In vitro kinetic characterization of inhibition of acetylcholinesterase by organophosphate and carbamate compounds in food animals

Organophosphate, dichlorvos (2,2-dichlorovinyldimethylphosphate, DDVP), and diazinon (DZN) as well as carbamates are currently found in contaminated water, dust, soil, agricultural waste, and elsewhere in the environment, and can be harmful after accidental or deliberate exposure. Intoxication by these compounds causes a generalized cholinergic crisis due to the inhibition of acetylcholinesterase (AChE), whose major physiological role in mammalian tissues is in nervous transmission. The half-maximal inhibitory concentration (IC₅₀) was found to be higher for carbaryl than DDVP and DZN, about 3–4 times in liver and about 4–6 times in muscles. The half time (t _1/2) of inhibition (8?µmol?L^?1 DDVP or DZN; 40?µmol?L^?1 carbaryl) ranged between 4 and 16?min and decreased as follows: pig?>?cattle?>?sheep for liver tissue, and cattle?>?sheep?>?pig for muscle. Rate constants of inhibition (k _i) ranged between 43?×?10^?3 and 168?×?10^?3?min^?1 for liver and from 46?×?10^?3 to 115?×?10^?3?min^?1 for muscles. Very little residual AChE activity (<6.1%) was seen in liver, but more was seen in muscle (<17%).     

Neurotoxicity and oxidative stress induced by permethrin in gills of the freshwater mussel Unio ravoisieri

Pyrethroids are contaminants found in the aquatic environment, and their toxicological effects on aquatic organisms have received extensive attention. However, the impact on freshwater bivalve of exposure to these chemicals is still largely unknown. Freshwater mussels Unio ravoisieri were exposed to two nominal permethrin (PM) concentrations C1?=?50?µg/L and C2?=?100?µg/L during 7 days. The measured concentrations of PM using gas chromatography (GC/ECD) in the treated aquariums were, respectively, 28.7–62.3?µg/L. Catalase (CAT), Glutathione-S-transferase (GST), and Acetylcholinesterase (AChE) activities, Glutatione (GSH) and Malondialdehyde (MDA) levels were determined in gills of U. ravoisieri. Significant increase in CAT activity by the lowest concentration and decrease by highest concentration were observed. Additionally, GST activity was increased in a concentration-dependent manner. However, statistically significant decrease in GSH levels (about 39%) was observed only at high concentration of this compound (100?µg/L). PM generated an increase in MDA levels reaching the highest value at the high concentration. AChE activity of mussel ranging from 51% inhibition at lowest concentration 50?µg/L to 89% inhibition at highest concentration 100?µg/L. The results indicated that oxidative stress and cell damage might be one of the main mechanisms of PM toxicity to freshwater mussels.     

Proteomics study of Southern Punjab Pakistani cobra (Naja naja: formerly Naja naja karachiensis) venom

Because of the potential of significant geographic variation in the compositions of snake venoms, much insight can be gained by examining individuals originating from diverse locations, and detailed characterization of any species can only be done by including animals from the entire range. Pakistani N. naja (formerly N. naja karachiensis) venom was decomplexed by applying diverse techniques and found a concoction of proteins (a total of 43 venomous proteins comprising 11 super families) from 6 to 200 kilodaltons in size. Among them three finger toxins (58%), phosphoplipses A₂ (19%), snake venom metalloproteinases (5%), l-amino acid oxidases (5%), helvepryns (3%), vespryns (2%), cobra venom factor (2%), 5′-nucleotidases (2%), venom nerve growth factor (2%), and Kunitz type serine protease inhibitor (2%) were included. It is the first report that this venom possesses significantly higher percentage of three finger toxins belonging to the cytotoxins (32% overall) and α‐neurotoxins (32% overall, predominately long chain neurotoxins, 24% overall) compared to N. naja from other geographic locations. Better understanding of intraspecific variations in venom can improve the development of location-specific anti-venoms. Furthermore, this pool of diverse toxins could potentially be a source of novel drug candidates for the treatment of muscarinic acetylcholine receptors mediated nervous disorders.     

Effects of bisphenol A on Gammarus fossarum and Lumbriculus variegatus in artificial indoor streams

The impact of bisphenol A (BPA) on Gammarus fossarum and Lumbriculus variegatus was studied in four artificial indoor streams (0, 5, 50 and 500?µg?L^?1 BPA, nominal) over 103 days in a pulse–dose exposure scenario (weekly BPA application). For G. fossarum populations at day 103, the proportions of juveniles and of breeding females from the highest BPA treatment were in tendency reduced. For individually exposed gammarid pairs an EC₁₀ of 17?µg?L^?1 BPA (nominal) for the proportion of reproductive females in the fourth brood was determined. During the first three broods, the largest brood size occurred at the highest BPA concentration, whereas in the fourth brood it decreased concentration-dependently (fourth brood EC₁₀?=?5?µg?L^?1 BPA, nominal). Effects on L. variegatus were a reduced population growth (103?d-EC₁₀ of 2?µg?L^?1 BPA, nominal) and an increase in dry weight and the number of segments in large, complete worms.     

Protective role of epigallocatechin 3-gallate against lead-induced toxicity in human neuroblastoma cells

Lead (Pb) is a heavy metal, known to induce oxidative stress and produce damage to the antioxidant defence system ultimately leading to cell death. Antioxidants such as epigallocatechin 3-gallate (EGCG), a green tea polyphenol, was shown to play a protective role during Pb-exposure. In this study, human SH-SY5Y neuroblastoma cells were exposed to different concentrations (0.01–10?µM) of Pb for 48?h to determine effects on the viability of cells. It was observed that IC₅₀ was at 5?µM and at this concentration the cells exhibited a significant increase in caspase-3 activity, an indicator of apoptosis at least by 10-fold and the decrease of 59.4% in glutathione (GSH) content. The total cellular prostaglandin-E₂ (PGE₂) level was found to be elevated at least 10-fold upon Pb exposure. However, the effects of Pb on cells pre-incubated with 50?µM EGCG followed by 5?µM Pb showed 40% inhibition in cell viability, 17.3% decrease in caspase-3 activity, 23% increase in GSH content, and 11.4% fall in PGE₂ levels when compared with cells exposed to Pb only. Data suggest that EGCG exerted a significant protection to cell viability in preventing cell death and elevation in levels of GSH in cells exposed to Pb. However, EGCG did not elicit any significant effect on release of PGE₂ indicating the nature of EGCG as an effective anti-apoptotic, antioxidant, and anti-inflammatory agent.     

Differential effects of UV-B radiation fluence rates on growth,photosynthesis, and phosphate metabolism in two cyanobacteria under copper toxicity

This work was undertaken to ascertain the impact of different fluence rates of ultraviolet-B (UV-B) radiation on two cyanobacterial biofertilizers, Phormidium foveolarum and Nostoc muscorum, growing under copper toxicity. Copper (2 and 5?µmol?L^?1) and high UV-B fluence rate (UV-B_H; 1.0?µmol?m^?2?s^?1) decreased the growth, pigment content, photosynthetic oxygen yield, phosphate uptake, and acid phosphatase activity in both the strains analyzed after 24 and 72?h of experiments, and combined exposure further enhanced the toxic effects. Respiration and alkaline phosphatase activities were stimulated appreciably. The damaging effect was shown on the order on pigments: phycocyanin?>?chlorophyll a?>?carotenoids, and on photosystems: whole chain photosynthetic reaction?>?photosystem II?>?photosystem I. Partial recovery in the photosystem II activity in the presence of artificial electron donors; diphenyl carbazide (DPC), hydroxylamine (NH₂OH), and manganese chloride (MnCl₂) pointed out the interruption of electron flow on the oxidation side of photosystem II. Unlike UV-B_H, low UV-B fluence rate (UV-B_L; 0.1?µmol?m^?2?s^?1), rather than causing damaging effect partially, alleviated the toxic effects of Cu. This study suggests that the cyanobacterium P. foveolarum is less sensitive against UV-B_H and excess Cu (2 and 5?µmol?L^?1), thus P. foveolarum may be used as a biofertilizer for sustainable agriculture.     

Seasonal changes in inorganic nutrient concentration in the alexandria western harbour (Egypt)

Surface and bottom water samples were collected on a monthly basis from six locations in the Alexandria Western Harbour between April 2002 and March 2003. Total nitrogen, ammonia, nitrite, nitrate, total phosphorus, reactive phosphate and silicate were analysed. The average content of total nitrogen ranged from 81.1 to 65.7?µmol?l^?1 in the surface and bottom waters, respectively, while ammonia ranged from 13.77 to 15.79?µmol?l^?1 in surface and bottom waters, respectively. Also, the average concentration of nitrite was relatively higher in surface waters than in bottom waters (0.89 and 0.61?µmol?l^?1, respectively). The results of this study also indicated a considerable temporal variation in nitrate concentrations which ranged from 1.12 to 13.83?µmol?l^?1. Total phosphorus displayed an irregular pattern throughout the year, ranging from 1.9 to 11.8?µmol?l^?1 in surface waters and from 1.7 to 9.1?µmol?l^?1 in bottom waters. The results of PO₄-P analysis showed higher values in surface waters (0.28–2.75?µmol?l^?1) than in bottom waters (0.10–1.70?µmol?l^?1). The average concentration of silicates was relatively lower in the surface than in the bottom waters (8.97 and 10.1?µmol?l^?1, respectively). The analysis of variance (ANOVA) among seasons and sites revealed significant differences for ammonia, total nitrogen and phosphate, while nitrate showed no significant differences among stations. Finally, silicate did not show any significant variance among sites and seasons (ANOVA, P?>?0.05).     

Morpho-histological and enzymatic alterations in earthworms Drawida willsi and Lampito mauritii exposed to urea,phosphogypsum and paper mill sludge

The effects of varying concentrations of urea, phosphogypsum and paper mill sludge (PMS) on the morphology, histology, tissue protein content, lipid peroxidation (LPX), activities of lactate dehydrogenase (LDH), acetylcholinesterase (AChE) and catalase in earthworms Drawida willsi and Lampito mauritii have been studied over an exposure period of 24?hr. Integumentary lesions, clitellar swelling and loss of pigmentations were found to be major morpho-pathological changes in the worms. Histology indicated cuticular damage, ruptured epithelium and muscle fibres with accumulation of cellular debris. Lowest tissue protein content (57.02?±?4.02?mg/g tissue) and highest LPX (0.113?±?0.04, 0.137?±?0.08?nmol/mg protein) were noticed in D. willsi at a high concentration of PMS, whereas highest tissue protein content was observed in L. mauritii (115.32?±?7.18?mg/g tissue) with the same treatment. In both the species, LDH activity was minimum at a high concentration of urea (0.172?±?0.02; 0.247?±?0.08?U/mg protein). AChE activity was highest (0.099?±?0.002?U/mg protein) at a high concentration of PMS in D. willsi, whereas catalase activity was the maximum (0.338?±?0.02?U/mg protein) at high concentrations of PMS in L. mauritii. The study indicated that morpho-histological and enzymatic alterations in these earthworms exposed to agrochemicals could be useful biomarkers to evaluate soil toxicity.     

Toxicokinetics,metabolism, and microsomal studies of chlorpropham in rats

A study on the toxicokinetic behavior, metabolism of chlorpropham, and its effect on cytochrome P₄₅₀ from liver microsomes was carried out in albino rats after a single and consecutive oral administration at 500?mg?kg^?1 body weight for 10 and 20 days. Chlorpropham was detected in the blood at 0.08?h (11.43?±?1.72?µg?mL^?1) reaching a maximum concentration at 2?h (30.90?±?2.55?µg?mL^?1) and a minimum at 48?h (1.95?±?0.20?µg?mL^?1) after a single oral administration of 500?mg?kg^?1. The absorption rate constant (K _a) was 0.66?±?0.48?h^?1. The Vd _area (18.01?±?2.78?L?kg^?1) and t _1/2 β (12.23?±?1.96?h) values suggested a wide distribution and long persistence of the compound in the body, respectively. The higher Cl_R (0.82?±?0.00?L?kg^?1?h^?1) compared to Cl_H (0.18?±?0.02?L?kg^?1?h^?1) value indicated that a major portion of chlorpropham was excreted through the urine (30%) compared to the faeces (2.81%). Chlorpropham residue was detected in all tissues of rat at 0.25?h while its metabolite, meta-chloroaniline was detected in liver, kidney, heart, lung, and spleen tissue at 0.25?h. Meta-chloroaniline was not detected in skeletal muscle, brain, fat, and stomach tissue at any time of the observation period. Maximum concentrations of chlorpropham and meta-chloroaniline were detected at 2?h (except in the spleen), and minimum concentrations of chlorpropham at 24 (heart, lung, spleen, skeletal muscle, and stomach) and 48?h (liver, kidney, brain, and fat tissue) respectively; and meta-chloroaniline at 24?h (except heart and spleen). The tissue half-life of chlorpropham in rat varied from 3.80 to 11.60?h. Repeated oral administration of chlorpropham at 500?mg?kg^?1 for 10 and 20 days caused an induction of the liver microsomal pellet of rat.     

Arachis hypogaea derived activated carbon steered remediation of Benzimidazole based fungicide adsorbed soils

Sorption characteristics of the Benzimidazole fungicide Carbendazim were assessed in seven different soils using batch equilibrium method and analysed by UV-Vis spectrophotometer. The values of adsorption co-efficient K_d ranged from 14.3 to 39.8?µg/mL depending upon unique physiochemical properties of soils. Negative values for Gibbs free energy (ΔG) proposed an exothermic and low interaction between Carbendazim and soil samples leading to physiosorption. Statistical analysis showed a negative correlation of soil pH and K_d (R²= ?0.80) and a positive correlation with organic matter (R^2?=?0.77). Activated carbon prepared from Arachis hypogaea (peanut shells) by acid activation for Carbendazim removal from soils was characterised by FTIR spectrometry, indicating the change in functional groups. The highest percentage removal observed was 70% in 5?ppm initial Carbendazim concentration while 65% in 7.5?ppm concentration. This method can be implied in agricultural soils as an efficient and cheap technique for removing the hazardous pesticides from the environment.     

Efficiency of Zn phytoextraction,biomass yield and formation of low-molecular-weight organic acids in S × rubens – a hydroponic experiment

The aim of the study was to estimate Zn phytoextraction and changes in biomass of S?×?rubens growing in modified Knop's solution with different levels of Zn addition (0.5, 1.0, 2.5 and 5.0?mM). Obtained results were correlated with secretion of selected low-molecular-weight organic acids (LMWOAs) in the rhizosphere, roots and leaves. An increase in Zn concentration in Knop's solution resulted in Zn accumulation in roots, shoots and leaves. The highest accumulation was observed for plants growing in 5?mM Zn, at concentration levels 4741.36?±?98.66, 1227.31?±?16.57 and 2241.65?±?34.90?mg?kg^?1 DW in roots, shoots and leaves, respectively. The bioaccumulation factor and the translocation factor for plants growing in 0.5, 1.0 and 2.5?mM Zn clearly indicate that this Salix taxon is an effective Zn accumulator. The general reduction of Salix biomass with an increase in Zn concentration in the solution was observed. In the rhizosphere, the total LMWOA concentration was almost 0.93?µmol?kg^?1 DM for control (Zn free) plants, while for 5.0?mM of Zn it was 4.9?µmol?kg^?1 DM. Increasing concentrations of acids were observed in roots (1.34 for the control and 5.57?µmol?kg^?1 DM for plants treated with 2.5?mM of Zn).     

Environmental human silver exposure

Environmental exposure to silver (Ag) was assessed in occupationally non-exposed adult human population by analyzing Ag in the hair (H?·?Ag) and whole blood (WB?·?Ag). H?·?Ag was analyzed in 311 (123 men, M; 188 women, W); while WB?·?Ag was determined in 235 of these individuals (90 M, 145 W). Women had more H?·?Ag than men (M 0.05 vs. W 0.076), whereas WB?·?Ag concentrations in men and women were not significantly different. A natural distribution of the median derivatives was utilized to generate the dataset to fit the logistic sigmoid curve to assess the current human body burden of environmental Ag population exposure for M and W separately. The H?·?Ag (µg?g^?1) below 0.0105 for M and 0.0145 for W, reflects low level of environmental Ag exposure. The adaptive physiological saturation phase followed where H?·?Ag rose rapidly, first for M and then for W in parallel with biological assay. Both parallel saturation curves converged and plateaued at 0.215 for M and 0.965 for W (µg?g^?1). The current level of human environmental Ag exposure is low, but cases of high Ag exposure occurred sporadically. In conjunction with the medical histories overt clinical neural toxicity may be expected for H?·?Ag at 4?µg?g^?1and higher. There were no significant correlation between the H?·?Ag and WB?·?Ag.     

Effects of lead on calmodulin content,proliferation and interleukin-2 in lymphocytes of mice

The mechanism inderlying lead (Pb)-induced toxicity was presumed to involve interaction between Pb²⁺ and calcium (Ca²⁺) associated with calmodulin-dependent systems. The aim of this study was to (1) examine the hypothesis that Pb alters calmodulin content and (2) assess the effects of Pb on proliferation and interleukin-2 (IL-2) levels in lymphocytes. The influence of Pb on calmodulin content in mouse lymphocytes was assessed by western blot, while the proliferation of lymphocytes was studied using MTT test, and IL-2 level by ELISA assay. The results showed that 100?µM PbCl₂ decreased calmodulin content, phytohemagglutinin (PHA)-induced proliferation, and IL-2 levels, but a lower concentration (1?µM PbCl₂) did not appear to be effective. Evidence indicates that IL-2 levels and lymphocyte proliferation might not be calmodulin content dependent. Our data suggested that Pb²⁺-induced toxicity was likely to be complex with multiple factors involved, including calmodulin, IL-2, and proliferation of lymphocytes.     

Speciation of mercury in South African coals

Total mercury (Hg_TOT) concentrations were determined by inductively coupled plasma mass spectrometry (ICP MS) for South African Highveld coals. The distribution of Hg in coals was investigated using a four-stage sequential leaching protocol and isotope dilution/gas chromatography coupled to ICP MS (ID-GC-ICP MS). The results show that Hg_TOT ranged from 144 to 303?µg?kg^?1 with a mean of 199?±?26?µg?kg^?1, while Hg_TOT leached from coals using different solvents ranged between 103 and 310?µg?kg^?1 (mean: 218?±?60?µg?kg^?1). Hg leaching rates of 53–78% were achieved in crushed coals. Hg⁰, Hg²⁺, and CH₃Hg⁺ were identified in all coals. CH₃Hg⁺ in studied coals ranged between 0.1 and 0.4 (mean: 0.2) µg?kg^?1. GC ICP MS chromatograms also showed unknown Hg peaks which were identified as other organomercury species such as ethylmercury. Modes of occurrence of Hg in coals were variable with the organic-bound (37–40%) and the sulfide-bound (37–39%) being the dominant mercury forms. Increasing the HCl concentration in the used protocol increased the amount of Hg leached (16%) during this step.     

In vitro antioxidant and antibacterial activities of certain 3-{4-(5-mercapto-1,3,4-oxadiazole-2-yl)phenylimino}-indolin-2-one derivatives

A new series of 5- or 7-substituted 3-{4-(5-mercapto-1,3,4-oxadiazol-2-yl)phenylimino}-indolin-2-one derivatives were screened for in vitro antioxidant and antimicrobial activities. The results of this study revealed that all synthesized compounds were significantly scavenged DPPH free radicals in a concentration-dependant manner. The IC₅₀ values of all test compounds were found to be between 18.34 and 50.54?µM. All compounds significantly inhibited growth of bacteria and most of the minimum inhibitory concentration values were between 20 and 100?mg?mL^?1 indicating potent to moderate antibacterial activity. In conclusion, 3-{4-(5-mercapto-1,3,4-oxadiazole-2-yl)phenylimino)-5 or 7-substituted indolin-2-ones derivatives exhibited moderate antioxidant and antimicrobial activities.     

Acute toxicity evaluation of triazophos,deltamethrin and their combination on earthworm,Eudrilus eugeniae and its impact on AChE activity

The acute toxicity of three formula grade pesticides namely, triazophos (an organophosphate, OP), deltamethrin (a pyrethroid) and combined pesticide (triazophos?+?deltamethrin) was determined in earthworm, Eudrilus eugeniae. They were exposed to different concentrations of these pesticides for 48 h by paper contact toxicity method. The LC₅₀ values for triazophos, deltamethrin and of combination were determined as 0.076, 0.031 and 0.065?μg/cm², respectively. To study the sublethal effect of these pesticides, E. eugeniae were exposed to 5% and 10% of LC₅₀ of pesticides for 48 h. Morphological alterations such as coiling, clitellar swelling, mucus release and bleeding followed by body segmentation were observed in exposed earthworm. Acetylcholinesterase (AChE) activity assayed in different regions of body segment exhibited a significant (p?<?0.05) decrease in its activity particularly in the pre-clitellar region as compared to other regions. The altered behavioural responses in pesticides exposed earthworms would have been due to decline in AChE activity of the nervous system.     

In vitro and in vivo inhibition of Ca2+-Mg2+-ATPase activity by cadmium in post larvae of Penaeus monodon

Ca²⁺-Mg²⁺-ATPase is a membrane-bound enzyme and is responsible for regulating cytosolic free calcium. In vitro and in vivo effects of cadmium were studied on Ca²⁺-Mg²⁺-ATPase activity in plasma membrane/mitochondrial fraction of Penaeus monodon post larvae. In vitro studies revealed a concentration-dependent decrease in enzyme activity with an IC₅₀ value of 11.02?µM. In vivo experiments were conducted by exposing the post larvae to 1/10th (0.12?ppm) and 1/5th (0.24?ppm) of LC₅₀ values of cadmium for 30 days. Both ATPase activity and metal accumulation were estimated in post larvae exposed to 0.12 and 0.24?ppm of cadmium at different intervals of 24?h, 48?h, 96?h, 10 days and 30 days. ATPase activity showed a gradual decrease in post larvae on exposure to both the sub-lethal concentrations with respect to their controls and the decrease was significant (p?相似文献   

Toxicity of endosulfan on growth,photosynthesis, and nitrogenase activity in two species of Nostoc (Nostoc muscorum and Nostoc calcicola)

Treatments of the cyanobacteria Nostoc muscorum and Nostoc calcicola with the insecticide endosulfan (5, 10, and 20?µg?mL^?1) inhibited growth, photosynthetic pigments, photosynthetic, and nitrogenase activities. The sensitivity of N. muscorum to endosulfan was higher than that of N. calcicola. The toxic effect of endosulfan was more pronounced on phycocyanin; however, a considerable reduction in chlorophyll a and carotenoids was also noticed. ¹⁴C-fixation appeared to be more sensitive to the insecticide than photosynthetic oxygen evolution. Endosulfan caused strong inhibition of photosystem (PS) II activity whereas PS I was least affected. The inhibition of PS II activity was partially restored by electron donors (DPC, NH₂OH, and MnCl₂) at low dose of endosulfan. Nitrogenase activity was significantly suppressed in both species by the endosulfan at high dose (20?µg?mL^?1). On the basis of our comparative analysis, N. calcicola was found to be endosulfan resistant and can be used in paddy fields for better productivity under pesticide stress.     

Inhibition of tetrachloromethane-induced cross-linking of microsomal fatty acids by the spin-trap α-phenyl- N - t -butylnitrone

The effect of the spin-trap α-phenyl-N-t-butylnitrone (PBN) on the formation of fatty acid dimers induced by the anoxic microsomal metabolism of tetrachloromethane was investigated in vitro, in particular, with respect to cross-linking of arachidonic acid with a second one (C₂₀–C₂₀) and of arachidonic acid with docosahexaenoic acid (C₂₀–C₂₂). PBN exerted no effect on anaerobic tetrachloromethane metabolism but inhibited dimer formation, half-maximally at a concentration of 0.1 mmol L^?1. Trichloromethyl octadecenoic acid was also found, but at levels 10-fold lower than the fatty acid dimers. The data suggest that the hepato-protective effect of PBN against tetrachloromethane-toxicity may be due to its ability to scavenge fatty acid radicals, resulting in stable, non-reactive entities, which do not undergo secondary reactions such as cross-linking. The data confirm and extend previous findings that crosslinking plays an important role in tetrachloromethane toxicity and similar cases of metabolic radical-induced toxicity.