Plasma Acetylcholinesterase Research Articles

Effects of chronic noise stress on spatial memory of rats in relation to neuronal dendritic alteration and free radical-imbalance in hippocampus and medial prefrontal cortex

Spatial memory is coordinated with different brain regions especially hippocampus (HIP) and medial prefrontal cortex (mPFC). Influence of noise stress on working and reference memory error in rats was evaluated by radial eight-arm maze experiment. Changes in the dendritic count were observed in the brain regions such as CA1, CA3 regions of HIP and layers II, III of mPFC. In order to understand the possible mechanism behind noise stress-induced changes, free radical status and acetylcholinesterase (AChE) activity in HIP and mPFC were evaluated. Plasma corticosterone level was also evaluated. Results obtained in this study showed that after noise-stress exposure, 100 dBA/4 h per day for 30 days, working and reference memory error increased significantly ( P < 0.05) when compared to control animals. Neuronal dendritic count in the HIP was reduced in the 2nd and 3rd order dendrites but not in the mPFC. Superoxide dismutase, lipid peroxidation, plasma corticosterone level and AChE activity were significantly increased in the 1 day, 15 days and 30 days stress groups animal significantly. Catalase and glutathione peroxidase activity were increased in the 1 day and 15 days noise-stress groups but decreased in the 30 days noise-stress group and GSH level was decreased in all the stress exposed animals. In conclusion, oxidative stress, increased AChE activity, reduced dendritic count in HIP, mPFC regions and elevated plasma corticosterone level which develops in long-term noise-stress exposed rats, might have caused the impairment of spatial memory.

Changes in liver and plasma acetylcholinesterase in rats with cirrhosis induced by bile duct ligation

Classical studies of cholinesterase activity during liver dysfunction have focused on butyrylcholinesterase (BuChE), whereas acetylcholinesterase (AChE) has not received much attention. In the current study, liver and plasma AChE levels were investigated in rats with cirrhosis induced after 3 weeks of bile duct ligation (BDL). BDL rats showed a pronounced decrease in liver AChE levels (approximately 50%) compared with sham-operated (non-ligated, NL) controls; whereas liver BuChE appeared unaffected. A selective loss of tetrameric (G4) AChE was detected in BDL rats, an effect also observed in rats with carbon tetrachloride-induced cirrhosis. In accordance, SDS-PAGE analysis showed that the major 55-kd immunoreactive AChE band was decreased in BDL as compared with NL. A 65-kd band, attributed in part to inactive AChE, was increased as became the most abundant AChE subunit in BDL liver. The overall decrease in AChE activity in BDL liver was not accompanied by a reduction of AChE transcripts. The loss of G4 was also reflected by changes observed in AChE glycosylation pattern attributable to different liver AChE forms being differentially glycosylated. BDL affects AChE levels in both hepatocytes and Kupffer cells; however, altered AChE expression was mainly reflected in an alteration in hepatocyte AChE pattern. Plasma from BDL rats had approximately 45% lower AChE activity than controls, displaying decreased G4 levels and altered lectin-binding patterns. In conclusion, the liver is an important source of serum AChE; altered AChE levels may be a useful biomarker for liver cirrhosis.

Effects of Piperonyl Butoxide on the Metabolism of DEF S,S,S-Tributyl Phosphorotrithioate) in Fingerling Channel Catfish

The present study was undertaken to investigate the significance of monooxygenases in bioactivation of DEF to a more effective anticholinesterase in fish. Channel catfish were exposed via the water column for 20 h to piperonyl butoxide (PBO) followed by a 4-h exposure to the organophosphate defolient DEF (concurrent with the PBO). Acetylcholinesterase (AChE) and aliesterases (ALiEs) activities were determined at 0 and 12 h after the exposure period. Inhibition of brain, liver, and plasma AChE activity by DEF was antagonized by PBO; muscle AChE was not inhibited by DEF. Piperonyl butoxide did not antagonize the inhibition of liver or plasma ALiEs by DEF. These results suggest that PBO retards the formation of the metabolite(s) of DEF that inhibit AChE, and that DEF is an effective inhibitor of ALiEs without metabolic activation.

Delivery of human acetylcholinesterase by adeno-associated virus to the acetylcholinesterase knockout mouse

The purpose of this work was to develop a gene delivery system that expressed acetylcholinesterase (AChE) for prolonged periods. An adeno-associated virus (AAV) expressing human AChE was constructed by cotransfecting three plasmids into HEK 293T cells. The purified vector expressed 0.17 μg AChE per 1 million viral particles in culture medium in 23 h, or 0.8 U/ml. The AAV/hAChE was injected into muscle of adult AChE knockout mice and into the brains of 3–6 week old AChE knockout mice. Intramuscular injection yielded plasma AChE levels approaching 50% of the AChE activity of wild-type mouse plasma. The highest AChE activity was found on day 3 post-injection. AChE activity declined thereafter to a constant 7% of normal. The decreased level was accompanied by the appearance of anti-human AChE antibodies, suggesting partial clearance of AChE from plasma by antibodies. Intrastriatal injection resulted in AChE expression in the striatum. No antibodies were detected in animals treated intrastriatally. Motor coordination was improved and the lifespan of intrastriatally-treated AChE knockout mice was prolonged. Human AChE was expressed in mouse brain for up to 7 months after intrastriatal injection of an AAV/hAChE construct. Gene-therapy to supply AChE to the striatum improved motor coordination and prolonged the life of mice genetically deficient in AChE, probably by reducing their susceptibility to spontaneous seizures. This supports the hypothesis that their seizures are induced by excess acetylcholine.

Anticholinesterase and Pharmacokinetic Profile of Phenserine in Healthy Elderly Human Subjects

To evaluate the safety, maximum tolerated dose (MTD), pharmacokinetics (PK), and pharmacodynamics (PD) of the acetyl-selective anticholinesterase, phenserine tartrate, in healthy elderly subjects. 32 healthy elderly volunteers received single oral doses of phenserine tartrate (5-20 mg). Physical and vital signs were monitored over the ensuing 24 hours. Analyses were performed on plasma samples to determine PK, and PD were assessed using an erythrocyte acetylcholinesterase (AChE) inhibition assay. No serious adverse events (AEs) occurred; the most common were headache and vomiting. The MTD of phenserine tartrate was 10 mg. The Cmax and AUC(0-24) of phenserine increased with dose, but neither were dose-proportional. Subjects receiving 10 mg of phenserine tartrate had a Cmax of 1.95 ng/mL at 1.5 hours, and the mean peak inhibition (Imax) of AChE was 26% (range: 18-34%) at 1.75 hours (tImax) following dosing. The half-life of AChE inhibition (tI1/2) was 11 hours. Evaluation of PK/PD relationships suggested a linear correlation between plasma phenserine concentration and AChE inhibition in the blood. Phenserine tartrate was safe and well tolerated when administered as a single oral dose of either 5 mg or 10 mg. An increase in the severity and frequency of AEs occurred at the 20 mg dose level.

1- N-Substituted thiocarbamoyl-3-phenyl-5-thienyl-2-pyrazolines: A novel cholinesterase and selective monoamine oxidase B inhibitors for the treatment of Parkinson's and Alzheimer's diseases

Twelve 1- N-substituted thiocarbamoyl-3-phenyl-5-thienyl-2-pyrazoline derivatives were synthesized and their biological interactions with human plasma and erythrocyte acetylcholinesterase (AChE) and butrylcholinesterase (BuChE) enzymes were assessed. Compounds 3i– 3l of newly synthesized N-substituted pyrazolines, which were presented as selective and irreversible MAO-B inhibitors in our previous report, were found to inhibit human erythrocyte and plasma AChE activities selectively and non-competitively suggesting that these compounds may interact with a region close to the peripheral site of the enzyme molecule which could shift the proper positioning of the catalytic center. Compounds 3e– 3h inhibited both AChE and BuChE activities of human erythrocytes, but the inhibitory potencies of these compounds towards BuChE were found to be higher than that of towards AChE. Inhibition was found to be non-competetive and reversible. These data suggested that newly synthesized N-substituted pyrazoline derivatives can be evaluated as both MAO-B and cholinesterase inhibitors which may have promising features in the treatment of Alzheimer's and Parkinson's diseases.

Plasma acetylcholinesterase as a biomarker of triazophos neurotoxicity in young and adult rats

The organophosphate pesticides exhibit their action by inhibiting acetylcholinesterase (AChE) enzyme in central and peripheral nervous system. They are known to affect the young animals to a greater extent, as their developing brain is more susceptible to their toxic effects. Besides inactivating acetylcholine at synaptic terminals AChE also plays an important role in neuronal growth and differentiation. A reduction in AChE activity in plasma has no known physiological function in causing brain or tissue damage, but if a good correlation between brain and plasma AChE inhibition exists, then circulating plasma AChE can be used as a reliable marker for detection of cholinesterase inhibitors. Therefore, the present investigation was designed to differentiate age and gender related neurotoxicity of an organophosphate pesticide-triazophos and to explore whether plasma AChE can serve as a biomarker of its neurotoxicity in young, i.e. post natal days 20 (PND 20) and adult rats i.e. post natal days 90 (PND 90) after single intraperitoneal administration in different doses.

Inherited and acquired interactions between ACHE and PON1 polymorphisms modulate plasma acetylcholinesterase and paraoxonase activities

The 5.5 Mb chromosome 7q21-22 ACHE/PON1 locus harbours the ACHE gene encoding the acetylcholine hydrolyzing, organophosphate (OP)-inhibitable acetylcholinesterase protein and the paraoxonase gene PON1, yielding the OP-hydrolyzing PON1 enzyme which also displays arylesterase activity. In search of inherited and acquired ACHE-PON1 interactions we genotyped seven polymorphic sites and determined the hydrolytic activities of the corresponding plasma enzymes and of the AChE-homologous butyrylcholinesetrase (BChE) in 157 healthy Israelis. AChE, arylesterase, BChE and paraoxonase activities in plasma displayed 5.4-, 6.5-, 7.2- and 15.5-fold variability, respectively, with genotype-specific differences between carriers of distinct compound polymorphisms. AChE, BChE and arylesterase but not paraoxonase activity increased with age, depending on leucine at PON1 position 55. In contrast, carriers of PON1 M55 displayed decreased arylesterase activity independent of the - 108 promoter polymorphism. Predicted structural consequences of the PON1 L55M substitution demonstrated spatial shifts in adjacent residues. Molecular modelling showed substrate interactions with the enzyme variants, explaining the changes in substrate specificity induced by the Q192R substitution. Intriguingly, PON1, but not BChE or arylesterase, activities displayed inverse association with AChE activity. Our findings demonstrate that polymorphism(s) in the adjacent PON1 and ACHE genes affect each other's expression, predicting for carriers of biochemically debilitating ACHE/PON1 polymorphisms adverse genome-environment interactions.

PLASMA B-ESTERASE ACTIVITIES IN EUROPEAN RAPTORS

B-esterases are serine hydrolases composed of cholinesterases, including acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), and carboxylesterase (CbE). These esterases, found in blood plasma, are inhibited by organophosphorus (OP) and carbamate (CB) insecticides and can be used as nondestructive biomarkers of exposure to anticholinesterase insecticides. Furthermore, B-esterases are involved in detoxification of these insecticides. In order to establish the level of these enzymes and to have reference values for their normal activities, total plasma cholinesterase (ChE), AChE and BChE activities, and plasma CbE activity were determined in 729 European raptors representing 20 species, four families, and two orders. The diurnal families of the Falconiforme order were represented by Accipitridae and Falconidae and the nocturnal families of the Strigiforme order by Tytonidae and Strigidae. Intraspecies differences in cholinesterase activities according to sex and/or age were investigated in buzzards (Buteo buteo), sparrowhawks (Accipiter nisus), kestrels (Falco tinnunculus), barn owls (Tyto alba), and tawny owls (Strix aluco). Sex-related differences affecting ChE and AChE activities were observed in young kestrels (2-3-mo-old) and age-related differences in kestrels (ChE and AChE), sparrowhawks (AChE), and tawny owls (ChE, AChE, and BChE). The interspecies analysis yielded a negative correlation between ChE activity and body mass taking into account the relative contribution of AChE and BChE to ChE activity, with the exception of the honey buzzard (Pernis apivorus). The lowest ChE activities were found in the two largest species, Bonelli's eagle (Hieraaetus fasciatus) and Egyptian vulture (Neophron percnopterus) belonging to the Accipitridae family. The highest ChE activities were found in the relatively small species belonging to the Tytonidae and Strigidae families and in honey buzzard of the Accipitridae family. Species of the Accipitridae, Tytonidae, and Strigidae families were characterized by a BChE contribution that dominated the total ChE activity, while in the species of the Falconidae family, AChE activity dominated. With the exception of the barn owl, CbE activity (eserine-insensitive alpha-naphthyl acetate esterase [alpha-NAE] activity) in all species was almost absent or very low. The values obtained in this study for ChE, AChE, and BChE activities and the AChE:BChE ratios for buzzard, kestrel, barn owl, and tawny owl provide a good estimate of the normal values in free-living individuals of these European species. They can be used as a baseline to evaluate the effect of anticholinesterase insecticides in the field.

Repeated dose 28-day oral toxicity study in Wistar rats with a mixture of five pesticides often found as residues in food: alphacypermethrin, bromopropylate, carbendazim, chlorpyrifos and mancozeb

Six dose groups of 8 male and female rats respectively received a daily dose equivalent to 0, 0.15, 0.006, 0.03, 0.15 or 0.3 mg/kg b.w./day chlorpyrifos (groups 1–6) and the last four dose groups (groups 3–6) received in addition daily doses equivalent to 18 mg/kg b.w./day alphacypermethrin, 30 mg/kg b.w./day bromopropylate, 45 mg/kg b.w./day carbendazim and 12.5 mg/kg b.w./day mancozeb for 28 days. Plasma acetylcholinesterase was significantly decreased in the groups 2, 5 and 6 males. Total white blood cell count was significantly lower in females of group 6. Total red blood cell count, haematocrite and haemoglobin concentration was significantly reduced in both male and female rats of groups 5 and 6. Relative liver weight was significantly increased in groups 3–6 male and female rats. Absolute thyroid gland weight was significantly increased in groups 3, 5 and 6 male rats and of groups 3–6 female rats, and relative thyroid gland weight was significantly increased in groups 2–6 male rats and of groups 3–6 female rats. Absolute thymus weight of groups 3–6 male and female rats and relative thymus weight of groups 3–6 male rats and groups 3 and 4 female rats was significantly decreased. A mild degree of centrilobular cell hypertrophy of the liver was seen in all male rats and of three female rats of group 6. In the thyroid gland follicular cell hypertrophy was present in one female in the control group and in six females and seven males of group 6. It was concluded that inhibition of acetylcholinesterase activity in plasma and brain by chlorpyrifos was not enhanced by co-administration of the other four pesticides. Effects were seen in liver, thyroid, thymus and blood in the combination groups. However, identification of the pesticide(s) responsible for these changes would require further studies of the individually pesticides as well as various combinations of the pesticides.

Metabolical effects of Folidol 600 on the neotropical freshwater fish matrinxã, Brycon cephalus

The neotropical freshwater fish matrinxã, Brycon cephalus (Günther, 1869), was exposed to 1/3 of 96h-LC50 of Folidol 600 (methyl parathion) for 96 h and allowed to recover for 24, 48, 96, and 192 h. Acetylcholinesterase (AChE), alanine aminotransferase (ALAT), aspartate aminotransferase (AAT), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), and glutamate dehydrogenase (GDH) were assayed in brain, liver, muscle, and plasma. Plasma and brain AChE activities were strongly inhibited (64% and 87%, respectively), remaining low in the brain during recovery. Liver ALAT decreased 59.4% and plasma ALAT increased 94.2%. This response, associated with GDH reduction during the recovery period, was attributed to impairment of amino acid metabolism and to liver damage. The increase of heart and plasma AAT suggested tissue injury. Liver and plasma LDH and MDH did not change during the exposure to Folidol, but liver MDH decreased 34% during recovery, indicating a failure of oxidative metabolism in such tissue. Hepatic glycogen and glucose levels decreased 80.4 and 55%, respectively, followed by glucose mobilization to the plasma. The hepatic and muscular profile of lactate after recovery suggested gluconeogenesis without tissue hypoxia. These results revealed that methyl parathion affects the intermediary metabolism of matrinxã and that the assayed enzymes can work as good biomarkers of organophosphorus pesticide contamination.

Acetylcholinesterase activity is affected by stress conditions in Paracentrotus lividus coelomocytes

The presence and function of acetylcholinesterase (AChE) in blood cells and plasma is well known. The enzyme activity is associated to red blood cell membranes, as well as to the perinuclear region of leucocytes, suggesting a role in the function of the latter. Recently coelomocytes, acknowledged progenitors of immune cells in vertebrate systems, have been shown to respond to stress conditions, both in laboratory experiments and field studies. Bearing in mind the similarity between coelomocytes and vertebrate blood cells, we investigated the presence of AChE activity in coelomocytes of the sea urchin Paracentrotus lividus characterized by specific inhibitors, in relation to cold stress induced in the laboratory. Exposure to stress was followed by electrophoretic analysis of AChE activity and Western blot analysis of AChE-like immunoreactivity. Moreover, quantitative analysis of the different molecular forms of AChE was carried out with the aid of a spectrophotometric method. The effects of stress on AChE activity were time dependent. In fact, the enzyme activity was detected by the catalytic activity in cleaving the substrate acetyl thiocholine iodide, which appears to be formed by different isoforms of the enzyme, compared to that of Electrophorus, which was used as a control. Western blot analyses showed the presence of different immunoreactive molecules, with molecular mass ranging from 265 kDa (corresponding to Electrophorus molecules) to about 180 kDa. According to quantitative analysis, the isoforms that were differentiated corresponded to an AChE with the capacity to cleave acetyl-β-methyl thiocholine iodide (the so-called true AChE, E.C. 3.1.1.7, corresponding to the 265 kDa band) and to a ChE with the capacity to cleave propionyl thiocholine iodide as well. The latter is an ancestral form of AChE, mainly present in invertebrates. Each of these forms was affected differently by varying time exposures.

Acetylcholinesterase and butyrylcholinesterase activities in brain and plasma of freshwater teleosts: cross-species and cross-family differences

Brain and plasma acetylcholinesterase (AChE; EC 3.1.1.7) and plasma butyrylcholinesterase (BChE; EC 3.1.1.8) specific activities were assayed in 16 freshwater teleosts belonging to four families: Cyprinidae, Percidae, Esocidae and Lotidae. Brain AChE activity varied among fish species approximately 15-fold, ranging from 138 to 2011 μmol/g per h. All cyprinids had higher brain AChE activity than other fish families. Plasma AChE activity was on average 100-fold lower than that in brain, varying from 1.2 to 18.6 μmol/ml per h. Plasma BChE activity was found only in cyprinids with the exception of the common and crucian carp, and sabrefish. It varied from 26 to 1083 μmol/ml per h. In bream ( Abramis brama) only 30% of specimens studied had BChE activity. The correlation coefficient values between activities of brain and plasma AChE, brain AChE and plasma BChE, plasma AChE and BChE were 0.67, 0.68 and 0.84, respectively. Cross-species and also cross-family differences in AChE and BChE activities among fish were demonstrated. Possible reasons for the differences are discussed.

Neuropsychiatric evaluation in subjects chronically exposed to organophosphate pesticides.

Long-term exposure to low levels of organophosphate pesticides (OP) may produce neuropsychiatric symptoms. We performed clinical, neuropsychiatric, and laboratory evaluations of 37 workers involved in family agriculture of tobacco from southern Brazil who had been exposed to OP for 3 months, and in 25 of these workers, after 3 months without exposure to OP. Plasma acetylcholinesterase activity levels of all subjects were within the normal range (3.2 to 9.0 U/l) and were not different between on- and off-exposure periods (4.7 +/- 0.9 and 4.5 +/- 1.1 U/l, respectively). Clinically significant extrapyramidal symptoms were present in 12 of 25 subjects, which is unexpected in such a population. There was a significant reduction of extrapyramidal symptoms after 3 months without exposure to OP, but 10 subjects still had significant parkinsonism. Mini-mental and word span scores were within the expected range for this population and were not influenced by exposure to OP. Eighteen of the 37 subjects (48%) had current psychiatric diagnoses in the first interview (13 with generalized anxiety disorder and 8 with major depression). Among the 25 subjects who completed both evaluations, the total number of current psychiatric diagnoses, after 3 months without using OP, dropped from 24 to 13 and the number of affected individuals with any psychiatric diagnosis dropped from 11 to 7. In conclusion, this study reinforces the need for parameters other than acetylcholinesterase activity to monitor for chronic consequences of chronic low-dose OP exposure, and it suggests that subjects have not only transient motor and psychiatric consequences while exposed, but may also develop enduring extrapyramidal symptoms.

Acupuncture on gnosia and acetylcholinesterase in senile dementia patients

Purpose To observe the effect of acupuncture on gnosia and acetylcholinesterase in patients with senile dementia.Methods Eight patients diagnosed with mild or moderate senile dementia were treated by acupuncture of Sishencong (Ex-HN 1), Shenmen (HT 7) and Taixi (Kl 3) for 1 month; gnosia was evaluated by Mini-mental state examination before and after the treatment; plasma acetylcholin esterase activity was measured by fluorier before and after the treatment.Results There was a significant difference in gnosia between pre- and post-treatment with acupuncture (P 0.05).Conclusion Acupuncture has a certain improving effect on gnosia in senile dementia; one month’s acupuncture treatment had little effect on plasma acetyl-cholinesterase activity.

GLUCOSE FEEDING EXACERBATES PARATHION-INDUCED NEUROTOXICITY

Excessive dietary intake of sugars could alter various biotransformation processes and the pharmacological and toxicological properties of numerous xenobiotics. In the present study, the effects of glucose supplementation were examined on the neurotoxicity of the organophosphorus (OP) pesticide parathion (PS) and its active metabolite, paraoxon (PO), a potent inhibitor of acetylcholinesterase (AChE). Rats (n = 6-12/treatment group) were given free access to tap water or 15% glucose (w/v) in tap water beginning 7 d prior to OP toxicant exposure. Food, caloric intake, and body weight were measured daily. Animals were challenged with either PS (4.5, 9, or 18 mg/kg, sc) or PO (0.3, 0.5, or 0.7 mg/kg, sc) and clinical signs of neurotoxicity (i.e., autonomic dysfunction, involuntary movements) were recorded daily for the following 13 d. Glucose feeding was associated with a dramatic drop (~50%) in feed intake and an increase (~20%) in total caloric consumption over the 7 d prior to OP exposure. Functional toxicity associated with PS exposure was increased in glucose-fed (GF) rats, but the glucose diet had no apparent effect on clinical signs of toxicity following PO treatment. Glucose feeding increased the magnitude of AChE inhibition in the frontal cortex and plasma at lower dosages (i.e., 4.5 and 9 mg/kg) 3 d following PS treatment. Time-course studies (3, 7, and 11 d after PS exposure, 18 mg/kg, sc) indicated significantly greater brain and plasma AChE inhibition in glucosefed animals at later time points. In contrast, glucose feeding had no effect on the degree of AChE inhibition following PO exposure. Neither liver microsomal oxidative desulfuration of PS, nor liver or plasma paraoxonase, nor liver or plasma carboxylesterase activities were measurably affected by glucose feeding. Downregulation of muscarinic receptors 7 d after PS exposure (18 mg/kg, sc) was more extensive in GF rats. It is postulated that excessive glucose consumption decreases the intake of other dietary components, in particular amino acids, limiting the de novo synthesis of AChE and consequent recovery of synaptic transmission. Due to the shorter duration of inhibition following PO exposure, spontaneous reactivation of AChE may be more important than de novo protein synthesis in recovery of function, and thus with the effects of glucose feeding on its toxicity. Individuals that derive a large proportion of their calories from sugars may be at higher risk of acute toxicity from organophosphorus pesticides such as PS.

Effects of Dissolved Copper on Select Hematological, Biochemical, and Immunological Parameters of Wild Rainbow Trout ( Oncorhynchus mykiss )

Rainbow trout (Oncorhynchus mykiss) were sampled from a creek in the western Sierra Nevada, Plumas County, CA, that receives run-off from a 40-ha copper (Cu) tailings pile. Reference sites included a site upstream of the Cu input and another site located on a nearby creek. Hepatic Cu concentrations were significantly elevated in trout from sites with elevated dissolved Cu concentrations compared with concentrations in trout from reference sites. Trout at the Cu-contaminated sites also exhibited decreased hematocrit (Hct), leukocrit (Lct), and percentage of lymphocytes in blood compared to trout from reference sites. The percentage of monocytes in blood and respiratory burst activity were affected by gender and age, respectively. Condition factor, percentage of neutrophils in blood, muscle glycogen and protein, and plasma acetylcholinesterase were not affected by dissolved Cu concentration or gender. Age also did not appear to be a factor. The data from this study support the use of immune system parameters to assess alterations in salmonids experiencing prolonged exposure to low-level Cu contamination and illustrate the variability in physiological responses of wild fish caused by demographic features. Overall, of the parameters measured, Hct, Lct, and percentage of lymphocytes in blood appeared to offer robust measures for assessing effects of metals on wild fish and did not appear affected by select demographic features.

The effect of community-acquired pneumonia on plasma esterases in older people.

Pneumonia is a major cause of morbidity and mortality in older people. The poor outcome of older pneumonia patients despite treatment is still not understood. The aim of this study was to examine the effect of community-acquired pneumonia on enzymes of drug metabolism in older people. Fifteen patients (median age 67 years) with a clinical and radiological diagnosis of community-acquired pneumonia and 14 healthy volunteers matched for age and gender (median age 75 years) were recruited. Plasma activities of benzoylcholinesterase, butyrylcholinesterase, acetylcholinesterase and aspirin esterase were determined spectrophotometrically at three time points in pneumonia patients--within 24 h of admission to hospital, 2 days later and 10 days later. Monocyte aryl hydrocarbon hydroxylase (AHH) activity was determined spectrofluorimetrically at the same time points. Enzyme activities were measured at one time point in healthy controls. Mean plasma benzoylcholinesterase activity was significantly lower in pneumonia patients on admission to hospital (mean +/- SEM 848 +/- 100) and after 10 days of treatment (mean +/- SEM 925 +/- 114) than in healthy controls (mean +/- SEM 1333 +/- 84, P < 0.05). Similarly, plasma acetylcholinesterase activity was significantly lower in pneumonia patients on admission (P = 0.007) and after 10 days of treatment (P = 0.01) than in controls. Butyrylcholinesterase activity was lower in pneumonia patients on admission (P = 0.029) than in healthy controls, but improved slightly after treatment so there was no longer a significant difference at 10 days compared with controls (P = 0.077). In contrast there were no significant differences in plasma aspirin esterase activity or induced monocyte AHH activity between pneumonia patients and healthy controls. The activities of benzoylcholinesterase (r = -0.536, P = 0.04), butyrylcholinesterase (r = -0.638, P = 0.01), acetylcholinesterase (r = -0.583, P = 0.022) and aspirin esterase (r = -0.624, P = 0.013) correlated inversely with the British Thoracic Society pneumonia poor prognostic index. The activities of several esterases are reduced in older pneumonia patients. Other enzymes including aspirin esterase and induced monocyte AHH activity are unaltered in pneumonia. There was a significant inverse relationship between the activities of all esterases studied and the British Thoracic Society pneumonia poor prognostic index.

An Unusual Case of Organophosphate Intoxication of a Worker in a Plastic Bottle Recycling Plant: An Important Reminder

A young man was sent to our emergency unit because he had suffered from vomiting and cold sweating for 2 days. At the time he was admitted, he had no acute abdominal pains or gastrointestinal symptoms, and a physical examination revealed nothing but a faster heart rate and moist, flushing skin. The patient had worked for 6 years at a plastic bottle-recycling factory, but none of his co-workers had the same symptoms. Nevertheless, because the plant also recycled pesticide bottles, we suspected organophosphate pesticide intoxication. The patient's plasma acetylcholinesterase level was checked, revealing 1498.6 microU/L (normal range: 2,000-5, 000) on the first day and 1,379 microU/L on the second day. Upon questioning, the patient recalled that one of his shoe soles had been damaged and that his foot had been wet from walking all day in rain collected on the factory floor on the day that his symptoms first occurred. We conducted a study in the change of preshift and postshift acetylcholinesterase levels among six of his co-workers on a rainy day. We used the Wilcoxon signed rank test to compare the preshift and postshift plasma acetylcholinesterase levels; no significant difference was revealed (p = 0.600), leaving contamination via the damaged shoe sole suspect. We reviewed the literature on organophosphate intoxication; pesticide bottle-recycling factories were reported to be at a low risk of organophosphate toxicity in the working environment. However, because the potential risk of intoxication is still present, protective equipment such as clothing, gloves, and water-proof shoes should be worn, and employees should be educated on the potential risks.

Association of azinphos-methyl with rat erythrocytes and hemoglobin

To investigate whether hemoglobin might serve as a biomarker of exposure to azinphos-methyl (AZM) encountered by agricultural workers, we exposed rats to [14C]azinphos-methyl ([14C]AZM). We administered single doses of 1.5 mg/kg, 3 mg/kg, or 6 mg/kg of [14C]AZM by gavage to rats and collected blood 3 days later. We found a dose-dependent association between radioactivity and erythrocytes and hemoglobin (measured by liquid scintillation spectrometry). In another experiment, we administered a single dose of 3 mg/kg [14C]AZM by gavage to rats and collected blood 3, 11, 15, and 22 days after administration. Radioactivity continued to be associated with erythrocytes and hemoglobin at all time-points. Brain and plasma acetylcholinesterase (AChE) activities were not significantly inhibited, nor did we observe signs of acute toxicity in any of the treated animals. Loss of radioactivity associated with erythrocytes and hemoglobin over the study period was consistent with the expected kinetics of erythrocyte turnover, indicating adduct stability. Approximately 0.49%, 0.43%, 0.39%, and 0.32% of the original radioactivity was recovered in the hemoglobin 3 11, 15, and 22 days, respectively, after administration. These data support our hypothesis that AZM may form a hemoglobin adduct and may be useful as a biomarker of AZM exposure.