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239:424-427, 1980.
 Am J Physiol Regulatory Integrative Comp Physiol
A. A. Marino, J. M. Cullen, M. Reichmanis, R. O. Becker and F. X. Hart
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 Integrative and Comparative Physiology American Journal of Physiology - Regulatory,
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Sensitivity to change in electrical environment:a new bioelectric effect
ANDREW A. MARINO, JAMES M. CULLEN, MARIA REICHMANIS,ROBERT 0. BECKER, AND FRANCIS X. HARTVeterans Administration Medical Center and Department of Orthopedic Surgery, State Universityof New York Upstate Medical Center, Syracuse, New York 13210; and Department of Physics,University of the South, Sewanee, Tennessee 37775MARINO, ANDREW
A.,
JAMES
M.
CULLEN, MARIA REICH-
MANIS,ROBERTO.BECKER,AND FRANCISX.HART.
Sensitiuity
to change in electrical environment: a new bioelectric effect.Am. J. Physiol. 239 (Regulatory Integrative Comp. Physiol. 8):R424-R427,1980.-The action of a 60.Hz, 5 kV/m electric fieldon erythrocyte parameters in mice was determined. No effectsattributable to the magnitude of the field were found, but atransition either from or to an environment containing the fieldcaused decreased red blood cell concentrations and decreasedhematocrits. The failure of others to observe effects on eryth-rocyte parameters following exposure to low-frequency electricfields may have been due to an inappropriate choice of durationof exposure.mice; 60-Hz electric field; erythrocytes; stressor
EXTREMELY LOW-FREQUENCY (ELF) electric and mag-netic fields-less than
100
Hz-have produced biologicaleffects in subjects ranging from amoeba to man
(6).
Therole of exposure duration is among the many unansweredquestions concerning such bioeffects. It might be sup-posed that longer exposure periods would result in eitherstronger effects, or increased probability of observing aneffect, but often this is not the case. Exposure of humanbeings to ELF fields for
1
day or less resulted in increasedleucocyte concentrations (5)) increased triglycerides (2))and altered psychological test performance (4). Also, ELFelectric fields of less than
1
kV/m have altered themotility of amoebas in
10
min (3), the calcium efflux fromcat cerebral tissue in
20
min
(1),
and the mitochondralfunction of guinea pig brain cells in 90 min
(11).
On theother hand, rats exposed to a 100 kV/m, 60-Hz electricfield for 15-20 days exhibited no change in 29 bloodparameters (10).There is, presently, little hope of satisfactorily under-standing the ELF-field effects on the basis of fundamen-tal biophysical principles. Two important reasons arethat the electrical constants of biological tissue such asthe dielectric constant and conductivity are not wellcharacterized, and that it is very difficult to directlymeasure the fields inside living organisms. ‘These prob-lems have prevented the calculation of unique or verifi-able internal field levels. For example, let us model amouse as a sphere of tissue of radius a, surrounded by alayer of skin of thickness b-a, and subjected to an applied
R424
electric field of &OS cz>t ith angular frequency ~3. Letthe conductivities (g ), dielectric constants (K), and fields(E) in each of the two regions be denoted, respectively,as follows: g,, gt; K, K,; E, Et. It can be shown thatEt = [
(1 +
Dz/a”)Cl + D&/a3]cos mt- [(I + D2/a3)C2 - D&/a3]sin ci>tE, = [(D& + D&)/r3 - Cl]cos atwhere+
[(DzCz
- D1Cl)/r3 - Cz]sin ut
Cl = R&/RI + S/RIC
2
= -SRz/(R1” +
Rz2)
S = 3wcob3Eo/2RI = g& + w&(KF, - 1) - acob3 (1 + K/2)
R2
= gs.&- WQ,D~(K, - 1) - gsb3/2D1= 3a&(K,gt - Ktg,)D2 = Z[W~~~~(K~
Kt)(Ws + Kt) + lgs -
gt)&s
+ &>I
2 = a3/[W2e02(2Ks +
Kt)’ + (2gs + gt)“]
and where a < r < b and ~0 is the permittivity of freespace. As can be seen in Table 1, the calculated fields inthe tissue and skin are strongly dependent on the as-sumed values of the tissue constants-no one set of whichcan be said to be correct to the exclusion of the others.More realistic geometries introduce even further varia-tions in the calculated fields.Despite the present inability to uniquely identify theinternal fields, Table 1 shows that the range of uncer-tainty is far below the levels associated with the knownmechanisms for ELF-field : tissue interaction-tissueheating and neural stimulation. It is clear, therefore, thatthere must exist an as yet unidentified mechanism.We think that, for now, the most fruitful approach tounderstanding most aspects of ELF bioeffects is at thesystemic level. We have suggested that ELF fields arebiological stressors and can elicit physiological changesthat are part of the classical stress syndrome (7, 12).During a stress response the body adapts to maintainhomeostatic conditions and this leads to strongly time-
 
SENSITIVITY TO CHANGE IN ELECTRICAL ENVIRONMENT
R425dependent alterations in peripheral blood composition(12). The exposure duration, therefore, could markedlyinfluence the likelihood of observing an effect. To explorethis possibility and to help provide the biological dataneeded to successfully frame a physical theory of ELFbioeffects we examined the erythrocyte component ofthe whole blood of mice in connection with Z-day expo-sures to an ELF electric field.
METHODS
We looked for changes in hematological parameters ofHa/ICR mice due to short-term exposure to a full-bodyvertical 60.Hz electric field of 5 kV/m.To ensure maximum statistical sensitivity every mousewas sampled twice, once after exposure to the field for 2days and once following a Z-day nonexposure period.
TABLE
1.
Calculated values of electric field and powerdissipated inside a mouse modeled mathematicallyas sphere of tissue surrounded by layer of skin
Tissue ElectricalConstants*Electric Field,V/m X
lo4
Power Dissipated,W/m3 X
10’
DielectricConduc-constanttivity, Tissue Skin Tissue Skinat-’ .m-’80 1 0.707 3.92 5.00 15480 0.33 2.08 4.49 14.3 66.58 x 10” 0.2 2.97 4.48 17.6 40.18 x 10” 0.02 10.1 61.9 20.4 7.66
Calculated values are strongly dependent on assumed values ofelectrical constants in each region. For simplicity, only
1
set of skinconstants has been considered. In all cases, a =
0.12
m and b =
0.15
m.Values in the skin depend on location; listed values pertain to the outerradius (r = b). * Skin electrical constants:
10, 0.1 SZ’-’ .
m-‘.
There were four consecutive experiments, two with malesand two with females. In each there were two groups: inone, the control period preceded the exposure period(nF ---) F) and in the other the pattern was reversed(F -+ nF).Our exposure facility has been described in detail (8).Briefly, the mice were housed individually in a nonme-tallic cage and vibration, light, light-dark cycle, and tem-perature were controlled. Two assemblies, each consist-ing of three pairs of shelves, were used; each shelf was ametal plate sandwiched between two layers of wood. Inthe assembly that housed the nonexposed mice the plateswere electrically grounded, and in the second assemblythe lower plate in each pair was energized and the upperplace grounded. We applied 1,590 V, which produced 5kV/m in the living space of each mouse in the exposureassembly; the 60-Hz field in the control assembly wasessentially zero. Except for the fields, the environment ofeach mouse was identical in all respects.On day 1 of each experiment the mice were dividedinto two groups and the electric field was applied to one-half the population. On day 3 the blood parameters weremeasured in each mouse and immediately thereafter theexposed and nonexposed groups were interchanged. Onday 5 the blood parameters were measured again and themice were killed.Blood was collected from the opthalmic vessels byinserting a 25-~1 capillary pipette into the orbit medial tothe eye. This procedure was performed in less than 20 sand did not produce any signs of trauma.Measurements were made using a Clay-Adams HA-5hematology analyzer. The freshly drawn blood was di-luted (1: 260) in a phosphate-buffered counting fluid con-taining an anticoagulant. This dilution was used for he-moglobin (Hb) determinations, and a second dilution was
TABLE
2. Means t SD of observed changes in hematologic parametersLn mace
ExperimentConditionRBC Hct Hb MCV MCH MCHC
1
2
‘1
2
1
2 1 2 1 2 1 2
A
Male control nF ---) nF (18)Female control nF + nF (20)
B
Male I F -+ nF
(16)
nF --) F (16)Male IIF --) nF (19)nF -+ F (19)Female IF ---) nF
(18)
nF --, F (18)Female IIF -+ nF (20)nF -j F (20)6.91
to.556.70to.336.14to.706.02to.746.99to.406.89to.626.41kO.566.41to.657.02kO.567.04to.547.03kO.556.96t,O.32*5.85 35.4t0.69” t4.65.7134.7t0.70*t4.66.36 40.7t0.46” t2.26.44 40.0t0.54* 23.76.15 37.1t0.72* zk3.76.00 37.1t0.71* 2k3.96.6541.8t0.52*t3.56.54 42.4t0.54*t3.440.0t3.538.8t2.040.8t3.440.4tl.B*33.6t4.333.0k4.537.0t2.8*37.2t3.4*35.4t4.8*34.6t5.9”39.3t3.4*38.5t3.3*13.3 12.721.2 *l.O*12.1 11.9to.7 to.6
NM
12.2 11.8to.5 t0.6*12.3 12.0to.6 t0.7*11.9 11.4to.9 &0.8*11.6 11.2tl.O &0.6*11.7 12.1k1.0 to.811.5 11.9to.8 to.557.0t2.157.5tl.157.2zk1.757.2t1.557.9zkO.857.8t1.557.6t1.657.5kl.859.4t1.459.9t1.457.6k1.057.6to.957.2to.757.3t1.257.7zk1.357.4tl.156.9t1.557.2t2.958.7-+1.5*58.5Ilr1.2*19.3 18.1t1.5*1.4*18.117.2t1.2zko.9*
NM
17.6 18.6tl.O t1.2*18.0 18.7t1.5 t1.2*18.6 18.7k2.0 t1.518.2 18.9t1.7t1.916.8 18.2t1.8t1.4*16.4 18.2k1.4 t1.4*33.3 31.2t2.7 k2.4”31.2 29.6t1.6 t1.5*NM30.2 32.0t1.6 &3.2*31.0 32.9k2.7
t1.9*
32.3 32.7t3.6 t3.231.5 33.0k3.3 k4.328.1 30.9t3.1 +2.7*27.3 31.0t2.6 t2.9*
A, no change in exposure condition;
B,
change in
exposure condition as indicated. NM, not measured.
No. of mice in each group is shown inparentheses. Units of each parameter are listed in text.
* P < 0.05.
of 00

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