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Cell Hydration as Cell Hydration as an Essential Cell Parameter for an Essential Cell Parameter for Estimating the Biological Effect of Estimating the Biological Effect of Electromagnetic Field Electromagnetic Field

Sinerik N. Ayrapetyan

UNESCO Chair - Life Sciences International Educational Center Yerevan, Armenia

To estimate the EMF effect quantitatively To estimate the EMF effect quantitatively we need a cellular parameter we need a cellular parameter characterizing cell functional state and characterizing cell functional state and serving as a common target serving as a common target for various environmental factors! for various environmental factors!

Cell Hydration Cell Hydration

!

?

PURPOSE

To prove the hypothesis on the cell hydration as a universal target for various environmental factors by the experimental data

a) water structure as a universal target through which the LF EMF signals are transferred to the cell metabolic cascade b) EMF sensitivity of cell volume and ion transporting systems in membrane responsible for its regulation.

To suggest a method for quantity estimation of EMF-induced cell hydration.

Present approaches to the dependency of cell hydration on the main ion transporting systems in membrane

[Na]i Na]i [Ca]i

H Y D R A T I O N

2 K+ 3 Na+

ATP cAMP

3 Na+ Ca+2

Receptor Channel Potential Channel

3Na+

cGMP

Ca+2

Environmental Factor

EMF Induced Water Structure Changing Effect on Ion Transporting Systems in Cell Membrane

Bathing aqua solution

[Na]i Na]i [Ca]i

H Y D R A T I O N

2 K+ 3 Na+

ATP cAMP

3 Na+ Ca+2

Receptor Channel Potential Channel

3Na+

cGMP

Ca+2

EMF

80 85 90 95 100 105

C SMF 4Hz 10Hz 20Hz 50Hz

G.% The Effect of SMF and EMF on The Effect of SMF and EMF on the Specific Electrical Conductivity of Distilled Water the Specific Electrical Conductivity of Distilled Water

Control Control

270 270 mT mT SMF SMF

0.2T, 60min 60min 50 50Hz, 30min Hz, 30min 4 4Hz, 30min Hz, 30min 0.2 0.2T, 30min T, 30min

12 12 mT mT EMF EMF

30 60 0.0 0.2 0.4 0.6 0.8 1.0

C

20 40 60 80 100

4 10 20 50

CONTROL

Quantity of E.coli Quantity of E.coli lon lon-

• HM9 mutant cells able to form

HM9 mutant cells able to form macrocolonies macrocolonies under the influence of SMF and EMF under the influence of SMF and EMF

CONTROL

Hz % min

150 100 50

A B The effect of magnetized physiological solution on The effect of magnetized physiological solution on 3H 3H-

• timidine

timidine involvement in DNA of spleen cells. involvement in DNA of spleen cells. A A-

• control; B

control; B-

• incubation of spleen slices in magnetized

incubation of spleen slices in magnetized physiological solution (270mT SMF) physiological solution (270mT SMF)

The effect of magnetized physiological solution on snail heart contractility and time dependency of heart sensitivity threshold on it. A-10, B-30 and C-60 min after applying.

physiological solution

physiological solution

physiological solution previously exposed to magnetic field

physiological solution physiological solution

physiological solution previously exposed to magnetic field

physiological solution

A B C

physiological solution previously exposed to magnetic field

The effect of 0.3T SMF treated physiological solution on The effect of 0.3T SMF treated physiological solution on sperm motility sperm motility

Time, min

20 40 60 80 100 120 140 160

Motility, %

20 40 60 80 100 120 140

Control SMF exposure

initial motility, 100%

The effect of magnetized physiological solution on snail neuronal volume and its electrical activity

A and C– normal physiological solution, B - physiological solution preliminary treated by SMF a – in normal physiological solution, b-after 5’, c-7’, d-10’, e-15’ incubation in magnetized physiological solution; f-3’, g-5’, h-7’ washed

49mV

a b

49mV 49mV 49mV 47mV 49mV

c d e

45mV

f g

53mV

h

20sec 40mV

Changes in Content of Cyclic Nucleotides and Changes in Content of Cyclic Nucleotides and 45

45Ca Influx of

Ca Influx of Helix Helix pomatia pomatia Ganglia Incubated in Either Control Solution or Ganglia Incubated in Either Control Solution or Physiologic Solution Previously Exposed to Physiologic Solution Previously Exposed to 25mT Static Magnetic Field 25mT Static Magnetic Field 164.9 ± 41.6b 79.1 ±9.5a Exposed 100.0 ± 11.6 100.0 ±9.6 Control

cGMP cAMP Solution Content of cyclic nucleotide

aP < 0.02, bP < 0.05

62.67 ± 9.00a Static 38.0 mT 74.33 ±9.22a Static 4.6 mT 100.0 ± 10.07

Percentage control 45 Ca influx Magnetic Field

aP < 0.05 compared to control

Bathing aqua solution

[Ca]i

H Y D R A T I O N H Y D R A T I O N

2 K+ 3 Na+

ATP cAMP

3 Na+ Ca+2

Receptor Channel Potential Channel

3Na+

cGMP

Ca+2

EMF cGMP Dependent Na:Ca Exchanger as Extrasensitive Target for EMF

Fast (A) and slow (B) inward Ca currents in snail neurons in Fast (A) and slow (B) inward Ca currents in snail neurons in normal physiological solution (1) and in normal physiological solution (1) and in solution preliminary exposed in SMF (2) solution preliminary exposed in SMF (2) A B

2

125 msec

2 1 1

3 msec

20nA

Ach Ach-

• induced membrane currents in an internally

induced membrane currents in an internally perfused perfused Helix Helix neuron. neuron. Ach Ach-

• induced current (10

induced current (10-5 M) A and C M) A and C-

• in control; B

in control; B-

• in a physiological

in a physiological solution previously exposed to magnetic field (27 solution previously exposed to magnetic field (27 mT mT). ).

1s

A

1nA

B C

Binding of [ Binding of [3H] Ouabain to H] Ouabain to Helix Helix pomatia pomatia Cell Membrane as a Cell Membrane as a Function of Concentration of Glycoside in Solutions with Function of Concentration of Glycoside in Solutions with Different Tonicity (x 10 Different Tonicity (x 108 molecules/mg dry weight) molecules/mg dry weight)

283?19.4 508?30.1 793?45.6 1x10-8 147?9.7 266?15.8 431?29.4 6x10-9 53.8?3.1 90.5?5.7 144?29.4 3x10-9 12.2?0.9 21.1?1.4 32.0?2.2 1x10-9 10.0?0.67 17.9?1.2 28.9?2.0 6x10-10 6.29?0.41 11.7?0.87 18.3?1.4 3x10-10 2.03?0.16 3.23?0.24 4.59?0.32 1x10-10

Hypertonic Isotonic Hypotonic Incubation medium Ouabain content (Mol)

*

The The ouabain

• uabain H

H3

3 (5x10

(5x10-9M) binding by different tissues of rats M) binding by different tissues of rats after after wholebody wholebody exposure to SMF expressed in percents as exposure to SMF expressed in percents as compared with control compared with control

The effect of chemotherapy and radiotherapy on water content The effect of chemotherapy and radiotherapy on water content

• f normal glandular and cancer tissues of breast cancer patients
• f normal glandular and cancer tissues of breast cancer patients

Hydration state of Hydration state of normal

normal tissues

tissues

A1 2 35% 1 65% B1 2 35% 1 65% C1 2 30% 1 70%

after chemotherapy after chemotherapy after radiotherapy after radiotherapy previously untreated previously untreated

A2 2 20% 1 80% B2 2 30% 1 70% C2 1 90% 2 10%

Hydration state of Hydration state of cancer

cancer tissues

tissues

previously untreated previously untreated after chemotherapy after chemotherapy after radiotherapy after radiotherapy

? 11,29% p<0.01,n=5 17564± 789.6 15782± 500.769

• uabain 10-6M

? 38% p<0.001,n=5 1547±50,49 1119.8±90,61

• uabain 10-7M

? 20,81% p<0.05,n=5 97.2±5.71 122.75±4.57

• uabain 10-8M

? 8,45% p<0.05,n=6 15±0.6716 16.328±1.21

• uabain 10-9M

Cancer tissues

? 11,75% p<0.05,n=5 13920.0±952.995 12456.0±519.91

• uabain 10-6M

? 33,67% p<0.05,n=5 1211.79±199.289 906.500±13.7949

• uabain 10-7M

? 27.36% p<0.001,n=5 92.100±3.4403 126.79±6.6359

• uabain 10-8M

? 30.3% p<0.001,n=6 10.740±0.588 15.418±1.6599

• uabain 10-9M

Normal tissues of a breast cancer patient

Effect % P SMF (DPM) Control (DPM) [ouabain] (M)

Changes of Radioactive Labeled Changes of Radioactive Labeled Ouabain Ouabain Binding by Binding by Normal Glandular and Cancer Tissues of Breast Cancer Patient und Normal Glandular and Cancer Tissues of Breast Cancer Patient under the er the Influence of SMF 0.2T in Different Concentrations of Influence of SMF 0.2T in Different Concentrations of Ouabain Ouabain in the Medium. in the Medium. Mean Values (DPM) Mean Values (DPM) ± ±Standard Deviations are Shown Standard Deviations are Shown

Changes of Changes of Ouabain Ouabain Binding by Breast Glandular Tissue of Binding by Breast Glandular Tissue of Healthy Women under the Influence of SMF 0.2T in Healthy Women under the Influence of SMF 0.2T in Different Concentrations of Different Concentrations of Ouabain Ouabain in the Medium. in the Medium. Mean Values (DPM) Mean Values (DPM) ± ± Standard Deviations are Shown. Standard Deviations are Shown. ?26,88%

p<0.001,n=6 7892±340.91 6220±327.1

• uabain 10-6M

n=6 616.42±74.6 640.87±52.64

• uabain 10-7M

n=6 64.53±4.45 65.4±8.21

• uabain 10-8M

n=6 6.772±0.26 6.91±0.63

• uabain 10-9M

Effect % P SMF (DPM) Control (DPM) [ouabain] (M)

CONCLUSION CONCLUSION

Cell hydration serves as a universal and extrasensitive cell parameter which can be used for estimating the beneficial or hazardous effects of EMF on cell or organisms.

QUESTION ? QUESTION ?

Which percentage of cell hydration has to be considered as overhydration (hazardous)?

ANSWER !

The percent of cell hyperhydration which is not accompanied by the increase of the number of

• ubain binding sites in the membrane should be

considered as the threshold of EMF hazardous doze.