Effect of the pulsed radiofrequency field on time course of hemoglobin deoxygenation.

<p>Time course of oxy hemoglobin (HbO₂) concentration, after a single 30 min pulsed radiofrequency electromagnetic field (PRF) exposure of Hb, under deoxygenating conditions. Concentration was determined by visible light spectroscopy at 560, 576, 630 nm. PRF exposure resulted in a significant (30.8±10.3)% reduction in oxyHb concentration, as compared to controls (14.6±1.3 µM vs. 21.1±2.5 µM, P<0.03, n = 5), suggesting an alteration in Hb solution properties that persisted after PRF signal was removed.</p

Effect of the static magnetic field on time course of hemoglobin deoxygenation.

<p>Hb deoxygenation after 10 min exposure to 186 mT static magnetic field (SMF) in 5M urea. No significant change in oxy/deoxy ratio was visible until the time of rapid deoxygenation, at approximately 40 min. The time of most rapid deoxygenation occurred approximately 10 min earlier for SMF treated samples (P<0.002 for 42 min</p

EMF effect on hemoglobin (Hb) visible light spectra.

<p>Spectra representative of the effects of pulsed radiofrequency (PRF) signal and 186 mT static magnetic field (SMF) on Hb visible light spectra during bHb)deoxygenation. Data shown are typical of samples drawn from a single tube for each EMF exposure condition, shown here at 150 min after a single 30 min EMF exposure. Deoxygenation of 100 µM Hb was carried out in 50 mM Hepes buffer (pH 7.2) using the reducing agent dithiothreitol (20 mM) at 22°C, and is characterized here by the passage of the spectrum from a two-peaked to one-peaked form. Deoxygenation occured at an earlier time for EMF exposed samples (traces with one peak), as compared to control samples exposed only to the ambient geomagnetic laboratory environment (trace with two peaks). The EMF effect was observable at the time of most rapid deoxygenation.</p

Addition of 5 M urea to reaction mixture.

<p>Addition of 5 M urea to the deoxygenation assay reduces the time required for deoxygenation and render the PRF effect more apparent. The maximal difference between PRF-treated and control samples occurred at 73 min, with PRF-treated samples showing a significant (70.4±9.7)% reduction in oxyHb concentration, as compared to controls (8.5±2.1 µM vs. 28.8±6.3 µM, P<0.03, n = 5), in contrast to the 30.8% reduction observed in the absence of urea (cf. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061752#pone-0061752-g002" target="_blank">Figure 2</a>).</p

Reaction plate showing that addition of 5M urea to reaction solution renders magnetic field effect more apparent.

<p>96-well spectrophotometer plate shown at 46 min after initiation of reaction of hemoglobin with dithiothreitol (DTT). Samples treated for 10 min with 186 mT static magnetic field (3 left-hand columns in plate) substantially completed (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061752#pone-0061752-g004" target="_blank">Figure 4</a>) DTT-induced deoxygenation before untreated samples began to lose O₂, rendering the magnetic field effect visibly apparent.</p