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The first paper of this series (Lindstrom et al. 1974), summarized a set of assumptions
governing the oral ingestion, uptake, and movement of highly lipid-soluble drugs and pes-
ticides, such as HEOD1 (dieldrin), in mammals. These assumptions allowed a preliminary
model for the pharmacodynamics of dieldrin to be constructed. As more quantitative ex-
perimental information becomes available, the model will be refined to include this infor-
mation. For example, quantitation of dieldrin, DDT2, etc., movement across the lumen-
mucosa-blood barrier has yet to be made. Information on the kinetics and thermo-
dynamics of dieldrin binding to proteins, lipo-proteins, and even classes of lipids is
generally still lacking.
The line of reasoning we chose to follow in constructing a working model of the hy-
pothesis of pharmacodynamics of dieldrin (or other highly lipid-soluble drugs as well) in
mammals parallels that of Bischoff and Brown (1966), Dedrick and Bischoff (1968),
Bischoff and Dedrick (1970), Bischoff et al. (1971), and Dedrick et al. (1973). These
workers began with fairly simple hypothetical models for the drugs thiopental and
methotrexate (MTX). As more a priori information became available, it was included in
the models, making them more complete and accurate in their predictive value. Thio-
pental and MTX are quite water-soluble and much less lipid-soluble than dieldrin or DDT.
Dieldrin has a partition coefficient of the order of l0 s for a peanut oil/water system, and
DDT has a partition coefficient of the order of 106 for the same system (Wauchope
1972). In view of this highly important difference, it is conjectured that the excellent and
11,2,3,4,10,10-hexachloro-6,7-epoxy-l,4,4a,5,6,7,8,8a-octahydro-l,4:5,8-endo, exo-dimethanonaph-
thalene
21,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane
Lindstrom et al. (1974) concluded their paper with examples of estimated dieldrin
distributions in a hypothetical rat and a hypothetical man using several modes of dieldrin
administration (e.g., ad lib dietary, a single capsule daffy, intravenous). All blood flow and
compartment parameters (including the dieldrin destruction rate coefficient) were as.
sumed to be constant in time. This paper presents several more examples of estimated
dieldrin distributions in a hypothetical rat when: 1) auto-induction of hepatic microsomes
is allowed, 2) the adipose compartment is allowed to grow linearly from some base value
in time, and 3) the model is compared particularly with experimental studies of dieldrin
pharmacokinetics.
Discussion
Case I. Role of auto-induction. Durham (1969) has posed the question: Do pesticides
acting as hepatic inducers affect their own metabolism? Because dieldrin is a potent
inducer of hepatic mixed function orddases (Ghazal et al. 1964, Gillett and Chart 1968)
which include enzymes involved in dieldrin metabolism (Matthews and Matsumura 1969),
it may, be postulated that dieldrin is capable of altering its own metabolism (e.g., auto.
induction) and that the time course of tissue concentrations and the levels of residues
should reflect effects related to increases in dietary exposure. If auto-induction is strong
(e.g., if/~vl, the coefficient of induction of specific dieldrin-metabolizing enzymes, is
large and positive), residues in tissues would be expected to be markedly lower, relative
to dosage, as dosage is increased. In the initial preliminary model (Lindstrom et al. 1974),
l~pt was assumed to be zero. It is necessary, however, to test the hypothesis that dieldrin
does affect its own metabolism; to that end, application of the model serves a very use.
ful function.
If the level of dieldrin intake influences the relationship of residue to exposure level,
then simulation of residues in rats exposed to several levels of dietary dieldrin at several
arbitrarily set values of/3p t should provide estimates of a value of/~p 1 yielding residues
comparable to those measured in actual feeding experiments. If this value of/3p I is large,
then auto-induction can be assumed to play a role in dieldrin pharmacokinetics; con-
versely, a low value for/~p I would suggest that auto-induction is a negligible factor.
Table I (Lindstrom et al. 1974) lists estimates of the blood lipid flow and compart-
mental lipid mass for each of the respective compartments in a hypothetical 300-g
mature male rat eating 20 g of food per day during a 12-hr interval. The model is assumed
to be strictly flow-limited and all compartments to remain constant except for induced
changes in lipid and enzymes in the liver. The parameters characterizing F, the hepatic
dieldrin metabolism coefficient, are listed in Tables II and III. The absence of some par-
ticular values in Table II arises solely because of the assumptions for this simulation that
/3p2, 3 and S = 0. Table III sets forth the values of/~pl for low (1), intermediate (10), and
strong'(100) induction by dieldrin of dieldrin-metabolizing enzymes, the values of three
dietary levels of exposure (~), and the model simulation of the adipose lipid concentra-
Table I. Compartment mass, blood flow, and other important parameters for a hypothetical 300-g mature male rat U,
Oo
Compartment Symbol Mass Lipid fraction Lipid mass Blood flow Blood lipid
x 7]~x fx Mx rate flow rate
(g) ( g lipid ~ (g lipid) qx Qx
\ g tissue / ( g blood
hr ) ( g blood
hr lipid )
>
co.
~'Pi g dieldrin Kcpi (1/hr) prote!,n. _ hr
)
g lipid - hr KDi \ g lipid
O
2 0.006 0.0 - 28 t~
3 0.012 0.0 - 1
aThese estimates were obtained from data supplied by Davis et al. (1973) and by Davis (1973).
',D
170 F.T. Lindstrom et al.
tions (CF) of dieldrin at the end of a 4-day feeding period for each of the combinations
~Pl and ~.
The relationship of the resultant residue to the level of exposure as affected by/~p 1 is
shown in Figure 1, a log-log plot of adipose lipid concentration vs. daily dietary dieldrin
intake. As/~p I increases, the relationship of residue to intake become increasingly curvi-
linear exhibiting slopes progressively less than 1.0, the slope obtained with no auto,
induction or interaction of dietary level with residue. For each value of/~p l' least squares
regression lines can be calculated according to a power law rule (y = k - xn), providing
estimates (Table IV) of the constants for Equation (1):
C F = k(~) n. (1)
Walker et al. (1969) described the "steady state" adipose tissue residues (C~) of dieldrin
in rats by a power law equation:
where C~ is the whole adipose tissue residue Oag/g tissue) and CD is the dietary dieldrin
concentration 0ag/g food). Based on the assumptions used in our simulation (20 g food
consumed per day, 63.7% of the adipose tissue as lipid), the data of Walker etal. (1969)
can be expressed as:
CF = 0.169~ ~ (3)
Comparison of the parameters of Equation (3) with those for the simulations in Table IV
indicates only a small auto-inductive effect. This view is supported further by examina-
tion of the predicted residues at "steady state" conditions (when dieldrin input equals
dieldrin excretion plus metabolism). Using the parameters of Table IV in Equation (1) it
is shown (Table III) that "steady state" adipose residues in mature male rats (Walker et al.
Table lII_ Comparison o f hypothetical rat model with data based upon
the experiment o f Walker et at (1969)
1969) are very close to those predicted by the model assuming that auto-induction is
weak (~p 1 = 1.0).
Although true "steady state" conditions in an animal can only be achieved under cir-
cumstances where the compartment lipid mass, blood flow, metabolic rate, etc. are con-
stant (which never actually occurs in a live animal), the residues achieved in the model
simulation approach "steady-state" conditions (Figure 2). Even though the discontinu-
ous administration of dieldrin (12 hr feeding, 12 hr rest) imposes cyclic variations in
blood and adipose residues, by day 15 the differential in residues in these tissues varies
little from one day to the next, at the same hour. If the dieldrin-fed rat is then switched
to a control ration, and the loss from the adipose compartment followed for another 20
days, an approximation of the daffy destruction rate can be calculated. For the case
shown in Figure 2 (tip z = 1.0; ~ = 41.67 #g dieldrin/hr), the apparent first-order rate of
loss is about 12% per day. Values reported in the literature range from 5% to 20% per
100 ...."
/
..-~
"0
.../
O. o,*" &
E o..'~
o.O"
e-
~z,. . o.."
..~
~5 .:.~
...--
2n o ~176176176176
o.~~176
'~ 10
0C].
(~ ~ /3p]. = 1.0
& -- #PZ = 1 0 . 0
X ~ /~p]. = 1 0 0 . 0
1 I ! ! I ! ! ! I ! ! I ! | ! ! ! | I
10 100 1000
Daily dieldrin intake (Mg dieldrin)
Fig. 1. Log plot of adipose tissue lipid phase dieldrin residue (~g dieldrin/gm lipid) vs. the
log of the daily dieldrin intake rate (/lg dieldrin/day). Time t = 40 days on dieldrin.
172 F.T. Lindstrom et al.
day; Walker et el. (1969) found a value of 11% per day. In model simulations of the
"feed-off" period for greater values of #p 1 and ~ (not shown) the decay curves become
increasingly curvilinear and have steeper initial slopes as the value of BpI increases, but
are parallel between various values of ~(Lindstrom et al. 1974).
It is important to point out that with the assumptions used to construct the lipid-phase
model of dieldrin pharmacodynamics, even the assumption of a weak (tip I -- 1.0) auto-
induction is associated with marked changes in both dieldrin-metabolizing enzymes and
hepatic lipids. Figure 3 shows the relative increases in hepatic constituents in a hypotheti-
cal mature male rat (300-g, feeding 12 hr per day,/~Pt = 1.0, ~ = 41.67/ag/hr) during a
30-day exposure to a 25-ppm dieldrin diet followed by a 20-day feed-off period. In addi-
tion to the critical information on the actual mass (or total activity) of the several
enzymes of dieldrin metabolism (Figure 3a) together with the rate constants of each
activity, also unknown in real terms are data on the turn-over rate of these proteins. In-
creases in total and microsomal lipid (Figure 3d and 3e) tend to offset increases in
dieldrin-metabolizing enzymes, by decreasing the concentration of dieldrin available for
reaction, and hence changes in F (Figure 30 are relatively minor. At higher values of#p l'
F is greatly affected bemuse changes in p 1 are relatively large compared to lipid changes.
Since activity changes following the cessation of exposure do not persist as long as do the
rnorphologlc and structural changes (Ortega 1970), increased mierosomal lipid may
persist relative to induced dieldrin-metabolizing enzymes during feed-off (simulated by
setting kcp 4 < kcp 1)" As with several aspects of this preliminary model, detailed data on
hepatic activities and rates of change are vital to a more explicit and sophisticated
treatment.
Case II. The role of growth (a f'trst look). The growth of an animal during exposure to
a highly lipid-soluble agent such as a chlorinated hydrocarbon introduces numerous
physiologic and biochemical factors which are essential in the development of an a priori
model of residue build-up and distribution. Even in the sexually mature male rat, some
tissues (e.g., muscle, adipose, skin, etc.) are still growing. Not only is the lipid composition
of some tissues undergoing dynamic alteration, but the blood flow (and lipid flow) rates
to the tissues are also changing with growth. However, the assemblage of realistic growth
parameters suitable for incorporation in the preliminary model of lipid-phase dieldrin
pharmacokinetics is a disparate step that the currently sparse literature on lipid corn-
~Pl k n
1 0.172 0.97
10 0.180 0.86
positions of growing animals does not seem to be able to reconcile. Nevertheless, studies
by Schemmel e t al. (1970a, b) on obesity in rats do provide an opportunity to evaluate
the influence of growth of a single compartment (e.g., adipose) on dieldrin distribution.
By summing the weight of adipose tissue for the seven selected tissues assayed and
plotting the log of this sum against the log of body weight, the data of Schemmel e t al.
(1970b) provide a good power rule relationship for either a low- or high-fat diet (Figure 4).
Whether or not these curves for the Osborne-Mendel strain of rats are peculiar to that
strain or are applicable to other strains (e.g., Wistar, from which most other parameters
of the rat model have been obtained) obviously limits the validity of simulation. Examina-
tion of the data of Schemmel e t al. (1970b) allows a simple estimate of 0.2 g adipose
tissue/day for an average linear growth rate for rats on the low-fat diet beginning at a
body weight of about 300 g and growing to a body weight of 400 g in about 70 days.
That the sum of the seven fat depots shows a linear log-log relationship raises some
interesting questions concerning the overall mechanics of growth of adipose tissue.
To simulate the effects of adipose tissue growth, a mature male rat weighing 300 g
(at time t = 0) and consuming about 20 g of food per day is assumed to 1) exhibit weak
auto-induction (tip 1 = 1.0), and 2) to maintain a constant fraction of the adipose tissue
as lipid (fir = 0.637). All other parameters were identical to those employed in the pre-
vious simulations. It is necessary to calculate the small increment of blood volume gained
by the addition of the new depot tissue. Assuming that this increment is proportional to
the blood supply of adipose tissue generally [about 2% by volume (Specter 1956)], so
that the incremental volume is about 2% of the daily adipose tissue weight gain, and
assuming that the blood lipid fraction remains constant (fB = 0.004 g lipid/g blood), an
estimated 1.6 • 10 - 5 g of lipid would be added daily to the blood lipid pool. Actually,
similar increments are occurring in muscle, skin, etc. consonant with the general growth,
but are ignored for this illustration.
If the blood lipid flow through the fat mass is constant (QF/MF = Rt), evaluation of
100 jBiood
r O.
~=
mE
"o . _ 1 0
"o._
=9
O. v
"0
-J
5 10 15 20 25 30 35 40 45 50 55 60
Time (days)
Fig. 2. Typical semilog plot of blood lipid and adipose tissue lipid phase dieldrin residue
vs. time for a 60-day hypothetical feed-up/feed-off experiment on a 300-g mature male
rat.
174 F.T. Lindstrom et al.
(d)
I(a) Active inducible
[ mocrosomalprotein mass (P1) Microsomal lipid mass (MMs)
r
004 e ~
9 1 7 4 9 9
9 o ~ 9
CN
C/ . . . . .
2610
,
20
,
30
,
40
ii
2610
(b) (e)
A Non-active, inducible,
L.
microsomal protein mass (P4) Total liver lipid mass (M L)
1.2
E1 .(~ 1.0
g O 9 1 4 9 1 4 9o ~ 9 9 9 9 9 9 9 9 9 9 9 9 9 9
oe o o o o e e 9 9
cq .8
~.6 9 9149 9 9 9 9 9 9 9 9 ~.6
'- .4 3.4
o 99 o
~.2 2
0
2 ' 6 "1'0 20 30 40 50 o ~ '~'lb ~ ~ ~
A
(c) Total micros9 5 (f)
protein mass 2; Pi (t)--P4(t) Total destruction rate coefficient (F)
i=1
e o e 9 I ~ ~ ~ e ~ 3 'oeeoeeoo
~ . 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9
~I.0 eo
9 9
9 0
~ .8 o oeoe 9 9 9 9 9 9 --.~ . 2
o~ O.
~ .6
"- .4 ~.1
=.2
0
2' 6 'lb 2b 3b ~ ~ ~' ~' 1'0 2b ab ~ s~
Time (days) Time (days)
Fig. 3. Linear plots of some of the important liver morphologic responses to the hypo-
thetical long time chronic dieldrin insult.
Applications of Dieldrin Distribution Model 175
These equations (4-6) are then assumed to def'me the time-dependent relationships of the
growing adipose and blood pool compartments. The simulation format is completed by
specifying that the dietary concentration is 25 ppm for the first 40 days and 0 for 20
days thereafter.
Dieldrin residues in blood and adipose lipid (Figure 5) are compared for constant and
increasing adipose tissue compartments. Substantial and significant differences are appar-
ent. Although residues rise for the f~rst few days to about the same levels, the oscillations
of blood residues in the growth model become increasingly dampened. During this time,
the dilution of growth constitutes less than 3% of the adipose lipid. At the end of 40 days
exposure, the residues in the lipid of the "growing" rat are much lower (55 as compared
to 65/ag/g lipid); however the body burden of the growing rat is correspondingly higher
(Keane and Zavon 1969) than that of the constant Case I. The continuous addition of
1000
1.905
A Total adipose (grns) = 467 (B.W')
0
Dietary fat 60% by wt. ~ 9
\ ./
O3
/
~100
E
g~
E
/.
~
0
10
Q.
1.406
0
Total adipose (gms) = 99 (B.W.)
Dietary fat 2.7% by wt.
li t I I I I I | II 1 I I I I LIII I I I ! I II I
0.01 0.1 1.0 10.0
Body Weight (B.W.) Kg
Fig. 4. Log-log plot of Osborne-Mendel strain rat adipose tissue as a function of the body
mass (Schemmel e t al. 1970b).
176 F.T. Lindstrom et al.
lipid mass not only dilutes the adipose residue, but retards redistribution to other tissues.
Even after 40 days a true "steady state" is not achieved in the growing rat; the mean
blood and adipose lipid dieldrin residues are still rising. Residues at day 60 are about
15/~g/g of lipid in the growing model and 5.5 #g/g of lipid in the constant animal (Case I),
further reflecting retention of dieldrin in adipose and prevention of its metabolism in the
liver.
The average total body burden at 40 days is about 3.2 mg for Case I and about 3.36 mg
for the growing adipose case. Whereas this difference is only 5% at day 40, by day 60 the
average body burdens have dropped to about 0.26 mg and 0.88 rag; the difference then is
about 240%. One obvious implication of this situation is that attempts to free dieldrin-
contaminated individuals of residues during a period of adipose growth, by metabolism
alone, may not be as effective as during times of no growth.
A second implication is that the effects of growth simulated in this limited fashion by
the preliminary model cannot account for disparities between the observations of Barren
and Walton (1971) and this model. Although the decay curves for the growing compart-
ment model are more curvilinear than those for the constant compartment case (not
easily seen in Figure 5 due to figure reduction), no combination of adipose compartment
growth rate, dieldrin-metabolism rate (KD 1) and induction coefficient (tip 1)' and enzyme
catabolic rate (kCp 1) could be postulated which produced a simulation with: a) a negative
slope to the residues (plotted vs. time) at the terminal portion of the exposure prior to
cessation of exposure; b) the experimentally observed residue level at that point; and
c) the observed residue level 20 days after exposure ceased. It would thus appear that the
hypothesis of a strictly all tissue flow-limited a priori model may be in error. However,
sparse literature data relating parameter~ to specific time portions of the simulation are
the primary limitation. For example, how was the proportion of lipid (fF) changing in
the adipose tissue of the experimental rats? Is the model in error or are subordinate
o 100 rBIood
~._~
-~. 10
"0
_.1 1.
5 10 15 20 25 30 35 40 45 50 55 60
Time (days)
Fig. 5. Comparison semilog plots of adipose and blood lipid phase dieldrin residues. Top
curves are for the constant body mass 300-g mature male rat. Bottom curves are for the
growing adipose and blood compartments (linear growth). Growth begins at t = 0 with
initial body mass being 300 g.
Applications of Dieldrin Distribution Model 177
assumptions inadequate? It would appear that this latter situation - the lack of appro-
priate subordinate assumptions and data - prevents validation of this model in every
instance. More probably, the system is neither flow-limited nor membrane-transport
limited, but is in fact a combination of both transport mechanisms (Dedrick e t al. 1973).
One can hypothesize that muscles high in phospholipid are so due to the presence of
proportionately greater amounts of sacrosomal and plasma membranes consistent with
greater metabolic activity, and therefore these muscles could be expected to be more
highly perfused. If the differential perfusion rate were sufficiently great, and if the lipid
composition differences are solely related to perfusion-controlling factors, then under
the regimen used to generate the data of Yadrick et al. (1971) (nine doses in a 13-day
period) it might be possible to simulate residue differences between tissues or muscles in
the preliminary model in some relationship to the phospholipid content. Attempts with
the preliminary model to generate simulated residues that demonstrate dependence on
perfusion rates beyond the first few hours after each dose have been unsuccessful. For
example, a three-fold difference in perfusion rate between two muscles having a similar
three-fold difference in percent phospholipid (and an inverse three-fold difference in
Liver 60 25 33 8
Kidney 50 24-27 28 9
Brain 97 45-47 3 2
Heart 27 (13-15) c 19 11
Adipose 600-900 20 11 4
Blood - - 30 7
total lipid) yields residues differing by 133% after one hour, less than 5% by the fifth
hour, and less than 1% after the full 13-day regimen. Thus, it would appear to be neces-
sary for perfusion rates to differ by at least an order of magnitude, and probably more, in
order to be effective in generating residue differences over the three-fold range found by
Yadrick et al. (1971). Since the muscles examined do not differ that greatly in phospho-
lipid concentration, it would appear that the association of high percentage phospholipid
with high residues may be related through some factor other than increased vasculariza-
tion or simple binding of dieldrin to phospholipid.
lipid dieldrin) vs. time plot. No realistic growth rate of adipose tissue alone could afford
simulation of such data. However, if the efficiency of absorption (0) from the ingestion
were to decline with age, then declining residues would occur in spite of constant food
intake. - --
Conclusions
Viewed from the point of the adipose tissue residues, the preliminary model of lipid
phase dieldrin pharmacokinetics behaves as if intracellular and inter-organ compart-
mentalization, whether by differential tissue uptake, selective binding, or " . . . orderly
active turnover...", is a factor of lesser significance than the flow-limiting considerations
inherent in this system. Excellent representations of adipose tissue residue accumulation
with exposure to dieldrin and decay with subsequent cessation of exposure under a
variety of dosage regimens can be simulated for the mature male rat. Auto-induction of
dieldrin metabolism appears to play a relatively minor role in the disposition of ingested
dieldrin. Current physiologic growth data is either lacking or inadequate to simulate
accurately the effects of growth on dieldrin lipid-phase residues, although the preliminary
model behaves generally as predicted from studies such as those of Keane and Zavon
(1969). It would appear also to be useful in evaluating certain hypotheses regarding even
some types of compartmentalization.
100.0
"O
=.
O
O.
~I0.0
E
e-
I.O
" ' 10 ,5 2'5 I 5o
On dieldrin (500/~g/12 hrs/day) Time (days) 30 Off dieldrin
Fig. 6. Semilog plots for two hypothetical cases of feed-up and feed-off, illustrating the
effect that auto-induction has on long time adipose tissue lipid phase dieldrin residues.
Barton and Walton's (1971) feed-off data (vertical bars) plotted for comparison.
Applications of Dieldrin Distribution Model 181
Acknowledgment
This research was supported by U. S. Public Health Service Grant ES 00040 from the
National Institute of Environmental Health Sciences. This paper is issued as Technical
Paper No. 3787 from the Oregon Agricultural Experiment Station.
References
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182 F.T. Lindstrom et al.