the injection of adrenalin. The time between the inje^ion and the
appearance of the maximum dilatation seemed in this? series to be
somewhat longer than with smaller doses. In three experiments
this period was one minute; in one, two minutes; in three, four
minutes; and in one, seven (average, three) minutes. In most, if
not in all of the animals in which the maximum dilatation appeared
late, there were rather marked symptoms produced by the adren-
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650
Effects of Adrenalin on Pupils of Rabbits.
aliiL These symptoms were those of severe prostration, during
which the animal's nose usually rested on the table. Occasionally,
however, the animal might lie sprawled out for a few minutes, the
respiration being deeper and more rapid than normally, the animal
taking no interest in its surroundings, and making no attempt to
move about. These symptoms were shown in varying degrees by
most of the rabbits which received the larger doses; i. e., Ko and
%o of a cubic centimeter per kilo. One rather striking effect of this
kind will be mentioned in the account of the next series.
Here, as also with %o of a cubic centimeter per kilo, the amount
of dilatation was fairly constant. It varied in the eight experi-
ments between 2.75 and 4.5 millimeters, with an average of 3.84
millimeters. A beginning recovery was first noted between five
and twenty (average, ten) minutes after the injection. Recovery
from dilatation was complete in from twenty-eight to sixty-five
(average, forty-five) minutes after the injection.
Adrenalin, %o of a Cubic Centimeter per Kilo of Body Weight. —
In this series of eleven animals, a superior cervical ganglion had been
removed twenty-four or more hours before the injection of adren-
alin, the dose here employed, 0.2 of a cubic centimeter per kilo of
body weight, being the largest used in any of the five series (table
V). In seven of the eleven experiments the maximum dilatation
TABLE V.
Adrenalin, 2/10 of a Cubic Centimeter per Kilo of Body Weight of a 1:1,000
Solution (=0.02 of a Milligram of Adrenalin per Kilo),
Namber of
Time between re-
Amount of dilata-
Time between in-
jection and first
Time between injec-
«xperiinent.
moval of ganglion
and experiment.
tion produced.
sign ot recovery of
pupil.
tion and complete
recovery of pupil.
45
24 hours
5.50 millimeters
5 minutes
38 minutes
43
24 hours
4.00 millimeters
4 minutes
45 minutes
29
4 days
4.75 millimeters
8 minutes
95 + + minutes
SI
4 days
6.25 millimeters
27 minutes
51 + + minutes
31
5 days
4.00 millimeters
30 minutes
no minutes
39
6 days
4.25 millimeters
14 minutes
170 minutes
36
8 days
3.25 millimeters
13 minutes
76 minutes
37
8 days
4.25 millimeters
8 minutes
40 minutes
35
8 days
41 minutes
145 minutes
40
9 days
5.7s millimeters
43 minutes
1 70 + + minutes
47
14 days
2.50 millimeters
28 minutes
105 + + minutes
Averages for all
eleven of the
experiments
4.36 millimeters
20 minutes
100+ minutes.
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Don B. Joseph. 661
was recorded within one to two minutes after the injection. In
the remaining four, this period was much longer, being respectively,
ten, fourteen, seventeen, and twenty-two minutes in length, so that
the average for the eleven animals, six and a half minutes, is con-
siderably longer than in any of the other series with smaller doses
of adrenalin.
I shall now describe in detail the conduct of the pupil and the
symptoms following the injection of adrenalin into the animal in
which the maximum dilatation appeared only after twenty-two
minutes.
A white female rabbit, weighing 1,630 grams, received very
slowly in an ear vein %o of a cubic centimeter per kilo of the adren-
alin solution. In three minutes the pupil had increased 2.5 milli-
meters in diameter. It continued to increase for a short time until
the total dilatation was 3 millimeters. Then serious symptoms
developed. The animal which had up to this time appeared restless,
sneezed repeatedly, brushed its nose with its paws, then jumped vio-
lently out of the box a number of times. The iris was very pale,
and the heart was very rapid. Now and then the symptoms nearly
disappeared and the animal sat quietly on its haunches. Respira-
tion was deep and slightly more rapid than normally. During the
attack the pupil became smaller and did not again dilate until the
animal quieted down, when, twenty-two minutes after the injection,
the point of maximum dilatation was reached.
The amount of dilatation produced in this series varied between
2.5 and 6.25 millimeters, with an average of 4.36 millimeters. In
only three of the eleven experiments was the maximum amount of
dilatation produced less than 4 millimeters, and in two of these it
was 3.25 and 3.5 millimeters respectively. The first sign of recov-
ery from dilatation was recorded between four and forty-three min-
utes after the injection, with an average of twenty minutes. In
five of the eleven experiments the maximum dilatation persisted for
from twenty-seven to forty-three minutes. This long continued
maximum dilatation was more prominent with this dosage than
with any other that was used. The return of the pupil to the
normal diameter was also far more gradual than after smaller doses
of adrenalin. The shortest interval between injection and com-
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652 Effects of Adrenalin on Pupils of Rabbits.
plete recovery of the pupil from dilatation was thirty-eight minutes,
while in one case the recovery was not complete in one hundred and
seventy minutes (about three hours). The average length of this
interval is over one hundred minutes. It cannot be given accu-
rately, for in several of the experunents the observations were dis-
continued for various reasons before complete recovery had taken
place.
In considering the experiments in each of the foregoing series, I
have thus far said nothing regarding those in which the injections
were given during the first twenty-four hours after removal of the
ganglion. By referring to tables I, III, and IV, and comparing in
each table the response of the pupil to adrenalin during the first
twenty-four hours after gangliectomy, with the response given at
a later time to the same dose of adrenalin, the following points will
be noted: Two injections (table IV), made four and six hours
respectively after the removal of the ganglion, gave, not a dilata-
tion, but a constriction of the pupil. In another experiment in the
same table (rabbit 44), an injection sixteen hours after gangliec-
tomy gave a dilatation of only 0.5 of a millimeter, which was fol-
lowed in three minutes by a constriction that made the pupil smaller
than normally. In still another animal (rabbit 4) an injection
given after twenty hours gave a dilatation of 3.5 millimeters; *. e.,
well up to the average for the whole series of injections given in
this table.
In brief, then, these few injections indicate that for the first few
hours after removal of the corresponding ganglion, the pupil is no
more sensitive to the action of adrenalin than is the pupil of a
normal animal; that within sixteen hours after gangliectomy, a
faint dilatation may be obtained, while somewhere between the
eighteenth and the twenty- fourth hour the sensitiveness rises rapidly
to nearly or quite that which is seen at any later time.
The following protocols, one taken from each of the series with
different doses of adrenalin, will serve to illustrate the course of
the action of adrenalin. As already stated, where two diameters
are given for a single pupil, the first is always the horizontal diam-
eter as the animal stands erect, and the second the vertical diameter.
In each protocol the pupil on the normal side serves as a control for
the one whose ganglion has been removed.
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Don B. Joseph.
663
Protocol No. i.
Rabbit 9, November 23, 1911. White Male; Weight, 1,790 Grams,
Right Superior Cervical Ganglion Removed Five Days Ago.
Time.
Size of right pupil in mm.
Size of left (control) pupil in mm.
I0:S3
4.0
4.5
4.0
4.5
11:00
4.0
4.5
4.0
4.5
11:05
4.0
4.5
4.0
4.5
11:14
Received 1/50
cc.
per kilo of body weight of adrenalin into left ear
vein.
ii:iS
6.5
4.0
4.5
11:18
5.5
4.0
4.5
11:23
4.0
4.5
4.0
4.5
11:35
4.5
4.0
4.5
11:56
4.0
4.5
4.0
4.5
12:20
4.0
4.5
4.0
4.5
Protocol No. 2.
Rabbit 23, December g, 1911, White Male; Weight, 1400 Grams.
Left Ganglion Removed Nineteen Days Ago.
Time.
Size of right pupil (
control) in mm.
Size of left pupil
in mm.
10^6
4.0
4.5
3.5
3.75
10:56
4.0
4.5
3.75
4.0
11:05
4.0
4.5
3.75
4.0
11:20
Received 1/30 cc.
per kilo of body weight of adrenalin into right ear
vein.
11:21
4.0
4.5
6.0
11:24
4.0
4.5
6.5
7.0
11:30
3.75
4.0
5.5
6.0
11:40
4.0
4.5
3.75
4.0
12:00
4.0
4.5
4.5
12:15
4.0
4.5
4.0
Protocol No. 3.
Rabbit 2. November 20, 1911, White Male; Weight, 1,315 Grams.
Left Superior Cervical Ganglion Removed Forty-Eight Hours Ago.
Time.
Size of right pupil (<
:ontrol) in mm.
Size of left pupi
in mm.
ii:zo
5.5
6.0
6.0
6.5
11:17
5.5
6.0
6.0
6.S
ii:2i
5.5
6.0
6.0
6.S
11:35
Receivedi/20(
cc. per kilo of body weight of adrenalin into right ear'vein.
11:35
5.5
6.0
lO.O
•
11:40
5.5
6.0
lO.O
11:46
6.0
9.0
11:52
6.0
6.5
8.5
12:00
6.0
6.5
8.0
12:10
6.0
6.5
7.5
8.0
12:29
6.5
7.0
7.5
12:50
6.0
6.5
6.5
7.0
1:15
6.5
7.0
1:28
6.0
6.5
6.5
7.0
1:45
6.0
6.5
7.0
2:08
6.0
6.5
7.0
4:1s
6.5
7.0
6.5
7.0
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654 Effects of Adrenalin on PupiU of Babbits.
Protocol No. 4.
Rabbit 48, January 17, 1912, White Female; Weight, 1,600 Grams,
Right Superior Cervical Ganglion Removed Fifteen Days Ago.
Time.
Size of right pupi
in mm.
Size of left (control) pupil in mm.
1:18
4.7s
5.7s
5.25
6.25
1:28
5.25
5.75
4.7s
5.5
1:33
4.5
5.0
4.7s
5.25
1:3s
Received i/io c.c. per
kilo of body weight of adrenalin into left ear vein.
1:36
7.5
8.5
4.75
5.75
1:39
9.0
9.25
4.5
5.0
1:4s
8.0
8.25
4.75
525
i:SS
7.25
8.0
4.75
5.5
2:07
7.0
4.75
5.0
2:30
5.75
6.0
4.75
5.5
3:13
5.2s
S.75
4.75
5.5
Protocol No. 5.
Rabbit 40. January 12, igi2. White Male; Weight, 1,675 Grams,
Right Ganglion Removed Nine Days Ago.
Time.
Size of right pupil in mm.
Size of left (control) pupil in mm.
10:44
3-75
4.25
3.75
4.10
10:52
3.75
4.0
4.5
5.0
10:56
Received 2/10 c.c per kilo of adrenalin into left
ear vein.
10:57
9.5
3.75
4.5
11:02
8.0
9.0
2.5
3.0
11:07
8.5
9.0
3.5
3.75
11:16
8.0
8.5
3.75
4.0
11:26
9.0
4.0
4.5
11:39
8.0
3.5
4.0
11:51
7.0
7.5
4.0
4.5
12:06
7.0
7.5
4.0
4.5
12:21
7.0
4.5
5.25
12:29
6.5
7.25
4.5
525
i:i5
5.75
6.25
4.25
5.0
2.04
5.0
5.5
4.0
4.75
3:14
4.75
5.75
4.0
4.5
In the following chart have been plotted the average figures ob-
tained from each series of animals from which one ganglion had
been removed twenty-four hours or more before the intravenous
injection of adrenalin. The figures show the comparative effects
of the diflferent doses of adrenalin upon the pupil. The amount of
dilatation is shown on the ordinates, taking the abscissa as the
normal diameter before the injection of adrenalin, while along the
abscissa is given the time in minutes after the injection.
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Don R. Joseph. 655
Text-fig. i. For description see text.
SUMMARY AND CONCLUSIONS.
It has been shown by these experiments that after the removal
of one superior cervical ganglion of a rabbit, %o of a cubic centi-
meter of a I : I, GOO adrenalin solution (=0.02 of a milligram of
adrenalin) per kilo of body weight injected intravenously, is prac-
tically the minimum amount that will produce in nearly all cases a
perceptible dilatation of the pupil. In one out of eight animals this
dose failed to give a definite effect, while in two others the effect
was slight.
Six experiments with injections of %o of a cubic centimeter of
adrenalin per kilo of body weight gave the following averages: a
maximum dilatation of 1.62 millimeters, which showed the first sign
of decreasing in four minutes and had disappeared entirely in ten
minutes after the injection.
Six experiments with %o of a cubic centimeter per kilo gave the
following averages: a maximum dilatation of 2.25 millimeters,
which began to disappear in eight minutes and had entirely disap-
peared in twenty-eight minutes.
For thirteen experiments with %o of a cubic centimeter of adren-
alin per kilo these averages were: a maximum dilatation of 3.61
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666 Effects of Adrenalin on Pupils of Babbits.
millimeters, a beginning recovery from dilatation in six minutes,
with complete recovery of the pupil in thirty-seven minutes.
With Ko of a cubic centimeter of adrenalin per kilo (eight ex-
periments) the averages were: a maximum dilatation of 3.87 milli-
meters, a beginning recovery in ten minutes, with complete recovery
of the pupil in forty-five minutes.
Finally, eleven experiments with %o of a cubic centimeter of
adrenalin per kilo gave averages as follows : a maximum dilatation
of 4.38 millimeters, a beginning recovery from dilatation in twenty
minutes, with complete recovery only after one hundred minutes
or more.
The time between the injection and the appearance of the maxi-
mum dilatation of the pupil varied somewhat according to the dose
of adrenalin. In the series of experiments with %o of a cubic centi-
meter of adrenalin per kilo of body weight the average for this
interval was one minute; in the series with Vso, 34o, and Ho of a
cubic centimeter the average length of this interval was about
three minutes, while the experiments with %o of a cubic centimeter
give us an average for this interval of six and a half minutes, with
individual instances in which the maximum dilatation was seen only
after fourteen, seventeen, and twenty-two minutes after the injection.
In those animals in which the maximum pupil dilatation was espe-
cially delayed, there were almost always more or less alarming
symptoms of general prostration for a short time after the injec-
tion, and the maximum dilatation appeared as these symptoms grad-
ually disappeared.
In other words, with an increase in the size of the dose of adren-
alin, there was a gradual increase in the following: (i) the time
between the injection and the appearance of the maximum dilata-
tion; '(2) the amount of dilatation produced; (3) the interval be-
tween the injection and the beginning of recovery from dilatation ;
and (4) in the total time between the injection and the return of the
pupil to its normal size.
After removal of the ganglion a certain time must elapse before
the increase in the sensitiveness develops. During the first ten to
fifteen hours there is practically no increase in the sensitiveness.
After eighteen hours a moderate effect can be obtained which rap-
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Don B. Joseph. 667
idly increases so that by twenty to twenty-four hours after the
removal of the ganglion any given dose of adrenalin produces prac-
tically as great a dilatation as it will give at any later time.
The experiments teach us that, on account of the individual varia-
tion in the degree of dilatation produced by a given dose of adrenin,
we may not determine with exactness from any given degree of dila-
tation of the pupil the quantity of adrenin injected into the blood
stream, and that we can not state with absolute exactness either
the smallest dose that will constantly cause a dilatation, or mention
the exact amount of adrenin which will in no case cause a dilatation
of the pupil. However, the results permit the general statement
that very small doses of adrenin on entering the circulation cause
a fairly prolonged definite dilatation of a deganglionized pupil.
Since this is true, there are then at least two ways in which
a rabbit from which one superior cervical ganglion has been pre-
viously removed may be used for determining qualitatively the
amount of adrenin present. If, for example, we have a solution
which may or may not contain adrenin, and upon injecting it obtain
a dilatation of the deganglionized pupil only while the pupil on the
normal side remains unchanged in size, we are justified in assuming
that the solution contains adrenin. Again, if after stimulation of
the peripheral end of a splanchnic nerve or other procedure upon
the animal body under conditions that exclude the possibility of
reflex effects, a dilatation of the deganglionized pupil results, while
its normal mate remains unchanged, the assumption is warranted,
though not absolutely proved, that some adrenin has been thrown
into the circulation.
This test for the presence of adrenin has the advantage over
other tests, that the pupil on the normal side will always act as a
control.
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TBANSPLANTABLE TUMOBS OF THE FOWL:
A NEGLECTED MATEBIAL FOB
CANCEB BESEABCH*
PEYTON ROUS, M.D., JAMES B. MURPHY, M.D., AND
W. H. TYTLER, M.D.
NEW YORK
The recognition that neoplasms exist in many species
of animals throughout the vertebrate world has given
to the tumor problem a broader biologic significance.
And at first glance it appears strange that so few animal
forms have been utilized for experimental cancer
research. Attention has centered on the new growths of
the rat and mouse, on a peculiar round-celled tumor of
the dog, transmissible by coitus, a dubious tumor of wild
hares, endemic in certain portions of Germany, and on
a thyroid growth of fishes, the exact nature of which is
still under dispute. There exist good reasons for this
limitation. The species employed for tumor work must
be available in quantity and capable of thriving under
laboratory conditions; and its "spontaneous" growths
must be transplantable. True tumors are rare in cats,
rabbits and guinea-pigs, and, while more frequent in
dogs, are difficult to study because of the number of
individuals which must be employed and the frequent
necessity that all shall be of one age and breed.
Some two years ago one of us transplanted a spindle-
celled sarcoma of the fowl, a very typical neoplasm,
metastasizing by means of its cells, and in other ways
regulated by their peculiarities. The growth has since
been observed in twenty-seven series of fowls. Becently
it has been successfully transmittea by the Berkefeld
filtrate of an extract of the fresh tumor-tissue in Ring-
er's solution* and by the dried or glycerinized tissue.
These results have led us to a study of other chicken-
• Prom the Laboratories of the Rockefeller Institute for Medical
Research.
1. Rous. Peyton: Jour. Exper. Med., 1910, xli, 696; 1911, xHl.
397 ; Thb Journal A. M. A., 1910, Iv, 1805 ; 1911, ivl, 198. Rous.
l\, nnd Murphy, James B. : Jour. Expor. Med.. 1912. xv. 119, 270.
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Iiirnorv. Work with them hat* not yet advanced to test?
for an extranH)n>* c-aunative agent; but it has disclosed
lh<* prc^-cncc In chickcnH of an abundant material well
adapted for cancer research. From a single firm selling
live [)oultrv in New York we have acquired during the
»jiHt Hcv<»n monthn twenty-seven chicken-tumors, a num-
>er greater than this institute was able to obtain of
tnouHc-tuniorH from all sources in the course of three
veaiH.^ Furthermore, the avian growths were encoun-
[vvvi] in a rehitively unfavorable stock, young for the
ino'^t part and supposedly healthy.
The existence of tumors of the fowl has been demon-
strated by the work of several investigators.^ The
variety of growths is much greater than among rats and
mice. We have been especially struck with the frequencv
of benign tumors. Among 353 primary neoplasms of
the mouse, Haoland* found but three which may be
termed benign — a melanoma, a mixed tumor and a
tlbromyoma uteri. There were 344 carcinomas and six
sarcomas.* Hut among our first twenty chicken-tumors
were a large, characteristic lipoma, a pure myxoma, and
several stationary growths, made up of blood-, or lymph-
sinuses and lym]>hoid tissue, which are perhaps best
thought of as nevi. Of malignant tumors there were an
osteosarconia with largi* blood-sinusi^, from one of which
the (o\\\ bled to death at operation, numerous round-
idled tumors, a connivtive-t issue growth producing car-
tilagt^ and bone, thnv spindkMH^UtHl sarcomas, a fibro-
sa rconui and an epithelioma, IVevious observers have
noted san'omata and carcinomata in about ei^ual pro|x>r-
tions. Hut the\ have dealt in large part with auti>psy
n^aterial and with gnnvths primary in the visivra:
whoivas our nHHU*\ls have to do with tumors <uiH^rfiriallv
>ituatiHK and for this rt^ason iH^me to hand during the
litV of the host.
Small bits of ea^h sfHmlantvus gnnvth were nnnovt^l
at o^HM\Hti^n\ and transt^lanted bv means of a tnx"'"ar.
r*fo Mv<vt'vni v^t a xHMu'ar site for t^e crufts has mUiV
MiiiH>ttar.vv, V>ua'*\ t'H^\ A\e'^' p^utNl dtvr^ in tV.e p^a-
4 Utt * -.i M K^' ' i.i S< K"!* I iv K';n"c; ^.< it^ l^:^Ti:x\ Taio -
K.-M >^ ^ " t l-'-l IV I
,\ Mil ■'*,• 'Vv* ' •'>' f!>»t» •'»,'^;*> !*^«'-i>^ f'^'-'x of T*S* '•••TJ
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toral muscle. The normal fowls taken as hosts were
young, and of the same variety as* the chicken with the
spontaneous growth. The general failure to transplant
avian growths is certainly attributable, in part, to lack
of observance of such points.®
Many attempts were made to transplant the first
tumors obtained, quite irrespective of the indications
afforded by their histologic appearance. More recently,
taught by negative results, we have resorted to pre-
liminary' autotransplantations, and have tried to prop-
agate further only such growths as proved autotrans-
plantable, or were found micoscopically to be malignant.
On much of the work it is too early to report. But
thus far two new tumors have been successfully trans-
ferred to, other hosts. One of these, a connective-tissue
growth forming cartilage and bone, is noteworthy,
because in the original host it appeared to be a
symmetrical, developmental anomaly connected with the
sternum. It is now in its fifth ^^tumor-generation."
The other growth is a spindle-celled sarcoma which had*
its primary seat amid the muscles of the upper leg, but
metastasized to the neck and gizzard. It is now growing
rapidly in a large proportion of the fowls constituting
the second "tumor-generation" (second consecutive series
inoculated).
Of special interest is the question as to whether our
spindle-celled sarcoma with a filterable cause is epidemic
among chickens. But in the new material it has not
even been encountered. True, several spindle-celled sar-
comas have been observed, including one that is trans-
plantable. But each has its distinguishing histologic
features. There remains the possibility, nevertheless,
that these various growths may have a common ctiuse, a
point which we hope to investigate in the near future.
Sixty-Sixth Street and Avenue A.
6. Rons: Notf 1. From a recent publication (Verhandl. d. Japan,
pntbolog. Gesellsch.. 1911, p. 114) It appears that A. Fujinaml and
K. Inamoto reported in Osaka, in 1910, their success in transplant-
ing a myxoma of the fowl. They make no mention of the influence
of Tariety of the host on the success of transplantation.
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Reprinted from The Journal of Vie American Medical Association
June /, J912, Vol. LVIIJ, pp, JttS? and 1683
Copyright, 1912