ABSTRACT
Hygiene hypothesis and sanitization are two important pivots of modern civilization. The drinking water should be free from urine and stool contamination. Coliform test is popular for understanding feces contamination. However, understanding urine contamination in drinking water is a difficult task. On the other hand, urine contamination can cause disease like leptospirosis. It occurs mainly in animals and infects humans through contaminated water, food, and soil and causes serious consequences. Rat urine is the most common source of such disease outbreaks. Further, sophisticated laboratories with high-end technologies may not be present at the site of disease outbreaks. In this context, we have proposed a spectrofluorimetric approach to screen urine contamination in water. The screening method can sense up to 156 nl/ml of rat urine
ABSTRACT
Hemolysate cholinesterase is currently recognized as the most preferred biomarker to detect acute organophosphorus poisoning. Direct visualization of cholinesterase activity on polyacrylamide gels is routinely practiced using acetylthiocholine as a substrate. Overnight incubation with the staining solution is required to understand the enzyme activity bands on gels. Therefore, the need arises to explore rapid detection methods, which can specifically detect hemolysate cholinesterase on polyacrylamide gels. Here, we have explored alternative substrates, such as 1-NA and 2-NA which might have the potential to behave as specific substrates for the detection of hemolysate cholinesterase activity on the gels. It is observed by the in silico studies that 1-NA bind at the active site of acetylcholinesterase akin to acetylcholine (ACh) with a better fitness score. Secondly, the hemolysate cholinesterase activity, as well as its inhibition by organophosphorus pesticides is understandable within 10 min using Fast Blue RR dye for the detection of 1-NA. The organophosphorus inhibited activity is regained in the presence of cholinesterase reactivator. Moreover, the enzyme activity bands formed using 1-NA proves the specificity of the substrate for hemolysate cholinesterase as in the presence of specific acetylcholinesterase inhibitors the band formation disappears. On the other hand, ATCh requires minimum 8-12 h staining time for detection of enzyme activity band following Karnovsky and Roots protocol. Our results prove that 1-NA is an alternative substrate of hemolysate cholinesterase which specifically detects the enzyme activity on gel rapidly. We recommend 1-NA for rapid detection of hemolysate cholinesterase activity on the gels.