Sensitive and reversible perylene derivative-based fluorescent probe for acetylcholinesterase activity monitoring and its inhibitor.

A reversible fluorescence probe for acetylcholinesterase activity detection was developed based on water soluble perylene derivative, N,N'-di(2-aspartic acid)-perylene-3,4,9,10-tetracarboxylic diimide (PASP). Based on the photo-induced electron transfer (PET), PASP fluorescence in aqueous is quenched after combining with copper ions (Cu2+). Acetylcholinesterase (AChE) is well known to catalyze the hydrolysis of acetylcholine (ATCh) to produce thiocholine, whose affinity is strong enough to capture Cu2+ by thiol (-SH) group from the complex PASP-Cu, resulting in the fluorescence signal of PASP recovers up to 90%. This optical switch is highly sensitive depended on the coordination and dissociation between PASP and Cu2+. We proposed its application for AChE activity detection, as well as its inhibitor screening. According to the change of fluorescence intensity, quantifying the detection limit of AChE was 1.78 mU·mL-1. Classical inhibitors, tacrine and organophosphate pesticide diazinon, were further evaluated for drug screening. The IC50 value of tacrine was calculated to be 0.43 µM, and the detection limit of diazinon was 0.22 µM. Both of these performances were much better than previous results, revealing our probe is sensitive and reversible for screening applications.

Effect of separate and combined exposure of selenium and diazinon on rat sperm motility by computer assisted semen analysis.

Effects of selenium (Se) and diazinon (DZN) on sperm motility parameters in rats were investigated. Male rats received a separate dose of Se (2mgkg-1 b.w., intraperitoneally, 5mgL-1, per os in drinking water), diazinon (20mgkg-1 b.w., intraperitoneally, 40mgL-1, per os in drinking water), and in combination (Se+DZN) with the same dosage as in the separate administration. 36h an intraperitoneal (i.p.) and after 90days of per oral (p.o.) exposure, thirteen parameters of sperm motility were evaluated using a Computer Assisted Sperm Analyzer (CASA). Almost all the evaluated sperm motility parameters significantly decreased in Se p.o. exposed groups. In the Se i.p. group decrease was noted only in beat cross frequency (BCF) and progressive motility. Significant decline in the sperm motility, progressive motility, BCF and increase in amplitude of lateral head displacement (ALH) were recorded after DZN i.p. administration. In DZN p.o. group, significant increase in ALH, velocity average path (VAP) and curvilinear velocity (VCL) but decrease in progressive motility and BCF was detected. Se+DZN i.p. administration caused a significant decrease in motility, progressive motility and BCF. Per oral administration of Se+DZN decreased all motility parameters except LIN, WOB and ALH. Sperm abnormalities increased in all experimental conditions. Se and DZN negatively affected sperm structure and function in separate doses or in combination. No protective effect of Se was observed.

Enhanced expression of hypothalamic nitric oxide synthase in rats developmentally exposed to organophosphates.

Nitric oxide synthase (NOS) is highly expressed in the hypothalamus, and nitric oxide (NO) specifically contributes to the regulation of neuronal activity within distinct hypothalamic regions. We studied the long-lasting effects of developmental exposure to low doses of organophosphate chlorpyrifos (CPF) and diazinon (DZN) on the expression of NOS in the hypothalamic subnuclei that subserve neuroendocrine, autonomic and cognitive functions. A daily dose of 1 mg/kg of either CPF or DZN was administered to developing rats during gestational days 15-18 or postnatal days (PND) 1-4. Brain sections from PND 60 rats were processed using NADPH-diaphorase (NADPH-d) and neuronal NOS (nNOS) immunohistochemistry. The number of labeled neurons and the optical density (OD) were assessed in the supraoptic (SON), paraventricular (PVN), medial septum, vertical limb, and horizontal limb of the diagonal band. Developmental exposure to organophosphates increased the number of labeled neurons and OD in different subnuclei in the hypothalamus without gender selectivity. The effect on OD was more pronounced and was significant for more cases. Prenatal exposure to CPF and DZN significantly increased the OD in all regions studied with the exception of PVN. Neonatal exposure to DZN also consistently increased OD in all studied subnuclei. For rats that treated with CPF during early postnatal period, this effect was statistically significant only for the SON and PVN. These findings suggest that overexpression of NOS in the hypothalamus may contribute to the mechanisms inducing or compensating for endocrine, autonomic and cognitive abnormalities after developmental exposure to organophosphates.

Antioxidant effect of selenium on lipid peroxidation, hyperlipidemia and biochemical parameters in rats exposed to diazinon.

Diazinon (DZN) is one of the most organophosphate insecticides that widely used in agriculture and industry. Selenium is generally recognized to be a trace mineral of great importance for human health, protecting the cells from the harmful effects of free radicals. Therefore, the present study was carried out to investigate the alterations in biochemical parameters, free radicals and enzyme activities induced by diazinon in male rat serum, and the role of selenium in alleviating the negative effects of DZN. Animals were divided into four groups of seven rats each; the first group was used as control. Groups 2, 3 and 4 were treated with selenium (Se; 200µg/kg BW), diazinon (DZN; 10mg/kg BW) and diazinon plus selenium, respectively. Rats were orally administered their respective doses daily for 30 days. Results obtained showed that DZN significantly induced thiobarbituric acid reactive substances (TBARS) and decreased the activities of glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) and the levels of reduced glutathione (GSH) in rat sera. Aminotransferases (AST, ALT), phosphatases (AlP, AcP) and lactate dehydrogenase (LDH) activities were significantly increased while acetylcholinesterase (AChE) activity was decreased due to DZN administration. Also, DZN treatment caused significant perturbations in lipids profile and serum biochemical parameters. On the other hand, Se alone significantly decreased the levels of TBARS, total lipids, cholesterol, urea and creatinine, while increased the activities of antioxidant enzymes and glutathione content, total protein (TP) and albumin. In addition, Se in combination with DZN partially or totally alleviated its toxic effects on the studied parameters. In conclusion, Se has beneficial effects and could be able to antagonize DZN toxicity.

Reduction of diazinon and dimethoate in apple juice by pulsed electric field treatment.

BACKGROUND: Organophosphorus pesticides (OPPs) are widely used in agricultural production in China, and residues of OPPs in agro-products and foods have become a public health concern. Chronic exposure to OPPs can result in potential immunosuppressive effects, cytotoxicity and mutagenicity. Pulsed electric fields (PEFs) have the potential to be used as an alternative to conventional techniques of food production. The aim of the present study was to investigate the influence of PEFs on the degradation of diazinon and dimethoate added to apple juice. RESULTS: PEF treatment significantly promoted the degradation of both pesticides (P < 0.05). The extent of degradation was strongly influenced by the electric field strength and treatment time, and maximum degradation of both diazinon (47.6%) and dimethoate (34.7%) was achieved by treatment at 20 kV cm⁻¹ for 260 µs. The degradation behaviour of the pesticides was described by Weibull and Hülsheger models, both of which successfully fitted the degradation of diazinon and dimethoate. In addition, the decline in the toxicity of samples was observed with a photobacterium bioassay. CONCLUSION: PEF treatment was effective in eliminating residues of diazinon and dimethoate spiked in apple juice and in mitigating sample toxicity.