Rats were randomized to one of four experimental groups (n = 6 in each group): (i) no exposure (no MPT was given), (ii) direct exposure (MPT, 150 mg/kg), (iii) sham gel (220 mg of gel; MPT, 150 mg/kg), and (iv) poly-Oxime gel (220 mg of gel; MPT, 150 mg/kg). Polymer concentration in the gel was 2% (w/w). The dorsal coat was clipped using a hair clipper under mild anesthesia (2.5% isoflurane) 24 hours before the dermal pesticide application, taking care not to damage the integrity of the skin. Unless specified otherwise, the total area of 10 cm2 was marked and used for dermal exposure experiments. In gel treatment groups, 220 mg of gel was uniformly applied 1 hour before the pesticide exposure (Fig. 3). Before exposure, 200 μl of blood was collected using retro-orbital puncture and evaluated for various parameters as an internal control. After 96 hours of post-exposure, blood was collected using cardiac puncture under anesthesia. After sacrificing the animals, organs such as brain, heart, lungs, and liver were harvested. Organ tissue was homogenized (Polytron PT-MR 2100, 15,000 rpm) in nine volumes of solution D [1 M NaCl, 1% Triton X-100, 0.01 M tris-HCl, 0.01 M EDTA (pH 7.4)] and incubated on ice for 1 hour, followed by centrifugation at 13,300 rpm for 45 min. The supernatant was used to quantitate active AChE. Whole blood diluted 1:1000 in phosphate buffer was used for AChE quantification. The activity of AChE in blood and organs was quantified by modified Ellman’s assay. In this assay, we used DTNB and ASChI, which is specific for AChE. For the colorimetric assay, according to Ellman’s method, reaction mixtures were made up in 0.1 mM phosphate buffer (pH 7.4) containing 0.5 mM DTNB and ASChI at a final concentration of 20 mM. The reaction was performed at 25°C and monitored at 405 nm.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.