94 resultados para Acetylcholinesterase (AChE)


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Background the Test-mate kit determines acetylcholinesterase (AChE, EC 3.1.1.7) and hemoglobin content of a drop of blood, displaying enzyme activities normalized to 25degreesC. Previous models produced inconsistent results at different temperatures. This report focuses on the current model, ChE 400, and two instruments of a previous OP model.Methods AChE activities were determined by the Ellman assay, using the three kits and a 96-well microplate reader Temperatures ranged from 10 to 37degreesC. Fetal bovine serum was the source of AChE.Results Normalized activities decreased below 20degreesC in the ChE model and below 25 C in the OP models. Activities of the same serum sample differed between the three Test-mate kits, ranging from 1.03 to 1.49 mumoles/min/ml. Percent errors were greater than with the microplate reader at all temperatures.Conclusions Neither we nor the manufacturer recommend the current Test-mate model for fieldwork. Nevertheless, there have been field measurements with Test-Mate kits, and we recommend that an enzyme activity standard be run in parallel with their use. (C) 2002 Wiley-Liss, Inc.

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The aim of this study was to investigate whether handling and acclimatization could affect the biomarker responses in oysters Crassostrea gigas. Adult oysters were sampled in a farming area, subjected to handling stress during two hours (shell cleaning and transport), and then acclimatized in laboratory for 2, 3 and 4 weeks. Groups of five oysters were sampled before and after the handling (T0 and T1, respectively), and after 2, 3 and 4 weeks acclimatization. During the acclimatization, water was renewed daily, food given twice a day and temperature and salinity maintained at 22 °C and 25 ppt, respectively. One group, in another tank, was kept in similar conditions and was exposed for 1 week to 0.1 % diesel after the 2-weeks acclimatization period. After exposure, gills were immediately frozen in liquid N 2 for biochemical analyses. Higher expression of heat-shock proteins (HSP70) was observed after handling, and after acclimatization periods of 3-week and 4-week, compared to the T0 group. The diesel exposed group did not show elevated levels of HSP70, when compared to the 3-week acclimatized group. The activity of glutathione S-transferase (GST) was unchanged after handling, but was lower after all acclimatization periods, compared to the T0 group. Exposure to diesel caused an increase in GST activity compared to the 3-week acclimatized group, but not compared to T0. The activity of catalase (CAT), acetylcholinesterase (AChE), and the MDA levels remained unchanged during the whole experiment. These results point to the need of a special care in laboratory and field experiments employing HSP70 and GST as biomarkers. (Supported by CNPq-CTPetro to ACDB.). © 2008 Published by Elsevier Ltd.

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Pós-graduação em Química - IBILCE

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The aim of this study was to determine the no-observable-adverse-effect concentration (NOAEC) for trichlorfon, an antiparasitic agent used in aquaculture, in Piractus mesopotamicus (pacu) using acetylcholinesterase (AChE) activity as an end point. Fish were exposed 24 h/d for 15 d to different concentrations of trichlorfon in tanks of water for which a curve of dissipation was previously determined. Analysis of trichlorfon in water and fish plasma using gas chromatography with electron capture detection (GC-ECD) enabled measurement of limit of detection (LOD) and limit of quantification (LOQ), respectively, to be 3 and 10 ppb. Thirty-six hours after trichlorfon dilution in water, the concentration was below the LOD, and data showed that plasma concentrations did not exceed the LOQ. Apart from the 6.25 g/L, all concentrations of trichlorfon significantly inhibited plasma and brain AChE activity compared to controls. The AChE activity levels returned to control values in 7 d. These data may be useful to determine the concentration of trichlorfon that destroys parasites without producing adverse effects in fish.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)