Toxicokinetics and bioavailability of indoxacarb enantiomers and their new metabolites in rats.

Indoxacarb is a chiral insecticide that consists of two enantiomers, S-(+)-indoxacarb and R-(-)-indoxacarb, of which only S-(+)-indoxacarb has insecticidal activity. Previous enantioselective toxicology studies of indoxacarb focused mostly on simple environmental model organisms. The lack of a toxicology evaluation of indoxacarb conducted in a mammalian system could mean that the extent of the potential health risk posed by the insecticide to humans is not adequately known. In this study, we reported on a new pair of enantiomers, S-IN-RM294 and R-IN-RM294, derived from the metabolic breakdown of S-(+)-indoxacarb and R-(-)-indoxacarb, respectively, in rats. The toxicokinetics of S-(+)-indoxacarb, R-(-)-indoxacarb, S-IN-RM294, and R-IN-RM294 in rats were evaluated to provide a more comprehensive risk assessment of these molecules. The bioavailability and excretion rates of both S-(+)-indoxacarb and R-(-)-indoxacarb were relatively low, which may be due to their faster metabolism and accumulation in the tissues. In addition, there were significant differences in the metabolism and distribution between the two indoxacarb enantiomers and their metabolites in vivo. S-(+)-Indoxacarb was found to be more easily metabolized in the blood compared with R-(-)-indoxacarb, as shown by the differences in pharmacokinetic parameters between oral and intravenous administration. Analysis of their tissue distribution showed that S-(+)-indoxacarb was less likely to accumulate in most tissues. The results obtained for the two metabolites were consistent with those of the two parent compounds. S-IN-RM294 was more readily cleared from the blood and less likely to accumulate in the tissues compared with R-IN-RM294. Therefore, whether from the perspective of insecticidal activity or from the perspective of mammalian and environmental friendliness, the application of optically pure S-(+)-indoxacarb in agriculture may be a more efficient and safer strategy.

Transcriptome Analysis and Identification of Cadmium-Induced Oxidative Stress Response Genes in Different Meretrix meretrix Developmental Stages.

Cadmium (Cd) is one of the major pollutants in the aquatic environment, and it can easily accumulate in aquatic animals and result in toxic effects by changing the metabolism of the body, causing a serious impact on the immune system, reproductive system, and the development of offspring. The clam Meretrix meretrix is one of the commercially important species that is cultivated in large-scale aquaculture in China. To elucidate the underlying molecular mechanisms of Cd2+ in the developmental processes, fertilized eggs and larvae of M. meretrix at different developmental stages were exposed to Cd2+ (27.2 mg L-1 in natural seawater) or just natural seawater without Cd2+ (control), and high-throughput transcriptome sequencing and immunohistochemistry techniques were used to analyze the toxic effects of Cd on larvae at different early developmental stages. The results revealed 31,914 genes were differentially expressed in the different stages of M. meretrix development upon treatment with Cd2+. Ten of these genes were differentially expressed in all stages of development examined, but they comprised only six unigenes (CCO, Ndh, HPX, A2M, STF, and pro-C3), all of which were related to the oxidative stress response. Under Cd exposure, the expression levels of CCO and Ndh were significantly upregulated in D-shaped and pediveliger larvae, while pro-C3 expression was significantly upregulated in the fertilized egg, D-shaped larva, and pediveliger. Moreover, HPX, A2M, and STF expression levels in the fertilized egg and pediveliger larvae were also significantly upregulated. In contrast, CCO, Ndh, HPX, A2M, STF, and pro-C3 expression levels in the postlarva were all downregulated under Cd exposure. Besides the genes with changes in expression identified by the transcriptome, the expression of two other oxidative stress-related genes (MT and Nfr2) was also found to change significantly in the different developmental stages of M. meretrix upon Cd exposure, confirming their roles in combating oxidative stress. Overall, the findings of this study indicated that Cd would interfere with cellular respiration, ion transport, and immune response through inducing oxidative stress, and changes in the expression of oxidative stress-related genes might be an important step for M. meretrix to deal with the adverse effects of Cd at different stages of its development.

Some evidence supporting the use of optically pure R-(-)-diniconazole: Toxicokinetics and configuration conversion on chiral diniconazole.

Diniconazole is a chiral pesticide that exists in two enantiomers, R-(-)-diniconazole and S-(+)-diniconazole, with the R-enantiomer being much more active than the S-enantiomer. Previous enantioselective toxicology studies of diniconazole focused mostly on simple environmental model organisms. In this study, we evaluated the toxicokinetics of the two diniconazole enantiomers in rats and mice to provide a more comprehensive risk assessment. The two enantiomers displayed clear differences in their stereoselective contents in vivo. The t1/2 of R-(-)-diniconazole was 7.06 ± 3.35 h, whereas that of S-(+)-diniconazole was 9.14 ± 4.60 h, indicating that R-(-)-diniconazole was eliminated faster in vivo. The excretion rates of R-(-)-diniconazole and S-(+)-diniconazole were 4.08 ± 0.50 % and 2.68 ± 0.58 %, respectively, indicating more excretion of R-(-)-diniconazole. S-(+)-diniconazole had a higher bioavailability than R-(-)-diniconazole (52.19 % vs. 42.44 %). S-(+)-Diniconazole was also found in relatively high abundance in tissues such as the stomach, large intestine, small intestine, cecum, liver, kidney, brain, and testes, with the abundance being 1.71-2.48-fold that of R-(-)-diniconazole. The selective degradation of both enantiomers in the tissues and their mutual conversion in vivo were not observed, and this could indicate that configuration conversion did not contribute to the differences in the content of enantiomers in the tissues. Instead, such differences were mainly caused by the differences in affinity of each enantiomer for the tissues. Furthermore, investigation of the interconversion between optically pure R-(-)-diniconazole and S-(+)-diniconazole monomers in soil revealed no interconversion. All of the above results indicated no interconversion between R-(-)-diniconazole and S-(+)-diniconazole in vivo and in the soil, and that S-(+)-diniconazole tends to have a greater potential to accumulate in vivo. Thus, if only R-(-)-diniconazole is used as a pesticide, the negative impact on mammals and the environment will be reduced, suggesting that in agriculture, the application of optically pure R-(-)-diniconazole may be a better strategy.

Assessment of toxic metal pollution in Yueqing Bay and the extent of metal-induced oxidative stress in Tegillarca granosa raised in this water.

Yueqing Bay is an important economic shellfish culture zone in Zhejiang Province, China. However, increased pollution in the water caused by toxic metals has led to the bioaccumulation of toxic metals in cockles such as Tegillarca granosa, and the consequence of toxic metal-associated toxicity in these animals. This study aimed to assess the concentration of toxic metals in the water and sediment in four different sites (Baisha, Qingjiang, Nanyue, and Wengyang) within Yueqing Bay and to evaluate the extent of metal bioaccumulation in T. granosa raised in the aquaculture farms located within the four sites, as well as the changes in biomarkers in T. granosa in response to the metals. The assessment was carried out at two different times of the year, January and July. The water and sediment samples taken from the aquaculture farms in Baisha (S1), Qingjiang (S2) and Nanyue (S3) were found to have a comprehensive toxic metal pollution index (Pc) <1, indicating that these farms were not polluted. However, the water and sediment samples taken from the aquaculture farm in Wengyang (S4) had a Pc between 1 and 2, indicating mild toxic metal pollution. The edible risk assessments (HQ) of T. granosa in all four farms were <1, and therefore, these cockles could be considered safe for human consumption. The toxic metal enrichment in T. granosa exhibited a strong correlation with the toxic metal content in the sediment. In all four farms, CAT and SOD activity levels in the visceral mass of T. granosa were higher than those found in the foot, and a significantly higher level of CAT activity was detected in July compared with January. Similarly, MDA and H2O2 contents in the visceral mass were also higher in July than in January. Tegillarca granosa individuals taken from S4 and S3 farms exhibited significantly higher levels of metallothionein (MT) mRNA and MDA compared with individuals from S1 and S2 farms. Furthermore, the levels of MDA and MT mRNA showed significant positive correlations with Cd, Cr, Hg, and Cu. Elevation of lipid peroxidation in these cockles coincided with increasing levels of endogenous antioxidants. The visceral mass of T. granosa and its MDA level could be used as a tissue indicator and a biochemical marker, respectively, for detecting toxic metal pollution. MT mRNA might also be used as a molecular marker of toxic metal pollution. The integrated biomarker response version 2 (IBRv2) values of the four aquaculture farms in Yueqing Bay showed the order S4 > S3 > S2 > S1, indicating that S4 had the most serious metal-induced stress. Furthermore, the IBRv2 values correlated with the Nemerow composite index (Pc) for all the cockles examined. Thus, as far as the contamination of aquaculture farms in Yueqing Bay by toxic metals is concerned, the aquaculture farm in Wengyang (S4) was mildly contaminated by toxic metals. However, the contamination was relatively low, presenting a low risk for the local population of T. granosa.

Identification of Novel Alkaloids from Portulaca oleracea L. and Characterization of Their Pharmacokinetics and GLP-1 Secretion-Promoting Activity in STC-1 Cells.

Six new alkaloids (compounds 1-6) were isolated from Portulaca oleracea L. The compounds were triple pair (1 and 2, 3 and 4, and 5 and 6) enantiomers, with 1, 3, and 5 in the R-configuration and 2, 4, and 6 in the S-configuration, and all could bind to SUR1 according to molecular docking analysis. Treatment of STC-1 cells with each compound led to an influx of intracellular Ca2+, eventually leading to the secretion of glucagon-like peptide-1 (GLP-1), with compound 3 giving the highest secretion, resulting in 24.3 ± 7.03% more GLP-1 than nateglinide-treated cells, suggesting that these alkaloids may be able to reduce blood glucose based on their ability to stimulate the release of GLP-1. Furthermore, compound 3 also exhibited slightly faster absorption than nateglinide, as shown by pharmacokinetic analysis conducted in rats. Therefore, the results showed that some purslane alkaloids (such as compound 3) had good pharmacological activity in vivo and may have preventive and therapeutic effects on diabetes.

Toxicokinetics of Two Oxathiapiprolin Enantiomers in Rats and Their Stereoselective Interaction with Oxysterol Binding Protein.

Oxathiapiprolin is a chiral fungicide, and it can affect the metabolism of the cholesterol compounds by inhibiting oxysterol binding protein (OSBP) to exert its fungicidal effect. The application of oxathiapiprolin in agriculture is widespread, and its residue in the environment is a threat to both human and animal health. The two oxathiapiprolin enantiomers differ in their fungicidal activity, biotoxicity, and degradation by environmental forces. However, their biotoxicity has not been reported in animals. The toxicokinetics of a pesticide should be a crucial component for the evaluation of its toxicity in vivo. In this study, we investigated the absorption, bioavailability, tissue distribution, and excretion of the two oxathiapiprolin enantiomers in rats to verify their toxicokinetic process in animals. An ultrahigh-performance liquid chromatography triple quadrupole tandem mass spectrometry (UHPLC-QQQ/MS) method was established to quantify the two oxathiapiprolin enantiomers in vivo. The two oxathiapiprolin enantiomers were found to have approximately the same absorption rate and bioavailability, and both were excreted mainly in the feces. The half-life of R-(-)-oxathiapiprolin was nearly twice that of S-(+)-oxathiapiprolin. R-(-)-oxathiapiprolin also had greater distribution than S-(+)-oxathiapiprolin in the liver, lungs, heart, spleen, kidneys, stomach, large intestine, small intestine, brain, and pancreas, supporting the notion that R-(-)-oxathiapiprolin could better bind with OSBP. The stereoselectivity of S-(+)-oxathiapiprolin in these tissues may be responsible for it being readily metabolized in vivo. The molecular docking technique was subsequently used to verify the more superior binding between R-(-)-oxathiapiprolin and OSBP compared with the binding between S-(+)-oxathiapiprolin and OSBP. The findings of this study could provide more reliable data for determining the toxicokinetics of a single enantiomer of oxathiapiprolin in animals, thereby providing some theoretical basis for its subsequent toxicological study.

Purification and characterization of two forms of methanol dehydrogenases from a marine methylotroph.

Two methanol dehydrogenases (MDHs), MDH1 and MDH2, were purified from a marine methylotroph, Methylophaga sp. strain 1. Both enzymes had very similar properties, including the same native molecular weight, sizes of subunits and substrate specificity. The N-terminal amino acid sequence of the alpha-subunit of MDH2 differed from that of MDH1 by having a histidine residue at a highly conserved glutamate position, but both sequences showed approximately 50% homology to the alpha-subunits of other MDHs. MDH1 had higher specific activity than MDH2 with respect to methanol and ethanol as a substrate. The two enzymes did not appear to be isoforms but that either MDH1 or MDH2 could be a mutant arising from spontaneous mutation.