Reversible and time-dependent inhibition of the hepatic cytochrome P450 steroidal hydroxylases by the proestrogenic pesticide methoxychlor in rat and human

Methoxychlor, a currently used pesticide, is demethylated and hydroxylated by several hepatic microsomal cytochrome P450 enzymes. Also, methoxychlor undergoes metabolic activation, yielding a reactive intermediate (M*) that binds irreversibly and apparently covalently to microsomal proteins. The study investigated whether methoxychlor could inhibit or inactivate certain liver microsomal P450 enzymes. The regioselective and stereoselective hydrox-ylation of testosterone and the 2-hydroxylation of estradiol (E₂) were utilized as markers of the P450 enzymes inhibited by methoxychlor. Both reversible and time-dependent inhibition were examined. Coincubation of methoxychlor and testosterone with liver microsomes from phenobarbital treated (PB-microsomes) male rats, yielded marked diminution of 2α- and 16α-testosterone hydroxylation, indicating strong inhibition of P4502C11 (P450h). Methoxychlor moderately inhibited 2β-, 7α-, 15α-, 15β-, and 16β-hydroxylation and androstenedi-one formation. There was only a weak inhibition of 6β-ydroxylation of testosterone. The methox-ychlor-mediated inhibition of 6β-hydroxylation was competitive. By contrast, when methoxychlor was permitted to be metabolized by PB-microsomes or by liver microsomes from pregnenolone-16α-car-bonitrile treated rats (PCN-microsomes) prior to addition of testosterone, a pronounced time-dependent inhibition of 6β-hydroxylation was observed, suggesting that methoxychlor inactivates the P450 3A isozyme(s). The di-demethylated methoxychlor (bis-OH-M) and the tris-hydroxy (ca-techol) methoxychlor metabolite (tris-OH-M) inhibited 6β-hydroxylation in PB-microsomes competitively and noncompetitively, respectively; however, these methoxychlor metabolites did not exhibit a time-dependent inhibition. Methoxychlor inhibited competitively the formation of 7α-hydroxytestosterone (7α-OH-T) and 16α-hydroxy-testosterone (16α-OH-T) but exhibited little or no time-dependent inhibition of generation of these metabolites, indicating that P450s 2A1, 2B1/B2, and 2C11 were inhibited but not inactivated. Methoxychlor inhibited in a time-dependent fashion the 2-hydroxylation of E2 in PB-microsomes. However, bis-OH-M exhibited solely reversible inhibition of the 2-hydroxylation, supporting our conclusion that the inactivation of P450s does not involve participation of the demethylated metabolites. Both competitive inhibition and time-dependent inactivation of human liver P450 3A (6β-hydroxylase) by methoxychlor, was observed. As with rat liver microsomes, the human 6β-hydroxylase was inhibited by bis-OH-M and tris-OH-M competitively and noncompetitively, respectively. Testosterone and estradiol strongly inhibited the irreversible binding of methoxychlor to microsomal proteins. This might explain the “clean” competitive inhibition by methoxychlor of the 6β-OH-T formation when the compounds were coin-cubated. Glutathione (GSH) has been shown to interfere with the irreversible binding of methoxychlor to PB-microsomal proteins. The finding that the coincubation of GSH with methoxychlor partially diminishes the time-dependent inhibition of 6β-hydroxylation provides supportive evidence that the inactivation of P450 3A isozymes by methoxychlor is related to the formation of M*.     

Interactive effects of selected pesticides on the two-spotted spider mite and its fungal pathogen Neozygites floridana

Benomyl affected populations of Tetranychus urticae by interfering with the pathogenic fungus, Neozygites floridana. Benomyl delayed but prolonged spider mite outbreaks. Few mites were infected with the pathogen when benomyl was used. Reductions in mite populations treated with fentin hydroxide were associated with a high incidence of N. floridana infection. Benomyl did not affect sporulation of N. floridana but appeared to inhibit conidial germination or growth of the fungus.

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Résumé Le bénomyl a modifié les populations de Tetranychus urticae Koch en interférant avec son champignon pathogène, Neozygites floridana (Weiser & Muma). Le bénomyl retardait mais prolongeait les pullulations de l'acarien. Peu d'acariens étaient infectés par le champignon quand on utilisait du bénomyl. Les réductions des populations d'acariens traitées avec l'hydroxyde de fentine étaient associées à un haut niveau d'infection par N. floridana. Le bénomyl ne modifiait pas la sporulation de N. floridana mais semblait inhiber la germination des conidies ou la croissance du champignon.

     

Residues and contaminants in medicinal herbs—A review

With the increasing popularity and use of medicinal herbs, their global demand has gained momentum. Developing countries, including China, India and South East Asian (SEA) countries, are the centres of origin and major global suppliers for most of these traditionally used medicinal herbs. One of the factors affecting the quality of these herbs is the contamination of heavy metals, mycotoxins, pesticide residues, polycyclic aromatic hydrocarbons (PAHs) and fumigants. These contaminants can accumulate during the cultivation, storage and processing of herbs and may have adverse effects on consumer health. There have been various reports regarding the presence of these contaminants in medicinal herbs. This review discusses the important contaminants of medicinal herbs, the frequency and magnitude of their occurrences, the potential causes of contamination and their regulatory limits in medicinal herbs. The major challenge in the international trade of medicinal herbs is the lack of common guidelines, regulatory measures and monitoring body to strictly enforce their regulation.     

Effect of pesticide exposure on the cholinesterase activity of the occupationally exposed tea garden workers of northern part of West Bengal,India

Context: Pesticide poisoning and related deaths are a global concern, but there is little information about its effect on the occupationally exposed tea garden workers of North Bengal.

Objective: This study investigates the level of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) in the blood of the tea garden workers at risk of exposure to a mixture of pesticides.

Materials and methods: The study sample consisted of pesticide exposed workers, non-exposed (control), smokers and alcoholics. AChE and BuChE activity was measured and tested for significance.

Results: Results showed that AChE activity was half in the pesticide exposed individuals than controls (p≤ 0.001). BuChE activity was also significantly decreased in the pesticide exposed individuals than controls (p≤ 0.001), while AChE and BuChE activity in smokers and alcoholics were not different from that of controls. However, significantly decreased AChE and BuChE activities were recorded in pesticide exposed workers compared to smokers and alcoholics.

Conclusions: The results indicated that the decrease in enzyme activities in tea garden workers was due to mixed pesticides (containing organophosphates) exposure. Age was not found to influence the enzyme activities. However, the gender had little effect on the enzyme activities but the effect was not so prominent.     

两种测定农药在大鼠和小鼠体内毒性蓄积的方法

目的了解有机磷酸酯和氨基甲酸酯类农药在大鼠和小鼠体内的蓄积毒性效应,建立评价这两类农药在大鼠和小鼠体内的蓄积毒性测定方法。方法采用2种评价方法-大鼠的剂量固定20 d蓄积法和小鼠的剂量递增法同时对这两类杀虫剂的蓄积毒性进行测试。结果所测定的这两类共8种杀虫剂蓄积系数均大于5,在机体内属于轻度蓄积。结论固定剂量法和剂量递增法均可用于大鼠和小鼠的体内蓄积毒性评价。常用的有机磷和氨基甲酸酯类农药蓄积性较弱。     

Amperometric biosensor based on enzymes immobilized in hybrid mesoporous membranes for the determination of acetylcholine

Acetylcholine sensor is successfully prepared by using immobilized enzymes, i.e., acetylcholinesterase and choline oxidase within separate hybrid mesoporous silica membranes with 12 nm pore diameter (F127M). The measurement was based on the detection of hydrogen peroxide produced by two sequential enzyme reactions. The determination range and the response time are 6.0–800 μM and within approximately 3 min, respectively. The sensor is very stable compared to free enzymes and 80% of the initial response was maintained even after storage for 80 days. These results show that two enzymes are successfully immobilized and well stabilized, and at the same time, two sequential enzyme reactions efficiently proceed within the separate hybrid mesoporous membranes. Further, we studied the possible detection of organophosphorus pesticides in terms of the inhibition of acetylcholinesterase activity, i.e., the decrease of current response, and demonstrated that the nanomolar concentrations of pesticide (DZN-oxon) can be detected with our sensor.     

Drin pesticides removal from aqueous solutions using acid-treated date stones

This work describes the potential applicability of chemically and thermally treated date stones for removing drin pesticides (aldrin, dieldrin and endrin) from aqueous solutions. The effect of several parameters, such as sorbent particle size, adsorbent dose, shaking speed, shaking time, concentration of pesticide solution and temperature, was evaluated by batch experiments. Pesticide determination was carried out using stir bar sorptive extraction and gas chromatography coupled with mass spectroscopy. Maximum removal efficiency (93%) was reached using 0.1 g of acid-treated date stones (ATDS) (63–100 μm) and 100 mL of aldrin (0.5 mg L⁻¹). The removal efficiency of drin pesticides decreased in the order of aldrin, dieldrin and endrin, and decreased as the temperature rose. Adsorption data were processed according to various kinetic models. Lagergren and Morris-Weber equations were applied to fit the kinetic results. The second order model was the most suitable on the whole, and intra-particle diffusion was found to be the rate-controlling the adsorption process. According to adsorption kinetic data, 3.5 h were considered as the equilibrium time for determining adsorption isotherms. Adsorption data were analyzed by the Langmuir, Freundlich and Dubinin–Radushkevich adsorption approaches. Experimental results showed that the Freundlich isotherm model best described the adsorption process. In addition, thermodynamic parameters such as ΔH, ΔS and ΔG were calculated. Negative values of ΔH and ΔG indicate the exothermic and spontaneous nature of pesticide adsorption on ATDS.     

“Entombed Pollen”: A new condition in honey bee colonies associated with increased risk of colony mortality

Here we describe a new phenomenon, entombed pollen, which is highly associated with increased colony mortality. Entombed pollen is sunken, capped cells amidst “normal”, uncapped cells of stored pollen, and some of the pollen contained within these cells is brick red in color. There appears to be a lack of microbial agents in the pollen, and larvae and adult bees do not have an increased rate of mortality when they are fed diets supplemented with entombed pollen in vitro, suggesting that the pollen itself is not directly responsible for increased colony mortality. However, the increased incidence of entombed pollen in reused wax comb suggests that there is a transmittable factor common to the phenomenon and colony mortality. In addition, there were elevated pesticide levels, notably of the fungicide chlorothalonil, in entombed pollen. Additional studies are needed to determine if there is a causal relationship between entombed pollen, chemical residues, and colony mortality.     

低农药残留量的乌龙茶种质资源筛选研究

以福建省武夷山茶叶科学研究所鸟龙茶种质资源圃120份资源为试验材料,依外部形态特征初步筛选出31份比较有希望的品种（系）。将联苯菊酯、甲氰菊酯、氯氰菊酯和噻嗪酮4种农药喷施于这31份资源,7d后采摘鲜叶,烘干固样。用气相色谱法分析检测31份品种（系）鲜叶中4种农药的残留量,筛选出低联苯菊酯、低甲氰菊酯、低氯氰菊酯和低噻嗪酮残留量的乌龙茶特异资源各5、4、3和7份。     

云杉蚜虫-天敌-杀虫剂相互作用控制模型的动态特性

使用微分方程与控制理论研究了使用杀虫剂对一类云杉蚜虫与其天敌相互作用模型的动态特性的影响．其中包括云杉蚜虫-天敌-杀虫剂相互作用控制模型正平衡点存在个数及正平衡点的稳定性与杀虫剂使用量的关系．研究结果表明：适当地使用杀虫剂可以在天敌数量不足的情况下控制害虫爆发的事件．