*2.4. H2O2Assay*

H2O2 level was determined by the cell permeant probe 2-7-dichlorodihydrofluorescin diacetate (DCFDA). PBMCs were incubated with 10 μM DCFDA in the dark at 37 ◦C for 20 min, pelleted at 600× *g* for 5 min, washed and resuspended in the assay buffer (100 mM potassium phosphate, pH 7.4, 2 mM MgCl2). An aliquot was used for protein determination. The H2O2 dependent oxidation of the fluorescent probe (507 nm excitation and 530 nm emission wavelengths) was measured by a Jasco FP6200 spectrofluorimeter (Jasco SRL, Cremella, Italy).

### *2.5. Data Analysis*

All presented data are means ± standard error of mean (SEM). Statistical difference was determined by Student' *t*-test. *p*-value of 0.05 was considered as statistically significant (\*\*\* *p* < 0.001; \*\* *p* < 0.01; \* *p* < 0.05).

Correlation plots for controls and MS have been performing with Excel Microsoft software using Pearson's correlation analysis. *P*-values less than 0.05 were considered statistically significant.
