Tissue‐specific in vivo inhibition of cholinesterases by the organophosphate fenthion in Oreochromis niloticus

This study was designed to elucidate the effect of the organophosphate fenthion exposure on cholinesterase‐specific activities in brain, liver, and kidney tissues of juvenile Oreochromis niloticus, and to define the best indicator tissue to fenthion exposure. The 96‐h LC₅₀ value was determined as 2.27 mg/L and fish were exposed to 20% of this concentration for 24‐, 48‐, and 96‐h. Recovery periods in similar durations were provided to evaluate the ChE activities. AChE and BChE activities were determined spectrophotometrically. The activities of these enzymes were significantly inhibited in all the tissues tested, liver was the most and kidney was the least affected tissues. The inhibition percentages of AChE and BChE were at similar levels in the liver while BChE was more affected in kidney. BChE was not detected in the brain. A significant positive correlation in ChE inhibitions was found among tissues, and the effect of fenthion on ChE activities was tissue specific. In general enzymes activities were not significantly recovered in 96‐h recovery period; however, an elevation in AChE inhibition was observed in brain. Based on the data of this study, the liver may be suggested as the best indicator tissue especially for phosphorothioate exposure. © 2009 Wiley Periodicals, Inc. Environ Toxicol 25: 391–399, 2010.     

The first evidence of cholinesterases in skin mucus of carps and its applicability as biomarker of organophosphate exposure

The presence of cholinesterase (ChE) activity in skin mucus of three carps, Cirrhinus mrigala, Labeo rohita, and Catla catla and its applicability as biomarker of the organophosphorus insecticide exposure were investigated. Biochemical characterization, using specific substrates and inhibitors, indicated that measured esterase activity in skin mucus was mainly owing to ChEs. Significant difference in the proportion of acetylcholinesterase and butyrylcholinesterase activities was observed in skin mucus of three carps. Enzyme kinetic analysis, using the substrate acetylthiocholine iodide revealed significantly high V_max value in C. catla compared to that in L. rohita and C. mrigala. In contrast, V_max value using the substrate butyrylthiocholine iodide was significantly high in C. mrigala than in L. rohita and C. catla. In vitro treatment of skin mucus of three carps, with the organophosphorus insecticide Nuvan®, showed strong inhibition of ChE activities. In vivo experiments conducted using C. mrigala and exposing the fish to the sublethal test concentrations (5 and 15 mg/L) of the insecticide also revealed significant inhibition of ChE activity in mucus. In C. mrigala, exposed to the sublethal test concentrations of the insecticide for 4 days and then kept for recovery for 16 days, mucus ChE activity recovered to the control level. Thus, ChE activity in skin mucus could be considered a good biomarker of the organophosphorus insecticide exposure to fish and a useful tool in monitoring environmental toxicity. © 2012 Wiley Periodicals, Inc. Environ Toxicol 29: 788–796, 2014.     

The kinetic study of the inhibition of human cholinesterases by demeton-S-methyl shows that cholinesterase-based titration methods are not suitable for this organophosphate

The organophosphorus insecticide, demeton-S-methyl (DSM), is considered as a good surrogate of the highly toxic nerve agent VX for skin absorption studies due to similar physico-chemical properties and in vitro percutaneous penetration profile. But, when skin distribution was estimated by measuring inhibition of cholinesterase activity, the results were poorly reproducible. The various grades of commercial DSM solutions were suspected to be the origin of the discrepancies. This hypothesis was tested by measuring inhibition of human acetyl- and butyrylcholinesterase by two commercial DSM solutions. The inhibition rate was independent on the enzyme concentration confirming pseudo-first order conditions. But complete inhibition of butyrylcholinesterase activity was achieved only when the DSM concentration was at least 1500-fold higher than the enzyme concentration. Besides, complete inhibition of acetylcholinesterase was never achieved. Mass spectrometry analysis of the inhibited butyrylcholinesterase adducts identified monomethoxyphosphorylated-serine, the aged product of inhibition by DSM or a derivative with a modified leaving group. Neither spontaneous reactivation nor aging of the dimethoxyphosphorylated-serine could account for the inhibition kinetics observed, suggesting an overly complicated kinetic scheme not compatible with the requirement of a titration experiment. In conclusion, cholinesterase-based analytical methods should be avoided for DSM titration in skin penetration studies.     

Subchronic intoxication with chlorfenvinphos, an organophosphate insecticide, affects rat brain antioxidative enzymes and glutathione level.

Organophosphate pesticides (OP) belong to the class of xenobiotics that are intentionally released to the environment. Toxicity of these compounds is mainly due to inhibition of acetylcholinesterase (AChE), but many authors postulate that OP in acute as well as in chronic intoxication disturb the redox processes, changing the activities of antioxidative enzymes and causing enhancement of lipid peroxidation in many organs. Epidemiological studies have demonstrated a relationship of certain human diseases with pesticide exposure and with changes in antioxidative enzymes. There is also evidence that oxidative stress is an important pathomechanism of neurological disorders such as Alzheimer disease and Parkinson disease, cardiovascular disorders and many others. The study objective was to investigate the activities of brain antioxidative enzymes and reduced glutathione level in rats subchronically intoxicated with chlorfenvinphos. In the rat brain the activities of such enzymes as superoxide dismutase, catalase, glutathione peroxidase and reductase were found to increase, while reduced glutathione level decreased in chlorfenvinphos intoxication. Based on experimental findings of this study, it can be suggested that subchronic administration of chlorfenvinphos leads to a change in the brain oxidative status and that the change occurs at a dose of 0.3 mg/kg/day, i.e., twice smaller than LOAEL level for rats.     

Role of socialization, stress and sex of chickens on response to anesthesia and on response to an organophosphate neurotoxicant.

The influence of socialization of chickens on response to exogenous substances in the presence and absence of stress was examined. Chickens of both sexes were habituated to human beings (socialized) by being talked to and offered food from the hand of the caretaker. After 19 w, half the socialized group and half the unsocialized group were subjected to stress (loud noises for 120 sec twice daily) for 2 w. At the end of this period, response to 37 mg pentobarbital/kg iv was evaluated by length of sleeping time. Males slept longer than females, and males that were socialized but not stressed slept significantly longer than other males. Stress and socialization did not significantly affect pentobarbital sleeping time in females. This indicated that response to pentobarbital was dependent on sex and on socialization, with the most notable effects occurring in socialized males. Socialized and nonsocialized, stressed and unstressed male and female chickens were also administered a single po dose of 360 mg tri-ortho-tolyl phosphate (TOTP)/kg. This organophosphorus ester induced a delayed neuropathy that caused ataxia in all the chickens. The ataxia was significantly more pronounced earlier in males than in females, although the sex difference became insignificant 18 d after dosing. Socialized chickens were ultimately more affected. Noise stress did not affect TOTP-induced ataxia. This indicated that response to an organophosphate neurotoxicant was also dependent on sex and socialization, with the most notable effects again seen in socialized males. Response to endogenous substances in chickens depends both on sex and on familiarity to humans.     

Thermoregulatory response to an organophosphate and carbamate insecticide mixture: testing the assumption of dose-additivity

Most toxicity data are based on studies using single compounds. This study assessed if there is an interaction between mixtures of the anticholinesterase insecticides chlorpyrifos (CHP) and carbaryl (CAR) using hypothermia and cholinesterase (ChE) inhibition as toxicological endpoints. Core temperature (T(c)) was continuously monitored by radiotelemetry in adult Long-Evans rats administered CHP at doses ranging from 0 to 50mg/kg and CAR doses of 0-150 mg/kg. The temperature index (TI), an integration of the change in T(c) over a 12h period, was quantified. Effects of mixtures of CHP and CAR in 2:1 and 1:1 ratios on the TI were examined and the data analyzed using a statistical model designed to assess significant departures from additivity for chemical mixtures. CHP and CAR elicited a marked hypothermia and dose-related decrease in the TI. The TI response to a 2:1 ratio of CHP:CAR was significantly less than that predicted by additivity. The TI response to a 1:1 ratio of CHP and CAR was not significantly different from the predicted additivity. Plasma and brain ChE activity were measured 4h after dosing with CHP, CAR, and mixtures in separate groups of rats. There was a dose-additive interaction for the inhibition of brain ChE for the 2:1 ratio, but an antagonistic effect for the 1:1 ratio. The 2:1 and 1:1 mixtures had an antagonistic interaction on plasma ChE. Overall, the departures from additivity for the physiological (i.e., temperature) and biochemical (i.e., ChE inhibition) endpoints for the 2:1 and 1:1 mixtures studies did not coincide as expected. An interaction between CHP and CAR appears to depend on the ratio of compounds in the mixture as well as the biological endpoint.     

Inorganic arsenic exposure affects pain behavior and inflammatory response in rat

Inorganic arsenic (iAs) contamination of drinking water is a worldwide problem associated with an increased risk for the development of various types of cancer and noncancerous damage. In vitro studies have suggested that iAs can modulate the activity of macrophages producing an over-expression of cyclooxygenase-2 (COX-2) and resulting in an increase in prostaglandin E(2) (PGE(2)) concentrations in endothelial cells. These effects may lead to an in vivo enhancement of inflammatory and pain responses. Our aim was to determine the effect of a single dose of arsenic or subchronic exposure to arsenic on pain behavior and tissue inflammation in rats. Rats were given a single dose of sodium arsenite (0.1, 1 and 10 mg/kg i.p.) or submitted to subchronic exposure to arsenic added to the drinking water for 4 weeks (0.1, 1, 10 and 100 ppm). Inflammatory pain was assessed by using the formalin and tail-flick tests, while inflammation was evaluated with the carrageenan model. Arsenite did not induce pain or significant inflammation by itself. In contrast, arsenite in both single dose administration and subchronic exposure increased not only the inflammatory process and the underlying hyperalgesic pain, but also induced a decrease in the pain threshold. Alterations in pain processing were dependent on the arsenic dose and the length of exposure, and the underlying mechanism involved an increased release of local PGE(2). These results suggest that inorganic arsenic exposure enhances pain perception and exacerbates the pathological state of inflammatory diseases.     

Ammonia response to constant exercise: differences to the lactate response

1. We evaluated the plasma ammonia response to constant exercise at different intensities. Ten healthy male volunteers were asked to perform constant exercise for 15 min at five different intensities: 80, 90, 100, 110 and 120% of their ventilatory threshold (VT). Blood concentrations of lactate, ammonia and hypoxanthine were measured during and after exercise. 2. The concentration of lactate increased continuously during exercise intensities equivalent to 100, 110 and 120% VT. Plasma ammonia began to increase at 6 min exercise and continued increasing during exercise at all five exercise intensities. Plasma hypoxanthine levels also increased continuously during exercise at all exercise intensities; however, they peaked at 5-10 min after exercise. The response of plasma ammonia and hypoxanthine increased with increasing intensities of exercise. 3. While the extent of the increase in lactate levels during exercise at 100, 110 and 120% VT was significantly higher than that at 80% VT, only the increase in ammonia and hypoxanthine levels at 120% VT were significantly higher than those at 80% VT. 4. In conclusion, the plasma ammonia response to constant exercise differed to the lactate and ammonia responses to short-term exhaustive exercise.     

Carvedilol affects the physiological and behavioral response to smoked cocaine in humans

The noradrenergic system is implicated in mediating some of the physiological effects of cocaine. The purpose of this study was to investigate whether treatment with an adrenergic blocker, carvedilol, which would be expected to attenuate the physiological effects of cocaine, would also attenuate the subjective and behavioral response to cocaine in humans. Twelve crack cocaine users participated in this double-blind, placebo-controlled outpatient study. Acute treatment with 50 mg of oral carvedilol attenuated the smoked cocaine-induced increases in heart rate, systolic and diastolic blood pressure. The number of cocaine self-administrations was lower under 25 mg carvedilol treatment condition compared with 50 mg carvedilol or placebo treatment conditions. The subjective responses to smoked cocaine deliveries were not affected by carvedilol treatment. These results suggest that acute treatment with carvedilol attenuates the physiological effects of smoked cocaine. The effects of carvedilol on cocaine self-administration need to be studied further.     

盐酸吲满氨酯及依色林对胆碱酯酶的抑制作用

用微量二硫-双-硝基苯甲酸(DTNB)法测定胆碱酯酶(ChE)活力,比较了盐酸吲满氨酯(TMDMC)与依色林抑制ChE的作用,对于电鳐AChE依色林表现为竞争性抑制,K_i=0.115 μmol·L~1,TMDMC表现为竞争与非竞争混合性抑制,K_i=3.870μmol·L~(-1),K_i=16.321μmol·L~(-1)依色林和TMDMC抑制小鼠脑匀浆ChE活力的IC_(50)分别为1.78和5.07μmol·L~1、抑制小鼠全血ChE活力的IC_(50)分别为19.52和3.60μmol·L~1,说明TMDMC与依色林虽属同类化合物,但它们的抑酶类型和对真,假性ChE的抑制强度是不同的。     

Faecal blood loss in response to exercise.

Recently qualitative tests have indicated that gastrointestinal bleeding during exercise may be an important contributory factor in sports anaemia. In six healthy men who walked 37 km on four consecutive days faecal haemoglobin content remained normal (reference range 0.10-2.53 mg/g faeces) with no significant differences between values. In 28 marathon runners who refrained from taking drugs or food containing blood the median faecal haemoglobin content increased by 0.42 mg/g faeces (95% confidence interval 0.12 to 0.83 mg/g) from 1.06 (0.86 to 1.31) mg/g before the race. In 13 runners who had taken drugs before the race the corresponding increase in the median faecal haemoglobin content was 0.87 (-0.03 to 2.20) mg/g from the value before the race of 0.93 (0.46 to 1.55) mg/g. Prolonged walking had no effect on gastrointestinal blood loss. Intense endurance exercise in the form of marathon running induced a significant but clinically unimportant increase. This may be exaggerated by the ingestion of drugs and assume importance in causing iron deficiency and sports anaemia. The use of drugs, particularly analgesics, by marathon runners should be actively discouraged.