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International Angiology 1999 June;18(2):127-30


language: English

A simple test to monitor oxidative stress

Cesarone M. R., Belcaro G., Carratelli M., Cornelli U. *, De Sanctis M. T., Incandela L., Barsotti A., Terranova R. **, Nicolaides A. ***

From the Cardiovascular Institute, Chieti University, PAP/PEA Institute, San Valentino (Pescara), Italy Angiology and Vascular Surgery and Clinical Trials Unit, Pierangeli Clinic, Pescara, Italy * Loyola University Medical School, Maywood Ill., USA ** Institute of Gerontology, University of Catania, Italy *** Irvine Laboratory, Department of Vascular Surgery, Imperial College London, UK


Back­ground. The role of oxy­gen free rad­i­cals is con­sid­ered impor­tant in the devel­op­ment of car­di­o­vas­cu­lar dis­ease. How­ev­er, until recent­ly deter­mi­na­tion of free rad­i­cals plas­ma lev­els and the ­effect of anti­ox­i­dant ther­a­py on these lev­els has been dif­fi­cult. The aim of the study was to deter­mine the oxi­da­tive ­stress and the ­effect of the anti­ox­i­dant com­pound ARD Ste­nov­it on this ­stress in nor­mal sub­jects and ­patients with inter­mit­tent claud­i­ca­tion after oral admin­is­tra­tion for one week.
Meth­ods. A port­able, free rad­i­cals (FRs) deter­mi­na­tion ­system (D-Roms test, Dia­cron, Gros­se­to, Italy) was used. This test is based on the abil­ity of tran­si­tion met­als to cat­a­lyse in the pres­ence of per­ox­ides with for­ma­tion of FRs which are ­trapped by an alchil­a­mine. The alchil­a­mine ­reacts form­ing a ­coloured rad­i­cal detect­able at 505 nm. The ­reagents util­ised are the cro­mo­gen (R1, an alchil­a­mine) and a pH 4.8 buf­fer (R2). Ten μl of hemol­y­sis-free serum are to 1 ml of R2 and to 10 μl of R1. The sam­ple is mixed, incu­bat­ed (1 min; 37°C) and read for opti­cal den­sity. After ­another min­ute, the sam­ple is read again. The aver­age delta A/min is mul­ti­pled by a K fac­tor and cal­cu­lat­ed using serum with ­defined value.
­Results. In nor­mal sub­jects the mean (±SD) lev­els of free rad­i­cals were 312±49 U.CARR (Car­ra­tel­li units) ­before treat­ment and 218±33 U.CARR after treat­ment (p<0.05). A ­decrease of at least 10% was detect­ed in every sub­ject. In ­patients with periph­er­al vas­cu­lar dis­ease the mean (±SD) lev­els of free rad­i­cals were 404±42 U.CARR. ­before treat­ment and 278±33 U.CARR after treat­ment (p<0.02). A ­decrease of at least 15% was detect­ed in every ­patient (medi­um value 31%).
Con­clu­sions. The D-Roms test pro­vides a sim­ple, inex­pen­sive and prac­ti­cal meth­od to iden­ti­fy sub­jects with a high level of oxi­da­tive ­stress and to dem­on­strate the ­effect of treat­ment. The com­pound ARD Ste­nov­it is effec­tive in reduc­ing cir­cu­lat­ing free rad­i­cals. Its ­action on the pro­gres­sion of ath­e­ros­cle­rot­ic dis­ease ­should be ­assessed in ­future stud­ies.

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