Journal of Applied Toxicology最新文献

The teratogenic potential of dapagliflozin, a sodium-glucose co-transporter 2 (SGLT2) inhibitor, widely used in managing type 2 diabetes, remains poorly characterized. This study investigates the dose-dependent developmental effects of dapagliflozin using a chick embryo model to provide critical insights into its safety during early developmental stages. Fertilized eggs were incubated under controlled conditions, and dapagliflozin was administered at concentrations of 1.0-2.5 mg/mL on embryonic Day 3. Morphometric analysis revealed significant reductions in body weight (up to 47%) in higher-dose groups compared to controls, with survival rates declining sharply (40% mortality) in the 2.5 mg/mL group. Histological examination demonstrated hepatic steatosis, pulmonary emphysema, and cardiac inflammation, alongside neural apoptosis and vascular abnormalities. Reactive oxygen species (ROS) levels, measured via DPPH (2,2-diphenyl-1-picrylhydrazyl), superoxide dismutase (SOD), and chloramphenicol acetyltransferase (CAT) assays, results from DPPH assay showed up to 94.7% of radical scavenging activity in heart tissue, while intracellular oxidative stress is confirmed by SOD and CAT assays, implicating oxidative stress as a central mediator of these abnormalities. This study provides the first comprehensive evidence of dapagliflozin's dose-dependent teratogenicity in a nonmammalian vertebrate model. The findings underscore the need for caution in prescribing dapagliflozin during pregnancy and warrant further investigations in mammalian systems to evaluate its potential implications for human health.

In recent years, one of the most striking consequences of overpopulation and consumption activities worldwide has been the rapid increase in environmental pollutants. Sodium pyrithione (NaPT) is an organosulfur compound commonly used for its antimicrobial and antifungal properties in industrial and personal care products. The primary objective of this study was to determine the impact of NaPT exposure on whole-body vitellogenin (Vtg) levels in adult male zebrafish, a highly relevant model for toxicological studies. As Vtg functions as a key estrogen-responsive biomarker, its induction serves as a critical indicator for assessing endocrine disruption, thereby justifying its selection as the primary endpoint. The adaptation period completed, stocked zebrafish were transferred to experimental aquariums and exposed to 1 and 5 μg/L concentrations of NaPT for 24, 72, 96 h, and 7 days. Commercial ELISA kits determined Vtg levels. The whole-body zebrafish Vtg analysis revealed that Vtg levels were increased at all time intervals, regardless of dose differences, compared to the control group (p < 0.05). Only at 24 h, low and high doses of NaPT had significantly different Vtg levels (p = 0.009). Biocidal products, among the environmental pollutants, have been found to affect time-dependent levels, especially in non-target organisms. The results suggested that NaPT exposure leads to significant alterations in Vtg expression, indicating its potential to interfere with endocrine function in fish. The observed endocrine-disrupting effects of NaPT directly underscore the potential risks marine pollution poses to ecosystem health and, consequently, to human well-being through shared environmental pathways.

Thiram is a dithiocarbamate fungicide that protects turf, fruits and ornamental plants. The purpose of this experiment was to ascertain the toxico-pathological effects of thiram induced toxicity in Cyprinus carpio. The fish were divided into four groups: Group A was kept as control, compared to groups B, C, and D exposed to 40, 80, and 120 μg/L thiram, respectively. The results indicated a significant (p < 0.05) reduction in body weight, while both the absolute and relative weights of gills increased significantly (p < 0.05) in fish exposed to a higher concentration of thiram. Hematological parameters showed significant (p < 0.05) reduction in RBCs, Hb, HCT, LYM, MCV, MCHC, and platelet counts, while neutrophils and leukocytes were significantly (p < 0.05) increased. Oxidative stress parameters (TBARS and ROS) were significantly (p < 0.05) increased, while antioxidant enzymes (GSH, POD, CAT, and SOD) were significantly (p < 0.05) decreased. Moreover, comet assay showed significantly (p < 0.05) higher percentile rate of DNA damage in gills tissues of treated fish exposed to 80 and 120 μg/L thiram compared to control fish. Moreover, results showed severe histopathological changes in the gills like aneurysm, degeneration of cartilaginous cores, disruption of primary lamellae, necrosis of lamellar pillars and atrophy of lamellae. Hence, thiram demonstrates severe detrimental effects on physical and hematological parameters, induces oxidative stress, affects anti-oxidant enzyme activity, causes DNA damage and leads to histopathological alterations in the gills tissues of the freshwater fish.

Microplastics (MPs) are emerging contaminants in aquatic ecosystems and are able to modify the bioavailability and toxicity of co-occurring chemicals. In the present study, it was investigated whether high-density polyethylene (HDPE)-MPs exacerbate the toxicological effects of the pyrethroid insecticide deltamethrin in juvenile common carp (Cyprinus carpio). Fish (n = 270; 25 ± 5 g) were randomly allocated to a 2 × 3 factorial design (three replicates per group; 15 fish per tank) and exposed for 30 days to sub-lethal deltamethrin (12.5 μg/L; 10% of 96-h LC₅₀) and/or HDPE-MPs (0, 350, or 700 μg/L; 200-250 μm). Hepatic detoxification- and antioxidant-related gene expression was quantified, together with redox status, oxidative damage, and plasma biochemical biomarkers of organ function and injury. Deltamethrin and MPs alone altered transcriptional and biochemical endpoints, while co-exposure (particularly with 700 μg/L MPs) produced the most pronounced responses, including stronger induction of detoxification/metal-binding genes, reduced overall antioxidant capacity, and marked increases in lipid peroxidation and protein oxidation. Plasma activities of AST, ALT, ALP, LDH, GGT, and CK increased, and butyrylcholinesterase was inhibited, indicating cellular membrane damage and multi-organ dysfunction. Changes in circulating metabolites and creatinine further supported hepatic and renal impairment, and significant interaction effects for multiple endpoints suggested potentiation of deltamethrin toxicity by MPs. Overall, HDPE MPs act as relevant co-stressors that intensify deltamethrin-driven toxicity in fish, underscoring the need for mixture-aware monitoring and risk assessment in contaminated freshwater environments.

This work presents the first demonstration of a tube-based droplet microfluidic implementation of the Ames test, bridging single-droplet resolution with regulatory genotoxicity testing. The Ames test is a cornerstone assay for detecting mutagenicity, but conventional plate- and well-based formats suffer from high reagent consumption, low throughput, and limited automation. We report a droplet-based microfluidic Ames test assay using Salmonella typhimurium TA98, combining nanoliter compartmentalization with multiparameter optical detection. Cell density screening identified an optimal inoculum range of 10⁶-10⁷ cells/mL that maximized sensitivity while limiting spontaneous revertants. Dose-response analysis with the reference mutagen 4-nitro-o-phenylenediamine (4-NOPD) revealed clear increases in the fraction of droplets with growth of revertants, followed by a cytotoxic suppression at ≥ 8 μg/mL. A threshold-based evaluation enabled robust quantification of stochastic mutation events at single-droplet resolution. Compared with the classical fluctuation assay, the microfluidic format reduced reagent consumption by > 90%, generated statistically powerful datasets within 48 h, and eliminated subjective scoring. This study establishes segmented-flow microfluidics as a scalable, sensitive, and resource-efficient platform for mutagenicity testing, with applications in regulatory toxicology, environmental monitoring, and high-throughput chemical screening.

Taizhou City in Zhejiang Province was once one of China's largest e-waste-polluted areas. We assessed the effects of perinatal exposure to multiple heavy metals and dioxins on serum steroid levels in children from this area. We recruited 42 pairs of mothers and infants, collected breast milk and blood samples from the children, and detected dioxins and heavy metals in the breast milk, as well as steroid hormones in the serum. We employed multiple analytical approaches, including multivariable linear regression (MLR) model and Bayesian kernel machine regression (BKMR) models, to investigate the relationship between heavy metals, dioxins, and the serum steroid hormone levels in these children. In a separate exposure model using MLR, total polychlorinated dibenzofurans (PCDFs) (β: -0.930, 95% CI: -1.837, -0.023) and total polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/DFs) (β: -0.578, 95% CI: -1.142, -0.013) were negatively associated with DHEA. Within the MLR mixed exposure model, a negative association was observed between cadmium (Cd) and progesterone (β: -0.225, 95% CI: -0.447, -0.004), while a positive association was found between 2,3,7,8-TeCDD (TCDD) and dehydroepiandrosterone (DHEA) (β: 0.841, 95% CI: 0.196, 1.485). However, based on the BKMR results, these associations were attenuated. Perinatal exposure to heavy metals and dioxins may disrupt sex hormone levels and may have an impact on later reproductive health.

A widely used antibacterial drug, triclosan (TCS), is becoming more commonly acknowledged as an emergent aquatic pollutant with the potential to disrupt hormones. The physiological, biochemical and histological consequences of long-term TCS exposure in Oreochromis niloticus were assessed in this study. For 15, 30 and 45 days, adult fish were exposed to three sublethal doses of TCS (0.146, 0.219 and 0.438 mg L^-1). To ascertain reproductive and hepatic responses, growth indices, hepatosomatic index (HSI), gonadosomatic index (GSI), plasma vitellogenin (VTG) levels and gonadal histopathology were evaluated. The results showed significant increases in HSI and decreases in GSI, indicating hepatic enlargement and gonadal suppression, along with concentration-dependent declines in body weight and total length. Biochemical investigations revealed significant changes in VTG synthesis, with males displaying a brief increase followed by suppression and females showing a dose and time-dependent reduction, indicating a dualistic, sex-specific endocrine response. Reproductive impairment was confirmed by histopathological testing, which revealed severe degenerative alterations in gonadal tissues, including necrosis, vacuolation, interstitial cell infiltration and follicular atresia. Potential effects on breeding fitness, offspring production and population stability are suggested by the incapacity of O. niloticus to tolerate TCS-induced stress during the spawning period. All of these results point to triclosan being a strong endocrine and cytotoxic drug that interferes with the hepatic and gonadal physiology of O. niloticus. The study highlights the critical need for more stringent regulation and monitoring of this molecule in aquatic habitats, as well as the ecological risk connected to persistent TCS contamination.

Sphaeranthus amaranthoides, a lesser-known species of the Asteraceae family, has shown promising medicinal properties, prompting investigation into its cytotoxic and pro-apoptotic effects on human cancer cells. This study evaluated aqueous, methanol, ethyl acetate, and hexane leaf extracts for selective anticancer activity. Normal Vero cells and cancer cell lines AGS (gastric) and HT-29 (colorectal) were exposed to Sa-EAE extract, and GC-MS analysis identified key bioactive compounds, including methyl 10,13-dimethyltetradecanoate, 3,7,11,15-tetramethyl-2-hexadecen-1-ol, and 1,3,5-triazine derivatives. The extract showed selective cytotoxicity against AGS cells while sparing normal Vero cells. Apoptosis was evidenced by mitochondrial-membrane depolarization (ΔΨm), cytochrome c release, activation of Caspase-9 and Caspase-3, LDH leakage, and modulation of the Bcl-2/Bax ratio. Apoptotic morphology was confirmed via AO/EB and PI staining. Flow cytometry revealed G₀/G₁ cell cycle arrest, with downregulation of Cyclin D1 and CDK4 and upregulation of p21 and p27. These findings indicate that Sa-EAE exerts dual effects of mitochondrial-mediated apoptosis and G₀/G₁ arrest in gastric cancer cells, with minimal toxicity to normal cells, supporting its potential as a therapeutic candidate for further in vivo validation.

Gold is used in different fields showing great promise in various biomedical applications like nanotechnology, oncology, and drug delivery. Meanwhile, its effects on some biological systems of male Wistar rats remain unclear. The aim of this study was first to investigate the impact of Allochrysine solution after its intraperitoneal administration to male Wistar rats on their nutritional status. Second, the biodistribution of gold in the liver, the testes, and the brain was investigated, and the 24-h urine samples were studied as well. Third, the biochemical evaluation of hepatic and renal functions was performed. Our results showed that gold altered the nutritional status of rats. The quantitative study allowed the measurement of very high levels of gold in 24-h urine samples, in the testes, in the liver, and in the brain. The biochemical study of blood parameters showed that gold induced hyperglycemia and toxicity in the liver (e.g., ALT, AST, ALP, and TB) of treated rats. The study of urine showed excretion of elevated urine output with high specific gravity. The absolute and relative creatinine clearance were significantly higher in the gold-treated group. Our results suggest that gold is capable of crossing the blood-brain barrier and the blood-testis barrier. In the brain, gold stimulated a significant weight loss and reduced food intake, leading to modification in their nutritional status. The hepatic and kidney tissues concentrated gold in order to eliminate the metal, via biliary and urinary excretion, respectively. These organs regulated gold concentration and enhanced its elimination to facilitate detoxification.