Combined exposure with microplastics increases the toxic effects of PFOS and its alternative F-53B in adult zebrafish.

Microplastics (MPs) can adsorb and desorb organic pollutants, which may alter their biotoxicities. Although the toxicity of perfluorooctane sulfonate (PFOS) and its alternative 6:2 chlorinated polyfluorinated ether sulfonate (F-53B) to organisms has been reported, the comparative study of their combined toxic effects with MPs on aquatic organisms is limited. In this study, adult female zebrafish were exposed to 10 µg/L PFOS/F-53B and 50 µg/L MPs alone or in combination for 14 days to investigate their single and combined toxicities. The results showed that the presence of MPs reduced the concentration of freely dissolved PFOS and F-53B in the exposure solution but did not affect their bioaccumulation in the zebrafish liver and gut. The combined exposure to PFOS and MPs had the greatest impact on liver oxidative stress, immunoinflammatory, and energy metabolism disorders. 16S rRNA gene sequencing analysis revealed that the combined exposure to F-53B and MPs had the greatest impact on gut microbiota. Functional enrichment analysis predicted that the alternations in the gut microbiome could interfere with signaling pathways related to immune and energy metabolic processes. Moreover, significant correlations were observed between changes in gut microbiota and immune and energy metabolism indicators, highlighting the role of gut microbiota in host health. Together, our findings demonstrate that combined exposure to PFOS/F-53B and MPs exacerbates liver immunotoxicity and disturbances in energy metabolism in adult zebrafish compared to single exposure, potentially through dysregulation of gut microbiota.

Serum uric acid and prognosis of ischemic stroke: Cohort study, meta-analysis and Mendelian randomization study.

INTRODUCTION: The role of serum uric acid (UA) levels in the functional recovery of ischemic stroke remains uncertain. To evaluate whether UA could predict clinical outcomes in patients with ischemic stroke. PATIENTS AND METHODS: A three-stage study design was employed, combining a large-scale prospective cohort study, a meta-analysis and a Mendelian randomization (MR) analysis. Firstly, we conducted a cohort study using data from the Nanjing Stroke Registry Program (NSRP) to assess the association between UA levels and 3-month functional outcomes in ischemic stroke patients. Secondly, the meta-analysis was conducted to integrate currently available cohort evidence. Lastly, MR analysis was utilized to explore whether genetically determined UA had a causal link to the functional outcomes of ischemic stroke using summary data from the CKDGen and GISCOME datasets. RESULTS: In the first stage, the cohort study included 5631 patients and found no significant association between UA levels and functional outcomes at 3 months after ischemic stroke. In the second stage, the meta-analysis, including 10 studies with 14,657 patients, also showed no significant association between UA levels and stroke prognosis. Finally, in the third stage, MR analysis using data from 6165 patients in the GISCOME study revealed no evidence of a causal relationship between genetically determined UA and stroke functional outcomes. DISCUSSION AND CONCLUSION: Our comprehensive triangulation approach found no significant association between UA levels and functional outcomes at 3 months after ischemic stroke.

Co-exposure to polystyrene microplastics and cypermethrin enhanced the effects on hepatic phospholipid metabolism and gut microbes in adult zebrafish.

Microplastics (MPs) can absorb environmental pollutants from the aquatic environment to cause mixed toxicity, which has received widespread attention. However, studies on the joint effects of MPs and insecticides are limited. As one of the most widely used pyrethroids, there was a large amount of residual cypermethrin (CYP) in water due to insufficient decomposition. Here, adult female zebrafish were exposed to MPs, CYP, and their mixtures for 21 days, respectively. After exposures, the MPs and CYP caused tissue damage to the liver. Hepatic triglyceride (TG) level increased significantly after MPs + CYP exposure, and the expression of genes about glycolipids metabolism was significantly altered. Furthermore, metabolome results suggested that MPs + CYP exposure resulted in increased content of some glycerophospholipid, affecting phospholipid metabolism-related pathways. In addition, through 16 s rDNA sequencing, it was found that MPs + CYP led to significant changes in the proportion of dominant phyla. Interestingly, Cetobacterium which increased in CYP and the co-exposure group was positively correlated with most lipid metabolites. Our results suggested that co-exposure to MPs and CYP enhanced the disturbances in hepatic phospholipid metabolism by affecting the gut microbial composition, while these changes were not observed in separate treatment groups. These results emphasized the importance of studying the joint toxicity of MPs and insecticides.

Novel PFOS alternative OBS inhibits body growth of developing zebrafish by triggering thyroid function disorder and osteoclast differentiation.

The extensive use of the perfluorooctane sulfonate (PFOS) alternative sodium p-perfluorous nonenoxybenzene sulfonate (OBS) has resulted in its widespread detection in the environment and enrichment in wildlife and humans. However, little is known about its potential toxicity, particularly in terms of body development. In this study, zebrafish embryos were acutely exposed to PFOS and OBS for a comparative developmental toxicity assessment. Both PFOS and OBS led to lower body weight and shorter body length, and the damaging effects of PFOS were more severe than those of OBS at the same exposure concentration. Biochemical assays of THs and transcription profiles correlated to the HPT axis demonstrated that OBS-induced body development inhibition resulted mainly from interference in THs synthesis, transfer, coupling with receptors, and conversion from T4 to T3, which was similar to the case of PFOS, except that the disruptive effects of OBS on thyroid function were more intense. Further transcriptome analysis showed that PFOS and OBS also promoted osteoclast differentiation, aggravating the inhibitory effects on body growth, and that PFOS had more obvious inhibitory effects than OBS. This study systematically explored the inhibitory effects of PFOS and OBS exposure on body development and tightly linked the toxic effects to thyroid function disorder and osteoclast differentiation. Our findings highlight that the health risks associated with OBS, an emerging substitute for PFOS, should not be ignored.

Bioaccumulation, tissue distribution, and maternal transfer of novel PFOS alternatives (6:2 Cl-PFESA and OBS) in wild freshwater fish from Poyang Lake, China.

As emerging alternatives to perfluorooctane sulfonate (PFOS), 6:2 chlorinated polyfluoroalkyl ether sulfonic acid (6:2 Cl-PFESA) and sodium p-perfluorous nonenox-benzenesulfonate (OBS) were frequently detected in the four freshwater fish species collected from Poyang Lake. Median concentrations of 6:2 Cl-PFESA and OBS in fish tissues were 0.046-6.0 and 0.46-5.1 ng/g wet weight, respectively. The highest concentrations of 6:2 Cl-PFESA was found in fish livers, whereas OBS was found in the pancreas, brain, gonads, and skin. The tissue distribution pattern of 6:2 Cl-PFESA is similar to that of PFOS. The tissue/liver ratios of OBS were higher than those of PFOS, suggesting that OBS has a greater tendency to transfer from the liver to other tissues. The logarithmic bioaccumulation factors (log BAFs) of 6:2 Cl-PFESA in three carnivorous fish species were greater than 3.7, whereas those of OBS were less than 3.7, indicating that 6:2 Cl-PFESA had a strong bioaccumulation potential. Notably, sex- and tissue-specific bioaccumulation of OBS has also been observed in catfish. Most tissues (except the gonads) exhibited higher OBS concentrations in males than in females. However, no differences were found for 6:2 Cl-PFESA and PFOS. Maternal transfer efficiency of OBS was higher than that of 6:2 Cl-PFESA and PFOS in catfish (p < 0.05), indicating that OBS presents a higher risk of exposure to males and offspring through maternal offloading.

Mitochondrial dysfunction in metabolic disorders induced by per- and polyfluoroalkyl substance mixtures in zebrafish larvae.

Several per- and polyfluoroalkyl substances (PFAS) have been linked to metabolic disorders in organisms. However, few studies have considered their combined effects, which would be more representative of PFAS occurring in the environment. In this study, zebrafish embryos were exposed to a mixture of 18 PFAS at three environmentally relevant concentrations for 5 days to assess their bioconcentration and metabolic consequences. The burdens of ∑PFAS in zebrafish larvae were 0.12, 1.58, and 9.63 mg/kg in the 0.5, 5, and 50 µg/L treatment groups, respectively. Exposure to the PFAS mixture accelerated hatching and larval heart rates, increased energy expenditure, and reduced ATP levels and glucose contents due to decreased feed intake and glucose uptake. Metabolomic analysis revealed that exposure to the PFAS mixture enhanced glycolysis but inhibited phospholipid synthesis, and significantly increased the expression of lipid metabolism related genes (srebf1, acox, and pparα), which indicated enhanced ß-oxidation. The significant changes in mitochondrial membrane potential, mitochondrial content, and the transcription of genes involved in the mitochondrial respiratory chain (mfn2, ndufs1, atp5fa1, and mt-nd1) and mitochondrial DNA replication and transcription (18rs-rrn, and polg1) suggested that exposure to the PFAS mixture could cause mitochondrial dysfunction and further disrupt glucose and lipid metabolic pathways, ultimately causing metabolic disorders in zebrafish larvae. These findings demonstrate the importance of assessing the metabolic effects of PFAS mixtures on early development in wildlife and humans.

Effects of PFOS, F-53B and OBS on locomotor behaviour, the dopaminergic system and mitochondrial function in developing zebrafish (Danio rerio).

Perfluorooctanesulfonic acid (PFOS) has widely been reported to persist in the environment and to elicit neurotoxicological effects in wildlife and humans. Following the restriction of PFOS use, 6:2 chlorinated polyfluorinated ether sulfonate (F-53B) and sodium p-perfluorous nonenoxybenzene sulfonate (OBS) have emerged as novel PFOS alternatives and have been detected in the environment. However, knowledge on the toxicological effects of these alternatives remains scarce. Using developing transgenic Tg(dat:eGFP) zebrafish, we evaluated the consequences of exposure to 0, 0.1 and 1 mg/l PFOS, F-53B and OBS on the dopaminergic system, locomotor behaviour and mitochondrial function. All compounds generally reduced locomotor activity under light conditions irrespective of exposure concentration. Exposure to OBS (at all concentrations), as well as PFOS and F-53B (at 1 mg/l), significantly reduced subpallial dopaminergic neuron abundance. PFOS also significantly reduced dat and pink1 expression irrespective of exposure concentration, while F-53B and OBS tended to reduce mitochondrial pink1 and fis1 expression across concentrations without reaching statistical significance. Mitochondrial function, in the form of reduced oxygen consumption rate and marginally inhibited ATP-linked oxygen consumption rate, was affected only in response to 1 mg/l PFOS. Together, PFOS and the emerging contaminants F-53B and OBS inhibit locomotion at similar concentrations, a finding correlated with decreased dopaminergic neuron numbers in the subpallium and decreased expression of pink1. These findings are relevant to wildlife and human health, as they suggest that PFOS as well as replacement compounds affect locomotion likely in part by negatively impacting the dopamine system.

Insights into the circadian rhythm alterations of the novel PFOS substitutes F-53B and OBS on adult zebrafish.

As alternatives to perfluorooctane sulfonate (PFOS), 6:2 Cl-PFESA (F-53B) and sodium p-perfluorous nonenoxybenzene sulfonate (OBS) are frequently detected in aquatic environments, but little is known about their neurotoxicity, especially in terms of circadian rhythms. In this study, adult zebrafish were chronically exposed to 1 µM PFOS, F-53B and OBS for 21 days taking circadian rhythm-dopamine (DA) regulatory network as an entry point to comparatively investigate their neurotoxicity and underlying mechanisms. The results showed that PFOS may affect the response to heat rather than circadian rhythms by reducing DA secretion due to disruption of calcium signaling pathway transduction caused by midbrain swelling. In contrast, F-53B and OBS altered the circadian rhythms of adult zebrafish, but their mechanisms of action were different. Specifically, F-53B might alter circadian rhythms by interfering with amino acid neurotransmitter metabolism and disrupting blood-brain barrier (BBB) formation, whereas OBS mainly inhibited canonical Wnt signaling transduction by reducing cilia formation in ependymal cells and induced midbrain ventriculomegaly, finally triggering imbalance in DA secretion and circadian rhythm changes. Our study highlights the need to focus on the environmental exposure risks of PFOS alternatives and the sequential and interactive mechanisms of their multiple toxicities.

Aberrant hepatic lipid metabolism associated with gut microbiota dysbiosis triggers hepatotoxicity of novel PFOS alternatives in adult zebrafish.

Perfluorooctane sulfonate (PFOS) has been reported to induce hepatotoxicity in wildlife and humans. Novel PFOS alternatives have been widely used following restrictions on PFOS, but little is known about their potential toxicity. Here, the first comprehensive investigation on the chronic hepatotoxicity and underlying molecular mechanisms of PFOS, 6:2Cl-PFESA (F-53B), and sodium p-perfluorous nonenoxybenzene sulfonate (OBS) was carried out on adult zebrafish through a histopathological examination, biochemical measurement, and multi-omics analysis. PFOS and its alternatives caused changes in liver histopathology and liver function indices in the order of F-53B > PFOS > OBS, which was consistent with their concentration in the liver. In silico modeling and transcriptional profiles suggested that the aberrant hepatic lipid metabolism induced by F-53B and PFOS was initiated by the action on peroxisome proliferator-activated receptor γ (PPARγ), which triggered changes in downstream genes transcription and led to an imbalance between lipid synthesis and expenditure. Gut microbiome analysis provided another novel mechanistic perspective that changes in the abundance of Legionella, Ralstonia, Brevundimonas, Alphaproteobacteria, Plesiomonas, and Hyphomicrobium might link to alterations in the PPAR pathway based on their significant correlation. This study provides insight into the molecular mechanisms of hepatotoxicity induced by PFOS and its novel alternatives and highlights the need for concern about their environmental exposure risks.

Concentration-dependent toxicokinetics of novel PFOS alternatives and their chronic combined toxicity in adult zebrafish.

The increasing use of perfluorooctanesulfonate (PFOS) alternatives has led to their release into the aquatic environment. This study sought to determine the effects of exposure concentration on the toxicokinetics of PFOS and its alternatives, including perfluorobutanesulfonic acid (PFBS), perfluorohexanesulfonic acid (PFHxS), chlorinated polyfluorinated ether sulfonate (F-53B) and sodium p-perfluorous nonenoxybenzenesulfonate (OBS) in adult zebrafish by exposure to mixtures of the five per- and polyfluoroalkyl substances (PFAS) at 1, 10, and 100 ng/mL for 28-day, followed by a 14-day depuration. PFAS predominantly accumulated in the blood and liver, and the bioconcentration factor (BCF) decreased in the order of F-53B > PFOS > OBS ≫ PFHxS > PFBS in whole-fish homogenates. The uptake rate constants and BCF of the short-chain PFAS (≤C6) positively correlated with increasing exposure concentration, while the long-chain PFAS (≥C8) exhibited a pattern of first increasing and then decreasing. A consistent increase in the elimination rate constants of short- and long-chain PFAS was observed with increasing exposure concentration. All PFAS form tight conformations with ZSA and ZL-FABP via hydrogen bonding as revealed by molecular docking analysis. Furthermore, chronic combined exposure to PFAS induced the occurrence of vacuolation and oxidative stress in the zebrafish liver. Our findings uniquely inform the concentration-dependent bioconcentration potential and health risks to aquatic organisms of these PFOS alternatives in the environment.

Spatiotemporal distribution, partitioning behavior and flux of per- and polyfluoroalkyl substances in surface water and sediment from Poyang Lake, China.

Thirty-five legacy and emerging per- and polyfluoroalkyl substances (PFAS) were analyzed in surface water and sediments collected from Poyang Lake, the largest freshwater lake in China. The Æ©PFAS concentrations ranged from 23 to 1000 ng/L in water dissolved phase, 1.3-9.8 ng/L in suspended particulate matters, and 0.26-2.9 ng/g dry weight in sediments. Short-chain and emerging PFAS were predominant in surface water and sediments, rather than legacy perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS). Hexafluoropropylene oxide dimer/trimer acid (HFPO-DA/TA), 6:2 and 8:2 chlorinated polyfluorinated ether sulfonic acids (6:2 and 8:2 Cl-PFESAs), 6:2 fluorotelomer sulfonate (6:2 FTS), and sodium p-perfluorous nonenoxybenzene sulfonate (OBS) were detected in all samples, indicating that these emerging PFAS have been widely produced and used in this region. The high concentrations of HFPO-DA/TA, 6:2 FTS, 6:2, 8:2 Cl-PFESAs, and OBS in sediments and their higher water-sediment distribution coefficients than those of predecessors (PFOA or PFOS) suggest that lake sediments could be an important long-term sink for these emerging alternatives. The positive matrix factorization model demonstrated that food packaging and textile treatments (50%) and fluoropolymer manufacturing (26% for alternative sources and 8.2% for legacy sources) were the two major sources of PFAS in Poyang Lake. The influx and outflux of total PFAS in Poyang Lake were 9.0 and 12.8 ton/year, respectively, and the OBS flux was estimated for the first time. The results provide insights into the environmental behavior and fate of emerging PFAS in freshwater ecosystems.

Comparative chronic toxicities of PFOS and its novel alternatives on the immune system associated with intestinal microbiota dysbiosis in adult zebrafish.

6:2 Chlorinated polyfluorinated ether sulfonate (F-53B) and sodium p-perfluorous nonenoxybenzene sulfonate (OBS) are widely used as perfluorooctane sulfonate (PFOS) alternatives in the Chinese market. Here, adult zebrafish were chronically exposed to 1 µM PFOS, F-53B, and OBS for 21 days to investigate the comparative immunotoxicity of these three per- and polyfluoroalkyl substances (PFAS). PFOS induced more severe oxidative stress in the liver than F-53B and OBS, and these three PFAS induced similar anti-inflammatory effects by repressing the expression of pro-inflammatory cytokines. The intestinal microbiota analysis showed that the relative abundance of Plesiomonas, Aeromonas, Cetobacterium, Shewanella, and Vibrio changed with the same trend in the three PFAS treatment groups. Furthermore, the PFAS increased the expression of hepcidin, muc, the immune-related genes mpo and saa, and decreased the expression of the tight junction-related gene occ in the intestine; moreover, villus height of the intestine was reduced after PFAS exposure, which indicated the functional disruption of the intestine. In particular, the significant correlation between the changed intestinal microbiota and liver and intestinal indicators also suggested the interaction between the immune system and intestinal microbiota. Taken together, our results indicate that exposure to PFOS and its alternatives F-53B and OBS can induce hepatic immunotoxicity associated with intestinal microbiota dysbiosis in adult zebrafish.

Pharmacokinetics of Azalomycin F, a Natural Macrolide Produced by Streptomycete Strains, in Rats.

As antimicrobial resistance has been increasing, new antimicrobial agents are desperately needed. Azalomycin F, a natural polyhydroxy macrolide, presents remarkable antimicrobial activities. To investigate its pharmacokinetic characteristics in rats, the concentrations of azalomycin F contained in biological samples, in vitro, were determined using a validated high-performance liquid chromatography-ultraviolet (HPLC-UV) method, and, in vivo, samples were assayed by an ultra-high performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) method. Based on these methods, the pharmacokinetics of azalomycin F were first investigated. Its plasma concentration-time courses and pharmacokinetic parameters in rats were obtained by a non-compartment model for oral (26.4 mg/kg) and intravenous (2.2 mg/kg) administrations. The results indicate that the oral absolute bioavailability of azalomycin F is very low (2.39 ± 1.28%). From combinational analyses of these pharmacokinetic parameters, and of the results of the in-vitro absorption and metabolism experiments, we conclude that azalomycin F is absorbed relatively slowly and with difficulty by the intestinal tract, and subsequently can be rapidly distributed into the tissues and/or intracellular f of rats. Azalomycin F is stable in plasma, whole blood, and the liver, and presents plasma protein binding ratios of more than 90%. Moreover, one of the major elimination routes of azalomycin F is its excretion through bile and feces. Together, the above indicate that azalomycin F is suitable for administration by intravenous injection when used for systemic diseases, while, by oral administration, it can be used in the treatment of diseases of the gastrointestinal tract.

Immunotoxicity of F53B, an alternative to PFOS, on zebrafish (Danio rerio) at different early life stages.

As an alternative to perfluorooctane sulfonate (PFOS), 6:2 chlorinated polyfluorinated ether sulfonate (F53B) has emerged in the Chinese market in recent years and has been frequently detected in the aquatic environment, but its ecological risk assessment is limited. In this study, zebrafish embryos and larvae were separately exposed to F53B, and their 96-h LC50 values were estimated to be 15.1 mg/L and 2.4 mg/L, respectively, suggesting that embryos were more resistant to F53B than larvae. The bioconcentration factor in larvae was basically higher than that of embryos, and the body growth of larvae was significantly affected by F53B rather than embryos, indicating that F53B may cause more severe toxicity to larvae. In addition to the excessive production of ROS and NO, the expression of many immune-related genes was increased in both embryos and larvae, but the number of dysregulated genes in larvae was more than that in embryos. Finally, the results of Point of Departure (PoD) indicated that the immunotoxicity of F53B was more sensitive to larvae than embryos at the molecular level. Our findings revealed the ecological risk of F53B by exploring the adverse effects of immunoregulation at different early life stages of zebrafish and indicated that the zebrafish larvae were more sensitive than embryos.

Maternal exposure to sodium ρ-perfluorous nonenoxybenzene sulfonate during pregnancy and lactation disrupts intestinal barrier and may cause obstacles to the nutrient transport and metabolism in F0 and F1 generations of mice.

Sodium ρ-perfluorous nonenoxybenzene sulfonate (OBS), a novel kind of perfluoroalkyl and polyfluoroalkyl compound, has been widely detected in the environment. The toxicity of OBS to living organisms has become a public concern. A growing body of research showed that maternal exposure to environmental pollutants caused intestinal and metabolic diseases that could be conserved across offspring. Here, female C57BL/6 mice were treated OBS at dietary levels of 0.0 mg/L (CON), 0.5 mg/L (OBS-L) and 5.0 mg/L (OBS-H) during the gestation and lactation periods. The results demonstrated that OBS treatment not only induced significant changes in the mucus secretion and ionic transport, but also disrupted the expression of antimicrobial peptides (AMPs) in the intestine of F0 and F1 generations. Additionally, OBS exposure altered bile acids metabolism and affected the transcriptional levels of critical genes involved in bile acids synthesis, signaling transfer, transportation and apical uptake. Together, all these results indicated that OBS exposure was perceived as a major stress by the intestinal epithelium that strongly affected the intestinal barrier function (including mucus, CFTR, AMPs, inflammation), and ultimately led to imbalance in the metabolism of bile acids (BAs). Moreover, we found that maternal OBS exposure had a more obvious toxicity effect on the male offspring in this experiment. Taken together, maternal OBS exposure during pregnancy and lactation had the intestinal and metabolism toxic effects on the dams and offspring, indicating that effects of maternal exposure on the toxicity of offspring could not be ignored.

Maternal exposure of mice to sodium p-perfluorous nonenoxybenzene sulfonate causes endocrine disruption in both dams and offspring.

The toxicity of certain novel perfluoroalkyl substances (PFCs) has attracted increasing attention. However, the toxic effects of sodium p-perfluorous nonenoxybenzene sulfonate (OBS) on the endocrine system have not been elucidated. In this study, OBS was added to the drinking water during the pregnancy and lactation of the healthy female mice at dietary levels of 0.0 mg/L (CON), 0.5 mg/L (OBS-L), and 5.0 mg/L (OBS-H). OBS exposure during the pregnancy and lactation resulted in the presence of OBS residues in the placenta and fetus. We also analyzed physiological and biochemical parameters and gene expression levels in mice of the F0 and F1 generations after maternal OBS exposure. The total serum cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) levels were significantly increased in female mice of the F0 generation. The androgen levels in the serum and the ovarian mRNA levels of androgen receptor (AR) also tended to increase after maternal OBS exposure in the F0 generation mice. Moreover, maternal OBS exposure altered the mRNA expression of endocrine-related genes in male mice of F1 generation. Notably, the serum TC and LDL-C levels were significantly increased in 8-weeks-old male mice of the F1 generation, and the serum high-density lipoprotein cholesterol (HDL-C) levels were decreased in 24-week-old male mice of the F1 generation. These results indicated that maternal OBS exposure can interfere with endocrine homeostasis in the F0 and F1 generations. Therefore, exposure to OBS during pregnancy and lactation has the potential toxic effects on the dams and male offspring, which cannot be overlooked.

6:2 Cl-PFESA has the potential to cause liver damage and induce lipid metabolism disorders in female mice through the action of PPAR-γ.

6:2 Cl-PFESA is a polyfluoroalkyl ether with high environmental persistence that has been confirmed to have significant adverse effects on animals. In this study, 6-week-old female C57BL/6 mice were exposed to 0, 1, 3 and 10 µg/L 6:2 Cl-PFESA for 10 weeks to estimate the hepatotoxicity of 6:2 Cl-PFESA and explore its underlying molecular mechanism. The results indicated that 6:2 Cl-PFESA preferentially bioaccumulated in the liver and induced hepatic cytoplasmic vacuolation and hepatomegaly in mice. In addition, serum metabolic profiling showed that 6:2 Cl-PFESA exposure caused an abnormal increase in amino acids and an abnormal decrease in acyl-carnitine, which interfered with fatty acid transport and increased the risk of metabolic diseases. Further experiments showed that 6:2 Cl-PFESA formed more hydrogen bonds with PPAR-γ than PFOS, Rosi and GW9662, and the binding affinity of 6:2 Cl-PFESA toward PPAR-γ was the highest among the ligands. 6:2 Cl-PFESA promoted the differentiation of 3T3-L1 cells by increasing PPAR-γ expression. Therefore, our results showed that 6:2 Cl-PFESA has the potential to induce liver damage and dysfunction in female mice, and this effect was achieved through PPAR-γ. This study is the first to reveal the hepatic toxicity of 6:2 Cl-PFESA in female mammals and provides new insights for subsequent in-depth research.

Developmental toxicity of the novel PFOS alternative OBS in developing zebrafish: An emphasis on cilia disruption.

In recent years, sodium p-perfluorous nonenoxybenzene sulfonate (OBS) has emerged as a substitute for PFOS with large demand and application in the Chinese market. However, little is known about potential developmental effects of OBS. In this study, zebrafish embryos were acutely exposed to different concentrations of OBS and the positive control PFOS for a comparative developmental toxicity assessment. OBS caused hatching delays, body axis curvature, neurobehavioral inhibition and abnormal cardiovascular development. These organismal effects were accompanied by change of development related genes expression profile, in which some cases were similar to PFOS. Overall, the toxic effects induced by OBS were generally milder than that of PFOS. Further investigation suggested that both OBS and PFOS disrupted ciliogenesis, evidenced by the ciliary immunostaining, changes in gene expression of kinesin family, dynein arm family and tubulin family members, as well as downregulation of the abundance of motor proteins including KIF3C, DYNC1H1 and DYNC1LI1. The influence of PFOS was stronger than that of OBS on ciliary genes and proteins. Molecular docking analysis revealed that both OBS and PFOS fitted into the motor proteins tightly, but binding affinity between OBS and motor proteins was lower than PFOS. Collectively, OBS and PFOS may act on ciliary motor proteins to interfere with ciliogenesis, leading to ciliary dysfunction and providing a novel probable action mode linked to developmental toxicity. This raises concerns regarding the health risks of the novel PFOS alternative OBS.

Comparison of toxicokinetics and toxic effects of PFOS and its novel alternative OBS in zebrafish larvae.

Sodium p-perfluorous nonenoxybenzene sulfonate (OBS), a novel alternative to perfluorooctanesulfonate (PFOS), is widely used in industry as a surfactant, firefighting foam and photographic material. The occurrence of OBS in the aquatic environment has been recently reported, but little information is available on its accumulation and toxic effects in aquatic organisms. In this study, zebrafish larvae (3 d post-fertilization) were subjected to OBS (10, 100 µg/L) and PFOS (10 µg/L) for a period of 48 h, followed by a 24 h of depuration period. The bioconcentration and depuration kinetics, oxidative stress and possible molecular mechanisms of OBS and PFOS were investigated in zebrafish larvae. Our results showed that the uptake and depuration of both OBS and PFOS fitted well with a first-order kinetic model. The uptake rate constant of OBS was similar to that of PFOS, but the depuration rate constant was much higher than PFOS with a half-life of 69.7-85 h for OBS and 222.2 h for PFOS. The calculated BCFs of OBS and PFOS were 238.0-242.5 and 644.2, respectively. In our acute toxicity assay, the enhanced expression of Nrf2 protein accompanied by the upregulation of CAT and SOD protein expressions indicated OBS and PFOS induced oxidative stress in zebrafish larvae, and the Nrf2-ARE signaling pathway was involved in this process. Collectively, OBS has a lower bioconcentration potential than PFOS, but its toxic effect on oxidative stress was comparable to PFOS in zebrafish larvae.

Spermidine improves gut barrier integrity and gut microbiota function in diet-induced obese mice.

Obesity is associated with impaired intestinal barrier function and dysbiosis of the gut microbiota. Spermidine, a polyamine that acts as an autophagy inducer, has important benefits in patients with aging-associated diseases and metabolic dysfunction. However, the mechanism of spermidine on obesity remains unclear. Here, we show that spermidine intake is negatively correlated with obesity in both humans and mice. Spermidine supplementation causes a significant loss of weight and improves insulin resistance in diet-induced obese (DIO) mice. These effects are associated with the alleviation of metabolic endotoxemia and enhancement of intestinal barrier function, which might be mediated through autophagy pathway and TLR4-mediated microbial signaling transduction. Moreover, spermidine causes the significant alteration of microbiota composition and function. Microbiota depletion compromises function, while transplantation of spermidine-altered microbiota confers protection against obesity. These changes might partly be driven by an SCFA-producing bacterium, Lachnospiraceae NK4A136 group, which was decreased in obese subjects and subsequently increased by spermidine. Notably, the change of Lachnospiraceae NK4A136 group is significantly correlated with enhanced gut barrier function induced by spermidine. Our results indicate that spermidine supplementation may serve as a viable therapy for obesity.