Search Results (102)

Aflatoxin B1 (AFB1) and patulin (PAT) are prevalent foodborne mycotoxins with hepatotoxic potential, but the hepatic effects of combined exposure remain largely unclear. This study investigated the hepatotoxic consequences of co-exposure to AFB1 and PAT using no-observed adverse effect levels (NOAELs) in C57BL/6 mice and low-cytotoxic concentrations in HepG2 cells selected by viability screening. Mice and cells were assigned to four groups: control, AFB1, PAT and AFB1 + PAT. Exposure to either toxin individually did not cause evident liver injury, whereas co-exposure significantly elevated alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, reduced liver index, and induced clear histopathological alterations. Co-exposure markedly aggravated oxidative stress, characterized by increased reactive oxygen species (ROS) and malondialdehyde (MDA) and decreased superoxide dismutase (SOD). In parallel, the levels of interleukin-6 (IL-6), interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) were elevated, together with the early fibrosis-related markers alpha-smooth muscle actin (α-SMA) and vimentin. The apoptotic response was characterized by increased Bcl-2-associated X protein (Bax) and reduced B-cell lymphoma-2 (Bcl-2), together with cysteine-dependent aspartate-specific protease-3 (caspase-3) activation. These findings indicate that co-exposure to AFB1 and PAT elicits hepatotoxicity through amplified oxidative stress, inflammation, and caspase-dependent apoptosis, supporting the need to further consider mycotoxin co-exposure in toxicological evaluation. Full article

The evaluation of the safety of chemical substances requires the identification of a safe dose, which has no adverse effects on humans. This is obtained through animal studies, with exposure prolonged for months. Repeated-dose toxicity is a term in toxicology and pharmacology referring to the highest tested dose of a substance, so-called No Observed Adverse Effect Level (NOAEL). Experimental data on NOAEL taken from the literature and the OpenFoodTox database (total n = 848). To speed up the processing of the enormous number of substances we are exposed to, in silico models are an attractive solution. Monte Carlo technique, incorporating the Las Vegas algorithm, was applied to develop models for repeated-dose toxicity in rats. Optimal descriptors were calculated using correlation weights for attributes of the Simplified Molecular Input Line Entry System (SMILES). Computational experiments were conducted 5 times, with splits obtained using the Las Vegas algorithm. Good predictive potential was observed for these models, with an average determination coefficient on the validation set of 0.77 ± 0.04. Full article

L-Ergothioneine (L-EGT), a naturally occurring thiol compound abundant mainly in edible fungi, is increasingly regarded as a potentially beneficial bioactive constituent. However, population-level exposure data remain limited. This study aimed to estimate background dietary exposure to L-EGT among Chinese residents, describe its distribution across population subgroups and regions, identify major food contributors, and characterize the risk using a margin of exposure (MOE) approach. Individual body-weight-normalized L-EGT intakes were estimated from published food concentration data and three-day dietary recalls of 42,218 participants. MOEs were calculated using a no observed adverse effect level (NOAEL) of 800 mg/kg bw/d obtained from subchronic toxicity studies. The mean dietary exposure to L-EGT was 0.043 mg/kg bw/d (MOE = 18,605) in the general population and 0.174 mg/kg bw/d (MOE = 4598) among consumers, with 95th percentile exposures of 0.244 mg/kg bw/d (MOE = 3279) and 0.644 mg/kg bw/d (MOE = 1242), respectively. MOE values were consistently above the safety threshold of 300 across all subgroups, with less than 0.3% of the total population and 1.3% of consumers aged 3–6 years falling below this value. These results indicate that current natural dietary exposure to L-EGT in China is low and does not raise safety concerns and provide essential baseline data for future studies on its health effects, optimal intake ranges, and long-term safety. Full article

This study investigated the chemical composition, antioxidant activity, and genotoxic potential of essential oils (EOs) obtained by hydrodistillation from aerial parts of four wild-growing Lamiaceae species in eastern Morocco: Spanish ziziphora (Ziziphora hispanica L.), felty germander (Teucrium polium L.), French lavender (Lavandula dentata L.), and topped lavender (Lavandula stoechas L.). Gas chromatography–mass spectrometry (GC-MS) analysis revealed eucalyptol (40.08%), thujopsene (11.25%), β-myrcene (15.82%), and fenchone (30.69%) as the major constituents in Z. hispanica, T. polium, L. dentata, and L. stoechas, respectively. Antioxidant capacity was evaluated using three complementary assays: 2,2-diphenyl-1-picrylhydrazyl radical scavenging, ferric reducing antioxidant power, and β-carotene bleaching. L. stoechas and L. dentata exhibited the strongest antioxidant activities, with IC₅₀ values ranging from 0.284 to 1.71 mg/mL across assays. Genotoxicity was assessed in rat leukocytes using the alkaline Comet assay at EO concentrations of 2.5, 5, and 10 µg/mL. All tested EOs induced statistically significant DNA damage compared to the negative control, though the extent varied by species and concentration; notably, L. stoechas at 2.5 µg/mL showed the lowest genotoxic impact. These findings highlight the dual potential of these EOs as natural antioxidants while underscoring the need for dose-dependent safety evaluation prior to therapeutic or industrial application. Given that DNA damage was detectable even at 2.5 µg/mL, a conservative practical recommendation is to keep EO levels below 2.5 µg/mL-equivalent in preliminary applications, pending further in vivo toxicology to establish NOAEL-based exposure limits. Full article

This study evaluated two formulations of L-carnitine, which were developed and impregnated in an oil-based self-emulsifying system (SEDDS), the first with tyrphostin AG17 and the second without the addition of tyrphostin AG17. The formulation with tyrphostin AG17 showed the presence of stable microvesicles up to 498 h after its preparation. To establish a robust safety profile in compliance with modern regulatory frameworks and the 3Rs principle (replacement, reduction, and refinement), a toxicological evaluation was conducted integrating an in silico quantitative structure–activity relationship (QSAR) analysis with confirmatory in vivo subchronic toxicity studies. The QSAR analysis, performed using the OECD QSAR Toolbox and strictly adhering to Organization for Economic Co-operation and Development (OECD) validation principles, predicted an acute oral LD50 of 91.5 mg/kg in rats, a value showing high concordance with the historical experimental data (87 mg/kg). Furthermore, computational modeling for repeated-dose toxicity yielded a no-observed-adverse-effect level (NOAEL) of 80.0 mg/kg bw/day, a no-observed-effect level (NOEL) of 60.4 mg/kg bw/day, and an ADI = 56 mg/day. These computational findings were substantiated by a 90-day subchronic toxicity study in male Wistar rats, where daily intragastric administration of tyrphostin AG17 at doses up to 1.75 mg/kg resulted in not statistically significant hematotoxic activity (p < 0.05), with a maximum cumulative dose over 90 days of 157.5 mg/kg. Collectively, these data indicate that tyrphostin AG17 combines high stabilizing efficacy with a manageable safety profile, supporting its proposed regulatory status as a functional food additive. Based on these results, it is concluded that tyrphostin AG17 shows promising characteristics for use as a stabilizer in food and other substances. Full article

Amino acid availability is central for the synthesis of macromolecules and numerous bioactive compounds. Amino acids are also involved in ATP production, cell signaling, and the epigenetic regulation of gene expression in human cells. From clinical and experimental studies, it appears that supplementation with specific amino acids may be relevant to correct for amino acid deficiency in the case of insufficient supply from dietary proteins with regards to the amounts needed for optimal metabolism and physiological functions. Clinical and experimental arguments suggest that amino acid supplementation may be indicated in specific situations under a specific nutritional context. However, it is essential not to overdose with excessive quantities of amino acids in supplements beyond the upper levels of safe intake (ULSI). In this narrative review, I recapitulate the protein and amino acid requirements for the general population and for subgroups of the population, and these requirements are compared to the usual consumption. Typical examples of clinical trials showing the benefits from amino acid supplementation in different physiological and pathophysiological contexts are presented together with results obtained from experimental studies. Parameters such as the no-observed-adverse-effect-level (NOAEL) values used to determine the ULSI for amino acid supplementation are defined, and values determined in clinical trials are given and discussed. Finally, prospects for future research in the field are proposed. Full article

Glyphosate (GLY), the active ingredient in widely used herbicides, was long considered specific to plants and bacteria, yet mounting evidence shows it can impair endocrine and reproductive functions in animals. Given its widespread use and environmental persistence, assessing its effects at regulatory-approved doses is critical. Here, adult female zebrafish (Danio rerio) were exposed for 21 days to different concentrations of dietary GLY at 0.5 mg/kg body weight/day (GLY0.5, acceptable daily intake, ADI), 5 mg/kg/day (GLY5), and 50 mg/kg/day (GLY50, no-observed-adverse-effect level, NOAEL). Our findings show that dietary GLY induces dose-dependent perturbations along the hepato-gonadal axis. At the highest dose, chronic stress responses were evident through elevated cortisol and cortisone, accompanied by hepatic glycogen accumulation and ferroptotic stress. Although follicle histology appeared normal, alterations in several genes involved in oocyte maturation and estrogen receptor signaling translated into reduced fertilization, revealing compromised gamete quality rather than overt follicular development abnormality. Likewise, the lowest dose triggered modifications in genes crucial for oogenesis without altering the follicle development, although in this case, potential compensatory mechanisms could have led to enhanced fertilization. GLY5 did not alter the number of fertilized eggs but significantly increased embryo mortality. Overall, dietary GLY disrupted hepatic metabolism, endocrine signaling, and reproduction in a non-monotonic manner, even at levels considered safe by EFSA. These findings highlight the need to reevaluate current safety thresholds with attention to female-specific reproductive risks. Full article

The present study aimed to assess the potential maternal toxicity of Ficus deltoidea var. kunstleri aqueous extract in pregnant rats, along with its impact on maternal hepatic drug metabolism and foetal skeletal development. Pregnant rats were divided into five groups and orally administered varying doses of F. deltoidea aqueous extract (0, 250, 500, 1000, and 2000 mg/kg body weight) from gestation day 6 to 20. Throughout the administration period, clinical observations, body weight, and food and water intake were monitored. On gestation day 21, the pregnant rats were sacrificed, and their vital organs and foetuses were collected for analysis. Gene expression related to hepatic drug metabolism was evaluated using the RT2 Profiler™ PCR array. Foetal external morphology was examined for abnormalities, and skeletal structures were stained with Alizarin Red to assess the effects of F. deltoidea aqueous extract on bone ossification during organogenesis. No maternal toxicity was observed, except for a significant increase in liver weight in the treated groups (p < 0.05). Analysis of 84 genes revealed significant changes in 15, 4, and 11 genes in the 250, 500, and 2000 mg/kg body weight groups, respectively. Notably, Gpx5 and Pkm, both phase II metabolising enzyme genes were downregulated in a dose-dependent manner. Despite some skeletal variations, the extract did not induce foetal external malformations or skeletal abnormalities. The significant increase in maternal liver weight, together with the downregulation of Gpx5 and Pkm, suggests an adaptive hepatic response to the extract rather than an adverse effect. These findings also suggest that F. deltoidea var. kunstleri aqueous extract does not cause embryo toxicity, foetal growth retardation, or developmental malformations, particularly in skeletal formation. The developmental no-observed-adverse-effect level (NOAEL) was determined to be >2000 mg/kg/day via oral administration. Further research is warranted to explore the synergistic interactions of genes involved in hepatic drug metabolism in response to the extract. Full article

Sulfoxaflor, an insecticide, acts on nicotinic acetylcholine receptors. It has a functional group similar to that of neonicotinoid insecticides, which are testicular toxicants. Recently, the adverse effects of sulfoxaflor on the testes have been reported in rats. This study aimed to address the lack of reports on sulfoxaflor administration in mice and its effects on the testes. ICR mice (3- and 10-week-old) were treated ad libitum with two different concentrations (10 and 100 mg/kg) of sulfoxaflor for 4 and 8 weeks. Histological analysis and real-time reverse transcription polymerase chain reaction were performed. Testis weights relative to body weights in the sulfoxaflor groups showed no significant difference compared to the control group. Testicular tissue in the sulfoxaflor groups was unchanged compared to that in the control group. The sulfoxaflor-treated group showed no significant differences in the mRNA expression of luteinizing hormone and follicle-stimulating hormone in the pituitary gland compared to the control group. Furthermore, no significant differences were noted in the mRNA expression levels of various gene markers in the testes between the sulfoxaflor-treated and control groups. These markers include those related to Leydig cells, testosterone synthesis, Sertoli cells, proliferating cells, meiotic cells, pachytene spermatocytes, round spermatids, apoptotic cells, antioxidant enzymes, oxidative stress factors, and mitochondrial function. In contrast to findings in rats, which showed testicular toxicity, sulfoxaflor administration at low concentrations did not adversely affect intratesticular cells in either mature or immature mice at the doses and time points examined. In the future, we would like to conduct research on high concentrations of sulfoxaflor by changing the administration method. Full article

Calcium nitrate tetrahydrate, used in fertilizers, wastewater treatment, and concrete admixtures, has limited toxicity data despite extensive industrial use. This study evaluated its repeated-dose and reproductive/developmental toxicity in Sprague Dawley rats following OECD TG 422, which combines TG 407 and 421 to extend dosing than TG 407 and reduce animal use compared with separate studies. Rats were administered 0, 100, 300, or 1000 mg/kg/day. Males were treated for 49 days and females from 2 weeks pre-mating to postpartum day 13; the recovery group was observed for an additional 2 weeks. Endpoints included clinical signs, body weight, food consumption, hematology, serum biochemistry, organ weights, histopathology, reproductive performance, and F1 development. No systemic toxicity was observed in F0 males. Minimal prostate atrophy occurred in high-dose males but was considered non-adverse due to limited severity. One high-dose female died on PPD 1, and high-dose F1 litters showed decreased litter size, increased post-implantation loss, and a reduced live-born index. Based on these results, NOAELs were cautiously assigned 1000 mg/kg/day for repeated-dose and male reproductive toxicity and 300 mg/kg/day for female reproductive and developmental toxicity. TG 422 efficiently characterized hazards while reducing animal use, though its limited duration and scope indicate the need for complementary studies. Full article

Coffee oil derived from spent coffee grounds of Coffea arabica is considered a novel food in the European Union (EU), requiring pre-market approval supported by comprehensive toxicological data. The effects of coffee oil on human health, particularly on blood parameters and liver enzymes, have been investigated in several studies. This review article summarizes the available toxicological literature on coffee oil, including its bioactive diterpenes cafestol and kahweol, which are known for their potential health effects. Considering the different modes of action of these two diterpenes, moderate consumption of coffee oil may be considered safe for healthy adults. Based on the changes in serum values in humans, this review provides initial estimations of LOAEL, NOAEL, and ADI for these diterpenes. The findings suggest that an intake of 225 mg of coffee oil per day might be considered safe assuming that coffee oil contains about 0.4% diterpenes. In summary, the assessment based on the published data indicates that (i) the consumption of coffee oil contained in any type of prepared coffee appears to be safe because the homeostasis of lipid levels in the blood is not significantly affected, and (ii) a low consumption of coffee oil as such might be acceptable but would require a refined risk assessment considering the exposure levels of the intended food product, which must be provided for novel food approval procedures. Full article

Background: The combined diphtheria–tetanus–acellular pertussis (three-component), Haemophilus influenzae type b (Hib, conjugate), and ACYW135 meningococcal (conjugate) vaccine (DTaP-Hib-MCV4) offers a promising alternative to single-component vaccines, potentially simplifying immunization schedules and improving vaccination coverage. Methods: We evaluated the safety, immunogenicity, and protective efficacy of DTaP-Hib-MCV4 in animal models. Acute and long-term toxicity studies were conducted in Sprague-Dawley (SD) rats with equal numbers of male and female animals. Immunogenicity was assessed in female NIH mice and SD rats using a three-dose regimen at 14-day intervals. Orbital blood was collected 14 days post-immunization to measure IgG titers against pertussis, diphtheria, tetanus, Hib, and meningococcal antigens. The protective efficacy was determined using potency tests for the pertussis, diphtheria, and tetanus components; passive protection studies for Hib; and serum bactericidal antibody (SBA) titers against A/C/Y/W135 meningococcal serogroups. Results: Acute and repeated-dose toxicity studies in SD rats showed no signs of abnormal toxicity or irritation at either high (three doses/rat) or low (one dose/rat) doses levels. The no-observed-adverse-effect level (NOAEL) for DTaP-Hib-MCV4 was established at three doses/rat after 8 weeks of repeated intramuscular administration and a 4-week recovery period. Specific IgG antibodies against all the vaccine components were detected in animal sera at both one and three doses/rat, with no evidence of immunotoxicity. Following two-dose primary immunization in murine models, the combined vaccine elicited robust antigen-specific antibody responses, with geometric mean titers (GMTs) as follows: 1,280,000 for pertussis toxin (PT); 761,093 for filamentous hemagglutinin (FHA); 1,159,326 for pertactin (PRN); 1,659,955 for diphtheria toxoid (DT); 1,522,185 for tetanus toxoid (TT); 99 for Haemophilus influenzae type b (Hib); and 25,600, 33,199, 8300, and 9051 for serogroups A, C, Y, and W135 of Neisseria meningitidis, respectively. In the rat models, three-dose primary immunization also elicited robust antigen-specific antibody responses. Protection studies demonstrated efficacy against pertussis, tetanus toxin, and diphtheria toxin challenges. In the Hib challenge study, none of the 10 animals given anti-DTaP-Hib-MCV4 antiserum developed bacteremia after the live Hib challenge (vs. 5814/0.1 mL in the negative control, p < 0.001). In addition, the SBA titers against meningococcal serogroups exceeded the protective threshold (≥1:8) in 92.2% of the immunized mice and 100% of the immunized rats. Crucially, the combined vaccine induced potent immune responses and protective efficacy, with antibody levels and protection against each component antigen comparable to or greater than those of the individual components: DTaP, Hib, and MCV4. Conclusions: These findings demonstrate that the DTaP-Hib-MCV4 combined vaccine is both safe and immunogenic, supporting its potential as a viable alternative to individual vaccines. This combined vaccine may streamline immunization programs and enhance vaccination coverage. Full article

This study rigorously evaluated the safety profile of dietary fiber extracted from cassava pulp, a promising functional food ingredient, through acute and 28-day sub-acute oral toxicity assessments in Wistar rats. This research hypothesized that cassava pulp fiber would exhibit minimal toxicity across a range of doses. In the acute study, rats received single oral doses of 175, 550, or 2000 mg/kg, while the sub-acute toxicity study involved daily doses of 250, 500, or 1000 mg/kg, with satellite groups included for reversibility assessment. Comprehensive monitoring encompassed clinical signs, mortality, body weight, food intake, hematological and biochemical parameters, relative organ weights, and detailed histopathological examination. Remarkably, no treatment-related mortality or overt clinical signs of toxicity were observed in either study. The LD₅₀ was higher than 2000 mg/kg for the acute study and the no-observed-adverse-effect level (NOAEL) was determined to be 2000 mg/kg for the acute study and 1000 mg/kg for the sub-acute toxicity study, indicating a high margin of safety. While statistically significant alterations were noted in some hematological, biochemical, and relative organ weight parameters, these changes were not considered toxicologically relevant. Notably, histopathological changes in the lungs were observed across all groups, including controls, warranting further investigation. These findings suggest that cassava pulp fiber is well tolerated at high oral doses, supporting its potential for safe application in food and nutraceutical formulations. However, the observed lung alterations necessitate further research to elucidate their etiology and clinical significance. Full article

Dietary exposure to a high dose of cadmium (Cd) ≥ 100 µg/day for at least 50 years or a lifetime intake of Cd ≥ 1 g can cause severe damage to the kidneys and bones. Alarmingly, however, exposure to a dose of Cd between 10 and 15 µg/day and excretion of Cd at a rate below 0.5 µg/g creatinine have been associated with an increased risk of diseases with a high prevalence worldwide, such as chronic kidney disease (CKD), fragile bones, diabetes, and cancer. These findings have cast considerable doubt on a “tolerable” Cd exposure level of 58 µg/day for a 70 kg person, while questioning the threshold level for the Cd excretion rate of 5.24 µg/g creatinine. The present review addresses many unmet challenges in a threshold-based risk assessment for Cd. Special emphasis is given to the benchmark dose (BMD) methodology to estimate the Cd exposure limit that aligns with a no-observed-adverse-effect level (NOAEL). Cd exposure limits estimated from conventional dosing experiments and human data are highlighted. The results of the BMDL modeling of the relationship between Cd excretion and various indicators of its effects on kidneys are summarized. It is recommended that exposure guidelines for Cd should employ the most recent scientific research data, dose–response curves constructed from an unbiased exposure indicator, and clinically relevant adverse effects such as proteinuria, albuminuria, and a decrease in the estimated glomerular filtration rate (eGFR). These are signs of developing CKD and its progression to the end stage, when dialysis or a kidney transplant is required for survival. Full article

Background: Epidemiological studies demonstrate that exposure to tobacco causes infertility. A reference cigarette contains up to 47 chemicals above the quantification level, of which acrylamide, benzopyrene, cadmium, ethylene oxide and lead are classified as known (category 1A), presumed (category 1B) or suspected (category 2) human reproductive toxicants due to their effects on fertility and sexual function. Methods: We collected toxicological information on these substances to establish their respective systemic-derived no-effect levels (internal doses predicted not to alter fertility). We also estimated the systemic exposure to these four substances by smokers consuming 20 cigarettes per day. Results: The risks (ratios between exposure and safe dose) were 0.23, 0.06, 0.18, 0.01 and 0.00002 for acrylamide, benzopyrene, cadmium, ethylene oxide and lead, respectively. The combined risk was 0.48. Conclusions: It was concluded that the changes in fertility resulting from the consumption of the substances in tobacco classified as toxic to fertility could not be explained by mechanisms with a toxicity threshold attributable to these five substances. No safe dose could be derived for tobacco use in persons seeking pregnancy; this applied to both active and passive smokers. Full article