Reconfiguring the online tool of SkinSensPred for predicting skin sensitization of pesticides.

Adverse outcome pathway (AOP)-based computational models provide state-of-the-art prediction for human skin sensitizers and are promising alternatives to animal testing. However, little is known about their applicability to pesticides due to scarce pesticide data for evaluation. Moreover, pesticides traditionally have been tested on animals without human data, making validation difficult. Direct application of AOP-based models to pesticides may be inappropriate since their original applicability domains were designed to maximize reliability for human response prediction on diverse chemicals but not pesticides. This study proposed to identify a consensus chemical space with concordant human responses predicted by the SkinSensPred online tool and animal testing data to reduce animal testing. The identified consensus chemical space for non-sensitizers achieved high concordance of 85% and 100% for the cross-validation and independent test, respectively. The reconfigured SkinSensPred can be applied as the first-tier tool for identifying non-sensitizers to reduce. animal testing for pesticides by 19.6%.

Lack of Acute Toxicity and Mutagenicity from Recombinant Epinephelus lanceolatus Piscidin Expressed in Pichia pastoris.

The antimicrobial peptide (AMP) piscidin was identified from Epinephelus lanceolatus and demonstrated to possess antimicrobial and immune-related functions. Supplementation of feed with recombinant Epinephelus lanceolatus piscidin (rEP)-expressing yeast pellets may minimize the excessive use of antibiotics and control pathogens in aquaculture or animal husbandry. However, before implementing rEP as a supplement, it is necessary to understand whether it harbors any toxicity. Since toxicological information on the topic is scarce, the present investigation was carried out to test whether rEP exhibits allergenic and/or toxic effects. In an oral acute toxicity test (OECD 425), Sprague Dawley (SD) rats were administered rEP dissolved in reverse osmosis water, yielding an LD50 > 5000 mg/kg (no observed animal death). The compound was therefore classified as non-toxic by oral administration. In an acute respiratory toxicity test (OECD 403), heads and noses of SD rats were exposed to liquid aerosol for 4 h (the highest concentration that could be administered without causing any animal death), and a lethal concentration (LC50) > 0.88 mg/L was obtained. The mass medium aerodynamics diameter (MMAD) of rEP aerosol particles was 8.18 µm and mass medium aerodynamics diameter (GSD) was 3.04, which meant that 25.90% could enter the airway (<4 µm) of a rat, and 58.06% (<10 µm) could be inhaled by humans. An ocular irritation test (OECD 405) with rEP powder was performed on New Zealand White (NZW) rabbits. Signs of irritation included conjunctival swelling and diffuse flushing 1 h after administration. The signs were less apparent after 24 h and disappeared after 72 h. The classification assigned to the powder was mild eye irritation. Skin sensitization was performed for a local lymphoproliferative test (OECD 442B) using BALB/c mice, with the highest soluble concentration of the rEP considered to be 100% test substance; formulations were diluted to 50% and 25%, and bromodeoxyuridine (BrdU) incorporation was used to measure the degree of lymphocyte proliferation. The stimulation indexes (SIs) were 1.06 (100%), 0.44 (50%), and 0.77 (25%), all of which were less than the cutoff value for a positive sensitization result (1.6). Negative response was also seen in the bacterial reverse mutation test (OECD 471), and no chromosomal effects on Chinese hamster ovary (CHO)-K1 cells were observed (OECD 487). Based on these six toxicity tests, rEP showed neither acute toxic effects in experimental animals nor mutagenicity. Thus, rEP can be considered safe for use in subsequent research on its application as a feed additive for poultry, cattle, or aquatic animals.

Prioritization of pesticides in crops with a semi-quantitative risk ranking method for Taiwan postmarket monitoring program.

A risk-based prioritization of chemical hazards in monitoring programs allows regulatory agencies to focus on the most potentially concerned items involving human health risk. In this study, a risk-based matrix, with a scoring method using multiple factors for severity and probability of exposure, was employed to identify the pesticides presented in crops that may pose the greatest risk to human health. Both the probability of exposure and the severity were assessed for 91 pesticides detected in a Taiwanese postmarketing monitoring program. Probability of exposure was evaluated based on the probability of consumption and evidence of pesticide residues in crops. Severity was assessed based on the nature of the hazard (i.e., the description of toxic effects), and the acceptable daily intake (ADI) reported by available toxicological reports. This study showed that the nature of the hazard and probability of consumption had the strongest contribution to risk score. Dithiocarbamates, endosulfan, and carbofuran were identified as the pesticides with the highest concern for human health risks in Taiwan. These pesticides should be monitored more frequently than others in crops during the postmarketing monitoring program. However, some uncertainties shall be noted or improved when this methodology is applied for risk prioritization in the future.

Activation of aryl hydrocarbon receptor reduces carbendazim-induced cell death.

Carbendazim inhibits microtubule assembly, thus blocking mitosis and inhibiting cancer cell proliferation. Accordingly, carbendazim is being explored as an anticancer drug. Data show that carbendazim increased mRNA and protein expressions and promoter activity of CYP1A1. In addition, carbendazim activated transcriptional activity of the aryl hydrocarbon response element, and induced nuclear translocation of the aryl hydrocarbon receptor (AhR), a sign the AhR is activated. Carbendazim-induced CYP1A1 expression was blocked by AhR antagonists, and was abolished in AhR signal-deficient cells. Results demonstrated that carbendazim activated the AhR, thereby stimulating CYP1A1 expression. In order to understand whether AhR-induced metabolic enzymes turn carbendazim into less-toxic metabolites, Hoechst 33342 staining to reveal carbendazim-induced nuclear changes and flow cytometry to reveal the subG0/G1 population were applied to monitor carbendazim-induced cell apoptosis. Carbendazim induced less apoptosis in Hepa-1c1c7 cells than in AhR signal-deficient Hepa-1c1c7 mutant cells. Pretreatment with ß-NF, an AhR agonist that highly induces CYP1A1 expression, decreased carbendazim-induced cell death. In addition, the lower the level of AhR was, the lower the vitality present in carbendazim-treated cells, including hepatoma cells and their derivatives with AhR RNA interference, also embryonic kidney cells, bladder carcinoma cells, and AhR signal-deficient Hepa-1c1c7 cells. In summary, carbendazim is an AhR agonist. The toxicity of carbendazim was lower in cells with the AhR signal. This report provides clues indicating that carbendazim is more potent at inducing cell death in tissues without than in those with the AhR signal, an important reference for applying carbendazim in cancer chemotherapy.