Significant Inhibition Of Acetylcholinesterase Activity Research Articles

Do magnetic fields related to submarine power cables affect the functioning of a common bivalve?

The aim of the study was to determine the effect of static magnetic field (SMF) and electromagnetic field (EMF), of values usually recorded near submarine cables, on the bioenergetics, oxidative stress, and neurotoxicity in the cockle Cerastoderma glaucum. Bivalves maintained a positive energy balance, but the filtration rate and energy available for individual production were significantly lower in SMF-exposed animals compared to the control treatment. No changes in the respiration were noted but ammonia excretion rate was significantly lower after exposure to EMF. Changes in the activities of antioxidant enzymes and the lipid peroxidation were not observed however, exposure to both fields resulted in increased protein carbonylation. After exposure to EMF a significant inhibition of acetylcholinesterase activity was observed. As the present study for the first time revealed the oxidative damage and neurotoxicity in marine invertebrate after exposure to artificial magnetic fields, the need for further research is highlighted.

Differential biochemical and behavioral responses induced by cocaine and benzoylecgonine exposure to the red swamp crayfish Procambarus clarkii

Cocaine (COC) and its main metabolite, the benzoylecgonine (BE), are the main illicit drugs measured in aquatic system worldwide, with concentrations up to hundreds of ng/L. Although their current environmental concentrations are low these molecules can induce adverse effects at sub-individual level in non-target organisms. In contrast, the information at individual and behavioral level are still scant. The present study aimed at investigating biochemical and behavioral effects induced by 14-days exposure to environmentally relevant concentrations (50 ng/L and 500 ng/L) of COC and BE towards Procambarus clarkii. At sub-individual level, the activity of antioxidant and detoxifying (superoxide dismutase - SOD, catalase - CAT, glutathione peroxidase - GPx and glutathione S-transferases - GST) enzymes, as well as the levels of lipid peroxidation (LPO), were measured as oxidative stress-related endpoints. We also measured the acetylcholinesterase (AChE) activity to check for neurotoxic effect of COC and BE. At individual level, the modulation of some behavioral tasks (i.e., response to external stimuli, changes in feeding activity and exploration of a new environment) were assessed. Although both COC and BE exposure did not induce an oxidative stress situation, a significant inhibition of AChE activity was noted, resulting in behavioral changes in crayfish exposed to COC only. Crayfish exposed to the higher COC concentration showed an increase in the boldness and a decrease in the feeding activity, suggesting that COC may act according to its psychotropic mode of action.

Fumigant toxicity and physiological effects of spearmint (Mentha spicata, Lamiaceae) essential oil and its major constituents against Reticulitermes dabieshanensis

This study evaluated the chemical components of spearmint essential oil, and determined individually the efficacy of spearmint EO and its major constituents, and their mutual binary combination against Reticulitermes dabieshanensis. We also evaluated the activities of esterases (ESTs), glutathione S-transferases (GST) and Acetylcholinesterase (AChE) enzymes in treated insects. GC–MS analysis showed that the major constituents of spearmint EO were carvone (52.25 %), limonene (19.78 %), and dihydrocarvone (11.1 %). In fumigant toxicity assay, the spearmint EO achieved a LC50 value of 0.194 μl/L. The three major constituents, carvone, dihydrocarvone, and limonene were most effective against R. dabieshanensis, with LC50 values of 0.074, 0.155, and 2.650 μl/L, respectively. The toxicity assay of binary mixtures of carvone + dihydrocarvone, carvone + limonene and limonene + dihydrocarvone in all the used ratios showed the three major constituents exhibited synergistic effects against R. dabieshanensis. Spearmint EO and its major constituents showed significantly stronger insecticidal efficacies at the high temperature, with rapid insecticidal action. The increased activity of ESTs and GST were observed, but with the decreased activity of AChE in all treatments. In vitro experiments, all treatments showed significant inhibition of AChE activity, except for dihydrocarvone, with IC50 values were 0.871, 2.405, 2.653 and 4.343 μl/mL for limonene, carvone, carvone + limonene and carvone + dihydrocarvone, respectively. The results showed that the insecticidal efficacy of spearmint EO can be attributed to the major component, possibly carvone, with strong AChE inhibition properties. Hence, spearmint EO and its bioactive constituents have the potential to be used as new environmentally safe insecticides for controlling R. dabieshanensis.

Microplastics alone or co-exposed with copper induce neurotoxicity and behavioral alterations on zebrafish larvae after a subchronic exposure.

Microplastics (MPs, <5mm) have been frequently detected in aquatic ecosystems, representing both health and ecological concerns. However data about the combined effects of MPs and other contaminants is still limited. This study aimed to evaluate the impact of MPs and the heavy metal copper (Cu) on zebrafish (Danio rerio) larvae development and behavior. Zebrafish embryos were subchronically exposed to MPs (2mg/L), two sub-lethal concentrations of Cu (60 and 125µg/L) and binary mixtures of MPs and Cu using the same concentrations, from 2-h post fertilization until 14 days post fertilization. Lethal and sub-lethal responses (mortality, hatching, body length) were evaluated during the embryogenesis period, and locomotor, avoidance, anxiety and shoaling behaviors, and acetylcholinesterase (AChE) activity were measured at 14 dpf. The results showed that survival of larvae was reduced in groups exposed to MPs, Cu and Cu+MPs. Regarding the behavioral patterns, the higher Cu concentration and mixtures decreased significantly the mean speed, the total distance traveled and the absolute turn angle, demonstrating an adverse effect on swimming competence of zebrafish larvae. Exposure to MPs and Cu, alone or combined, also affected avoidance behavior of zebrafish, with larvae not reacting to the aversive stimulus. There was a significant inhibition of AChE activity in larvae exposed to all experimental groups, compared to the control group. Moreover, a higher inhibition of AChE was noticed in larvae exposed to MPs and both Cu+MPs groups, comparatively to the Cu alone groups. Our findings demonstrate the adverse effects of MPs, alone or co-exposed with Cu, on fish early life stages behavior. This study highlights that MPs and heavy metals may have significant impacts on fish population fitness by disrupting locomotor and avoidance behaviors.

Effects of nickel oxide nanoparticles on survival, reproduction, and oxidative stress biomarkers in the marine calanoid copepod Centropages ponticus under short-term exposure.

Excessive use of nickel oxide nanoparticles (NiO NPs) in various industrial and commercial products can lead to various negative effects in human and environmental health due to their possible discharge into the environment. Nerveless, information about their ecotoxicological effects on marine organisms are lacking. Copepods are good ecotoxicological models because of their high sensitivity to environmental stress and their key role in the marine food webs. In this study, 48h acute tests were conducted on the marine planktonic copepod Centropages ponticus to assess lethal and sublethal toxicities of NiO NPs. The results revealed LC50 (48h) of 4mg/L for adult females. Aggregation and settling of NiO NPs were observed at concentrations ≥ 2mg/L. Exposure to sublethal concentrations (≥ 0.02mg/L for 48h) had significant negative effects on reproductive success in C. ponticus. Egg production after 24h and 48h decreased by 32% and 46%, respectively at 0.02mg/L and 70% and 82%, respectively, at 2mg/L. Hatching success was reduced by 70% and 79% at 2mg/L for eggs produced after 24h and 48h respectively. Antioxidant enzymatic activity increased significantly with NiO NP concentration and time, indicating that NiO NPs can cause oxidative stress in C. ponticus even under short-term exposure, while significant inhibition of acetylcholinesterase activity at 2mg/L after 48h suggests neurotoxic effects of NiO NPs.

Ginkgo biloba ameliorates fluoride toxicity in rats by altering histopathology, serum enzymes of heme metabolism and oxidative stress without affecting brain mGluR5 gene

Background: To evaluate the therapeutic potential of Ginkgo biloba extract (GBE) in experimental model of fluorosis. Objectives: To study the protective effect of GBE in fluoride toxicity by assessment of oxidative stress, serum biochemical parameters, acetylcholinesterase (AChE) activity, histopathology and brain mGluR5 gene expression. Materials and Methods: Fifteen adult male Wistar rats were randomly assigned to 5 groups (n = 3 rats in each group). Group 1 (control) received water, Groups 2–5 were treated with 100 ppm of sodium fluoride for 30 days, while the Groups 3, 4 and 5 were GBE treated with 50 mg/kg, 100 mg/kg and 200 mg/kg body weight for 15 days, after sodium fluoride treatment for 30 days. Results: Elevated serum delta aminolevulinic acid dehydratase and delta aminolevulinic acid synthatase levels in fluoride intoxicated rats were ameliorated by various doses of GBE treatment. Elevated serum glutathione and decreased oxidized gluatathione levels observed in fluoride intoxicated rats were also ameliorated by GBE treatment but effectively at 100 mg/kg dose. Reduced AChE activity of hippocampus in fluoride-induced toxicity was reverted by 50 mg/kg of GBE whereas other doses (100 and 200 mg/kg) caused significant inhibition of AChE activity in comparison with fluoride group. Fluoride group rats showed significant reduction of mGluR5 gene expression levels whereas in all GBE treatment groups those changes Were not significantly reverted. GBE treatment to fluoride intoxicated rats almost reverted the degenerative changes in liver and kidney caused by fluorosis. Conclusion: The present study concluded beneficial effects of GBE in experimental model of fluorosis.

Learning and memory impairments associated to acetylcholinesterase inhibition and oxidative stress following glyphosate based-herbicide exposure in mice

Numerous clinical and epidemiological data have reported the deleterious effects of glyphosate on learning and memory. The ability of this herbicide to cross the blood-brain barrier may have adverse effects on the structure and various functions of the nervous system. This study was conducted to highlight the effects of Glyphosate-based herbicide (GBH) on these two functions in mice treated daily with 250 or 500 mg/kg following acute (unique administration), subchronic (6 weeks) and chronic (12 weeks) treatments. The integrity of learning and memory was assessed by using a specific behavioral test battery: Novel object recognition, Y-maze and passive avoidance tasks. The acetylcholinesterase (AChE) and anti-oxidant enzyme activities, especially superoxide dismutase (SOD) and peroxidase (PO) were evaluated. Our results indicated that unlike acute treatment, both subchronic and chronic exposure to GBH decreased discrimination index and the step-through-latency indicating recognition and retention memory impairments, respectively. In contrast, only chronic exposure affected working memory manifested by decreased spontaneous alternation. Furthermore, our results showed also a prominent decrease in AChE, SOD and PO specific activities within the brain of treated mice following repeated exposures.This study demonstrates that GBH induced numerous cognitive abnormalities referred to different forms of memory likely associated with a significant inhibition of AChE activity and oxidative stress induction.

Phytochemical analysis and evaluation of antioxidant and anti-acetylcholinesterase activities of Euphorbia dendroides L. (Euphorbiaceae) latex

The aim of this study was to evaluate the antioxidant and anti-acetylcholinesterase properties and phytochemical constituents of the latex from Euphorbia dendroides L. (Euphorbiaceae) growing wild in Sicily. Phytochemical analysis revealed that into E. dendroides latex the triterpenoids were the most abundant among the identified compounds. Furthermore, a high content of polyphenols mainly as phenolic acids, was found. The antioxidant and free-radical scavenging properties, by several in vitro assays such as DPPH, TEAC and FRAP, have been evaluated. The results showed that E. dendroides latex has significant antioxidant activity, as measured by DPPH assay (2927.01 ± 98.03 µmols of Trolox equivalent (TE)/100g FW). Reactivity towards ABTS radical cation and ferric-reducing antioxidant power (FRAP) values were 7580.95 ± 97.65 µmols of TE/100g FW and 4383.13 ± 95.30 μmol of TE/100g FW, respectively. The latex exhibited also significant inhibition of acetylcholinesterase activity with an IC50 value of 4.46 µg/mL (C.L. = 2.002–9.947). Furthermore, Brine shrimp (Artemia salina) cytotoxicity bioassay showed that the larvae viability was significantly affected at higher concentrations than those capable to induce significant antioxidant and anti-acetylcholinesterase effects (LD50 25 µg/mL). The results suggest that polyphenols and terpenoids can contribute significantly to antioxidant and anti-acetylcholinesterase activities of E. dendroides latex.

Constant exposure to environmental concentrations of the antifouling biocide Sea-Nine retards growth and reduces acetylcholinesterase activity in a marine mysid

Sea-Nine (4,5-dichloro-2-n-octyl-4-isothiazoline3-one; DCOIT) antifoulant has been widely used owing to its broad spectrum of biocide activity against major fouling organisms. In this study, several physiological parameters of a marine mysid were analyzed upon exposure to sublethal environmental concentrations (1 and 100 ng L−1) of Sea-Nine in two exposure conditions, intermittent (weekly; once per week) and constant (daily; once per 24 h) exposure, for 4 weeks. In both experimental conditions, growth retardation, acetylcholinesterase (AChE) activity, glutathione S-transferase (GST) activity, and number of newborn juveniles as second generation, together with their survival were measured. Morphometric parameters of total body, antennal scale, exopod, endopod, and telson were significantly retarded by 22%, 14%, 13%, and 24%, respectively, by daily exposure to 100 ng L−1 Sea-Nine for 4 weeks. Significant inhibition of AChE activity was observed at week 4 in the 100 ng L−1 daily Sea-Nine-exposed groups, whereas no significant GST activity was measured at the same experimental conditions. Inhibition of AChE activity would be associated with impairment of cholinergic system and may adversely modulate growth parameters of the mysid. The number of newly hatched juveniles from females that were exposed daily to 100 ng L−1 Sea-Nine was significantly lower than that of the control. Although no significant differences were observed between survival percentages of newborn juveniles for 30 days, mortality (NOEC and LC50) increased in the surviving offspring from the 100 ng L−1-exposed 1st generation of mysids. These findings suggested that constant exposure to Sea-Nine has detrimental effects on the growth parameters of marine mysids with inhibition of AChE activity.

Enhanced cholinergic-tone during the stress induce a depressive-like state in mice

Major depressive disorder has a heterogeneous etiology, since it arises from the interaction of multiple factors and different pathophysiological mechanisms are involved in the symptomatology. This study aimed to investigate the role of the cholinergic system in the susceptibility to stress and, consequently, in the depression-like behavior. C57BL/6 mice were treated with Physostigmine (PHYS), an acetylcholinesterase (AChE) inhibitor, and were submitted to the social defeat stress. For the behavioral evaluation of the locomotor activity, anxiety-like and depression-like behaviors the open field, elevated plus maze, sucrose preference, social interaction and forced swim were used. Hippocampus and prefrontal cortex samples were collected for evaluation of AChE activity, as well as blood samples for analysis of serum cortisol levels. Our results showed that 15 min after the injection of PHYS there was a significant inhibition of AChE activity in the hippocampus and in the prefrontal cortex. On the other hand, in the end of the experimental design, day 12, there was no difference in AChE activity levels. Inhibition of AChE and exposure to the stress led to an increase in cortisol levels. Animals that received PHYS and were exposed to stress showed less social interaction and greater learned helplessness, anhedonia and anxious-like behavior. Taken together, our findings suggest that increasing the cholinergic tone shortly before stress induction impacts on the ability to cope with upcoming stressful situations, leading to a depressive-like state.

Zearalenone induced embryo and neurotoxicity in zebrafish model (Danio rerio): Role of oxidative stress revealed by a multi biomarker study

In the present study, we evaluated the zearalenone induced adverse effects in zebrafish embryos using various endpoints like embryo toxicity, heart rate, oxidative stress indicators (reactive oxygen species (ROS), lipid peroxidation (LPO), Nitric oxide (NO)), antioxidant responses (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase enzyme (GST) and reduced glutathione (GSH), metabolic biomarkers (lactate dehydrogenase (LDH) and Nitric oxide (NO)), neurotoxicity (acetylcholinesterase (AChE)), genotoxicity (comet assay and acridine orange staining (AO)) and histological analysis. In this study, four concentrations 350, 550, 750 and 950 μg/L of ZEA were chosen based on LC10 and LC50 values of the previous report. The results shows that ZEA induces developmental defects like pericardial edema, hyperemia, yolk sac edema, spine curvature and reduction in heart rate from above 550 μg/L exposure and the severity was increased with concentration and time dependent manner. Significant induction in oxidative stress indices (ROS, LPO and NO), reduction in antioxidant defence system (SOD, CAT, GPx, GST and GSH) and changes in metabolic biomarkers (LDH and AP) were observed at higher ZEA exposed concentration. Neurotoxic effects of ZEA were observed with significant inhibition of AChE activity at higher exposure groups (750 and 950 μg/L). Moreover, we also noticed DNA damage, apoptosis and histological changes in the higher ZEA treatments at 96 h post fertilization (hpf) embryos. Hence, in the present study we concluded that oxidative stress is the main culprit in ZEA induced developmental, genotoxicity and neurotoxicity in zebrafish embryos.

Aluminium-induced acute neurotoxicity in rats: Treatment with aqueous extract of Arthrophytum (Hammada scoparia)

ObjectiveTo study the antioxidative and protective properties of aqueous extract of Hammada scoparia (H. scoparia) against the effects of sub-chronic aluminium (Al) intoxication on mnemonic process and some neurochemical markers. MethodsAl was administered intraperitoneally (50 mg/kg body weight, three times a week), and H. scoparia and malic acid were given orally by gavage at a daily dose (100 mg/kg body weight) to rats for 90 days. ResultsAl caused significant short-term and long-term memory disturbances, a decrease in locomotor activity, a significant inhibition of acetylcholinesterase activity in brain and a significant depletion of antioxidant enzymes (catalase, glutathione reductase and glutathione peroxidase) and glutathione. It significantly increased lipid peroxidation levels in cerebrum and cerebellum. However, treatment with H. scoparia extract protected efficiently the neurological functions of intoxicated rats by considerably increasing antioxidants levels and decreasing production of thiobarbituric acid reactive substances by 4.26% compared to untreated group. We noted some controversial results with malic acid. It showed some positive results but it was not as efficient as H. scoparia extract. Current results were consistent with histopathological observations including neurodegeneration and vacuolated cytoplasm (spongiosis) in Al treated sections when H. scoparia and malic acid treated sections showed marked neuroprotection signs. ConclusionsThis study strongly suggested that H. scoparia extract could possibly restore the altered neurological capacities and antioxidant power in rats, and it could even be a good alternative to chelating agents or other chemical medicines against Al-induced neurotoxicity.

Chemically Contaminated Eel Fed to Pregnant and Lactating Mouse Dams Causes Hyperactivity in Their Offspring.

This study examined whether perinatal exposure to polluted eels (Anguilla anguilla L.) induces changes in the locomotor activity of offspring mice across lifespan (post-natal days (PNDs) 47 - 329), using the open field and the home cage activity tests. Dams were exposed during gestation and lactation, through diets enriched in eels naturally contaminated with pollutants including PCBs. Analysis of the eel muscle focused on the six non-dioxin-like (NDL) indicator PCBs (Σ6 NDL-PCBs: 28, 52, 101, 138, 153 and 180). Four groups of dams (n = 10 per group) received either a standard diet without eels or eels (0.8 mg/kg/day) containing 85, 216, or 400 ng/kg/day of ϵ6 NDL-PCBs. The open field test showed that early-life exposure to polluted eels increased locomotion in female offspring of exposed dams but not in males, compared to controls. This hyperlocomotion appeared later in life, at PNDs 195 and 329 (up to 32 % increase, p < 0.05). In addition, overactivity was observed in the home cage test at PND 305: exposed offspring females showed a faster overall locomotion speed (3.6 - 4.2 cm/s) than controls (2.9 cm/s, p <0.05); again, males remained unaffected. Covered distances in the home cage test were only elevated significantly in offspring females exposed to highest PCB concentrations (3411 ± 590 cm vs. 1377 ± 114 cm, p < 0.001). These results suggest that early-life exposure to polluted eels containing dietary contaminants including PCBs caused late, persistent and gender-dependent neurobehavioral hyperactive effects in offspring mice. Furthermore, female hyperactivity was associated with a significant inhibition of acetylcholinesterase activity in the hippocampus and the prefrontal cortex.

Methyl jasmonate enhances memory performance through inhibition of oxidative stress and acetylcholinesterase activity in mice.

Current research effort focuses on the development of safer natural compounds with multipronged mechanisms of action that could be used to ameliorate memory deficits in patients with Alzheimer's disease, as cure for the disease still remains elusive. In this study, we evaluated the effect of methyl jasmonate (MJ), a naturally occurring bioactive compound on memory, acetylcholinesterase activity and biomarkers of oxidative stress in mice. Male Swiss mice were treated with intraperitoneal injection of MJ (10-40 mg/kg) alone or in combination with scopolamine (3mg/kg) once daily for 7 days. Thirty minutes after the last treatment, memory functions were assessed using Y-maze and object recognition tests. Thereafter, acetylcholinesterase activity and levels of biomarkers of oxidative stress were assessed in mice brains using standard biochemical procedures. MJ significantly enhanced memory performance and reversed scopolamine-induced cognitive impairment in mice. MJ demonstrated significant inhibition of acetylcholinesterase activity suggesting increased cholinergic neurotransmission. It further decreased malondialdehyde concentrations in mouse brain indicating antioxidant activity. Moreover, MJ significantly increased glutathione levels and activity of antioxidant enzymes (catalase and superoxide dismutase) in mice brains. The increased oxidative stress; evidenced by elevated levels of malondialdehyde and decreased antioxidant defense systems in scopolamine-treated mice was attenuated by MJ. The results of this study suggest that MJ may be useful in conditions associated with memory dysfunctions or age-related cognitive decline. The positive effect of MJ on memory may be related to inhibition of oxidative stress and enhancement of cholinergic neurotransmission through inhibition of acetylcholinesterase activity.

Cylindrospermopsin induces neurotoxicity in tilapia fish (Oreochromis niloticus) exposed to Aphanizomenon ovalisporum

Cylindrospermopsin (CYN) is a cytotoxic cyanotoxin produced by several species of freshwater cyanobacteria, such as Aphanizomenon ovalisporum. CYN is a tricyclic alkaloid known for its ability to inhibit both protein and glutathione synthesis, and the alteration of different oxidative stress biomarkers in mammals and vertebrates. Although the liver and kidney appear to be the main CYN targets for this toxin, it also affects other organs. In fish, there is no evidence about the neurotoxicity of CYN yet. In the present study, we aimed to study the potential neurotoxicity of CYN, based on the measure of Acetylcholinesterase (AChE) activity, lipid peroxidation (LPO) levels and histopathological studies in brain of tilapia (Oreochromis niloticus) subchronically exposed to repeated concentrations of 10μg CYN/L by immersion in an A.ovalisporum culture for 14 days. The results showed significant inhibition of AChE activity and increases in LPO levels, as well as relevant histopathological alterations in the brain of fish (O. niloticus) subchronically exposed to the toxin. Moreover, we also investigated the potential recovery of these parameters by subjecting the fish to two depuration periods (3 and 7 days) in clean uncontaminated water, showing a recovery of the biochemical parameters since 3 days of depuration, and being necessary 7 days to recover the histopathological changes. In order to support these results, CYN was detected and quantified by enzyme-linked immunosorbent assay (ELISA) in brain of all the exposed fish and the effects of the depuration periods were also observed. Based on these results, it was demonstrated for the first time the neurotoxicity of CYN and its presence in brain of tilapia fish subchronically exposed to CYN.

Measure of Oxidative Stress and Neurotoxicity Biomarkers in Donax trunculus from the Gulf of Annaba (Algeria): Case of the Year 2012

Aims: The purpose of our study was to evaluate three biomarkers in an edible species, Donax trunculus(Mollusca, Bivalvia) associated with the environmental pollution in the Annaba gulf. The biomarkers selecte d were the antioxidant enzyme catalase (CAT), the detoxifying enzyme glutathione S -transferase (GST) and the neurotoxicity marker acetylcholinesterase (AChE). Place and DurationofStudy:Samples were collected monthly in 2012 at two sites in the Annaba gulf (Northeast Algeria): El Battah a relatively clean site and Sidi Salem, a polluted site. Methodology: Mollusc bivalves ( D. trunculus) with the same shell length (25±1mm) were collected monthly from the two sampling sites, transferred to the laboratory and dissected the same day. The mantle was dissected and samples were prepared for biomarker analyses.The enzymes activitieswere determined by using standard methods. Results: Biochemical data showed a significant inhibition of AChE activity and a significant increase in both CAT and GST activities in D. trunculusat the site of Sidi Salem compared to El Battah. These changes are discussed in relation to environmental factors and suggested an influence of site quality on the health of D. trunculus. Conclusion: The measurements of biomarkers inD. trunculusreveals differences between the two sites in relation to their pollution state confirming previous reports. The OriginalResearch Article

Investigation of landfill leachate toxic potency: An integrated approach with the use of stress indices in tissues of mussels

The present study investigates the harmful impacts of landfill leachate release and/or disposal into the marine environment, as well as its ability to induce lethal and pre-pathological alterations in marine organisms, such as the mussel Mytilus galloprovincialis. In specific, mortality test (96h), performed first in order to estimate leachate lethal endpoints, showed increased levels of mussel mortality after exposure to leachate higher than 0.5%, v/v (96h LC50=0.526%, v/v), while the exposure to 0.01 and 0.1% (v/v) of leachate showed negligible levels of mortality (96h LC10=0.167%, v/v). Furthermore, the estimation of lysosomal membrane integrity in hemocytes of exposed mussels (Neutral Red Retention Time assay) showed increased levels of lysosomal destabilization in cells of mussels exposed to sub-lethal concentrations of leachate (0.01, 0.1 and 0.5%, v/v) for 4 days. In order to exclude parameters, such as mussel mortality and cell death, which could interfere with the obtained results, leachate at final concentrations of 0.01 and 0.1% (v/v) were finally used for the estimation of a battery of stress indices in target tissues of mussels, such as hemolymph, gills and digestive gland. According to the results, leachate-exposed mussels showed a significant inhibition of acetylcholinesterase activity, increased levels of nuclear abnormalities, as well as increased levels of metallothionein, superoxide anion and lipid peroxides (in terms of malondialdehyde equivalents) in each tissue tested. The results of the present study clearly indicate leachate-induced lethal effects as well as the ability of leachate to induce disturbances on different levels of organism function before mortality occurs.

The protective effect of nimodipine, a calcium antagonist, and its influence on soman clearance in the anaesthetized rabbit.

The effect of pretreatment with a vasoactive compound, nimodipine, on soman intoxication in peripheral organs of rabbits was studied by measuring changes in the cholinesterase and acetylcholinesterase activity and by measuring clearance of soman in blood using gas chromatography/high resolution mass spectrometry. In animals receiving soman only, initial blood concentrations were approximately 100 ng mL-1 and were still detectable after 5 min. The clearance rate of soman in blood markedly increased following nimodipine pretreatment such that soman was below the detection limit (0.002-0.003 ng mL-1) in all samples. Soman injection caused a significant inhibition of the acetylcholinesterase activity in serum, and in brain. In rabbits pretreated with nimodipine, no significant inhibition of acetylcholinesterase activity occurred after soman injection. In view of the effects of nimodipine on soman clearance and on the acetylcholinesterase and cholinesterase inhibition during soman intoxication, we suggest that nimodipine has profound circulatory effects, which during soman intoxication, increase the vascular perfusion through the body and thereby increase the detoxifying capacity.

Biomonitoring of environmental pollution on the Algerian west coast using caged mussels Mytilus galloprovincialis:

Abstract An active biomonitoring study was carried out on the Algerian west coast usingwild reference mussels ( Mytilus galloprovincialis ) sampled from the Kristel (K)site and transplanted in net cages during one month (between May and June2007) to Oran Harbour (OH) and Mostaganem Harbour (MH), areas characterisedby high levels of urban and industrial pollution. The biological response ofthe mussels was evaluated by their condition index and the use of a generalstress biomarker (evaluation of lysosomal membrane stability: the neutral redretention time (NRRT) method), a genotoxic effects biomarker (determination ofmicronuclei (MN) frequency) and a neurotoxic effects biomarker (determination ofthe acetylcholinesterase (AChE) concentration).Compared to the K reference specimens, OH and MH caged mussels presenteda significant decrease of NRRT in lysosomal haemocytes (56.45±26.48 minand 67.25±22.77 min, respectively) (78±16.97 min for K mussels), an MNfrequency respectively 7.3 and 9 times higher in the haemocytes and the gillcells of the OH caged mussels, and 7.2 and 6.4 times higher in the twotissues of the MH caged mussels. Significant inhibition of AChE activity wasnoted in the gills (16.93±3.1 nmol min

Atrazine‐induced alterations in rat erythrocyte membranes: Ameliorating effect of vitamin E

Erythrocytes are a convenient model to understand oxidative damage to the membranes induced by various xenobiotics. The objective of the present study was to investigate the propensity of atrazine to induce oxidative stress and its possible attenuation by vitamin E. Experimental animals were orally administered atrazine (300 mg kg(-1) body weight, daily) and vitamin E (100 mg kg(-1) body weight, daily) for a period of 7, 14, and 21 days. Erythrocyte membranes were prepared and analyzed for acetylcholinesterase (AChE) activity, lipid peroxidation (LPO), and lipid composition. Susceptibility of erythrocytes to atrazine exposure was further investigated in terms of morphological alterations by scanning electron microscopy (SEM). Results indicate that atrazine exposure caused a significant inhibition of AChE activity and induction of oxidative stress in terms of increased malondialdehyde (MDA) levels. Atrazine treatment significantly decreased total lipid, cholesterol, and phospholipid content of erythrocyte membranes. SEM revealed varying degrees of distortion depending on duration of atrazine exposure. However, administration of vitamin E ameliorated the oxidative stress and changes in the erythrocyte membranes induced by atrazine.