United States
 Environmental Protection
 Agency
 Health Effects Research
 Laboratory
 Research Triangle Park NC 27711
 Research and Development
EPA/600/S1-85/016  Sept. 1985
 Project Summary
 Dosimetry for In-Vitro  Chick
 Brain Calcium-Ion  Efflux
 Experiments  by  Numerical
 Methods  Based  on  Zonal
 Harmonic  Expansions

 Guillermo Gonzalez and James C. Nearing
  The full report discusses the calcula-
tion of the electric field and power den-
sity distribution in chick brain tissue
inside a test tube, using an off-center
spherical model. The off-center spheri-
cal model overcomes many of the limita-
tions of the concentric spherical model
and permits a more realistic modeling of
the brain tissue as it sits in the bottom of
the test tube surrounded by buffer solu-
tion. The effect of the unequal amount
of buffer solution above the upper and
below the lower surface of the brain is
evaluated.
  The field  distribution is obtained in
terms of a rapidly converging series of
zonal harmonics. A  method that per-
mits the expansion of spherical harmon-
ics about an off-center origin in terms of
spherical harmonics at the origin is
developed to calculate in closed form
the electric field distribution.
  Numerical results  are presented for
the adsorbed power density distribution
and scaling ratios at carrier frequencies
of 50 MHz.  100 MHz, and 147 MHz. It
is shown that the off-center spherical
model yields scaling  ratios in the brain
tissue that  lie between the extreme
values predicted by the concentric and
isolated spherical models.
  This Project Summary was developed
by EPA's Health Effects Research Lab-
oratory, Research Triangle Park, NC, to
announce key findings of the research
project that is fully  documented in a
separate report of the same title (see
Project Report ordering information at
back).
Introduction
  A problem of considerable importance
to the understanding of biological phe-
nomena produced by electromagnetic
radiation is the mechanism of interaction
of amplitude-modulated radio-frequency
[RF] waves on calcium-ion efflux from in
vitro chick brain. Experiments have shown
that when amplitude is modulated with
extremely low frequency [ELF] sinusoidal
waves, RF radiation  can enhance the
calcium-ion  efflux from  in vitro chick
brain tissue.
  In order to better understand the effects
of amplitude-modulated RF radiation on
calcium-ion efflux, the electromagnetic
fields and absorbed power density distri-
bution in the chick brain must be accu-
rately determined. Concentric spherical
models have been used to calculate the
field and power density distribution in the
chick brain. These models have provided
much valuable  information about the
dosimetric aspects of the calcium-ion
efflux experiments. The models have also
been  used to develop scaling ratios to
determine the required incident power
density that produces calcium-ion efflux
at other carrier frequencies.
  This study overcomes the limitations of
the concentric spherical models. Concen-

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trie spherical models do not adequately
represent the chick brain half immersed
in the buffer medium, because the brain
as it sits in the bottom of the test tube is
surrounded by a layer of buffer solution
with a thickness that differs above and
below  the brain.  Concentric  spherical
models use a uniform layer of buffer solu-
tion to surround the brain. To determine
the dosimetric effects of the non-uniform
layer,  an off-center spherical  model  is
used to approximate the physical geom-
etry of the brain/buffer combination. The
off-center spherical model represents a
significant refinement to the concentric
spherical model, since the brain can be
modeled as a continuous, smooth, spher-
ical  surface  surrounded  by  unequal
amounts of buffer solution on its upper
and lower surfaces. The electric field and
absorbed power density distributions, and
scaling ratios for  carrier  waves of 50,
100, and 147 MHz are calculated using a
quasi-static approximation. This approx-
imation leads to a field representation in
terms  of a convergent series  involving
spherical harmonics.


Procedure
  The schematic of the experimental set-
up is shown in Figure 1. The chick brain
tissue sits at the bottom of the test tube
and is surrounded by a non-uniform layer
of buffer solution.
  The analytical model for the off-center
spherical model of the brain inside a test
tube is shown in Figure 2, along with the
direction of the incident electric field. This
model represents the geometry in Figure
1.1 n fact, except for the flat-topped buffer
region, the model is reasonably accurate.
The radii a and b, and the off center dis-
tance d, are selected so that the brain
volume occupies 0.3 ml, and the buffer
volume, 1 ml. Also, the limiting case as h
approaches zero is analyzed.
  The mathematical formulation that fol-
lows is by no means a simple extension of
the concentric spherical case. To the best
of the  authors' knowledge, exact solu-
tions to off-center spherical geometries
of the type shown in Figure 2 in terms of
zonal  harmonic expansions have not
appeared in the literature.
  The  mathematical formulation of the
problem follows.  The electric  field, as-
suming quasi-static conditions, can be
expressed as

               E = -V0

where  is the electrostatic potential. The
quasi-static approximation is  valid be-
cause of the dimensions, frequencies,
and electrical properties of the materials
involved. In our calculations, the relative
complex permittivity of the brain tissue at
the typical carrier frequencies  used in
               -1.42 cm -
   Buffer
   Solution
                             0.24 cm
calcium-ion studies are 100-J175 at 50
MHz, 72-J94.9 at 100 MHz, and 66-J69.2
at 147 MHz; for the buffer, the values are
71-J580 at 50 MHz, 71-J290 at 100 MHz,
and71-j200at147MHz.
  The  externally applied field, which
makes an angle of 90° with the z-axis
(i.e., JE = E0Uy),  produces a  potential
which can  be expressed in  spherical
coordinates in the form
                                                  V.It = -
                           Test Tube
                                         or
                                         where
          V.x, = -
   Brain
   Hemisphere
Figure 1.    Schematic of the  chick brain
            inside a test tube containing a
            buffer solution.
     Y5m (ft 0) = cos m0 P'S (cos 0)

Pi! (cosfl) are associated Legendre poly-
nomials.
  The potential in regions I, II, and III sat-
isfies Poisson's equation, subject to the
appropriate  boundary conditions. The
charge distributions at r. = a and n> = b
produce potentials V« and Vb of the form
Figure 2.    Off center spherical model for the chick brain.

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       V. =
        r°    rn
  Va,,n =\      \
                                     ()nYnm(0a, 0)  ( ra <
                                                                            (D
              Va out =

                                                              > 3
. and
       Vb =
               b ,n

              Vbou, =

                                         )" Y£m ( ft,, 0 )  ( rb <  b
                                       rb
 V, = V., in + Vb, out + V.,,


 V,, = V., out  + Vb, out + V.,»


 Vln = V., ln + Vb, in + V.,«
        /a, in
•r    r
 Ln=Qt.m=(
 I
 and
        b, ou, =
n

s=m

oo



 n

 s=m
              r
              »-s=
                                                                            (2)
 where anm and ftnm are constants to be
 determined from the boundary conditions.
 In terms of (1) and (2), the potentials in the
 three regions are given by
                       (3)

                       (4)

                       (5)
   The form selected for the potentials in
 (1)and(2) satisfy the continuity conditions
 at r,  = a  and  rb  = b. Therefore, the
 boundary conditions that  remain  to be
 satisfied at the interface are the continuity
 of the normal component of the current
 density at r, = a and rt> =  b.
   In order to apply the boundary condi-
 tions  at the surface  of  the off-center
 sphere (i.e., at rb =  b), we need to expand
 the solution V., m in terms of the coord-
 inates rb, 0b and 0, and the solution Vb, out
 in terms of the coordinates ra, 0a, and 0.
 The necessary expansions are
                                   (6)
                      , 0)


                       (7)
                             where
                                         and
  (n-s) I (s+m) I

 d'"n(s-m)l
(n-m) I (s-n) I
  Applying the boundary conditions to
(3), (4), and(5), and using (6) and(7), a set
of linear equations for trie n=j (j=0,1,...)
and  m=k (k=0,1,..,n) coefficients (i.e., ctjk
and /3|k) are obtained.
  Fixing the maximum n to be Nmax, the
set of linear equations is solved for a(k and
/3|k. The results are then inserted into (1)
to (5) to calculate the potential,  electric
field, and absorbed power density distri-
bution in the three regions.


Results and Discussion
  The absorbed power density distribu-
tions, assuming E0=1 V/m, were calcu-
lated for several cases; the most impor-
tant  was for a brain volume of 0.3 cm3,
buffer volume of 1 cm3, and h=0.05 cm.
Therefore,  a=0.677 cm,  b=0.415  cm,
d=0.212 cm, and  t=0.474 cm. Also, the
limiting cases as h and d approach zero
are considered. For h — 0 (i.e., when the
spheres touch at the bottom) the dimen-
sions  are  a=0.677  cm,  b=0.415  cm,
d=0.262 cm, and t=0.524 cm.
  Figures 3 and 4 graphically illustrate
the absorbed power density distribution
produced by  an incident  carrier wave,
with E0=1 V/m, at 147 MHz for the cases
where h=0.05 cm andd — 0, respectively.
Inthe figures, each line represents a con-
stant absorbed power density level. The
                                         Figure 3.    Absorbed power density distri-
                                                     bution at 147 MHz. The dimen-
                                                     sions are 8=0.677 cm, b=0.415
                                                     cm. d=O.212 cm. h=O.OS cm and
                                                     t=0.474 cm. The number scale
                                                     (0 to 9) is linear, where 9 repre-
                                                     sents an absorbed power density
                                                     of 0.5 mW/m3 for an incident
                                                     field of Eo=7 V/m.
                            Figure 4.   Absorbed power density distri-
                                       bution at 147 MHz for a cen-
                                       tered spherical model (d=O). The
                                       dimensions are a=0.677  cm,
                                       b=O.415 cm. andt=h=0.262 cm.
                                       The  number scale (0 to 9) is
                                       linear,  where 9 represents an
                                       absorbed power density of 0.5
                                       mW/m3 for an incident field of
                                       Ea=1  V/m.
                            number scale is linear, ranging from 0 to
                            9, where 9 represents an absorbed power
                            density of 0.5 mW/m3. For example, the
                            value of 2 represents an absorbed power
                            density of  (2/9)0.5=0.111  mW/m3. In

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Figure 4, the absorbed power density dis-
tribution in the brain region is constant at
a value of 0.109 mW/m3.
  Absorbed power density patterns sim-
ilar to those shown in Figures 3 and 4 are
produced at 50 and 100 MHz, except that
the absorbed power levels are smaller.
  It is significant to observe from Figure 3
that the absorbed power density distribu-
tion in the brain region is not uniform; the
bottom  of the chick brain has a higher
absorbed power density than the top.
Also, in Figures 3 and 4, the surface fields
around the chick brain exhibit a signifi-
cant variation from the bottom to the top.
  Scaling relations were calculated  for
the off-center spherical model. Figure 5
gives  the values of the scaling factor, P,
(50)/Pi(147), for points along the z-axis
inside the spherical  brain between the
top (i.e., z=0.415 cm) and  bottom (i.e.,
z= -0.415 cm). Figure 6 gives the values of
the scaling factors for points on the out-
side surface of the brain between the top
(i.e., £=0) and bottom (i.e., 8=ir) from 0 <
0< it. The scaling ratios at a given fre-
quency are continuous at the top and bot-
tom of the brain-buffer interface. Thisfol-
lows from the fact that the electric field is
tangential and continuous at the top and
bottom brain-buffer interface.

Conclusions and
R ecommendations
  The off-center spherical  model was
used  to calculate  the electric field and
absorbed power density distribution in
chick brain tissue inside a test tube. The
calculations were made with typical di-
electric data used in previous analytical
studies for carrier frequencies of 50 MHz,
100 MHz, and 147 MHz. In the experi-
mentally realistic case where the chick
brain is surrounded by lesser amounts of
buffer solution at the bottom, the pattern
of absorbed power densities showed  a
marked concentration of absorbed power
near  the bottom.  The absorbed power
density within the brain showed only
gradual variations with position. Outside,
at the surface of the brain, the absorbed
power density distribution varied by fac-
tors of 6 to 10.
  The scaling ratios were calculated as a
function of position for points inside, and
on the outside surface of the brain. The
off-center spherical model  revealed that
the scaling ratio inside the brain tissue is
not constant; the maximum change (h=0
case) from the top of the brain to the bot-
tom was approximately 11% for P45Q)/
PrflOO), and 19% for Pi(50)/Pi(147). This
contrasts with the concentric spherical
       7,-
 £    6	
                                                                     d=O
                                                                     h=.OS cm
                                                                     h=O
       0.415       0.249       0.083   0  -0.083

                                   z-axis (cm)
             -0.249
-0.415
Figure 5.    Scaling ratio values for P> (50)/P\ (147) along the z-axis of the brain.
        0     18    36    54    72    90   108   126   144   162   180

                                Theta (degrees)

Figure 6.   Scaling ratio values for P\ (50)/P\ (147) along the outside surface of the brain as a
           function of Q.
model, which predicts  no change. As
expected, the scaling ratios near the top
are essentially identical to the values
predicted  by the concentric spherical
model; at the bottom, the values become
closer to those predicted by an isolated
spherical  model. Thus, the off-center
spherical model yields  internal scaling
ratios that lie between the extreme values
predicted by the concentric and isolated
spherical models.
  Recent  experimental studies have
shown that the penetration of radioactive
calcium-ions in chick brain tissue was no
greater than  1  mm from  the  surface.
Thus, it may be concluded that surface
field values are of paramount importance.
The profiles for the surface scaling ratios
are symmetric for the concentric spheri-
cal case, with the peak value occurring at
a point on the surface (0=90°), where the
external electric field is normal to the sur-
face. However, the off-center spherical
model  yielded somewhat lower  peak
values whose position shifted toward the
bottom (0=*99°). In addition, the surface
ratios were similar at the top of the brain
for all the models, but the values at the
bottom varied considerably, with the h=0
case approaching the value given for an
isolated sphere.
  The scaling ratio results have provided
new insights into the interpretation of the
calcium-ion efflux phenomena. The exper-
imentalists can use the information pre-
sented  in this study to interpret their
results and  to formulate theories about
calcium-ion efflux phenomena.
                                                                             S. GOVERNMENT PRINTING OFFICE:1985/559-l 11/20680

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     Guillermo Gonzalez and James C. Nearing are with University of Miami. Coral
       Gables. FL 33124).
     Ronald J. Spiegel is the EPA Project Officer (see below).
     The complete report, entitled "Dosimetry for In-Vitro Chick Brain Calcium-Ion
       Efflux Experiments by Numerical Methods Based on Zonal Harmonic Expan-
       sions, " (Order No. PB 85-227 528/AS; Cost: $8.50, subject to change) will be
       available only from:
             National Technical Information Service
             5285 Port Royal Road
             Springfield. VA 22161
             Telephone: 703-487-4650
     The EPA Project Officer can be contacted at:
             Health Effects Research Laboratory
             U.S. Environmental Protection Agency
             Research Triangle Park. NC 27711
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
Official Business
Penalty for Private Use $300

EPA/600/S1-85/016
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