Naslov
Important parameters and considerations for in vitro reactivation assays of OP-inhibited cholinesterase

Autori
Zorbaz, Tamara ; Maček Hrvat, Nikolina ; Šinko, Goran ; Kovarik, Zrinka

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
16th Medical Chemical Defence Conference ˝100 years after the first use of sulfur mustard - re-emerging threats of chemical warfare agents and current state of medical research˝ / - München, 2017

Skup
16th Medical Chemical Defence Conference ˝100 years after the first use of sulfur mustard - re-emerging threats of chemical warfare agents and current state of medical research˝

Mjesto i datum
München, Njemačka, 05.04.2017. - 06.04.2017

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
AChE, organophosphates, reactivators, reactivation assay parameters

Sažetak
The risk of organophosphate (OP) poisoning is still present, currently mostly as occupational hazard in agriculture (i.e. pesticides), but there is also justified concern about the potential abuse of OP compounds as nerve agents. Standard oxime reactivators (2-PAM, HI- 6, TMB-4, obidoxime) lack antidotal efficiency and/or the optimal pharmacological characteristics ; therefore, new reactivators of OP-inhibited acetylcholinesterase are being synthesized. In order to evaluate their efficiency as reactivators, many research groups are employing reactivation tests based on the spectrometric Ellman assay. Although the method seems simple, the determination of reactivator efficiency and evaluation of kinetic constants are mostly incomparable among laboratories. Therefore, we addressed the need to standardize the reactivation assay with an evaluation of important factors for experimental design, such as side-reactions between reactants: enzyme, oxime, the Ellman reagent, and substrate. Additionally, concentrations of all of the reactants should be taken into consideration and presented in detail, e.g. there has been significant difference in the results depending on the concentration of the inhibited enzyme in the reactivation mixture, which has not been emphasized until now. Finally, the presented results can differ significantly depending on the evaluation of reactivation constants and overall data analysis. This study was meant to emphasize the important steps in the determination of reactivation parameters and suggest a uniform experimental design, data analysis and result presentation with an aim to achieve better agreement and comparability between the results of different laboratories, and overall a more efficient evaluation of in vitro reactivation potency.

Izvorni jezik
Engleski

Znanstvena područja
Kemija

POVEZANOST RADA