Effects of Oxime K203 and Oxidative Stress in Plasma of Tabun Poisoned Rats

The highly toxic nature of tabun has been known for many years, but there are still serious limitations to antidotal therapy. In this study, we used rats as an experimental model to evaluate the efficiency of bispyridinium para-oxime K203 as therapy against tabun poisoning as well as to examine if induction of oxidative stress is linked to organophosphate toxicity. K203 showed high potency in counteracting tabun poisoning. Either alone or in combination with atropine, this oxime significantly increased cholinesterase activity at 0.5 and 1 h compared to untreated rats poisoned with tabun. Simultaneous measurements of markers of oxidative stress (lipid peroxidation and superoxide dismutase) showed that tabun poisoning, but also therapy (oxime alone or oxime plus atropine) applied immediately after tabun poisoning, could generate free radical species that may cause oxidative stress in rats. (doi: 10.5562/cca1811

Determination of Total Polyphenol Content in Food with the Flow-Injection

U ovom radu predložena je modificirana automatizirana metoda ubrizgavanja u protok za određivanje sadržaja ukupnih polifenola u namirnicama bazirana na Folin-Ciocalteauovoj reakciji u 0,5 mol L-1 NaOH. Metoda omogućuje automatiziranu analizu različitih uzoraka brzinom protoka 55 uzoraka na sat uz upotrebu galne kiseline kao standarda. Primjenom predložene metode na konkretne uzorke (bijelo i crno vino, zeleni, indijski te čaj od lipe, metvice i kamilice i bistri voćni sokovi od crnog ribiza i višnje), određen je njihov “indeks ukupnih polifenola” s većom repetibilnošću za razliku od ranije objavljenih metoda, manje ovisno o razrjeđenju uzorka. U odnosu na “batch” metodu, ova je metoda visoko tolerantna prema najčešćim interferentima (SO2, reducirajući šećeri i askorbinska kiselina). Rezultati dobiveni predloženom metodom pokazali su relativno slaganje s onima dobivenim referentnom Folin-Ciocalteauovom metodom.This paper describes an optimised flow-injection method for the determination of total polyphenol in food based on the Folin-Ciocalteau reaction in 0.5 mol L-1 NaOH. The method allows different types of samples to be analysed automatically at a rate of 55 samples per hour by using gallic acid as standard. By applying the proposed method to real samples (white and red wines, green, Indian, lime-tree, mentha and chamomile teas, and blackberry and cherry juices), their total polyphenol indices were determined with a higher reproducibility than obtained by earlier methods, whatever the dilution used. This method is highly tolerant towards the most common interferences (SO2, reducing sugars, and ascorbic acid) associated with the batch method. The results obtained by the proposed method relatively agree with those obtained using the referent Folin-Ciocalteau method

Use of enhanced flow-injection method for the determination of the total polyphenol contetnt in foods

Cilj ovog rada bio je razvijanje kontinuirane metode ubrizgavanja u protok za određivanje sadržaja ukupnih polifenola u namirnicama. Metoda omogućuje automatsku analizu različitih uzoraka brzinom protoka 55 uzoraka h-1 uz upotrebu galne kiseline kao standarda. U odnosu na "batch" metodu, ova metoda je visoko tolerantna prema najčešćim interferentima (SO2, reducirajući šećeri i askorbinska kiselina). Rezultati dobiveni predloženom metodom pokazali su relativno slaganje sa onima dobivenim referentnom Folin-Ciocalteau metodom.The aim of this research was the development of a continuos, flow-injection method for the determination of total polyphenol content in foods. The method allows different types of samples to be analysed automatically at a rate of 55 samples per hour by using gallic acid as standard. The method is highly tolerant towards the most common interferences (SO2, reducing sugars and ascorbic acid) that occur with its batch counterpart. The results provided by the proposed method relatively agree with those obtained using the referent Folin-Ciocalteau method

Use of enhanced flow-injection method for the determination of the total polyphenol contetnt in foods

Cilj ovog rada bio je razvijanje kontinuirane metode ubrizgavanja u protok za određivanje sadržaja ukupnih polifenola u namirnicama. Metoda omogućuje automatsku analizu različitih uzoraka brzinom protoka 55 uzoraka h-1 uz upotrebu galne kiseline kao standarda. U odnosu na "batch" metodu, ova metoda je visoko tolerantna prema najčešćim interferentima (SO2, reducirajući šećeri i askorbinska kiselina). Rezultati dobiveni predloženom metodom pokazali su relativno slaganje sa onima dobivenim referentnom Folin-Ciocalteau metodom.The aim of this research was the development of a continuos, flow-injection method for the determination of total polyphenol content in foods. The method allows different types of samples to be analysed automatically at a rate of 55 samples per hour by using gallic acid as standard. The method is highly tolerant towards the most common interferences (SO2, reducing sugars and ascorbic acid) that occur with its batch counterpart. The results provided by the proposed method relatively agree with those obtained using the referent Folin-Ciocalteau method

A conjugate of pyridine-4-aldoxime and atropine as a potential antidote against organophosphorus compounds poisoning

A conjugate of pyridine-4-aldoxime and atropine (ATR-4-OX) was synthesized and its antidotal efficiency was tested in vitro on tabun- or paraoxon-inhibited acetylcholinesterase (AChE) of human erythrocytes as well as in vivo using soman-, tabun- or paraoxon-poisoned mice. Its genotoxic profile was assessed on human lymphocytes in vitro and was found acceptable for further research. ATR-4-OX showed very weak antidotal activity, inadequate for soman or tabun poisoning. Conversely, it was effective against paraoxon poisoning both in vitro and in vivo. All animals treated with 5 % or 25 % LD(50) doses of the new oxime survived after administration of 10.0 or 16.0 LD(50) doses of paraoxon, respectively. Based on the persistence of toxicity symptoms in mice, the atropine moiety had questionable effects in attenuating such symptoms. It appears that ATR-4-OX has a therapeutic effect related to the reactivation of phosphylated AChE, but not to receptor antagonization

Effects of Oxime K203 and Oxidative Stress in Plasma of Tabun Poisoned Rats

The highly toxic nature of tabun has been known for many years, but there are still serious limitations to antidotal therapy. In this study, we used rats as an experimental model to evaluate the efficiency of bispyridinium para-oxime K203 as therapy against tabun poisoning as well as to examine if induction of oxidative stress is linked to organophosphate toxicity. K203 showed high potency in counteracting tabun poisoning. Either alone or in combination with atropine, this oxime significantly increased cholinesterase activity at 0.5 and 1 h compared to untreated rats poisoned with tabun. Simultaneous measurements of markers of oxidative stress (lipid peroxidation and superoxide dismutase) showed that tabun poisoning, but also therapy (oxime alone or oxime plus atropine) applied immediately after tabun poisoning, could generate free radical species that may cause oxidative stress in rats. (doi: 10.5562/cca1811

Evaluation of HI-6 oxime: potential use in protection of human acetylcholinesterase inhibited by antineoplastic drug irinotecan and its cyto/genotoxicity in vitro

The function of acetylcholinesterase (AChE) is the rapid hydrolysis of the neurotransmitter acetylcholine (ACh), which is involved in the numerous cholinergic pathways in both the central and the peripheral nervous system. Therefore, AChE measurement is of high value for therapy management, especially during the course of intoxication with different chemicals or drugs that inhibit the enzyme. Pyridinium or bispyridinium aldoximes (oximes) are able to recover the activity of the inhibited enzyme. Since their adverse effects are not well elucidated, in this study the efficiency of HI-6 oxime in protection and/or reactivation of human erythrocyte AChE inhibited by the antineoplastic drug irinotecan as well as its cyto/genotoxicity in vitro were investigated. HI-6 was effective in protection of AChE and increased its activity up to 30%; the residual activity after irinotecan inhibition was 7%. Also, it reactivated the enzyme previously inhibited by 50% irinotecan (4.6 µg/ml) applied at ¼ of the IC50 value. The tested concentrations of HI-6 exhibited acceptable genotoxicity towards white blood cells, as estimated by the alkaline comet assay, DNA diffusion assay and cytogenetic endpoints (structural chromosome aberrations and cytokinesis-block micronucleus assay). The results obtained warrant the further investigation of HI-6 in vivo, as well as its development for possible application in chemotherapy

Tenocyclidine treatment in soman-poisoned rats - intriguing results on genotoxicity versus protection

This study aimed to evaluate the antidotal potency of tenocyclidine (TCP) that probably might protect acetylcholinesterase (AChE) in the case of organophosphate poisoning. TCP was tested alone as a pretreatment or in combination with atropine as a therapy in rats poisoned with ¼ and ½ of LD50 of soman. Possible genotoxic effects of TCP in white blood cells and brain tissue were also studied. Results were compared with previous findings on the adamantyl tenocyclidine derivative TAMORF. TCP given alone as pretreatment, 5 min before soman, seems to be superior in the protection of cholinesterase (ChE) catalytic activity in the plasma than in brain, especially after administration of the lower dose of soman. Plasma activities of the enzyme after a joint treatment with TCP and soman were significantly increased at 30 min (P < 0.001) and 24 h (P = 0.0043), as compared to soman alone. TCP and atropine, given as therapy, were more effective than TCP administered alone as a pretreatment. The above therapy significantly increased activities of the enzyme at 30 min (P = 0.046) and 24 h (P < 0.001), as compared to controls treated with ¼ LD50 of soman alone. Using the alkaline comet assay, acceptable genotoxicity of TCP was observed. However, the controversial role of TCP in brain protection of soman-poisoned rats should be studied further

Procjena oksima K203 kao antidota pri otrovanju tabunom

We studied bispyridinium oxime K203 [(E)-1-(4-carbamoylpyridinium)-4-(4-hydroxyimino methylpyridinium)- but-2-ene dibromide] with tabun-inhibited human acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in vitro, and its antidotal effect on tabun-poisoned mice and rats in vivo. We compared it with oximes K048 and TMB-4, which have proven the most effi cient oxime antidotes in tabun poisoning by now. Tabun-inhibited AChE was completely reactivated by K203, with the overall reactivation rate constant of 1806 L mol-1 min-1. This means that K203 is a very potent reactivator of tabun-inhibited AChE. In addition, K203 reversibly inhibited AChE (Ki = 0.090 mmol L-1) and BChE (Ki = 0.91 mmol L-1), and exhibited its protective effect against phosphorylation of AChE by tabun in vitro. In vivo, a quarter of the LD50 K203 dose insured survival of all mice after the application of as many as 8 LD50 doses of tabun, which is the highest dosage obtained compared to K048 and TMB-4. Moreover, K203 showed high therapeutic potency in tabun-poisoned rats, preserving cholinesterase activity in rat plasma up to 60 min after poisoning. This therapeutic improvement obtained by K203 in tabun-poisoning places this oxime in the spotlight for further development.Proučavali smo bispiridinijski oksim K203 [(E)-1-(4-karbamilpiridinij)-4-(4-hidroksiiminometilpiridinij)- but-2-ene dibromid] u uvjetima in vitro - studirajući njegove interakcije s ljudskom acetilkolinesterazom (AChE) i butirilkolinesterazom (BChe) inhibiranim tabunom te u uvjetima in vivo - određivanjem njegova antidotskog učinka na miševe i štakore otrovane tabunom. Radi usporedbe uključili smo rezultate dobivene s oksimima K048 i TMB-4 kao najučinkovitijim oksimima kod otrovanja tabunom. K203 je potpuno reaktivirao AChE inhibiranu tabunom sa sveukupnom brzinom reaktivacije od 1806 L mol-1 min-1 što ga svrstava u najučinkovitije reaktivatore AChE inhibirane tabunom. K203 je reverzibilno inhibirao AChE (Ki = 0,090 mmol L-1) i BChE (Ki = 0,91 mmol L-1) pokazujući svoja in vitro zaštitna svojstva od inhibicije tabunom. Terapija dozom K203 od ¼ njegove LD50 omogućila je preživljavanje svih miševa nakon otrovanja dozom tabuna od 8,0 LD50. Time je K203 pokazao bolju učinkovitost u usporedbi s K048 ili TMB-4. K tome, K203 je značajno zaštitio štakore od otrovanja tabunom kompenzirajući toksični učinak tabuna na aktivnost kolinesteraze i do 60 min nakon trovanja. Pokazano poboljšanje terapeutske učinkovitosti K203 ističe ovaj oksim pretečom za daljnji razvoj antidota u otrovanju tabunom