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 FieldElectromagnetic 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 HydrationCell 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]iNa]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

EnvironmentalFactor

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

Bathing aqua solution

[[Na]iNa]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 Waterthe Specific Electrical Conductivity of Distilled Water

ControlControl

270270 mTmT SMFSMF

0.2T, 60min60min 5050Hz, 30minHz, 30min

44Hz, 30minHz, 30min0.20.2T, 30minT, 30min

12 12 mTmT EMFEMF

30 600.0

0.2

0.4

0.6

0.8

1.0 C

0

20

40

60

80

100

4 10 20 50

CONTROL

Quantity of E.coli Quantity of E.coli lonlon-- HM9 mutant cells able to form HM9 mutant cells able to form macrocoloniesmacrocolonies under the influence of SMF and EMFunder the influence of SMF and EMF

CONTROL

Hz

%

min

150

100

50

0

A

B

The effect of magnetized physiological solution on The effect of magnetized physiological solution on 3H3H--timidinetimidine involvement in DNA of spleen cells. involvement in DNA of spleen cells.

AA--control; Bcontrol; 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 solutionphysiological

solution

physiological solution previously exposed to magnetic field

physiological solutionphysiological

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 motilitysperm motility

Time, min

0 20 40 60 80 100 120 140 160

Mot

ility

, %

0

20

40

60

80

100

120

140

ControlSMF 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 4545Ca Influx of Ca Influx of Helix Helix pomatiapomatia 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 Field25mT 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 mT74.33 ±9.22aStatic 4.6 mT100.0 ± 10.07 0

Percentage control 45 Ca influxMagnetic Field

aP < 0.05 compared to control

Bathing aqua solution

[Ca]i

H Y D R A T I O NH 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

13 msec

20nA

AchAch--induced membrane currents in an internallyinduced membrane currents in an internally perfusedperfused HelixHelix neuron. neuron. AchAch--induced current (10induced current (10--55 M) A and CM) A and C--in control; Bin control; B--in a physiological in a physiological

solution previously exposed to magnetic field (27solution previously exposed to magnetic field (27 mTmT). ).

1s

A

1nA

B C

Binding of [Binding of [33H] Ouabain to H] Ouabain to Helix Helix pomatiapomatia 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 10Different Tonicity (x 1088 molecules/mg dry weight)molecules/mg dry weight)

283? 19.4508? 30.1793? 45.61x10-8

147? 9.7266? 15.8431? 29.46x10-9

53.8? 3.190.5? 5.7144? 29.43x10-9

12.2? 0.921.1? 1.432.0? 2.21x10-9

10.0? 0.6717.9? 1.228.9? 2.06x10-10

6.29? 0.4111.7? 0.8718.3? 1.43x10-10

2.03? 0.163.23? 0.244.59? 0.321x10-10

Hypertonic Isotonic Hypotonic

Incubation medium Ouabain content(Mol)

*

The The ouabain ouabain HH3 3 (5x10(5x10--99M) binding by different tissues of rats M) binding by different tissues of rats after after wholebodywholebody exposure to SMF expressed in percents as exposure to SMF expressed in percents as

compared with controlcompared with control

The effect of chemotherapy and radiotherapy on water content The effect of chemotherapy and radiotherapy on water content of normal glandular and cancer tissues of breast cancer patientsof normal glandular and cancer tissues of breast cancer patients

Hydration state of Hydration state of normalnormal tissuestissues

A12

35%

165%

B12

35%

165%

C1230%

170%

after chemotherapyafter chemotherapy after radiotherapyafter radiotherapypreviously untreatedpreviously untreated

A2220%

180%

B2230%

170%

C2

190%

210%

Hydration state of Hydration state of cancercancer tissuestissuespreviously untreatedpreviously untreated after chemotherapyafter chemotherapy after radiotherapyafter radiotherapy

? 11,29%p<0.01,n=517564±789.615782±500.769ouabain 10-6M

? 38%p<0.001,n=51547±50,491119.8±90,61ouabain 10-7M

? 20,81%p<0.05,n=597.2±5.71122.75±4.57ouabain 10-8M

? 8,45%p<0.05,n=615±0.671616.328±1.21ouabain 10-9M

Cancer tissues? 11,75%p<0.05,n=513920.0±952.99512456.0±519.91ouabain 10-6M

? 33,67%p<0.05,n=51211.79±199.289906.500±13.7949ouabain 10-7M

? 27.36%p<0.001,n=592.100±3.4403126.79±6.6359ouabain 10-8M

? 30.3%p<0.001,n=610.740±0.58815.418±1.6599ouabain 10-9M

Normal tissues of a breast cancer patient

Effect %PSMF

(DPM)Control(DPM)

[ouabain](M)

Changes of Radioactive LabeledChanges of Radioactive Labeled OuabainOuabain Binding by Binding by Normal Glandular and Cancer Tissues of Breast Cancer Patient undNormal Glandular and Cancer Tissues of Breast Cancer Patient under the er the

Influence of SMF 0.2T in Different Concentrations ofInfluence of SMF 0.2T in Different Concentrations of OuabainOuabain in the Medium. in the Medium. Mean Values (DPM) Mean Values (DPM) ±±Standard Deviations are ShownStandard Deviations are Shown

Changes ofChanges of OuabainOuabain 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 ofDifferent Concentrations of OuabainOuabain 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=67892±340.916220±327.1ouabain 10-6M

0n=6616.42±74.6640.87±52.64ouabain 10-7M

0n=664.53±4.4565.4±8.21ouabain 10-8M

0n=66.772±0.266.91±0.63ouabain 10-9M

Effect %PSMF

(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

oubain binding sites in the membrane should be considered as the threshold of EMF hazardous doze.