## 3 Results
### Effect of Combined Magnetic Fields on ROS Generation by Neutrophils
#### 3.2.2 Water-Mediated Effect of Magnetic Fields on Neutroph  
Results presented in the sections above clearly demonstrated the effect of combined static and alternating magnetic fields on ROS generation by peritoneal neutrophils. However, it remains unclear whether the CMF act directly on the molecular mechanisms responsible for the production of ROS, or indirectly through water. Similar water-mediated effect of physical exposures in the form of microwaves at 36 GHz and 42 GHz has already been investigated in our laboratory both using biological and engineering models (capacitor discharge) [14; 15]. To check the ability of aqueous medium to mediate the effect of CMF on the ROS generation by neutrophils, we carried out a number of studies using similar cell chemiluminescence method. Water samples were first exposed to the CMF then they were added to neutrophil suspension, and luminol-enhanced chemiluminescence was measured after stimulation with fMLF. Figure 4: Effect of combined magnetic fields with static component of 60 \(\mu\)T on intensity of neutrophil chemiluminescence. On Y-axis—intensity of chemiluminescence (maximum value) in %, relative to control. Data is presented as M \(\pm\) SD. *—\(p<\) 0.05 vs. control group. Figure 5: Effect of combined magnetic fields on kinetics and intensity of chemiluminescence of neutrophils incubated under CMF with static component of 60 \(\mu\)T and alternating component of 100 nT with sinusoidal signal at: (**a**) frequency of 12.6 Hz; (**b**) frequency of 48.5 Hz. The settings of the magnetic field induction device for water CMF exposure were similar to the described in the previous sections and corresponded to static component of 60 uT and alternating component of 100 nT with sinusoidal modulation of 12.6 or 48.5 Hz. Table 3 shows that the exposure of \"Experimental\" sample to the CMF with alternating component of 12.6 Hz caused an increase of the cell chemiluminescence intensity by approximately 65% relative to \"Control\" (water sample incubated outside the magnetic field induction device in the GMF) (\(p\) = 0.002167). Meanwhile, no significant differences from \"Control\" sample were observed for the treatment with the CMF with alternating component of 48.5 Hz (\(p\) = 0.9372). Noteworthy, samples obtained after addition of water pre-exposed to the CMF with alternating component of 12.6 Hz demonstrated more pronounced stimulating effect on the intensity of chemiluminescence (water-mediated effect) compared to direct neutrophil suspension exposure to the CMF with the same frequency (Table 2). An increase in chemiluminescence amounted to 66% with the indirect effect of the CMF versus a 36% increase with the direct effect of the CMF on neutrophil suspension. 2.3 Water-Mediated Activation of Neutrophils with Samples Exposed to Magnetic Fields, Dilution, and Intensive Mechanical Treatment  
In this part of the study, \"Experimental\" sample (pre-incubated under CMF at 12.6 Hz or 48.5 Hz) was subjected to serial dilutions and shaking under the GMF or \"magnetic vacuum\". The CMF exposure was performed in the magnetic field induction device with the settings identical to the ones described in the previous sections and corresponding to magnetic field with static component of 60 uT and alternating component of 100 nT. Alternating component corresponded to sinusoidal signal of 12.6 Hz or 48.5 Hz. To confirm the relevance of individual procedures of the serial dilution method, vigorous shaking was carried out in three versions (detailed protocol is described in the relevant section of \"Materials and Methods\") and the results are shown in Table 3:  
1. \"GMF at 1099 dilution\"--dilution and shaking were performed under the GMF (outside the magnetic field induction device). 2. \"Magnetic vacuum\" at 109 dilution\"--dilutions were made under the GMF (outside the magnetic field induction device), while shaking was made inside the device under the so-called \"magnetic vacuum\" (~10 nT). 3.