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MICROCOPY RESOLUTION TEST CHART
NATIONAL BUREAU OF STANDARDS - 1963
MICROCOPAL BUREAU O

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Orun-Pullna
Submitted to: Journal of the
Amer. Veterinary Radiology Soc):
† APR 27 1988
CFSTI PRICES
CESTI PRICESP
H.C. $ 1.00; MN_.ID
LATE SOMATIC EFFECTS OF RADIATION EXPOSURE
* MASTER
1
.
IN LABORATORY MICE*
G. E. Cosgrove
Oak Ridge, Tennessee
RELEASED FOR ANNOUNCEMENT

The report w preparada a nocom al domnunat sponsor worth Mathar the Waited
motor, mor the Comminalom, nor any person nottu on h all of the Commission
A. Muu my warrut or representation, apewond or implied with repect to the noch
rhoy, bons platum, or whose of the tutoraation contaba la dis report, or that the one
olan t w appar , mother or powma dinclou la theroport may not Intranya
pentrumly owned rights or
3. A . m uamuita wa runot be the wm ole or for den gamlehny trom the
une a un baloration, apparatus, method, or procu dimelound in the report
-LEGAL NOTICE -
IN NUCLEAR SCIENCE ABSTRACTS
magna or contractor of the Commission, or employee of che contructor, to the stunt that
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denomination, or provide recono to, ay tumormation perinant to do apoyant or contract
when the Coundation, or Me employment with me contractor.
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*Research sponsored by the U. 8. Atomic Energy Commission under contract
with Union Carbide Corporation.
.
.
Send Proof to:
DR. G. E. COSGROVE
Biology Division
Oak Ridge National Laboratory
P. 0. Box Y
Oak Ridge, Tennessee 37831
.
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INTRODUCTION
The effects of radiation injury are usualiy divisible into two temporal
categories, acute and delayed. The amount of information on the acute effects
of ariation in mammals 18 zruch greater than the amount of information on
delayed effects, in spite of tue importance of the latter. The more obvious
reasons for this include the long time necessary for such studies and the
corresponding greater expense. Even so, information on the delayed effects
of radiation in mammals 18 accumulating rapidly. The nature and description
of the delayed effects which follow are based largely on my experience witia
Irradiated mouse populations. Studies of the pattern of delayed radiation
injury in mice can be very informative but the degree of applicability to
higher animals and man is not clear at present. Summaries of delayed radiation
effects can be found in a number of good radiobiology texts; some with general
biological orientation (e.g., 1), while others are more concerned with human
radiation effects (e.8., 2). Long-term studies of laboratory nice exposed to
radiation have been done on animals injected with Srº (5), exposed to an
atomic explosion (8), or exposed to whole body x-rays (4), as well as many others.
In this article some interesting and well-studied delayed radiation effects in
nice will be discussed.
MATERIALS AND METHODS
In our laboratory, several different strains of mice have been used
in experiments designed to study late somatic effects of radiation. One suche
experiment will be discussed more fully here (comprovata).
were obtained
Female 103F, [(101 x c3H/Anf)F, ) mice from Cumberland View Farms, Clinton,
obase
Tennessee. Wher-the-ntoa wapo* I2 weeks Ahose weighing 22 to 26 gm
were randomized and given whole-body exposures of 350 to 1300 r of 250-kvp
X rays, with physical factors of 30 ma, 80 cm target-to-mouse distance, inherent
filtration of 0.1 mm Al, 3 mm Al added filtration, HVL of 0,55 mm Cu, and dose
rate of 80 to 90 r/min. The 'animals exposed to 1300 r were given an 1.p.
injection of 9 mg. of AET (S, 2-aminoethylisothiouranium) 15 minutes prior to
irradiation. During irradiation, the mice were rotated at 6 to 8 rpm in a
secondo circular Lucite exposure cage. Other animals from each shipment were
maintained as unirradiated controls. Animals dying before 60 days were not
considered in the analysis of long-term effects. After introduction into the
experiment, animals were housed 10 per cage, with Purina laboratory chow and
drinking water freely available. As deaths occurred, animado from adfecent
Lager Were Houel.ly pooled.to keep the number of animatot the cage between
Sant to. Animals from selected cages of each experimental group were checked
at intervals for graying of the fur and opacities of the optic lens. Graying
was estimated as described previously (6), being graded 0 to 4 for each of
five locations along the back of the animal from forehead to rump and for one
other location on the flank. Grade O indicated no change in the normal agouti
brown hair of the mouse; grade 4 indicated all white hair with no residual
pigmentation. The intermediate grades 1, 2, and 3 were based on the observer's

estimates of the mixtures of normal color, gray and white. The six location
figures were totaled to obtain a single value representing over-all graying
with a range of 0 to 24. The lens was observed with a siit lamp, and the extent
of opacification of the lens cortex was graded 0 to 100% as described elsewhere
(3). At death, necropsies were performed on all animals except those that
were too decomposed or eaten Kess than 10%)ctmanimats were udsuitable for
graco exemitiation for these TESPORT The head-190-Hot routtuety opened beat
worseraufned if the previoue behavior_of_tha animal or some external change.
indiented the possible existence of the intracrandet locion. If the animal
was not decomposed, specimens of major organs and diseased areas were taken
for histological examination, approximately 20% of the animals in each group
being thus examined. Histological preparations were routinely prepared by
paraffin embedding and hematoxylin and eosin (H&E) stainingo cocasionally
specie insteino-vere-toed. Findings entered on individual animal cards., prepared at
the time the animal was originally put in the experiments were to per transcribed
to IBM cards for data processing with subsequent statistical analysis of
survival and incidence of lesions. In this experiment, group sizes were
from 100-200 mice. This is a specific example of the experimental design often
used by us. Many changes in design can be made; e.g., age at exposure, type
of radiation, and use of shielding, protective or recovery agents among others.
· were
UIN
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RESULTS
.
Survival. In the experiments described above, 350 and 500 r whole-body
exposures resulted in no acute radiation deaths, 1.e., these doser were 1D/30
days. Exposure to 750 r whole-body and 1300 r whole-body with use of AET ( the ar
radioprotective chemical resulted in approximately 50% mortality within 30
days, i.e., LD50/30 day. Table 1 and Fig. 1 illustrate the survival data of
the above groups. Lite shortening was proportional to dose except for the
overlap in 350 r and 500 r exposure groups. The mean age at death of the
unirradiated control group was 866 days and this was reduced to 460 days
in the group exposed to 1300 r and treated with AET. Average life shortening .
calculated in days/r decreased with increasing dose from .64 days/r in the group
exposed to 350 r to .31 days/r in the group exposed to 1300 r (Table 1).
Neoplasms of reticular tissues - leukemia and lymphoma. The 10 F, 18
a low leukemia/lymphoma strein of mouse. Tumors of reticular tissues were
found in about 9% of unirradiated controls (Table 2). Irradiation up to (T-2)
750 r whole-body exposure levels increased the incidence of thymic lymphoma,
induced some myeloid leukemia and had no consistent effect on the other
lymphomas. By comparison, the RF mouse in uss in this laboratory is a high : C
leukemia/lymphoma strain of mouse and induction of thymic and myeloid forms by
radiation is pronounced while the other lymphomas fall in incidence in irradiated
groups (Table 3). There are sex differences in incidence in the RF strain.' (T-3)
In both strains the mean age at death with thymic lymploma 18 early (~ 10 months),
with myeloid leukemia about 13 months, and with the other lymphomas nearly '-,
coincides with the mean age at death from all causes.pl (Tables 2-3).
Non-reticular neoplasms. In 1C3F mice ovarian tumors are induced by
radionice exposures et LD go levels or below. The incidence in unirradiated controls
is 30%, reaches a maximum of 73% arter 350 r exposures, and declines with
higher exposures (Table 4). Marmory tumors are induced by radiation but lung (T-4)
tumors decrease in incidence in irradiated groups (Table 4).
Degenerative changes. Glomerulosclerosis is uncommon in unirradiated
103F, nice (2%) and in radiation exposures below the LD 50/30 (1-4%). With
750 r the incidence increases to 25% and with 1300 r reaches 72% (Table 4).
Graying of the fur occurs after irradiation of the agouti-brown 1c3F, mouse
and the severity is related to the radiation exposure level (Table 4). After
exposure to 750 r or more the hair eventually turns white.
Opacity of the optic lens (cataract) also is induced by irradiation and the
severity and rate of progression are dose dependent as illustrated in Fig. 2. (F-2
There are many other radiation effects which are of lesser importance or
not 80 obvious which will not be mentioned here.
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DISCUSSION
Survival.
When a group of mice is exposed to radiation the number
and duratior of radiation as weil as on the age, physiology, and physical
condition of the mice. In recent years, animals have been enabled to survive
.
C
.
om
or therapeutic measures. The survivors of the post-radiation 30-day period
of acute lethality usually have a latent period of many months before
appreciable amounts of mortality begin again. When deaths appear in the group
after this latent period, the shape of the group mortality curve tends to mimic
that of a mortality curve for unirradiated control animals of the same species
but occurs earlier in time (Fig. 1). Within certain limits, there is correlation
between original radiation dose and time onset of the late mortality pattern.
This is reflected in the earlier mean age at death with higher exposures,
but the life shortening effect per R of exposure is less with higher exposures
(Table 1). The group mortality curve can be considerably affected when radiation
induces a marked increase in the frequency of a disease having a mean age at
death far from the norm for the group.
UNT
Induction of disease. The same types of disease appear in irradiated as
in control mouse groups but the incidence may be very different in the 2 groups
(Tables 2,3,4). Some of the diseases appearing in higher incidence in
irradiated groups are very important causes of death; these include infections,
neoplasms, and vascular lesions. Interesting non-lethal lesions include skin 33
atrophy, greying of hair, and degeneration of the lens of the eye.

Reticular neoplasms. The leukemias and lymphomas (reticular neoplasms)
are so prominent in long-term studies of irradiated animals that they are
usually considered separately from the reinainder of the neoplasms. In long-term
studies of irradiated mouse groups there are two forms of reticular neoplasms
which are commonly increased in incidence by radiation (7); these are thymic
lymphoma and myeloid (granulocytic) leukemia (Table 3). Thymic lymphoma
starts in the thymus with rapid enlargement due to proliferation of immature
lymphoid cells. These cells may appear in many other organs, especially liver,
spleen, and lymph nodes after the initial thymic phase. In some mice with
thymic lymphoma, there is a true leukemic syndrome with the presence of
III
abnormal lymphoid cells in the peripheral blood.
Myeloid (granulocytic) leukemia is a disease with marked spleen and liver
enlargement due to infiltration by immature cells of the granulocytic series
(Fig. 3). There is lesser but widespread involvement of lymph nodes and
other organs. The peripheral blood usually contains immature granulocytic
(F-3)
cells.
The other common forms of reticular tumors in mice are reticulvin cell
sarcoma and lymphosarcoma. They appear as solid tumor masses, usually multiple,
in a variety of organs, but predominantly in liver, spleen and lymph nodes
(Fig. 4). The principal cells are usually polymorphous reticulum cells or
F-4) VI
abnormal lymphoid cells.
These cells rarely appear in the peripheral blood.
These diseases are not increased in incidence in most strains of mica after
irradiation, and in fact, the incidence may be less (Table 3).
Non-reticular neoplasms. Non-reticular neoplasms include those of lung,
liver, kidney, ovary, mammae, uterus, skin, bone, and other organs. They
usually originate in parenchymal cells or supportive tissues and the effect of
.
10
radiation on their incidence is not uniform (Table 1). The spontaneous
incidence and the radiation effect on incidence very in different strains of mice.
The ovaries and mammae of the female mouse are very susceptibie to
post-radiation tumor formation. In most strains of mice the uterus, however,
is not a common site of radiation-induced tumors. Tumors of the ovary in mice
occur in several histologic variants including granulosa cell, luteoma, mixed
cell, and cystic timors (Fig.56). Many are hyperfunctional with
corresponding effect on other endocrine target organs. Mouse marmary tumors may
develop raywhere along the bilateral milk line. These tumors may be single or
multiple and usually present as a subcutaneous lump. Most are tumors of
epithelial tissue, but a low incidence of sarcoma of the breast also occurs.
tumor
They undergo complications such as hemorrhage, partial infection, and
ulceration of the overlying skin.
A corresponding susceptibility to radiation tumorigenesis in sexual organs
18 not seen in the nale mouse.
Spontaneous lung tumors in mice are usually adenomas (Fig. 7). In many (F-7)
strains of mice, lung tumors are not increased in incidence by external
radiation but are increased by inhaled 1sotopes which lodge in the respiratory
system. After isotopic or whole-body irradiation, lung tumors, in common with
many other types of tumors, appear earlier.
In mice, other tumors which appear in increased incidence in Irradiated
groups are tumors of the liver, kidney, adrenal, bone, skin, subcutaneous tissue
(Fig. 8), and orbital gland (8). In supraletbally irradiated mouse . (F-8)
populations protected against or treated for ordinarily lethal doses, there
is a marked reduction in neoplasm. incidence in the long-term survivors
(Tables 2,4). The acute radiation damage to the cells may amalan survival of
the cells but renders these cells or their progeny incapable of responding to the

.
11
stimuli for mi excessive growth characteristic of neoplasia.
Degenerative changes. The glomerulus of the kidney contains a capillary
network susceptible to late degeneration after doses of radiation approximating
the LD.co or greater (Table 4). This lesion has been noted in irradiated mice,
rats, guinea pigs, hamsters, and pigs. It consists of an increase in the
interstitial substance of the glomerulus with a resultant loss of cellularity
and narrowing of capillaries which progresses to an atrophic, scarred structure.
Involvement is not uniform in all glomeruli, but when enough of them have
become non-functional the animal dies in renal failure. This is a very common
cause of death in very heavily irradiated animals. A glomerulosclerotic kidney
is shrunken, dark, and has a finely pitted surface on gross examination. .
Glomerulosclerosis is a direct radiation effect, since the occurrence of the
disease can be abolished by shielding the kidneys. It is probably a normal
disease of aging in unirradiated mice, but in most strains occurs in very low
incidence and at a much later time.
Greying of fur (Table 4) and ocular cataracts are delayed radiation
effects which have much in common radiobiologically. The severity and rapidity
of onset of both are dose dependent (Fig. 2). They are easily observable
external indications of previous irradiation. They are non-lethal and illustrate
how damage proliferating cells is found much later by a delayed effect.
Reparable and irreparable damage. The delayed manifestations of radiation
llowed
discussed here illustrate the difference between reparable and irreparable
damage occurring eubooghen to exposure to radiation. The early damage seems
to undergo repietr in survivors; after a latent period of several months the
animals indicating that apparent repair was not complete.
.
an...
"...
SUMMARY
.
Mice exposed to sub-lethal, mid-lethal, or supra-lethal doses of X-rays
show group survival and disease incidence that are different from that seen
in unirradiated controls. Mean survival time of irradiated groups 18
shortened in grouportion to radiation dose.
Thymic lymphoma, myeloid leukemia,
ovarian, breast and certain other tumors appear in greater incidence and at :)
an earlier mean age in irradiated groups. Degenerative diseases such as
glomerulosclerosis, ocular lens cataract, and greying of the hair are likewise
raldation induced. These late changes are manifestations of the irreparable
component of radiation injury.
;
RIIERENCES
1. Bacq, z. M. and P. Alexander. Fundamentals of radiobiology, N. Y.,
Pergamon, 555 pp., 1961.
2. Behrens, C. F. and E. R. King. Atomic medicine, Baltimore, Williams
& Wilkins; 766 pp., 1964.
3. Christenberry, K. W., A. C. Upton, and J. W. Conklin. Recording lens
opacities, Arch. Ophthalmol. 72: 667-669, 1964.
4. Cosgrove, G. E., A. C. Upton, C. C Congdon, D. G. Doherty, K. W. Christenberry,
and D. G. Gosslee. Late somatic effects of x-radiation in mice treated
with AET and isologous bone marrow. Radiat. Res. 21: 550-574, 1964.
5. Finkel, M. P., B. 0. Biskis, and G. M. Scribner, The influence of strontium-90
upon life span and neoplasms of mice. In. Progress in Nuclear Energy,
series VI, Vol. 2, Bugher, J. G., J. Cour saget, and J. G. Loutit, eds.
New York, Pergamon, pp. 199-209, 1959.
6. Moshman, J. and A. C. Upton, Depigmentation of hair as a biological radiation
dosimeter. Science 119: 186-187, 1954.
7. 'Upton, A. C., F. F. Wolff, J. Furth, and A. W. Kimball, A comparison of the
induction of myeloid and lymphoid leukemias in x-radiated Rif mice.
Cancer Res. 28: 842-848, 1958.
8. Upton, A. C., A. W. Kimball, J. Furth, K. W. Christenberry, and w. H.:.:
Benedict, Some delayed effects on atom-bomb radiation in mice.
Cancer Res. 20: 1-62 (supplement), 1960.
...
LP
15
LEGENDS FOR FIGURES
Fig. 1
Cumulative mortality curves for 1c,f, female mice vith various .. 1
exposures to 250 kvp x-rays at 3 months of age. :
Fig. 2
The development of cataracts in 1cgt, female mice after various
whole-body x-ray exposures at 3 months of age.
715. 3
Myeloid leukemia in an RF mouse. Note the enlarged liver (L)
and spleen (s) (x2).
Fig. 4
Reticulum cell sarcoma in a 29 mo. old LAF, mouse.
Liver, spleen and lymph nodes are involved (x2).
Fig. 5
Left ovarian tumor (T) in a 10 f, mouse 19 months following
700 r of W. B. X-ray exposure. There is also uterine enlargement with
a fibroid tumor (F) (x5).
Fig. 6
Histologic appearance of an ovarian tumor (T) in an RF mouse dying
19 months after a 400 rad dose of 14 mev neutrons. (Uterus=t). (x10).
Fig. 9
Histologic appearance of a papillary tumor (T) of the lung in an RF
mouse 13 months after a 400 rad dose of 14 mev neutrons. (X32)
.. Fig. 8
Subcutaneous sarcoma in an LAF, mouse 15 months after a whole baly
X-ray exposure of 500 r. Note the greying of the fur on the head.
Table 1.
The Life Shortening Effect of Various Exposures to Whole-Body
X-Ray Giveu to 10 F, Female Mice at 3 Months of Age
-
Life Shortening in Survivors
days
days per R
Exposure
(R)
Mean age at Death
(days)
86€
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: 350
217
500.
633
233
750
549
317
1300 rt AET*
460
406
*Aminoethylisothiouronium given 1.p. 15 min. prior to radiation.
...
-

.
in minimo
Table 2. The Incidence of Various Forms of Leukemia/Lymphoma in 1C3F, Female Mice.
Whole-Body
X-ray Bcposure
Thymic Lymphoma
Incidence Mean age at death
(
. (days)
Myeloid Leukemia
Incidence Mean age at death
(%).
Other Lymphoma
Incidence Mean age at death
(%)
(days)
(days)
252
846
350
490
748
500
288
381
620
750
265
591
1300 r + AET
525
434
-
-
-

VL
AT
Table 3. The Incidence of Various Forms of Leukemia/Lymphoma in RF Mice as Related to Sex and
Radiation Exposure (Dete from Upton, stairs 1958 7 ).
Whole-Body
Ser
X-re
Thymic Lymphoma
ncidence Mean age at death
(model
Myeloid Leukemia
Incidence Mean age at death
(beproting
Other Lymphoma
Incidence Mean age at
()
The paths Ice
Male
0
15.5
14.4
20.6
Female
w
20.4
Male
150
10.2
♡
16.3
16.7
13.3
11.0
10.4
19.2
Male
300
be
10.3
17,7
Female
300
8.7
to
15.6
Male
450
8.5
Ô
10.8
15.8
of fur
Table 4. Incidence (%) of Certain Diseases and Degree of Graying A
in 1C3f, Female Mice Surviving Various Exposures to Whole-Body
X-ray Given at 3 Months of Age.
Exposure
Ovarian
Tumor
UNU
Mammary
Timor
Glomerulosclerosis
Lung
Tumor
Degree of Grayin
of Fur at 720 da
(range 0-24):1
O:
21
350
500
56
54
750
1300 r + AET

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