industrial areas. This study aimed to quantify the concentrations of essential and potentially toxic elements in blood and to examine the potential associations between multiple elements exposures, growth determinants, and liver and kidney function biomarkers in children living in several industrial areas in Dhaka, Bangladesh. The blood distribution of 20 trace elements As, Ag, Bi, Br, Cd, Co, Cr, Cu, I, Mn, Hg, Mo, Ni, Pb, Se, Sb, Tl, V, U, and Zn, growth determinants such as body mass index and body fats, blood pressure, liver and kidney injury biomarkers including serum alanine aminotransferase and alkaline phosphatase activities, serum calcium, and creatinine levels, blood urea nitrogen, and hemoglobin concentrations, and glomerular filtration rate were measured in 141 children, aged six to 16 years. Among these elements, blood concentrations of Ag, U, V, Cr, Cd, Sb, and Bi were measured below LOQs and excluded from subsequent statistical analysis. This comprehensive study revealed that blood concentrations of these elements in children, living in industrial areas, exceeded critical reference values to varying extents; elevated exposure to As, Pb, Br, Cu, and Se was found in children living in multiple industrial areas. A significant positive association between elevated blood Tl concentration and obesity (ß = 0.300, p = 0.007) and an inverse relationship between lower As concentration and underweight (ß = -0.351, p < 0.001) compared to healthy weight children indicate that chronic exposure to Tl and As may influence the metabolic burden and physical growth in children. Concentration-dependent positive associations were observed between the blood concentrations of Cu, Se, and Br and hepatic- and renal dysfunction biomarkers, an inverse association with blood Mo and I level, however, indicates an increased risk of Cu, Se, and Br-induced liver and kidney toxicity. Further in-depth studies are warranted to elucidate the underlying mechanisms of the observed associations. Regular biomonitoring of elemental exposures is also indispensable to regulate pollution in consideration of the long-term health effects of mixed-elements exposure in children.

Metabolism of bilirubin by human cytochrome P450 2A6.

The mouse cytochrome P450 (CYP) 2A5 has recently been shown to function as hepatic "Bilirubin Oxidase" (Abu-Bakar, A., et al., 2011. Toxicol. Appl. Pharmacol. 257, 14-22). To date, no information is available on human CYP isoforms involvement in bilirubin metabolism. In this paper we provide novel evidence for human CYP2A6 metabolising the tetrapyrrole bilirubin. Incubation of bilirubin with recombinant yeast microsomes expressing the CYP2A6 showed that bilirubin inhibited CYP2A6-dependent coumarin 7-hydroxylase activity to almost 100% with an estimated K(i) of 2.23 µM. Metabolite screening by a high-performance liquid chromatography/electrospray ionisation mass spectrometry indicated that CYP2A6 oxidised bilirubin to biliverdin and to three other smaller products with m/z values of 301, 315 and 333. Molecular docking analyses indicated that bilirubin and its positively charged intermediate interacted with key amino acid residues at the enzyme's active site. They were stabilised at the site in a conformation favouring biliverdin formation. By contrast, the end product, biliverdin was less fitting to the active site with the critical central methylene bridge distanced from the CYP2A6 haem iron facilitating its release. Furthermore, bilirubin treatment of HepG2 cells increased the CYP2A6 protein and activity levels with no effect on the corresponding mRNA. Co-treatment with cycloheximide (CHX), a protein synthesis inhibitor, resulted in increased half-life of the CYP2A6 compared to cells treated only with CHX. Collectively, the observations indicate that the CYP2A6 may function as human "Bilirubin Oxidase" where bilirubin is potentially a substrate and a regulator of the enzyme.

Visual outcomes and patient satisfaction after presbyopic lens exchange with a diffractive multifocal intraocular lens.

PURPOSE: To investigate the visual outcomes, severity of symptoms, and patient satisfaction after refractive lens exchange (RLE) with a diffractive multifocal intraocular lens (IOL). METHODS: A nonrandomized, unmasked, retrospective chart review study was performed. Patients who underwent RLE with ZMA00 (Abbott Laboratories) were identified from a hospital database. Eyes with preoperative uncorrected distance visual acuity or corrected distance visual acuity 20/20 or better were included. The study cohort comprised 45 eyes from 29 patients. Monocular uncorrected and distance-corrected visual acuity at distance, 67 cm, and 30 cm were measured 6 months postoperatively. A patient questionnaire assessing visual symptoms (halo, night glare, and starburst) and satisfaction with visual performance was administered. RESULTS: Six months postoperatively, mean uncorrected visual acuity (logMAR) was -0.10±0.13, 0.43±0.25 at 67 m (intermediate), and 0.18±0.05 at 30 m (near). Mean distance-corrected visual acuity at these distances was -0.02±0.06, 0.40±0.21, and 0.17±0.02, respectively. Twenty-seven patients completed the questionnaire. Patients reported postoperative halos (78%), night glare (26%), and starbursts (48%). All bilateral RLE patients were spectacle-free at all distances, whereas 50% of unilateral RLE patients required spectacles postoperatively. Bilateral RLE patients with habitual spectacle use preoperatively were the most satisfied with their postoperative visual performance. CONCLUSIONS: Refractive lens exchange with the ZMA00 is an option for presbyopic correction; however, significant glare, halo, and starburst issues are subjectively reported.

Dissolved organic carbon reduces uranium toxicity to the unicellular eukaryote Euglena gracilis.

The influence of dissolved organic carbon (DOC), in the form of Suwannee River fulvic acid (SRFA), on uranium (U) toxicity to the unicellular eukaryote, Euglena gracilis (Z strain), was investigated at pH 6. In a background medium without SRFA, exposure of E. gracilis to 57 µg L(-1) U resulted in a 50% reduction in growth (IC(50)). The addition of 20 mg L(-1) DOC (as SRFA), reduced U toxicity 4 to 5-fold (IC(50) increased to 254 µg L(-1) U). This reduction in toxicity was also evident at more sensitive effect levels with a 10% reduction in growth (IC(10)) occurring at 5 µg L(-1) U in the background medium and at 17 µg L(-1) U in the SRFA medium, respectively. This amelioration of toxicity with the addition of SRFA was linked to a decrease in the bioavailability of U, with geochemical speciation modelling predicting 84% of U would be complexed by SRFA. The decrease in bioavailability of U in the presence of SRFA was also evident from the 11-14 fold reduction in the cellular concentration of U compared to that of E. gracilis in the background medium. Stepwise multiple linear regression analyses indicated that UO(2)(2+) alone explained 51% of the variation in measured U toxicity to E. gracilis. Preliminary U exposures to E. gracilis in the presence of a reactive oxygen species probe, suggest exposure to ≥60 µg L(-1) U may induce oxidative stress, but this endpoint was not considered to be a sensitive biological indicator.

Genotoxicity assessment of per- and polyfluoroalkyl substances mixtures in human liver cells (HepG2).

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous, toxic, and persistent environmental chemicals of concern that have been widely detected in all environmental matrices including human biological fluids. Although humans are exposed to complex mixtures of PFAS, it remains uncertain whether the co-exposure to PFAS mixtures could induce genotoxic damage in humans. Hence, this study evaluated the combined genotoxicity of PFAS mixtures in a human cell line system. To assess the possible genotoxic damage caused by human exposure to PFAS and their mixtures, we investigated their potential to induce cytotoxicity (cell viability) and genotoxicity (DNA damage) in a human liver cell line (HepG2). The selected PFAS include perfluorononanoic acid (PFNA), perfluorooctane sulfonic acid (PFOS), perfluorodecanoic acid (PFDA), perfluorooctanoic acid (PFOA), and perfluorohexane sulfonic acid (PFHxS). The interaction toxicities of these PFAS in binary mixtures were also determined using the additive index approach. The results revealed that exposure to PFNA, PFOS, PFDA, PFOA, and PFHxS singly and in binary mixtures induced a concentration-dependent decrease in cell viability. The additive index values indicated that the binary mixtures of PFOS + PFNA, PFOS + PFDA, and PFOS + PFOA displayed synergistic interaction, whereas the binary mixtures of PFOS + PFHxS, PFOA + PFNA, PFOA + PFDA, and PFOA + PFHxS behaved additively. Using the alkaline Comet assay, the potential of PFAS and their mixtures to induce DNA damage was evaluated based on a 1:1 ratio of the concentration of respective compounds required to produce a 1/10th of effective concentrations causing 50 % inhibition in cell viability (EC50). The results revealed that exposure to PFNA, PFOS, PFDA, PFOA, and PFHxS singly and in binary mixtures (PFOS + PFNA, PFOS + PFDA, PFOS + PFOA, PFOS + PFHxS, PFOA + PFNA, PFOA + PFDA, and PFOA + PFHxS) caused a moderate increase in cellular DNA damage, but no dose-response relationship was observed. Overall, this study indicates that the tested PFAS causes a concentration-dependent decrease in cell viability and only a modest increase in cellular DNA damage under these conditions.

Combined effects of mixed per- and polyfluoroalkyl substances on the Nrf2-ARE pathway in ARE reporter-HepG2 cells.

Although the Nrf2-ARE pathway plays a critical role in cellular protection against toxicity and oxidative stress from environmental chemical stressors, the association between exposure to per- and polyfluoroalkyl substances (PFAS) mixtures and the changes of Nrf2-ARE pathway remains largely unexplored. This study evaluated the potential of PFAS to induce the Nrf2-ARE pathway as individual compounds and as binary, ternary, and multicomponent mixtures in the ARE reporter-HepG2 cells and compared the mixture toxicity data to the predictions by concentration addition (CA) model. The toxicological interactions between PFAS mixture components were also determined by the model deviation ratio (MDR) between the CA predicted and mixture toxicity values. The induction of the Nrf2-ARE pathway was quantified using the luciferase system, and the endpoint assessed was the concentration that induced an induction ratio (IR) of 1.5 (ECIR1.5). The results showed that exposures to both individual and mixed PFAS induced the Nrf2-ARE pathway in ARE reporter-HepG2 cells. Based on the MDRs, the combinations with PFOS showed synergistic interactive effects, while the combinations with PFOA showed additive effects. These results indicate that the CA model underestimated the mixture toxicity of PFAS with PFOS co-exposures and may have health risk assessment implications.

Concentrations of essential and toxic elements and health risk assessment in brown rice from Qatari market.

Twenty-two brown rice varieties available in the Qatari market were analyzed for essential and toxic elements by ICP-MS. Found concentrations (µg/kg) were: As: 171 ± 78 (62-343), Cd: 42 ± 60 (4-253), Cr: 515 ± 69 (401-639), Pb: 6 ± 7 ( 1 in million, may possibly be > 1 in 10,000 based on conservatively high brown rice consumption rates of 200 g/d or 400 g/d in Qatar. These elevated risks may be applicable to specific population subgroups with diabetic conditions who consume only brown rice. Non-cancer risks are mainly derived from Mn, V, Se, and Cd with a hazard index > 1 from some brown rice samples.

Toxicity assessment of historical aqueous film-forming foams (AFFFs) using cell-based assays.

Aqueous film-forming foam (AFFF) has historically contained high concentrations of long-chain per-and polyfluoroalkyl substances (PFAS), which have been linked with adverse health outcomes. However, the toxicity of historical AFFFs remains largely unknown, presenting uncertainties in their risk assessment. This study assessed the toxicity of historical AFFFs by exposing human liver cells (HepG2) to various dilutions of 3M Light Water AFFF or Ansulite AFFF (0.001%, 0.002%, 0.005%, 0.009%, 0.019%, 0.038%, 0.075%, 0.15%, and 0.3%) for 24 h. The effects of the two AFFF formulations on the cell viability, intracellular reactive oxygen species (ROS) production, Nrf2-ARE activity, and DNA damage were assessed by CellTiter 96® Aqueous One Solution Cell Proliferation Assay (MTS kit), dichlorofluorescein diacetate assay, luciferase assay, and alkaline Comet assay, respectively. The results revealed that the two brands of AFFFs tested were toxic to HepG2 cells at dilutions lower than the recommended 3% application formulation. Specifically, exposure to 3M Light Water AFFF or Ansulite AFFF induced a dilution-dependent decrease in cell viability, increased intracellular ROS production, and increased Nrf2-ARE activity. However, except for the highest concentration (lowest dilution) of 3M Light Water AFFF tested (0.038%.), both 3M Light Water AFFF and Ansulite AFFF did not significantly induce cellular DNA damage. Overall, 3M Light Water AFFF was more toxic than Ansulite AFFF. The findings from this study provided valuable in vitro toxicity data that may better inform the health risk assessment of these historical AFFFs.