Absorption, distribution, metabolism, and elimination of epoxiconazole enantiomers in lizards (Eremias argus).

Epoxiconazole (EPX) is a world widely used chiral triazole fungicide in the agriculture field. The excessive application of this triazole may cause damage to lizards. However, limited information is known about the toxicokinetics of EPX on lizards. Our study aimed to investigate the enantioselective absorption, distribution, metabolism, and elimination (ADME) of EPX in lizards following low and high dose exposure (10 and 100 mg kg-1 bodyweitht (bw)). The results demonstrated that (+)-EPX was easier absorbed than (-)-EPX in lizard plasma. Both (+)-EPX and (-)-EPX were detected in the liver, gonad, kidney, skin, brain, and intestine, with (+)-EPX preferentially distributed in these tissues. The elimination of (-)-EPX was faster than that of (+)-EPX in lizard liver and kidney in the high dose groups. Chiral conversion was found between EPX enantiomers in lizard skin. Simultaneously, five metabolites including M2, M4, M10, M18 and M19 were detected in lizard liver and kidney after EPX enantiomers exposure. The relative concentrations of M2, M4, and M10 were higher in the liver and kidney of (-)-EPX groups than those produced from (+)-EPX groups. The metabolic enzymes CYP3A4 and SULT1A1 primarily mediated enantioselective metabolism of EPX. The conclusions drawn from this study significantly enhance our understanding of the enantioselective behaviors of chiral triazole fungicides in reptiles, offering essential guidance for assessing the risks associated with different enantiomers of triazole fungicides.

Temperature-Dependent Bioaccumulation, Metabolism, and Hepatotoxicity of Flufiprole in Lizards (Eremias argus).

As a phenylpyrazole insecticide, flufiprole is an important substitute for fipronil in the agricultural field of China. However, its bioaccumulation and metabolism in terrestrial organisms especially in the lizards living in the agricultural area have rarely been investigated. As an ectothermic animal, lizards are also sensitive to temperature changes. Considering global warming, this study measured bioaccumulation, metabolism, and hepatotoxicity of flufiprole in the Chinese native lizard (Eremias argus) under different temperature stresses. Lizards exposed to flufiprole-contaminated soil adsorbed flufiprole through the skin and flufiprole was preferred to accumulate in lizard liver and brain. The oxidation product fipronil sulfone was the main metabolite of flufiprole in both lizard liver and human liver microsomes, which were mainly metabolized by lizard CYP3A19 or human CYP3A4. The fipronil sulfone concentration increased with increased temperature in lizard tissues. In addition, more serious oxidative damage was shown under higher temperature as the glutathione (GSH), malondialdehyde (MDA), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels in lizards increased with increased temperature after flufiprole exposure. Flufiprole exposure also induced lizard liver lesions, and these lesions became more serious in the higher-temperature groups. This study provided new insights into the risk assessment of flufiprole in lizards under global warming.

Developmental toxicity of TCBPA on the nervous and cardiovascular systems of zebrafish (Danio rerio): A combination of transcriptomic and metabolomics.

Tetrachlorobisphenol A (TCBPA), a widely used halogenated flame retardant, is frequently detected in environmental compartments and human samples. However, unknown developmental toxicity and mechanisms limit the entire understanding of its effects. In this study, zebrafish (Danio rerio) embryos were exposed to various concentrations of TCBPA while a combination of transcriptomics, behavioral and biochemical analyzes as well as metabolomics were applied to decipher its toxic effects and the potential mechanisms. We found that TCBPA could interfere with nervous and cardiovascular development through focal adhesion and extracellular matrix-receptor (ECM-receptor) interaction pathways through transcriptomic analysis. Behavioral and biochemical analysis results indicated abnormal swimming behavior of zebrafish larvae. Morphological observations revealed that TCBPA could cause the loss of head blood vessels. Metabolomic analysis showed that arginine-related metabolic pathways were one of the main pathways leading to TCBPA developmental toxicity. Our study demonstrated that by using omics, TCBPA was shown to have neurological and cardiovascular developmental toxicity and the underlying mechanisms were uncovered and major pathways identified.

Twenty-four hours of Thiamethoxam: In vivo and molecular dynamics simulation study on the toxicokinetic and underlying mechanisms in quails (Coturnix japonica).

Neonicotinoids is the most widely used insecticide, its contamination has led to sustained bird population declines. However, the toxicokinetic and underlying mechanisms of neonicotinoid toxicity in birds are largely unknown. Thiamethoxam (TMX), as a representative neonicotinoid insecticide, is now widely detected in most environmental medium and animal bodies. In this study, 5 mg/kg body weight TMX (potential environmental intake level) were orally administrated to male Japanese quails (Coturnix japonica). We found a rapid absorption, distribution, metabolism and elimination of TMX in quails in a period of 24 h, with the main metabolite, clothianidin (CLO), being extensively distributed and rapidly eliminated from tissues as well. The maximum plasm concentration of CLO was consistent with wild birds. Metabolomics analysis and followed determination of liver enzymes mRNA expression indicated the rapid metabolism was mediated mainly by CYPs and GSTs that involved riboflavin metabolism and glutathione metabolism pathways upon TMX exposure. Molecular dynamic simulation showed the strongest binding interaction in quail CYP2H1-TMX and CYP3A12-CLO complexes among a set of CYPs-substrate. The present study elucidated toxicokinetic and underlying metabolic mechanisms of TMX in quails at environmentally-relevant concentration, the findings would facilitate the understanding of potential risks of TMX and its metabolites to birds.

Toxicological effects of acute prothioconazole and prothioconazole-desthio administration on liver in male Chinese lizards (Eremias argus).

Prothioconazole (PTC) is a high effective systemic fungicide, and one of its major metabolites is prothioconazole-desthio (PTC-d). Because of its wildly use in the farmland of China, the local eco-toxicological effects of PTC as well as PTC-d are needed to be concerned. This study investigated hepatoxicity of Chinese lizards (Eremias argus), a local non-target organism, after single dose oral treated (100 mg kg-1 BW) through pathological, enzyme activity and gene expression analysis. PTC treatment caused ballooning and PTC-d treatment led to macrovesicular steatosis of hepatocyte. The elevation of serum indexes, including the activities of aspartate aminotransferase (AST), alkaline phosphatase (ALP) and alanine aminotransferase (ALT), further confirmed the hepatic injury. PTC and PTC-d treatments altered oxidative status reflected by the inhibition of superoxide dismutase (SOD) activity , meanwhile, the stimulation of catalase (CAT) activity, glutathione peroxidase (GPx) activity and malondialdehyde (MDA) content. The mRNA expression changes of apoptosis-related factors and cytokines genes, including Bax, Bcl-2, TNF-α, NF-κB, Caspase-3 and Nrf2, deeply uncovered the potential mechanism of hepatotoxicity caused by PTC and PTC-d. In brief, the results indicated that both of these two compounds altered oxidative status, then were likely to trigger caspase-3 by affecting the ratio of pro- and anti-apoptotic factors which belong to intrinsic apoptosis pathway. Specifically, more serious impacts were induced by PTC-d than its parent compound. This study is the first to provide specific insight into potential hepatotoxicity resulted from PTC and PTC-d in male Chinese lizards.

Integrative analysis of transcriptomics and metabolomics reveals the hepatotoxic mechanism of thiamethoxam on male Coturnix japonica.

Thiamethoxam (TMX), a representative neonicotinoids, is widely used for seed coating. The consumption of TMX-coated seeds posed threat to birds during crop sowing. The hepatotoxicity of TMX has been reported in mammals, however, no clear evidence showed TMX-induced toxic effects on bird liver. In this study, male Japanese quails (Coturnix japonica) were exposed to 20 or 200 mg/kg TMX-treated bird feed for 28 days. Results showed that Clothianidin (CLO), a TMX metabolite preferred to accumulate in quail plasma and liver, and inflammatory cell infiltration was found in quail livers. Oxidative stress-related biological processes were significantly enriched in both TMX treatment groups through transcriptomics analysis. Moreover, integrative analysis of transcriptomics and metabolomics indicated ferroptosis and DNA damage was implicated in hepatotoxicity caused by high- and low-concentration of TMX exposure, respectively. High-dose TMX treatment decreased CAT activity and GSH concentration and increased expression of the ferroptosis-related gene. In addition, the up-regulation of 8-OHdG concentration and DNA repair-related genes expression demonstrated low-dose TMX triggered oxidative DNA damage. The present results highlight the toxicity of TMX to bird livers and contribute to a better understanding of the TMX toxic mechanism in birds.

Acute temperature adaptation mechanisms in the native reptile species Eremias argus.

Reptiles are sensitive to temperature changes as ectotherm animals. The climate warming may pose more serious threat to reptiles. Although the behavior effect and reproduction biology have been well studied, little information is available about the adaptation mechanisms of reptiles to temperature stress. In this study, the native Chinese species, Eremias argus were incubated at 15 (cold stress), 25 (control group) and 35 °C (thermal stress) for 24 h. The transcriptome and metabolome technology were applied to investigate the molecular regulation mechanisms of lizards to acute temperature changes. The CIRBP and HSPA8 were hub genes in response to temperature adaptation. The increased expression of PER gene in lizard circadian rhythm is associated with tyrosine metabolism after cold or thermal stress. The poly-unsaturated fatty acids in female lizard liver were significantly increased with up-regulation of FASN and ACACA genes after thermal stress, which proved the disruption of fatty acid biosynthesis pathway in corporation with the altered body weight. The cortisol and testosterone were important steroid hormones in response to temperature changes especially in male lizard liver. The increased CIRBP gene expression in lizard gonads suppressed the KDM6B gene, which regulates the testis development and may induce sex reversal in male lizard after thermal stress. The adaptation responses of lizards to temperature stress may threaten the health status of wild population.

Development of QSAR models for evaluating pesticide toxicity against Skeletonema costatum.

Nowadays, the emergence of pesticides and its application in agriculture greatly improved the crop quality and food production. However, the resulted ecological problem caused by the widespread pesticide residues attracted more and more attention since the pesticides were harmful to most living organisms. Regulatory agencies such as Environmental Protection Agency (EPA) and European Chemicals Agency (ECHA) stipulated that a comprehensive pesticides risk assessment was essential and also underscored the application of computation method in evaluating pesticides. The present study aimed to use the Quantitative Structure-Activity Relationship (QSAR) method to establish models for quantitatively and qualitatively predicting the toxicity of pesticide against Skeletonema costatum. The regression model was developed using the Genetic Algorithm plus Multiple Linear Regression method and the classification model was established based on the Random Forest algorithm, respectively. Various internal and external validation metrics suggested that the obtained regression model was of good fitness (R2=0.722), robustness (QLOO2=0.653) and external predictive ability (QFn2:0.719-0.776, CCC = 0.878). The classification could correctly predict 79.4% of pesticides in the training set and 69.7% in the validation set. The relatively high sensitivity value of the classification model indicated its good performance in identifying high-toxic pesticides. It could be concluded from the selected modelling descriptors that molecular weight and polarizability impacted the toxicity the most. The atom-type E-state descriptors generally contributed negatively to the pesticide toxicity which verified the negative influence of molecular hydrophilicity. Moreover, the lipophilic, carbon-type, charge related descriptors demonstrated the important influence of lipophilicity and polarity on pesticide toxicity. The models presented in this work could be used to pre-evaluate the toxicity of pesticides within the applicability domain, thus focusing resources on the high-toxic pesticides and assessing the environmental risk of pesticides quickly and economically.

Enantioselective accumulation, elimination and metabolism of fenbuconazole in lizards (Eremias argus).

The enantioselective accumulation, elimination and metabolism of fenbuconazole in lizards were determined following a single-dose (25 mg/kgbw) exposure to racemic or enantiomeric fenbuconazole. Accumulation of fenbuconazole was found in lizard fat with rac-form > enantiopure enantiomers. The enantiomer fractions (EFs) were higher than 0.5 in the blood, while EFs were less than 0.5 in the liver, brain, skin and stomach. There was conversion from (+)-fenbuconazole to (-)-fenbuconazole in lizard liver and conversion from (-)-fenbuconazole to (+)-fenbuconazole in lizard liver and blood. The results showed that enantioselective accumulation appeared in lizards, but the direction varied among blood and different tissues. The elimination half-lives (t1/2) of (+)-fenbuconazole were higher than those of (-)-fenbuconazole in the blood and liver, suggesting that (-)-fenbuconazole eliminated faster than (+)-fenbuconazole in these tissues. In addition, both (+)-fenbuconazole and (-)-fenbuconazole eliminated faster in the liver and stomach exposed to racemate than those exposed to enantiopure enantiomers. On the contrary, the form of racemate decreased the elimination rate of fenbuconazole in lizard fat. Synergistic elimination may occur when two enantiomers coexisted in lizard liver and stomach, while the racemate produced antagonistic elimination in lizard fat. Simultaneously, three metabolites, RH-6467, RH-9029&RH-9030 and keto-mchlorophenol, were discovered in lizard liver. Only two metabolites, RH-6467 and RH-9029&RH-9030, were found in lizard blood. RH-9029&RH-9030 were the major metabolites. The discovered enantiomers of (+)-fenbuconazole metabolites were different from those of (-)-fenbuconazole. The findings of this study may provide a better understanding of the enantioselective behaviors of chiral triazole fungicides in reptiles.

Modeling pesticides toxicity to Sheepshead minnow using QSAR.

Nowadays, the environmental risk caused by the widespread use of pesticides and their ubiquitous residuals has received more and more attention in academia and regulatory agencies. Due to the large number of pesticides used in agriculture and their adverse effects on all living organisms and the numerous end-points, it is necessary to employ the in silico tools to quickly highlight hazardous pesticides. In this study, we have evaluated the toxicity of pesticides against Sheepshead minnow with the Quantitative Structure-Activity Relationship (QSAR) approach. The models for the specific-type (insecticides, herbicides and fungicides) as well as the general-type (combing all the specific-type pesticides and some microbicides, nematicides, etc.) pesticides were developed using the Genetic Algorithm and the Multiple Linear Regression method, subsequently validated with various metrics. The validation results suggested that the obtained models were highly robust, externally predictive and characterized by a broad applicability domain. Considering the modeling descriptors, the toxicity of pesticides would increase with the lipophilicity and decrease with the polarity and hydrophilicity. Most electrotopological state descriptors contribute negatively to the toxicity, while the influence of topological structure descriptors mainly depends on the physiochemical information they encode. The models proposed in this paper would be useful in filling the data gaps, prioritizing and then focusing experiments on more hazardous pesticides.

Polyurethane foam induces epigenetic modification of mitochondrial DNA during different metamorphic stages of Tenebrio molitor.

The present work investigated the changes in DNA methylation pattern of Tenebrio molitor mitochondria genome at different development stages, which was fed with polyurethane foam as a sole diet. Polyurethane foam could influence the global methylation levels in mitochondria DNA of Tenebrio molitor. Different leves of 5-methylcytosine appeared at CpG and non-CpG sites of Tenebrio molitor mtDNA while they were fed with polyurethane foam: 10 CpG and 49 non-CpG sites at larval stage, 4 CpG and 31 non-CpG sites at pupa stage, 7 CpG and 56 non-CpG sites at adult stage in general. Moreover, we observed the decreased levels of ATP generation with the mitochondria DNA methylation variation. The results demonstrated that mitochondria DNA gene could be methylated in response to environmental pollutants to modulate stage-specific functions. Moreover, mtDNA methylation of polyurethane-foam-feeding Tenebrio molitor existed discrepancy in the developmental stage. The tentative methylation mechanism of mtDNA might be that polyurethane foam induced oxidative stress and increased the permeability of mitochondrial membranes, which resulted in transmethylase entry into mitochondria.

Comparative toxicokinetics and tissue distribution of prothioconazole and prothioconazole-desthio in Chinese lizards (Eremias argus) and transcriptional responses of metabolic-related genes.

Prothioconazole (PTC) is a widely used triazolinthione fungicide with low toxicity and short residual period. However, its desulfurization metabolite, prothioconazole-desthio (PTC-d), is more persistent and has higher toxicity in terrestrial animals. In this study, the toxicokinetics (TK) and tissue distribution of PTC and PTC-d in Chinese lizards (Eremias argus) were measured following single oral dose (100 mg kg-1 body weight) treatments. TK parameters indicated that PTC was more rapidly absorbed than PTC-d, as indicated by its shorter time to reach peak concentrations in most tissues. Furthermore, the relative bioavailability of PTC in lizards was lower than that of PTC-d. Compared with PTC, PTC-d preferentially accumulated in lizards, as reflected by longer half-life of PTC-d. During the distribution process, PTC-d generated in vivo was transported from other tissues and was deposited in the skin and tail, where PTC-d may be excreted by exuviation or tail detachment. Preferential enrichment of S-enantiomer of both PTC and PTC-d were observed in all tissues. Hepatic cytochrome P450 gene expression measurement revealed that cyp1a5 and cyp3a28 exhibited the strongest responses in both treatment groups. In addition, the opposite responses of cyp2k4 in different treatment groups may indicate that this enzyme caused differences in the rates of metabolism of the two chemicals. This study compared the TK profile of PTC and its desulfurization metabolite PTC-d in lizards and demonstrated that the desulfurization of PTC could increase its ecological risk due to the higher bioavailability and persistence of PTC-d.

Myclobutanil accumulation, transcriptional alteration, and tissue injury in lizards (Eremias argus) treated with myclobutanil enantiomers.

Enantioselective toxicokinetics, accumulation, and toxicity of myclobutanil were investigated by oral exposure of myclobutanil enantiomers to lizards. After a single oral administration, the absorption half-lives ( [Formula: see text] ) and elimination half-lives (t1/2k) were in the range of 0.133-14.828 and 3.641-17.682 h, respectively. The absorption and elimination half-lives of (+)-myclobutanil showed no significant differences from those of (-)-myclobutanil in lizard blood, whereas preferential enrichment of (-)-enantiomer was observed in the liver, fat, skin, intestine, lung and kidney. In the bioaccumulation experiments, the residue of (-)-myclobutanil was detected in most tissues at 7, 14, and 28 days, while (+)-myclobutanil was found only in lizard skin, at a concentration lower than that of (-)-myclobutanil. Thus, (-)-myclobutanil was preferentially accumulated in lizards. The transcriptional responses of metabolic enzyme genes indicated that cytochrome P450 1a1 (cyp1a1), cyp2d3, cyp2d6, cyp3a4 and cyp3a7 played a crucial role in the metabolism of (+)-myclobutanil, whereas cyp1a1, cyp2d3, cyp2d6, cyp2c8, and cyp3a4 contributed to the metabolism of (-)-myclobutanil. The difference in metabolism pathways may be a reason for the enantioselectivity of myclobutanil in lizard. Myclobutanil also affected the expression of antioxidant enzyme genes, and the (+)-myclobutanil treatment might produce higher oxidative stress in lizard liver when compared with its antipode. Hepatic histopathological changes such as hepatocellular hypertrophy, nuclear pyknosis, vacuolation, and non-zonal macrovesicular lipid accumulation were observed in the liver of lizards for both (+)-myclobutanil and (-)-myclobutanil treatments. Thus, myclobutanil could affect lizard liver upon multiple exposure. The findings of this study provide specific insights into the enantioselective metabolism and toxicity of chiral triazole fungicides in lizards.

Enantioselective Distribution, Degradation, and Metabolite Formation of Myclobutanil and Transcriptional Responses of Metabolic-Related Genes in Rats.

Myclobutanil (MT), a chiral fungicide, can be metabolized enantioselectively in organisms. In this work, the associated absorption, distribution, metabolism and transcriptional responses of MT in rats were determined following a single-dose (10 mg·kg-1 body weight) exposure to rac-, (+)- or (-)-MT. The enantiomer fractions (EFs) were less than 0.5 with time in the liver, kidney, heart, lung, and testis, suggesting preferential enrichment of (-)-MT in these tissues. Furthermore, there was conversion of (+)-form to (-)-form in the liver and kidney after 6 h exposure to enantiopure (+)-MT. Enrichment and degradation of the two enantiomers differed between rac-MT and MT-enantiomers groups, suggesting that MT bioaccumulation is enantiomer-specific. Interestingly, the degradation half-life of MT in the liver with rac-MT treatment was shorter than that with both MT-enantiomer treatments. One reason may be that the gene expression levels of cytochrome P450 1a2 ( cyp1a2) and cyp3a2 genes in livers treated with rac-MT were the highest among the three exposure groups. In addition, a positive correlation between the expression of cyp2e1 and cyp3a2 genes and rac-MT concentration was found in livers exposed to rac-MT. Simultaneously, five chiral metabolites were detected, and the enantiomers of three metabolites, RH-9090, RH-9089, and M2, were separated. The detected enantiomers of (+)-MT metabolites were in complete contrast with those of (-)-MT metabolites. According to the results, a metabolic pathway of MT in male rats was proposed, which included the following five metabolites: RH-9089, RH-9090, RH-9090 Sulfate, M1, and M2. The possible metabolic enzymes were marked in the pathway. The findings of this study provide more specific insights into the enantioselective metabolic mechanism of chiral triazole fungicides.

The body burden and thyroid disruption in lizards (Eremias argus) living in benzoylurea pesticides-contaminated soil.

Dermal exposure is regarded as a potentially significant but understudied route for pesticides uptake in terrestrial reptiles. In this study, a native Chinese lizard was exposed to control, diflubenzuron or flufenoxuron contaminated soil (1.5 mg kg-1) for 35 days. Tissue distribution, liver lesions, thyroid hormone levels and transcription of most target genes were examined. The half-lives of diflubenzuron and flufenoxuron in the soil were 118.9 and 231.8 days, respectively. The accumulation of flufenoxuron in the liver, brain, kidney, heart, plasma and skin (1.4-35.4 mg kg-1) were higher than that of diflubenzuron (0-1.7 mg kg-1) at all time points. The skin permeability factor of flufenoxuron was more than 20-fold greater than that of diflubenzuron at the end of exposure. However, the liver was more vulnerable in the diflubenzuron exposure group. The alterations of triiodothyronine (T3) and thyroxine (T4) level after diflubenzuron or flufenoxuron exposure were accompanied with the changes in the transcription of target genes involved not only in hypothalamus-pituitary-thyroid (HPT) axis (sult, dio2, trα and udp) but also in metabolism system (cyp1a and ahr). These results indicated that flufenoxuron produced greater body burdens to lizards through dermal exposure, whereas both diflubenzuron and flufenoxuron have the potential to disturb metabolism and thyroid endocrine system.

Stereoselective degradation and thyroid endocrine disruption of lambda-cyhalothrin in lizards (Eremias argus) following oral exposure.

The disturbance of the thyroid system and elimination of chiral pyrethroid pesticides with respect to enantioselectivity in reptiles have so far received limited attention by research. In this study, bioaccumulation, thyroid gland lesions, thyroid hormone levels, and hypothalamus-pituitary-thyroid axis-related gene expression in male Eremias argus were investigated after three weeks oral administration of lambda-cyhalothrin (LCT) enantiomers. In the lizard liver, the concentration of LCT was negatively correlated with the metabolite-3-phenoxybenzoic acid (PBA) level during 21 days of exposure. (+)-LCT exposure induced a higher thyroid follicular epithelium height than (-)-LCT exposure. The thyroxine levels were increased in both treated groups while only (+)-LCT exposure induced a significant change in the triiodothyronine (T3) level. In addition, the expressions of hypothalamus-pituitary-thyroid axis-related genes including thyroid hormone receptors (trs), deiodinases (dios), uridinediphosphate glucuronosyltransferase (udp), and sulfotransferase (sult) were up-regulated after exposure to the two enantiomers. (+)-LCT treatment resulted in higher expression of trs and (-)-LCT exposure led to greater stimulation of dios in the liver, which indicated PBA-induced antagonism on thyroid hormone receptors and LCT-induced disruption of thyroxine (T4) deiodination. The results suggest the (-)-LCT exposure causes higher residual level in lizard liver while induces less disruption on lizard thyroid activity than (+)-LCT.

Enantiomerization and stereoselectivity in bioaccumulation of furalaxyl in Tenebrio molitor larvae.

Furalaxyl is a chiral pesticide and widely used in modern agriculture as racemate mixture. The enantiomerization and enantioselecive bioaccumulation by a single dose of furalaxyl to Tenebrio molitor larvae under laboratory conditions were studied using a high-performance liquid chromatography tandem mass spectroscopy method based on a ChiralPAK IC column. Our results showed that a significant enantiomerization (interconversion between R-enantiomer and S-enantiomer) was observed in Tenebrio molitor larvae under R- or S-furalaxyl exposure. Though the two furalaxyl enantiomers exhibited low-capacity of bioaccumulation in Tenebrio molitor larvae, bioaccumulation of rac-furalaxyl was enantioselective with a preferential accumulation of S-furalaxyl at 10mg/kg dosage exposure. In addition, enantiomerization and enantioselective degradation of the two enantiomers was not observed in wheat bran. These results showed that enantioselectivtiy of furalaxyl enantiomers was an important process combined with degradation, metabolism and enatiomerization in organisms.

Acylamino acid chiral fungicides on toxiciepigenetics in lambda DNA methylation.

Acylamino acid chiral fungicides (AACFs) are low-toxicity pesticides and considered as non-carcinogenic chemicals to laboratory animals. Though AACFs have potential toxicological effects on mammals by non-genotoxic mechanisms, the toxicoepigenomics of AACFs has not been documented. In this article, we explored toxiciepigenetics of metalaxyl, benalaxyl and furalaxyl through epigenetics research on lambda DNA under different concentration exposure. The toxicoepigenomic difference of stereoisomers was examined also. Our results showed that AACFs would affect methyltransferase activity resulting in modulating DNA methylation levels and pattern. The LOAEL of R-metalaxyl and S-metalaxyl were 30 mM and 0.3 mM, respectively. The LOAEL of (R, S)-benalaxyl and (R, S)-furalaxyl were 0.3 Mm and 30 mM, respectively. A significant dose-response effect between (R, S)-benalaxyl and global methylation level was observed. Global methylation level was more susceptible to S-enantiomer compared to R-enantiomer, which indicated enantiomers of AACFs have the enantioselectivity in toxiciepigenetics. Moreover, the dependence of the methylation inhibition on the chiral center of metalaxyl may suggest a considerable specificity of the compound of AACFs for DNA methyltransferases. The inhibition effect between R-enantiomer and S-enantiomer of AACFs on DNA methylation levels generated in this study is important for low-toxicity pesticides toxicoepigenomics evaluation.

Tissue distribution, metabolism and hepatic tissue injury in Chinese lizards (Eremias argus) after a single oral administration of lambda-cyhalothrin.

Lambda-cyhalothrin (LCT) is a widely used pyrethroid with neurotoxicity. However, little is known about the toxicokinetics of LCT in reptiles. In this study, the absorption, distribution, metabolism and excretion of LCT in Chinese lizards (Eremias Argus) were determined following a single dose (10 mg kg-1) treatment. In the liver, brain, gonads and skin, LCT levels peaked within several hours and then decreased rapidly. However, the concentration of LCT gradually increased in the fat tissue. More than 90% of the LCT dose was excreted in the faeces. One LCT metabolite, 3-phenoxybenzoic acid (PBA), was detected in lizard plasma and tissues. PBA preferentially accumulates in the brain and plasma. The half-life of PBA in the brain was 3.2 days, which was 35.4-fold greater than that of LCT. In the plasma, the concentration of PBA was significantly higher than that of LCT. The bioaccumulation of LCT in tissues was enantioselective, and the enantiomeric fractions (EF) ranged from 0.72 to 0.26. The preferential accumulation of enantiomers changed according to exposure time, but the reasons behind this phenomenon were not clear. For pathological analysis, vacuolation of the cytoplasm and large areas of necrosis were observed in the liver sections after 168 h of dosing. The liver tissues exhibited both decreases in the hepatosomatic index and histopathological lesions during the exposure period. In this study, the effect concentration of LCT in lizards was 200-fold lower than its LD50 value used in risk assessments for birds. These results may provide additional information for the risk assessment of LCT for reptiles and indicate that birds may not be an ideal surrogate for reptile toxicity evaluation.

[Hypoxia reduces the proliferation and occludin expression of primary sertoli cells].

OBJECTIVE: To investigate the effect of hypoxia on the proliferation and occludin expression of primary rat Sertoli METHODS: We constructed a primary Sertoli cell system by two-step enzymatic digestion in 18 -22 days old Wistar rats and identified it by oil red O and immunofluorescence methods. We randomly divided the Sertoli cells into five groups to be cultured in oxygen at the concentrations of 20%, 15%, 10%, 5% and 1%, respectively, for 6, 12, 24, 48 and 72 hours. We detected the proliferation of the Sertoli cells by CCK-8 assay, determined the expression of occludin by Western blot, and analyzed the differences among the five groups. RESULTS: Oil red O staining revealed red lipid droplets in the cytoplasm of the Sertoli cells, and immunofluorescence showed the positive expression of the FasL protein, with the purity of Sertoli cells over 95% in vitro. Compared with the 20% normoxic group, the proliferation of the Sertoli cells was gradually reduced in the 15% and 10% hypoxia groups, and significantly declined in the 5% and 1% groups (P < 0.01). At 12 hours, the expression of occludin began to decrease with the prolonging of time and reduction of oxygen concentration (P < 0.01). CONCLUSION: Hypoxia suppresses the proliferation of Sertoli cells and reduces the expression of occludin. It could be inferred that hypoxia could damage the integrity of blood-testis barrier and spermatogenesis of the testis.