To study the possibility that ion-protein interactions can be affected by magnetic fields with various frequencies and magnetic flux densities. Specifically, Ca2+-calmodulin-dependent myosin phosphorylation, in a cell-free preparation is examined from the viewpoint of searching for mechanisms of action and studying the effectiveness of various magnetic field signals used clinically.
The Ca2+-calmodulin-dependent myosin light chain phosphorylation is a well-studied biochemical enzyme system, which appears to depend upon ion binding.
The reaction was initiated by adding 32P ATP to the medium.
One of the experiments (Field 1) was performed to replicate a previous study (Shuvalova, L.A., Ostrovskayam M.V., Sosunov, E.A., Lednev, V.V. (1991). Weak magnetic fields tuned to the parametric resonance condition change the rate of calcium-calmodulin dependent myosin phosphorylation. Reports of the Academy of Sciences. USSR 317:227-231).
| Exposure | Parameters |
|---|---|
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Exposure 1:
8–20 Hz
Exposure duration:
continuous for 5 min
Simultaneously exposed to AC and DC magnetic fields as well as exposure to AC and DC magnetic fields separately.
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Exposure 2:
Exposure duration:
continuous for 5 min
DC magnetic fields in ambient range
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Exposure 3:
Exposure duration:
continuous for 5 min
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Exposure 4:
27 MHz
Modulation type:
pulsed
Exposure duration:
continuous for 5 min
pulsed radiofrequency field
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Exposure 5:
80–180 Hz
Exposure duration:
continuous for 5 min
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Exposure 6:
Exposure duration:
continuous for 5 min
gradient magnetic field
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- |
| Frequency | 8–20 Hz |
|---|---|
| Type | |
| Waveform | |
| Exposure duration | continuous for 5 min |
| Additional info | Simultaneously exposed to AC and DC magnetic fields as well as exposure to AC and DC magnetic fields separately. |
| Additional info | Reference article: Shuvalova, L.A., Ostrovskayam M.V., Sosunov, E.A., Lednev, V.V. (1991). Weak magnetic fields tuned to the parametric resonance condition change the rate of calcium-calmodulin dependent myosin phosphorylation. Reports of the Academy of Sciences. USSR 317:227-231. |
| Exposure source | |
|---|---|
| Chamber | Special designed Plexiglas chamber maintained at 37°C. |
| Setup | Two orthogonal Helmholtz pairs of square coils were used to apply vertical and horizontal DC magnetic fields and third pair of circular Helmholtz coils was mounted horizontally to apply AC field. The exposure volume was 100 µL of samples poured in Eppendorf tubes. |
| Measurand | Value | Type | Method | Mass | Remarks |
|---|---|---|---|---|---|
| magnetic flux density | 20.9 µT | effective value | measured | - | - |
| Frequency | |
|---|---|
| Type | |
| Exposure duration | continuous for 5 min |
| Additional info | DC magnetic fields in ambient range |
| Exposure source |
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|---|---|
| Setup | Same set-up as described in E1 but the tube was placed in a µ-metal box |
| Additional info | The magnetic fields were applied alternatively in the vertical and horizontal directions inside the shield. |
| Measurand | Value | Type | Method | Mass | Remarks |
|---|---|---|---|---|---|
| magnetic flux density | 10 µT | minimum | measured | - | 20 µT, 30 µT, 40 µT, 50 µT, 60 µT, 70 µT, 80 µT, 90 µT, 100 µT, 120 µT, 130 µT, 140 µT, 150 µT, 160 µT, 170 µT and 180 µT |
| Frequency | |
|---|---|
| Type | |
| Exposure duration | continuous for 5 min |
| Additional info | static magnetic field |
| Additional info | Reference article: Markov, M.S. (2000). Magnetic and electromagnetic fields- a new frontier in clinical biology and medicine. Millennium International Workshop on Biological Effects of Electromagnetic Fields. Crete, Greece, 365-372, ISBN # 960-86733-0-5. |
| Exposure source |
|
|---|---|
| Chamber | Same as in E1. |
| Setup | The magnets were configured with the opposite poles facing one another. One magnet was kept stationary while the other one was moved towards the first ones direction to get different magnetic fields. |
| Measurand | Value | Type | Method | Mass | Remarks |
|---|---|---|---|---|---|
| magnetic flux density | 5 mT | minimum | measured | - | 10 mT, 15 mT, 20 mT, 25 mT, 30 mT, 35 mT, 40 mT, 45 mT, 50 mT and 55 mT |
| Frequency | 27 MHz |
|---|---|
| Exposure duration | continuous for 5 min |
| Additional info | pulsed radiofrequency field |
| Additional info | Reference article: Markov, M.S., Muesham, D.J., Pilla, A.A. (1994 c). Modulation of cell free myosin phosphorylation with pulsed radio frequency electromagnetic fields. In: Allen, M.J., Cleary, S.F., Sowers, A.E., eds. Charge and Field effects in Biosystems 4. World Scientific, pp- 274-288. |
| Exposure source |
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|---|
| Measurand | Value | Type | Method | Mass | Remarks |
|---|---|---|---|---|---|
| magnetic flux density | 160 µT | effective value | measured | - | - |
| magnetic flux density | 200 µT | effective value | measured | - | - |
| Frequency | 80–180 Hz |
|---|---|
| Type | |
| Waveform | |
| Exposure duration | continuous for 5 min |
| Additional info | therapeutic electromagnetic field |
| Additional info | Therapeutic electromagnetic field applied at 80 pulses/sec, 100 pulses/sec, 120 pulses/sec, 140 pulses/sec, 160 pulses/sec and 180 pulses/sec. |
| Modulation type | cf. additional info |
|---|---|
| Additional info |
half sine wave pulsating signals were applied. |
| Exposure source |
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|---|
| Measurand | Value | Type | Method | Mass | Remarks |
|---|---|---|---|---|---|
| magnetic flux density | 5 mT | unspecified | measured | - | 10 mT, 15 mT, 20 mT and 25 mT |
| Frequency | |
|---|---|
| Type | |
| Exposure duration | continuous for 5 min |
| Additional info | gradient magnetic field |
| Exposure source |
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|---|---|
| Setup | Samples were positioned at a distance of 0, 5, 10 and 15 mm from the device. |
| Additional info | MagnaBloc device consisting of 4 NdFeB permanent magnets arranged with alternative polarity in a 2 x 2 array. |
No parameters are specified for this exposure.
The cell-free myosin phosphorylation model system is sensitive to a wide range of magnetic fields that differ in magnetic flux density, repetition rate, and frequency. It appears that the key target is in the binding of the calcium ion to the existing binding sites in the calmodulin structure. Except for the ambient range DC signals, all other signals initiate "resonance"- or "window"-like responses. Therefore, myosin phosphorylation assay may be tool for examining the "window hypothesis".
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