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LIBRARY

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THE AMERICAN MUSEUM

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NATURAL HISTORY

PROCEEDINGS

OF THE

SOCIETY FOR

EXPERIMENTAL BIOLOGY AND MEDICINE

VOLUME X

1912-1913

EDITED BY THE SECRETARY

NEW YORK
1913

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CONTENTS:

SCIENTIFIC PROCEEDINGS (50th—54th meetings):

Communications of the fiftieth meeting, October 16, 1912.
Communications of the fifty-first meeting, December 18, I912 .
Communications of the fifty-second meeting, February 19, 1913
Communications of the fifty-third meeting, April 16, 1913.
Communications of the fifty-fourth meeting, May 21, 1913
Recapitulation of the names of the authors and of the titles of the
communications ; d é :

EXECUTIVE PROCEEDINGS (50th—54th meetings)

REGISTER OF NAMES AND ADDRESSES OF THE MEMBERS

List OF OFFICERS . , :

CLASSIFIED List OF MEMBERS. :

INDEX OF THE SCIENTIFIC PROCEEDINGS.

PEATE te

SCIENTIBIC se ROCEEDINGS:

ABSTRACTS OF THE COMMUNICATIONS.
Fiftieth meeting.

Cornell University Medical College. October 16, 1912. Dr. Lee
in the chair.

1 (697)
A note on the mode of infection in epidemic poliomyelitis.
By SIMON FLEXNER and PAUL F. CLARK.

[From the Laboratories of The Rockefeller Institute for Medical
Research.|

Both in experimental and human epidemic poliomyelitis the
virus has been repeatedly demonstrated in the tonsils, in the nasal
mucous membrane, and in nasal washings, both from fatal and
acute cases. As the experimental disease can also be produced by
intranasal swabbing with the active virus it seems probable that
the nasal mucosa is one at least of the sources of the virus in the
outside world and also the means of its entrance to the body.
The marked viability of the virus under adverse conditions such as
drying, low temperature, etc., must also be considered as making
for a fairly well founded theory of the nasal route as one path
of the virus to and from the body.

The precise manner in which microédrganisms enter the body
through mucous membranes is difficult to establish. Because we
can produce experimental poliomyelitis by the application of the
active virus to the nasal mucous membrane, we have in this disease
a means of determining whether the virus so applied first enters
the blood stream and through this the central nervous system or
whether it ascends directly along the lymphatics that unite the
nasal mucosa with the central meninges. In experimental polio-
myelitis produced by any method of injection it is well known that
the virus is present throughout the central nervous system. But
after an intranasal injection, can the virus be demonstrated equally

early in all regions of the cord?
I

2 SCIENTIFIC PROCEEDINGS (50).

In order to answer this question, the nasal mucous membrane
of a Macacus rhesus monkey was swabbed lightly with a portion
of ground cord from a recently paralyzed monkey. The monkey
was killed at the end of 48 hours and the following portions of the
central nervous system were removed separately and aseptically:
(1) the olfactory lobes with small portions of the adjacent brain
substance, (2) the medulla, and (3) pieces of the cord at different
levels including the cervical and lumbar enlargements. These
different portions were injected separately as suspensions into
the brain and peritoneal cavity of three other Macacus rhesus
monkeys.

The monkey injected with the suspension of the olfactory lobes
came down in a manner typical of poliomyelitis in monkeys with
definite prodromal symptoms on the ninth day, paralysis on the
tenth and death on the twelfth day. At autopsy, lesions char-
acteristic of the disease were observed throughout the cord.

The two other monkeys remained entirely well and have never
shown any symptoms of paralysis.

The result of this experiment is definite. The virus of polio-
myelitis passes from the nasal mucous membrane to the olfactory
lobes and adjacent parts of the brain before it reaches the medulla
or cord. This distribution is what we would expect were the
ascent by the direct lymphatic path and not by the blood stream.
Were the dissemination by the latter route we should expect
early localization in those parts of the cord and medulla that
possess an especial affinity for the virus.

2 (698)
Effects on meningeal tuberculosis of the local injection of foreign
leucocytes.
By WILFRED H. MANWARING.
[From the Rockefeller Institute for Medical Research.]

In a previous report,! it was shown that the use of foreign
leucocytes as a local therapeutic agent in experimental meningeal
infections in dogs is limited by the toxicity of foreign leucocytes

1Vol. IX, p. 117.

Loca INJECTION OF FOREIGN LEUCOCYTES. 3

for these animals. Rabbit leucocytes injected into the meningeal
cavities of dogs invariably cause death. A single injection of
horse leucocytes, however, can be safely made in normal dogs,
although on repeating the injection, or on injecting horse leu-
cocytes for the first time into meninges already the seat of an
inflammatory lesion, death results.

The injection of horse leucocytes into the cerebral meninges of
dogs, simultaneously with the inoculation of the meninges with
tubercle bacilli, causes a slight delay in the development of the
paralytic symptoms in about half of the treated animals. This
delay, however, is very slight when compared with the great
prolongation of the latent period previously observed,! after treat-
ment with homologous leucocytes.

In the same report, it was shown that foreign leucocytes are
much less toxic for monkeys. Both rabbit leucocytes and horse
leucocytes can be safely injected into the meningeal cavities of
these animals.

The study of the therapeutic control of meningeal tuberculosis
in monkeys is made difficult by extra-dural leakage, when the
inoculations and treatments are made by the method of lumbar
puncture. The inoculations and treatments, in the later experi-
ments herein reported, were therefore made through a permanent
wax-trephine? opening in the skull.

The injection of foreign leucocytes into the meningeal cavities
of monkeys, either simultaneously with the inoculation with
tubercle bacilli, or subsequent to the inoculation has thus far
given no definitely positive prophylactic or curative effects. Ina
small group of monkeys, however, inoculated and treated by the
method of lumbar puncture, the repeated injection of rabbit
leucocytes was associated with a considerable prolongation of the
latent period in one of the treated monkeys, and by a complete
prevention of the subsequent tuberculosis in a second monkey.
The work with monkeys is being continued.

1 Jour. Exp. Med., 1912, XV, pp. I-13.
2 Jour. Exp. Med., 1912, XV, p. 3.

4 SCIENTIFIC PROCEEDINGS (50).

3 (699)
The importance of calcium in relation to growth.
By FRANCIS H. MCCRUDDEN.
[Rockefeller Institute Hospital, New York.)

In certain cases of retarded development there is faulty
skeletal development and disturbed calcium metabolism. The
bones are frail and easily fractured; large quantities of calcium
are lost through the feces, and the urine is almost free from
calcium. It seems probable that the retarded skeletal develop-
ment is due to the lack of calcium salts available for bone growth.
Other cases of retarded development show no such disturbances
of calcium metabolism and the bones are of normal solidity. In
these cases there is a more fundamental absence of the ‘‘ tendency
to grow”’ rather than any lack of material for growth.

4 (700)

The pyramid tract in the Canadian porcupine (Erethizon dorsatus
Linn.).

By SUTHERLAND SIMPSON.

[From the Physiological Laboratory, Medical College, Cornell Uni-
versity, Ithaca, N. Y.]

The nerve fibers of the pyramid tract pass caudalwards from
their cells of origin in the motor area of the cerebral cortex and
are homo-lateral until the lower part of the medulla oblongata is
reached. At this level, in the majority of mammals that have
been examined, most of the fibers decussate and take up a position
in the lateral column of the spinal cord on the opposite side, just
ventral to the posterior horn, forming the crossed lateral pyramid
tract. <A few fibers remain uncrossed and are found in the lateral
column on the same side, constituting the direct lateral pyramid
tract.

In man and the anthropoid apes a second uncrossed tract is
found in the ventral column—the direct ventral pyramid tract.
In mammals lower in the scale than the anthropoids, it is generally
held that this direct ventral tract is not represented at all.

THE PyrAMID TRACT IN THE CANADIAN PORCUPINE. 5

This disposition of the pyramid tract fibers is not found in all
mammals however. In the guinea-pig (v. Bechterew, Reveley),
mouse (v. Lenhossek), rat (Flechsig, King) and squirrel (Weigner),
and in the monotremes and marsupials (K6lliker and Ziehen), the
crossed fibers run in the dorsal and not in the lateral column of
the cord.

In the spring of the present year, I obtained some full-grown
porcupines with the object of investigating the course of the
pyramid tract fibers in this animal. The left motor cortex was
located by electrical stimulation and then extirpated in the usual
way. At the end of about a fortnight after the operation in each
case the animal was killed, the brain and cord removed, stained
by the Marchi method and sectioned at all levels. A full descrip-
tion of the resulting degenerations will be published later; in this
preliminary communication only the most important points will
be mentioned.

In the internal capsule, pes pedunculi, pontine bundles and
upper levels of the medulla oblongata, the fibers occupy the usual
positions and no special comment is called for, but in the lower
part of the medulla oblongata the arrangement is peculiar, and
nothing similar, so far as I know, has been hitherto described in
any other animal.

At the decussation of the pyramids most of the degenerated
fibers cross the median raphe, pass backwards through the gray
matter, and take up a position in the dorsal column of the opposite
side occupying the ventral portions of the fasciculi of Goll and
Burdach. A few of the crossed fibers curve outwards and enter
the opposite lateral column of the cord. A very considerable
number of fibers, however, remain uncrossed and are continued
into the cord, forming a comparatively large and compact bundle
in the ventral column extending along the margin of the anterior
median fissure. Some uncrossed fibers are also found in the
dorsal column of the same side but these are very scanty.

In this animal, then, the fibers of the anterior pyramid on
entering the spinal cord are divided into four fasciculi, two crossed
and two direct, viz: the crossed dorsal pyramid tract, the crossed
lateral pyramid tract, the direct ventral pyramid tract, and the
direct dorsal pyramid tract. Of these the crossed dorsal and

6 SCIENTIFIC PROCEEDINGS (50).

direct ventral tracts are much larger than the other two and can
be traced as far as the lower sacral segments. The crossed lateral
and direct dorsal seem to disappear in the upper thoracic region.
Arranged according to the number of fibers which they contain,
the order is crossed dorsal, direct ventral, crossed lateral, direct
dorsal.

5 (701)

Interpolated extra-systoles, of frequent occurrence, in an other-
wise normal human heart.

By M. DRESBACH and S. A. MUNFORD.

[From the Physiological Laboratory, Cornell University,
Ithaca, N. Y.]

In the extensive literature upon extra-systoles about twenty
writers have described the type known as the interpolated beat
of the ventricle. Although such beats have frequently been pro-
duced experimentally, their occurrence clinically is comparatively
rare.1 Almost without exception they have been observed in
cases showing gross lesions of the heart. The present case is of
interest because the interpolated contractions occur persistently
and frequently in a heart which, aside from a slow sinus rhythm,
is otherwise normal. It furnishes a striking example of an organ
on the borderland between a physiological and a strictly patho-
logical condition.

T., a Chinese student at Cornell University, is in good health.
He is able to take vigorous exercise, such as running, tennis, etc.
About two years ago a cardiac irregularity was discovered in his
routine physical examination. Prior to that time he was wholly
ignorant of any disturbance in his heart, and is at no time conscious
of the heart’s contractions. Polygraphic records have frequently
been made during the past two years and the organ has been found
beating normally on two occasions only, and but for a short time.

The tracings show extra-systoles of the ventricle which occur
with considerable regularity on some days, but usually they appear
at varying intervals. The normal rate of the dominant rhythm

1 Mackenzie, Lewis, Wenckebach, and others.

INTERPOLATED EXTRA-SYSTOLES. 7

averages about 60. It may go as low as 50 during rest. The
extra-systoles average about 35 per minute during rest and 45
after vigorous exercise. The ectopic beat follows the normal
contraction by a period averaging 14second. The pause following
the extra-systole is variable; occasionally it is fully compensatory.
The dominant rhythm is but slightly disturbed, as a rule. Often
it is strikingly constant, and it is seldom more variable than the
rhythm of many normal hearts. This is remarkable in view of the
fact that in the tracings there is good evidence of retrograde
contractions of the auricle which beats prematurely in response to
the heterogenetic stimulus. Evidence for the retrogression is
found in the absence of the ‘‘a’”’ wave at certain cycles. In place
of the ‘‘a’’ wave there is often seen a wave which precedes the
normal auricular systole by 3/30 to 5/30 of asecond. This prema-
ture wave is also preceded by a wave practically synchronous with
the extra-systole of the ventricle.

The interpretation of the curves has been uncertain because
of the unusual character of the phlebograms and cardiograms
and because of the difficulty in establishing proper time relations,
While several explanations may be offered for the type of irreg-
ularity exhibited, the evidence points to the one given as plausible,
but not altogether satisfactory.

Physiologically this heart is interesting because of the following
characters:

1. It very rarely has periods in which its action is absolutely
normal. These periods are of short duration.

2. The normal sinus rate is slow.

3. Functionally the heart meets all ordinary demands made
upon it.

4. There is a bathmotropic disturbance which causes persistent
interpolated extra-systoles.

5. The number of extra-systoles is not decreased, except
slightly in a few tracings, by an increase in the heart rate due to
exercise. The tendency is for the number to rise with acceleration
of the normal beat.

6. The pause following the extra-systole is variable. Occa-
sionally it is compensatory.

7. There is strong evidence that the extra-systole of the

8 SCIENTIFIC PROCEEDINGS (50).

ventricle is often followed by a premature beat of the auricle.
When the evidence of such retrograde contractions is best, the
indicated time of backward conduction is 7/30 to 8/30 of a second.

8. There is no evidence of lengthened As—Vs intervals except
at the extra-systoles.

9. There is a fairly fixed time relation between the normal
and abnormal ventricular systoles.

10. There is often a considerable delay in the appearance of
the carotid pulse resulting from the extra-systole.

11. There is a marked lack of synchronism between the carotid
pulse and the “‘c’’ wave. The latter may precede the former by
1/20 of a second.

12. The site of the abnormal stimulus is either in the ventricle
or some part of the conduction system, e. g., the node, or some
lower point.

6 (702)

On the differences in the effects of stimulation of the two vagus
nerves on rate and conduction of the dog’s heart.

By ALFRED E. COHN.

[From the Hospital of the Rockefeller Institute for Medical Research,
New York.|

The effects obtained in the action of dogs’ hearts on stimulating
the peripheral stumps of both vagus nerves were studied in fifty-
four experiments. The dogs were anesthetized with ether without
adjuvant, artificial respiration was maintained by the Meltzer-
Auer method, and registration was accomplished by the galvano-
metric method. When the chest was opened, curves of auricular
and ventricular contractions were also inscribed. Faradic stimula-
tion was employed. The secondary coil of an inductorium, fed
by a 2-volt dry cell, placed arbitrarily at 50 mm., was the source
of the current.

On stimulating the right vagus nerve, the usual effect was
obtained; both auricles and ventricles ceased to beat, generally
throughout the period of stimulation. Occasionally the ventricles
escaped from inhibition, but then the impulse to contraction arose

EFFECTS OF STIMULATION OF Doc’s HEarrT. 9

and spread in an abnormal manner; the structures normally con-
cerned with these functions remained inhibited.

When the left vagus nerve was stimulated in the same dog, with
a current of the same strength, a difference from the effect of
stimulating the right nerve was observed in 88 per cent. of the
fifty-four experiments. The auricles did not cease to beat, they
were merely slowed,—sometimes 100 or more beats. In one group,
normal ventricular contractions ceased entirely in twenty-four
cases. In a second of twenty-four other cases, a ventricular
contraction followed every second, occasionally every third, fourth
or more auricular beats, the mechanism being one of incomplete
dissociation. Ina third group, the only effect of stimulation was
an increase in the time occupied in conduction from auricles to
ventricles.

In the first group when the left vagus was stimulated, as has
been stated, normal ventricular activity ceased, but abnormal
activity occasionally continued. The rate of the abnormal ven-
tricular contractions differed from that of the slowed auricles and
complete As—Vs dissociation resulted. Similar ventricular activity
occurred also when the right vagus was stimulated, but the auricles,
as is usual, ceased to contract.

The explanation offered for the phenomena occurring on left
vagus stimulation is that the main effect consists in depressing
the conduction system. In the first group, the ventricles cease to
beat, because, on account of the great depression of conduction,
they receive no impulses from the auricles. If the ventricular
muscle is irritable, abnormal stimuli may be formed and an
idioventricular rhythm result. When a slighter degree of depres-
sion of conduction has taken place, every second or third beat
passes along the A—V bundle and initiates a ventricular contrac-
tion. A still slighter degree of depression results in a mere
lengthening of the time of conduction.

The mechanism which results from stimulating the right vagus
nerve and the negative chronotropic effect of the left nerve are
not discussed at present.

10 SCIENTIFIC PROCEEDINGS (50).

7 (703)

The creatine content of muscle under normal conditions.
Its relation to the urinary creatinine.

By VICTOR C. MYERS and Morris §S. FINE.

[From the Laboratory of Pathological Chemistry, New York Post-
Graduate Medical School and Hospital.|

Though an unusual amount of attention has been devoted to
urinary creatinine since the introduction of Folin’s simple colori-
metric method for its estimation in 1904, relatively little considera-
tion has been given to the supposedly related muscle creatine.
The few figures which have been published giving the content of
muscle creatine have simply served to indicate that the creatine
content of vertebrate muscle is in round numbers .4 per cent.,
though figures for different animals have shown individual results
varying from .3~-.5 per cent. We have estimated the creatine
concentration of muscle in a number of the common laboratory
animals, rabbit, dog, cat, guinea pig, monkey, and likewise in
man, in a few instances, where we have been able to obtain good
samples at autopsy. Our muscle analyses on 20 normal rabbits
and 5 dogs have yielded very uniform figures and indicate that the
creatine concentration of rabbit muscle is .52 per cent. and of the
dog muscle, .37 per cent. Our data on the muscle of the other
species are as yet insufficient to warrant special comment. This
uniformity recalls the constancy of the daily creatinine elimination
first noted by Folin.

It is a curious fact that the creatinine coefficient of the rabbit
is fully a third higher than that generally found in man and various
experimental animals, dog, pig, and guinea pig. In forty rabbits,
the average coefficient has been found to be 14.3 while for a healthy
man, the normal coefficient is about 9, and for the dog, the average
is probably a little lower (for 3 animals it was found to average 8.4).
Comparing these values with those for the creatine concentration
of the muscle (rabbit .52, man .39 and dog .37 per cent.), an
interesting, and, as we believe, more than an accidental relation-
ship is revealed.

CREATINE OF MuscLE UNDER NorMAL CONDITIONS. Tor

In view of the lack of proof of the connection of creatine and
creatinine in metabolism, our original experiments were planned
in an endeavor to ascertain whether in a given animal, in this case
the rabbit, there was a constant relationship between the total
creatine of the body and the daily creatinine. This ratio was
ascertained in a series of one growing and ten adult rabbits. It
was not absolutely constant, and appeared to vary with the
weight of the animal, being 53.3 : 1, in an animal of 1.39 kilos
and 44.7 : 1 where the weight was 2.13 kilos. For five animals of
nearly the same weight, 1.9-2.1 kilos, comparatively constant
ratios, 44.7, 45.0, 44.4, 44.4, and 44.9 : I were obtained.

In a further attempt to ascertain any relationship between
body creatine and urinary creatinine, the figures for the creatine
concentration of the body were arranged in order of the creatinine
coefficients. The first five adult animals with an average coeffi-
cient of 13.6 had an average body creatine concentration of .170
per cent., while the second five animals with an average coefficient
of 15.0 had a creatine concentration of .193 per cent.

Our fasting experiments below indicate that the amount of
creatine remaining in the body after starvation bears a relation-
ship, not only to the amount of creatine lost in the urine during this
period, but also to the amount of the creatinine elimination.

An interesting observation with regard to the creatinine, in the
series of eleven normal animals, is that the amount of its average
daily elimination followed that of the body weight in every case.
This is in accord with Folin’s original statement that body weight
appears to be the most important factor in determining the amount
of its elimination.

It is not supposed that these experiments offer any definite
proof of the origin of the urinary creatinine from muscle creatine
or some common precursor substance, but in view of the fact
that no data have been adduced to show such a relationship, these
experiments may be of value as advancing this hypothesis.

12 SCIENTIFIC PROCEEDINGS (50).

8 (704)

The creatine content of muscle during starvation and its relation
to urinary creatine.

By VICTOR C. MYERS and Morris S. FINE.

[From the Laboratory of Pathological Chemistry, New York Post-
Graduate Medical School and Hospital.|

There has recently arisen some little discussion with regard
to the creatine concentration of muscle during starvation. Data
have been submitted to show that inanition in the rabbit results in
an increase in the content of muscle creatine, and it has further
been argued that there is an actual increase in creatine formation.
In other quarters, it has been claimed that starvation (experiments
with the dog) produces a marked decrease in the percentage con-
tent of muscle creatine.

From nine experiments which we have already completed on
starving rabbits, we can agree with both of these observations as
regards an increase or a decrease in the content of the muscle
creatine. In three experiments, a decided increase in the content
of creatine in muscle has been found; in one the per cent. of
creatine was normal, while in five it was even more strikingly
below normal. In addition to the increased or normal creatine
content of muscle, the first four animals mentioned showed an
increased percentage content of creatine in the body. This we are
not inclined to ascribe to an increased creatine formation.

The reason for this increased or decreased content of body and
muscle creatine appears to be satisfactorily explained by our
results. The animals having an increase in the content of muscle
creatine eliminate a comparatively small amount of creatine in the
urine, while those in which the muscle and total body creatine
has been considerably depleted, eliminate an equally large amount
of creatine in the urine. In other words, the content of muscle
creatine during starvation is dependent upon the amount of and
the rate of creatine excretion in the urine. Just why some animals
eliminate creatine more rapidly than others, we are as yet unable
to explain, although we assume that this is connected with the
length of the fast and the state of nutrition of the animal. The

REVERSAL OF THE CARDIAC MECHANISM. 13

rate of the loss of creatine appears to closely parallel that of the
total nitrogen.

Still an added factor in this loss of creatine from the body is
the excretion of creatinine. In experiments where the creatine
of the urine plus that of the tissue does not entirely account for
the creatine which should normally be present in the body, it is
found that a considerable amount of creatinine has been eliminated
in the urine, e. g., in the case of a comparatively long fast. When
this creatinine in terms of creatine is added to the creatine of the
tissue and urine, this total exceeds the total normal body creatine
by about 10 per cent., this excess probably representing the amount
of creatine and creatinine formation. It would seem probable
from these data that during starvation, the creatine storehouse was
depleted not only by a loss of creatine in the urine, but also by the
loss of creatinine.

It has been assumed for some time that the creatine appearing
in the urine during starvation and in various pathological condi-
tions was derived from the creatine of the muscle, and measured
the amount of muscle disintegration, though so far as the authors
are aware this point has never been conclusively demonstrated.
In our experiments, we have found that when the weight of the
creatine excreted in the urine was added to the weight of creatine
still remaining in the body after the period of starvation, the
amount of creatine was only slightly below that which would
have been found in the body had the animal been killed prior to
starvation. This would seem to demonstrate that the creatine
appearing in the urine in starvation was derived from the creatine
of the muscle.

9 (705)

Reversal of the cardiac mechanism.
By HORATIO B. WILLIAMS and HENRY JAMES.

[From the Physiological Laboratory of Columbia University, New
York.|

The subject of investigation, H. M., came under observation at
the Vanderbilt Clinic July 15, 1912. He has had a persistent

14 SCIENTIFIC PROCEEDINGS (50).

diarrhea for a year with several watery stools a day. Otherwise
he has been well until three months before admission to the clinic.

During these three months and subsequently, he has suffered
from attacks of dizziness of increasing severity. In one of these
attacks he fainted and fell in the street. Muscular effort increases
the giddiness. He has had a good deal of headache. He is very
drowsy and is annoyed by numbness and tingling of the hands and
feet.

Until the onset of symptoms he had been a heavy drinker. He
denies lues and gonnorrhea. The Wassermann test was negative.
The blood and urine are normal. The heart is not enlarged.
The sounds are faint but clear, and there are no murmurs. The
third sound has been distinctly audible at most examinations and
has been recorded graphically. The pulse is regular, 40 per
minute. Systolic bloodpressure varies between 95 mm. and 110
mm. and diastolic pressure is usually about 65 mm. The electro-
cardiograms show that the heart beat is initiated by the ventricles,
the auricular beat succeeding the ventricular after the usual con-
duction interval. This is evidenced by the presence of an inverted
P between R and T in leads IJ and III. In lead I, P is of very
small amplitude, but can be clearly distinguished on close inspec-
tion and stands in the same relation to R and T as in the other
leads.

This interpretation of the electrocardiograms has been con-
firmed beyond reasonable doubt by examination with Roentgen
rays and fluoroscope. The contractions of the auricle can be
distinctly seen.

We are not yet ready to offer an explanation for the peculiarly
slow intrinsic rhythm of the auricles which permits the ventricle
to initiate the beat. Neither atropin nor digitalis has any measur-
able effect on the condition. The subject is still under observation.

Rote oF Lrporps IN NARCOSIS. 15

10 (706)

The réle of lipoids and particularly lecithin in narcosis.
By B. KRAMER.

[From the Physiological Laboratory of Cornell University Medical
College, New York City.]

Following the discovery of anesthetic properties of ether by
Jackson and Morton in 1846, numerous theories appeared in the
literature which aimed to explain this important phenomenon.
Of these the well-known Meyer-Overton theory is the only one that
has survived the test of time. It reads as follows: ‘‘The narcotiz-
ing substance enters into a loose physio-chemical combination
with the lipoids of the cells, perhaps with the lecithin, and in
doing so changes their normal relationship to other cell constituents
through which an inhibition of the entire cell chemism results.”
Evidently this theory casts no light upon the nature of the altera-
tions in cell chemism that follow. It remained for Verworn,
Mansfeld, Biicher and Heaton not only to demonstrate the nature
of these changes in cell chemism but also to show that the anes-
thetic state itself is in all probability dependent upon these
alterations.

Reicher, who demonstrated the constant presence of lipoidemia
after narcosis, explained this as being a protective mechanism; the
lipoid molecules acting as amboceptors uniting with anesthetic
and thus protecting the more vitally important brain lipoids.
Nerking attempted to prove this experimentally by injecting
various quantities of a I to 20 per cent. lecithin emulsion in normal
salt solution intravenously, intraperitoneally, intraspinally or sub-
cutaneously in animals which had been narcotized or were about
to be anesthetized and claimed to have shown that ‘‘ The injection
of lecithin has an undoubted influence upon the duration and after
effects of anesthesia in that it shortens its duration, brings about a
more rapid return to consciousness and eliminates unpleasant after
effects.”

A careful analysis of Nerking’s work shows it to be scarcely
worthy of serious consideration, owing to its lack of exactness in the
dosage of the narcotic and the indefiniteness of the criteria used.

16 SCIENTIFIC PROCEEDINGS (50).

Nevertheless, the importance of his conclusions, if confirmed, is
quite evident if this could be done with the above mentioned
experimental defects eliminated.

In order to accomplish this all anesthetics were given intra-
venously, a 5 or Io per cent. ether solution in normal salt solution
being used, and this was injected at definite rates; the lecithin
emulsion was also injected directly into the vein. The criteria
used were the appearance and disappearance of the corneal reflex,
or, when this did not occur, the loss of cutaneous pain sensation
was used. Other criteria were the time when the animal first
raised its head, when it first assumed the erect posture, and when
all ataxia disappeared. The results obtained seem to point over-
whelmingly to the following conclusions:

1. That the intravenous injection of 5 to 30 c.c. of a 5 or 10
per cent. emulsion of lecithin, depending upon the size of the
animal used, does not interfere with the induction of anesthesia and
that this can be accomplished as readily in animals thus injected
as in controls.

2. That in the majority of experiments, lecithin has no effect
upon the rapidity with which the various phenomena which
indicate the animal’s recovery from the effects of anesthetic appear,
in fact, in most cases, it retards their appearance.

The above experiments do not seem to bear out Reicher’s
assumption as to the cause of the lipoidemia and the explanation
of this phenomenon still remains an open question.!

II (707)

Habit and its relations to the nervous system in the earthworm.
By ROBERT M. YERKES.

[From the Psychological Laboratories, Harvard University.]

This is a preliminary report of an investigation now in progress,
the purpose of which is (a) to demonstrate whatever ability the
earthworm may have to acquire habits of a certain order; (0)
to discover the characteristics of any habits which appear; (c) to

1 The complete report of this work will appear in the Journ. of Exp. Med. for
February, 1913.

HaBiIT IN THE EARTHWORM. 17

enumerate and evaluate the various external and internal influences
on habit-formation; (d) to ascertain the degree of permanency of
the habits and (e) to discover their relations to the anterior ganglia
(brain).

By means of a T-shaped maze constructed from plate glass,
specimens of the manure worm, Allolobophora fetida, were tested.
The maze was placed with the stem directed toward the light.
Across one of the arms a piece of sandpaper was placed and, just
beyond it, a pair of electrodes. The other arm was left open so
that the worm might escape to an artificial burrow. The worms
were driven into the T by light and the chief motive for escape
therefrom was the tendency to avoid light. It was the purpose
of the test to demonstrate (a) any ability which the manure worm
may possess to acquire a direction habit and (0) to associate the
tactual experience of contact with sandpaper with the electrical
shock which regularly followed the tactual stimulus in case the
worm continued to move forward after reaching the sandpaper.

Trials were made in daily series varying in number from five to
twenty. The five-trial series were found, on the whole, most
satisfactory.

Referring now exclusively to the results obtained for a single
worm which has been under observation since October, 1911, the
following data may be presented. (1) Allolobophora is capable of
acquiring certain definite modes of reaction. (2) Modifications
appear as the result of from twenty to one hundred experiences.
(3) The behavior is extremely variable because of variations in
external conditions and in the condition of the worm itself. (4)
There is a tendency to follow the mucous path through the appa-
ratus but this is not sufficiently strong or constant to yield perfect
results. (5) The following are the chief modifications which have
been noted: (a) Increased readiness to enter the apparatus and
to desert it for the artificial burrow; (b) apparent ‘‘recognition”’
of the artificial burrow which is used as ‘‘exit tube”’; (c) a gradual
increase in the number of avoidances of the sandpaper and of
contact with the electrodes as a result of the ‘‘warning’’ influence
of the sandpaper; (d) the disappearance of the early tendency to re-
trace the path through the stem of the T; (e) the similar disap-
pearance of the tendency to turn back after progressing well

18 SCIENTIFIC PROCEEDINGS (50).

toward the exit tube. (6) The correct performance of a thoroughly
ingrained habitual act, of the kind studied in this investigation
is not dependent upon the “‘brain”’ (portions of the nervous system
carried by the five anterior segments), since the worm reacts appro-
priately within a few hours after its removal. (7) As the “‘brain’”’
regenerates, the worm exhibits increased initiative, its behavior
becomes less automatic, more variable. (8) Within four weeks
after the operation the regenerated segments appear superficially
complete and the worm naturally burrows in a mixture of earth
and manure. (9) Two months after the removal of the “brain,”
during the last four weeks of which period no training was given,
the habit had completely disappeared from worm No. 2, the
subject to whose responses this paper is devoted, and in its place
there appeared a tendency to turn in the opposite direction to
that demanded in the training. (10) Systematic training for two
weeks resulted in the partial re-acquisition of the original direc-
tion-habit.

The general results which have just been stated are subject to
modification in the light of additional data. To the experimenter,
it seems that the particular individual which has been longest
under observation is in many respects exceptional. It is perfectly
clear, however, from results obtained with six individuals that
important modifications in behavior appear as the result of train-
ing. It is equally certain that direction-habits are not readily
acquired.

It is the purpose of the experimenter in the continuation of
the investigation to pay especial attention to the relation of the
nervous system to modifications of behavior.

12 (708)

Preliminary note on the action of glucose upon the amount of
epinephrin in the blood.
By ISAAC OTT and JOHN C. SCOTT.
[From the Laboratory of Physiology, Medico Chirurgical College,
Philadelphia.]}
The modern theory of experimental diabetes is that a glyco-
secretory center is located in the medulla, from which impulses

HORMONES OF THE SECRETION OF MILK. 19

pass down the cord, emerge in the splanchnics, and go to the liver
to increase the transformation of glycogen into glucose. It is
held by some that as the splanchnics contain the secretory fibers
of the adrenals, that these impulses from the glyco-secretory center
increase the amount of epinephrin, which mobilizes the glycogen
of the liver and thus produces a diabetes. It is well known that
epinephrin is a stimulant of the sympathetic nerves, hence it is a
stimulant of the splanchnics. We have found the injection of
glucose per jugular in the cat increases the amount of epinephrin
in the blood, as shown by the intestinal strip of the rabbit. Hence
we have here a circle: epinephrin stimulates the secretory nerves
of the adrenals to produce epinephrin, which via the glycogen of
the liver produces more glucose, which in its turn generates more
epinephrin. We have also found the other sugars to increase the
amount of epinephrin in the blood. Pilocarpin, skatol and indol
also augment the amount of epinephrin in the blood.

13 (709)

Preliminary note on the inhibitory and synergistic hormones of
the secretion of milk.

By ISAAC OTT and JOHN C. SCOTT.

[From the Laboratory of Physiology, Medico Chirurgical College,
Philadelphia.|

We have studied on the goat the hormones which inhibit the
milk secretion. We have found that the ovary inhibits the action
of infundibulin, pineal gland, corpus luteum and thymus upon the
secretion of the mammary gland. Pancreas, spleen, iodothyrin-
parathyroid and adrenalin also inhibit the action of indundibulin.
Orchitic extract increases the activity of infundibulin, thus being
a synergistic agent. Egg albumen does not inhibit the action of
infundibulin.

20 SCIENTIFIC PROCEEDINGS (50).

14 (710)
Heterochromosomes in mammals.
By 'H. E. JORDAN.
[From the Department of Anatomy, University of Virginia.]

Heterochromosomes have now been reported for the male germ
cells of the following mammals: man and rat (Guyer, ’10);
armadillo (Newman and Patterson, ’10); opossum (Jordan, ’11);
guinea-pig (Stevens, ’11); and bat (Jordan, ’12). Winiwarter and
Sainmont, ’09, report a longitudinally split ‘‘monosome”’ in the
oécyte of the cat.

A comparative study of mammalian spermatogenesis reveals
the absence of typical heterochromosomes in mongoose,* cat,
squirrel, rabbit and pig. Heterochromosomes are clearly present
at synapsis and prophase in the primary spermatocytes of the
following forms: white mouse, sheep, horse, mule, dog and bull.
Regarding dog, rabbit, and the monkey, the evidence is not yet
decisive.

At certain stages the heterochromosomes (chromosome-nu-
cleoli) appear single (accessory; monosome), at others double or
bipartite. The latter appearance suggests a pair of idiochromo-
somes; but the body is more probably a split accessory.

The absence of discernible heterochromosomes in the male,
and their conspicuous presence in the female, of the cat indicates
their presence in one or the opposite sex in all forms. If this
hypothesis can be supported by evidence from the odcytes of
mongoose, squirrel, pig, rabbit and similar forms, cogent additional
confirmation is given to the idea of a special significance of het-
erochromosomes, probably in connection with the determination
of sex.

A simple explanation of sex-determination suggested by these
and other facts — and one in apparent accord with a large body of
experimental and cytological data— would seem to be to regard

* This material was collected at the temporary Marine Biological Station of the
Carnegie Institution of Washington at Montego Bay on the expedition to Jamaica,

B. W. I., in February and March, 1912, under the directorship of Dr. Alfred G.
Mayer.

DETERMINATION OF GLYCOGEN IN THE LIVER. 21

the heterochromosome-complex or ‘‘ X-element’”’ (Wilson), con-
tributed by the spermatozo6én, as an inhibitor to male sex.
Regarded in terms of Mendelian concepts, however, an apparent
contradiction results in that the presence of a determiner (inhibitor
to maleness) would here have to be recessive to its absence. But
in terms of a quantitative interpretation two X-elements in the
zygote would prevent, one X-element permit, the development of
male sex. Similarly with respect to the phenomenon of sex-
limited heredity: the X-element may act as the inhibitor in the
female to the male-limited character.

15 (711)

A comparison of chemical with microchemical methods for the
determination of varying amounts of glycogen in the liver.

By G. Y. RUSK.

[From the Hearst Laboratory of Pathology and Bacteriology, Uni-
versity of California.]

In collaboration with Dr. F. P. Gay, a study has been made of
the glycogen in the livers of 22 rabbits, comparing the chemically
determined amount with the histological appearance with a view
to finding the value of the latter for comparing small differences in
glycogen content. Pfliiger’s method was employed as far as the
conversion of glycogen to glucose and for the quantitative estima-
tion of the latter Bertrand’s method was used. For the histological
picture, Best’s carmine method and Langerhans’ modification of
Ehrlich’s iodine method were employed.

The following table gives the comparative results. The chem-
ical factors are reduced to a common denominator, viz., the amount
of copper which is reduced by 100 gm. of liver, and arranged from
the highest to lowest amounts. The histological results are placed
ina parallel column. While in the main there is faily close correla-
tion, yet there are two striking discrepancies (No. 23 and No. 24).

This study presents, so far as we can determine, the first
attempt to correlate chemical and microchemical findings with a
view to utilizing the latter for comparing slight differences of
glycogen in the liver. The results do not warrant the assumption

22

SCIENTIFIC PROCEEDINGS (50).

of parallel results between the chemical and microchemical results;
hence the histological method is not available for accurate com-
parative work.

Liver of
rabbit
number

23
20
22

18

Chemical deter-
mination of
glycogen (cop-
per gms. per
roo g. liver).

13.940
11.072
5.688
5.014

4.172
3.612

3-609

3.098
2.604
2.380
2.371

2.317
2.161
2.152
1.895
-978
-942
-836
-720
.560
-484
-366

Liver of
rabbit
number

TABLE.

Histological amount and distribution of glycogen.

Throughout lobule; most marked about central

vein and in single layer of cells at periphery.

Very marked in inner two-thirds of lobule;
trace in outer one-third.

Inner half of lobule shows well-marked glyco-
gen; also single layer about periphery.

Much the same distribution as No. 21 but less
in quantity.

Same general distribution but slightly less.

Slightly more diffuse in distribution but less
intense.

Narrow border about central vein shows glyco-
gen well marked.

Somewhat less.

Rather diffuse and pale in inner third.

Somewhat less.

In thin strip of 1 to 2 cells about central and
sublobular veins is well-marked glycogen.

Trace.

Trace.

Trace.

Trace.

Trace.

None.

None.

None.

None.

None.

None.

SCIENTIFIC PROCEEDINGS.

ABSTRACTS OF COMMUNICATIONS.
Fifty-first meeting.

The Rockefeller Institute for Medical Research, December 18, 1912.
President Ewing in the chair.

16 (712)

The effect of strychnin in cardiectomized frogs with destroyed
lymph hearts; a demonstration.

By S. J. MELTZER.

[From the Department of Physiology and Pharmacology of the
Rockefeller Institute.]

In several communications we have reported that the injec-
tion of solutions of strychnin, morphin or acid fuchsin in cardiec-
tomized frogs is liable to bring on convulsions of these animals.
The lymph hearts continue to beat for a while after the cardiec-
tomy. But since the lymph hearts assist the circulation only
by emptying their contents into veins, it seemed to be evident
that the removal of the blood heart eliminates also the circulatory
function of the lymph hearts. I have therefore assumed that
the above mentioned alkaloids reach the central nervous system
by way of the lymph spaces, which are connected throughout
the body, and which are capable of serving as a path for dis-
tribution by means of a peripheral mechanism. In a recent
paper by Abel (Jour. of Pharmacology, III, 581, 1912) in which
our facts were confirmed and in which it was admitted that the
activity of the posterior lymph hearts can not come into con-
sideration, the statement was made that ‘‘the appearance of
convulsions in the experiments of Meltzer and his pupils with
acid fuchsin, morphin and strychnin depends entirely on the
integrity of the anterior lymph hearts.’’ This statement is sup-
ported by a report of experiments in which, after destruction of

23

24 SCIENTIFIC PROCEEDINGS (51).

the anterior lymph hearts, in addition to cardiectomy the alkaloids
under discussion did not bring on any convulsions. I shall not
enter here into a discussion of the entire subject. I merely wish
to let you witness some indisputable facts. You see here a series
of frogs from whom the thoraco-abdominal viscera have been
removed, and in addition, the lymph hearts were destroyed by
cauterization. All these animals were injected about an hour
ago with strychnin; the injections were made in some animals
into the dorsal lymph sac and in others into the femoral sacs of
both thighs. You see that they respond to a tap with a tetanus.
This shows definitely that the injected strychnin reached the
central nervous system of these completely eviscerated frogs
without the help of the anterior lymph hearts. But you see also
that the trays holding the frogs are kept over ice. This is done
because at the room temperature, I could not be sure of the
success of my demonstration. It is possible that Abel’s observa-
tions were made in May, when such experiments are apt to fail.

17 (713)

Pulmonary lesions by intra-bronchial insufflation of cultures of
B. megatherium. With a demonstration.

By MARTHA WOLLSTEIN and S. J. MELTZER.

[From the Laboratories of the Rockefeller Institute for Medical
Research.]

The production of experimental pneumonia in dogs by means
of intra-bronchial insufflation of bacterial cultures has now been
carried out in several series of investigations. In the first series
Lamar and Meltzer produced lobar pneumonia by insufflation of
cultures of a virulent pneumococcus. In a second series we
produced lobular pneumonia by insufflation of a virulent strepto-
coccus and of the influenza bacillus. Besides the differences in the
gross appearances of the lesions both pneumonias offered the
following distinguishing points: In the lobar pneumonia of the
virulent pneumococcus there was a mortality of about 16 per
cent.; even in the non-fatal cases there was bacteremia present
in the first twenty-four hours; the exudate was rich in fibrin
and the framework of the lungs was invariably free from leucocytic

INTRA-BRONCHIAL INSUFFLATION OF B, MEGATHERIUM. 25

infiltration. In the cases of broncho-pneumonia produced by
the streptococcus or by the influenza bacillus there was no mor-
tality; at no time was there a bacteremia; the exudate contained
very little fibrin, but the framework of the lungs was invariably
and often very intensely infiltrated with leucocytes.

In a third series of experiments, which has not yet been
published, we studied the effects of insufflation of a non-virulent
pneumococcus. This organism also caused the development of a
lesion which macroscopically resembled the lesion of lobar pneu-
monia produced by the virulent pneumococcus. However, there
was no mortality, no bacteremia, and an exudate comparatively
poor in fibrin; the framework of the lung was here again practically
free from leucocytic invasion.

The results obtained with the non-virulent pneumococcus led
us to a study of the intra-bronchial insufflation of such a non-
pathogenic organism as the B. megatherium. Ten experiments
were made. The insufflation brought on a definite pneumonic
lesion in every case. From the gross appearance the lesion has
to be considered as that of lobar pneumonia of a milder type.
The entire process is essentially similar to the one caused by the
non-virulent pneumococcus, but its course is even milder. The
development is slower, the reaction is less intense and the resolu-
tion occurs sooner; there is practically no fibrin at all in the
exudate and bacteria can only occasionally be cultivated from
the exudate of 24 hours’ duration. There is little involvement
of the framework.

We show you here several lungs in the various stages of the
inflammatory process; the lesions were produced by the intra-
bronchial insufflation of twenty cubic centimeters of an eighteen
hour old bouillon culture of the B. megatherium.

26 SCIENTIFIC PROCEEDINGS (51).

18 (714)

The development of experimental pneumonia under direct obser-
vation of the lungs in the living animal.

By MARTHA WOLLSTEIN and S. J. MELTZER.

[From the Laboratories of the Rockefeller Institute.]

Experimental pneumonia produced by intra-bronchial insuf-
flation of virulent pneumococci may develop quite rapidly. Thus
in the experiments of Lamar and Meltzer it was observed that
in one instance seven hours after the injection, nearly complete
consolidation affecting the greater part of one lobe was already
present. We therefore now made several experiments in which
the lungs were under direct observation continuously for several
hours after the insufflation of the culture. The procedure
has been as follows. The dog was anesthetized by the cone
method, and a tube introduced into a bronchus as deep as it
could be pushed; the pneumococcus culture was then injected.
Immediately after, the tube was withdrawn, so that its lower
end was just above the bifurcation, and the arrangement made
for continuous intratracheal insufflation. The dog received about
“half ether’? and was continually under complete anesthesia,
The thorax was now widely opened transversely and about two
thirds of three or four of the lower ribs on the right side removed;
the right lower lobe, which is usually the seat of the inflammation,
was now exposed to full view. We shall not enter here upon
details. It may suffice to state that we were able to watch the
successive stages from the earliest signs of engorgement to complete
hepatization. We were also able to establish changes in the ausculta-
tion phenomena in the consolidated parts, in some instances even
clearly suggesting tubular breathing. We intended to demonstrate
such experiments, and we prepared several animals for this purpose.
The experiments were successful indeed, but we did not take the
lateness of the hour of the meeting sufficiently into account. We
prepared the experiments too early and all the animals died before
the meeting began. The acute infection, the great loss of heat,
the severe operation, and the long-lasting anesthesia are severe
factors to contend with. So far five hours has been the longest

DIFFERENCES IN Toxic EFFECTS OF ETHER AND CHLOROFORM. 27

time we could keep them alive. We show here the lungs of one
of these dogs. Death occurred in this instance about four hours
after the injection. The quantity of the injected pneumococcus
culture was quite large—about twenty cubic centimeters. About
two thirds of the right lower lobe is consolidated; as you see, it
is very dark and firm, and the surface shows numerous small
patches of fibrin.

19 (715)

Differences in the toxic effects of ether and chloroform, as
observed under intratracheal insufflation.

By T. S. GITHENS and S. J. MELTZER.

[From the Department of Physiology and Pharmacology of the
Rockefeller Institute.]

In studying the toxic effects of chloroform and ether, when
administered by the method of intratracheal insufflation, we
observed various differences in the course of the intoxication
brought on by excessive doses of these anesthetics. We shall
discuss here, however, only the differences in the toxic action of
these drugs upon the functions of respiration and blood pressure.
At the outset we have to point out, that when administering the
anesthetics by the insufflation method one of their dangerous
effects is here eliminated; it is the danger which is bound to result
from a partial or complete paralysis of the respiratory function.
Under the method of insufflation, life remains safe even when the
animal is completely curarized. Observations may be carried on,
therefore, even after spontaneous respiration is completely abol-
ished. On the other hand, intratracheal insufflation carried on
with ordinary, permissible air pressure, does not cause apnea,
that is, the individual continues to carry on its own spontaneous
respirations, which on tracings are easily distinguishable from the
infrequent partial interruptions of the continuous insufflation of
air.

The observations which we wish to report here briefly are as
follows. When using ether, a certain dose may be administered
which is amply sufficient to keep the animal completely anesthe-
tized, while respiration and blood pressure may remain practically

28 SCIENTIFIC PROCEEDINGS (51).

unimpaired for several hours. When this anesthetic dose is
exceeded, the first striking effect is upon the spontaneous respira-
tion which may be rapidly abolished. For instance, if complete
anesthesia has been accomplished by a dose which we term “%
ether’”’ or ‘‘34 ether” and now ‘“‘full ether’’ is turned on, the
respiration may stop completely within one to five minutes. At
this stage blood pressure is not impaired. When, however, “full
ether’’ is continued the blood pressure begins to come downs
The descent is very gradual and slow. It is rarely less than one
hour, in some cases it may be even several hours, before the
pressure reaches the dangerous stage. At that stage the blood
pressure may not be above 20 or 25 millimeters of mercury, and
the pulse pressure also considerably reduced. However, even at
this stage when the ether is turned off, the pressure may begin to
rise at once; although some time has to pass before the respiration
returns, and there is still another interval before reflexes and
consciousness return. The duration of the returning ascent is
shorter than the duration of the descent. It is, however, not
absolutely necessary to shut off the ether completely; a reduction
to ‘‘half ether”’ is, as a rule, soon followed by a return to a degree
of blood pressure which is sufficient to obviate danger.

The ready abolition of the respiration by ether is a practically
important phenomenon; it may serve as an indication that the
etherization has entered the toxic zone. It is a danger signal,
and since hours may pass before the real danger will be arrived at,
it is a safe and very valuable signal.

It is different with chloroform. In doses which are un-
doubtedly in excess of the anesthetic dose, respiration and blood
pressure go down practically together, and this in a comparatively
short time. When the administered dose is only slightly in excess
of the reliably anesthetic dose, the impairment of respiration does
not set in as early as under ether intoxication; but when this sets
in, blood pressure begins to fall also, although the respiration may
in some cases cease fifteen or twenty minutes before the fall of
blood pressure reaches the danger point. Respiration, perhaps,
resists chloroform intoxication slightly longer than that of ether,
while blood pressure is affected much more rapidly and profoundly
by chloroform than by ether. The zone separating the safe

THE RELATION oF LEucocyTic Extract To Bopy FLuips. 29

anesthetic and danger lines is narrow, and there is no reliable
danger sign.
20 (716)
The relation of leucocytic extract to body fluids.
By WILFRED H. MANWARING.

[From the Laboratories of the Rockefeller Institute for Medical
Research.|

In a previous communication! it was shown that a com-
paratively strong bactericidal substance can be extracted from
horse leucocytes. This substance loses its bactericidal power, in
whole or in part, if it is mixed with homologous or foreign sera,
with pathological exudates, with cerebro-spinal fluid, with the
products of aseptic tissue autolysis, or with most of the products
obtained by the bacterial decomposition of tissues. It also loses
its bactericidal power if it is mixed with the products obtained by
the prolonged autolysis of leucocytes themselves.

With sera that are in themselves bactericidal, not only is there
a loss of the bactericidal power of the leucocytic extract in such
mixtures, but there is also a destruction or inhibition of the bac-
tericidal power of the serum itself. This gives the phenomenon
of two bactericidal substances, an active serum and an active leu-
cocytic extract, added to each other, producing a non-bactericidal
substance, a good culture medium for bacteria.

An analysis of the antibactericidal action of serum shows that
it is due to the combined effects of three factors: (1) the anti-
bactericidal power of the serum colloids, (2) the antibactericidal
power of sodium chloride and the other neutral diffusible serum
components, and (3) the antibactericidal action of the diffusible
serum alkalies.

Alkalies are very strongly antagonistic to the leucocytic
bacteriolysin. The addition of 1/200 per cent. NaOH to leucocytic
extract is usually sufficient to completely inhibit its bactericidal
action. Acids, on the other hand, apparently have little or no
antibactericidal effect.

This antibactericidal power of serum and tissue fluids can not
be overcome by increasing the amount of leucocytic extract in the

1 These PROCEEDINGS, Vol. IX, 1912, p. 74.

30 SCIENTIFIC PROCEEDINGS (51).

mixtures, except when the serum and tissue fluids are tested in
minute quantities or greatly diluted. If tested in the full strength
in which they appear in the animal body, no amount of leucocytic
extract added to them is able to overcome or to exhaust their
antibactericidal power.

Attempts to lessen the antibactericidal power of serum and
body fluids by neutralizing their alkalinity with boric acid,
acetic acid and other weak acids, are occasionally partially
successful. Acidulation of the non-bactericidal mixtures occa-
sionally restores part of their bactericidal power. Inno experiment
thus far done however has more than a quarter of the original
bactericidal power been restored by this means.

The mechanism of the antibactericidal actions of serum and
tissue fluids has not been determined.

21 (717)
On the lysis of tubercle bacilli.

By WILFRED H. MANWARING and J. BRONFENBRENNER.

[From the Laboratories of the Rockefeller Institute for Medical
Research.]

It was pointed out by Koch, and has since been confirmed by
others, that an animal suffering from a chronic local tuberculous
lesion is more resistant than the normal to inoculation with
tubercle bacilli.

This heightened resistance has been studied by numerous
workers. Calmette, for example, found that tubercle bacilli,
injected into the subcutaneous tissues of tuberculous cattle, soon
showed marked involution and degenerating forms, but that the
bacilli did not completely disappear from the site of the inocu-
lation and the neighboring lymph glands by the end of 120 days.
A rapid lysis or destruction of the bacilli was not observed by
these workers.

Recently Beycke and Much! and others have applied the Pfeiffer

1 Beycke and Much, Beitrag z. Klinik f. Tuberk., 1910, XV, p. 277+

Much and Leschre, ibid, 1911, XX, p. 405.

Kraus and Hofer, Deutsch. med. Wochenschr., 1912, XXXVIII, p. 1227; Wiener
klin. Wochenschr., 1912, XXV, p. I112.

Tue ROLE OF PHAGOCYTOSIS IN INVOLUTING ORGANS. 31

technique to astudy of this phenomenon. They have found that
tubercle bacilli, injected into the peritoneal cavities of tuberculous
guinea pigs, undergo rapid degeneration and a rapid decrease in
number.

They interpret these changes asa rapid lysis of bacilli. If this
interpretation is correct, it will necessitate a revision of current
ideas concerning the resisting power of the tubercle bacillus.

We have repeated and extended their work. We have found
that tubercle bacilli, injected into the peritoneal cavities of
tuberculous guinea-pigs, will occasionally give the degeneration
forms and the non-staining forms they describe; and that, under
certain conditions, there may be a complete disappearance of the
bacilli from the peritoneal fluids within as short a period of time
as three hours.

Whether this disappearance is due to an actual lysis of the
tubercle bacilli, or to other causes, we have not yet determined.
As evidence in favor of lysis we have the observation that all of
the normal control guinea pigs injected intraperitoneally with the
test suspensions of tubercle bacilli have died from a fulminating
type of visceral tuberculosis, within a period of from three to four
weeks, while most of the tuberculous guinea pigs, receiving the
same test doses, have; survived for a longer period of time. A
few of these tuberculous guinea pigs however have died within
twenty-four hours after the intraperitoneal tests, suggesting an
anaphylactic reaction.

We have obtained a similar rapid disappearance of tubercle
bacilli from the peritoneal cavities of tuberculous rabbits, from
tuberculous rats, and from tuberculous dogs. The mechanism of
the phenomenon is now under investigation.

22 (718)
The role of phagocytosis in involuting organs.
By Max Morse.
[From the Boardman Laboratories, Trinity College, Hartford, Conn.]

In order to clear the way for a study of the transfer of protein
in the involuting tail of the tadpole, it was necessary to reéxamine
the rdle which has been ascribed by Metschnikoff, Barfurth,

32 SCIENTIFIC PROCEEDINGS (51).

Bataillon, Mercier and many others to the leucocytes in producing
the phenomenon. Looss believes on histological ground, that
if leucocytes play any part in the process of tissue atrophy and
absorption, they play a minor and secondary one rather than a
primary réle. The process of atrophy begins prior to the invasion
of the leucocytes into the muscle of the tail and this has been
described by Mercier and others, who hold that phagocytosis is
the principal factor. Moreover, it has been described for other
involuting organs, such as in the metamorphosis of insects, the
absorption of the gills of amphibia, etc. It has been suggested
for the involution of the mammalian uterus, likewise, so that it
may be said that investigators are in accord in observing a dis-
solution of the muscles and other tissues in atrophying organs prior
to the advent of leucocytes.

It is to be expected that if phagocytosis plays any important
role in the inception of the process of absorption of tissue in the
larva of the frog, the blood would show an increase in the total
number of leucocytes during the stages of metamorphosis and
moreover there would be an increase in polymorphonuclear
leucocytes during these stages, to compensate for the drainage
of these cells into the muscles. In order to examine this point,
smears were made of the blood from the larve of the bull-frog,
Rana catesbiana and from the western pickerel frog, Rana areolata.
Thirty specimens were used, the blood being permitted to flow into
a capillary tube from a lesion in the heart and then blown upon a
slide, dried in the air and stained with Wright’s stain. A large
number of hematocytometer counts were made upon fresh blood,
but this method of estimation was abandoned on account of the
unsurmountable difficulty of recognizing the different kinds of
young corpuscles. As Freidsohn has shown, the various sorts of
leucocytes together with the erythrocytes take their origin from
cells more or less similar in appearance in the earlier stages, this
common stage resembling the large mononuclear leucocyte of the
human blood.

The following table gives the summary of the differential counts
made upon the smears:

Polymorphonuclear leucocytes are in slight advance in absorb-
ing individuals over those not yet metamorphosing, but there is a

THE ROLE oF PHAGOCYTOSIS IN INVOLUTING ORGANS. 33

Polymorpho- | Basophiles,} Eosinoph., | Large M., | Small M.,

nuclear, percent.| per cent. per cent. per cent. per cent.
INDSOFDING sista sucls.o.s emp evens 9.8 4.2 6.5 36.1 42.4
Non-absorbing.......... 8.6 4.7 7.0 20.6 59.0

Adults) revniccdiene one ave 18.3 6.2 0.4 13.2 61.2

much greater number of these cells in the adult. Of basophiles,
there are more in the adult than in the larval stages and more in
the non-absorbing than in the absorbing animals, although the dif-
ference is slight. Eosinophiles occur in about the same relative
numbers as the basophiles, but they are rare in adults. The case
is more difficult to analyze in the large and small mononuclear
types, for doubtless these classes have been made to include young
leucocytes of the foregoing kinds. Inasmuch, however, as his-
tological sections of the tail of the metamorphosing larva show
few cells of either of these classes, it is permissible to rule them
out of the consideration as phagocytes.

From this evidence, it seems fair to conclude that phagocytosis
is not the primary factor involved in the involuting tadpole’s
tail and, since the process of involution everywhere involves a
breaking down of tissue before leucocytes have entered the tissue,
it may be concluded that these blood cells nowhere are of impor-
tance in explaining the process of tissue absorption. Exception
may be made in the case of certain types of carcinoma, which,
however, have not been satisfactorily studied from this point of
view.

Evidence will be presented later that the process of involution
concerns autolysis, as has been suggested in the case of mammalian
tissues. Evidence will also be submitted to show that the transfer
of tissue involves a closed circuit and that there is no increase in
total nitrogen in the excreta during the process of metamorphosis,

PAPERS MENTIONED.

BATAILLON, E. ’o1. Recherches anatomiques et expérimentales sur la méta-
morphose des Amphibiens anoures. Annales de l'Université de Lyon, Tome 2.

BARFURTH, D. ’876. Die Riickbildung des Froschlarvenschwanzes und die sogen-
nanten Sarcoplasten. Arch. fiir mikr. Anat., Bd. 28, S. 35 ff.

FREIDSOHN, A. ‘10. Zur Morphologie des Amphibienblutes, zugleich ein Beitrag
zur Lehre von der Differenzierung der Lymphocyten. Arch. fiir mikr. Anat.,
Bd. 75, S. 435:

Looss, A. ‘89. Uber Degenerations-Erscheinungen im Thierreich, besonders

34 SCIENTIFIC PROCEEDINGS (51).

tiber die Reduction des Froschlarvenschwanzes und die Verlauf derselben
auftretenden histolytischen Processe. Preisschriften gekrént und heraus-
gegeben von der fiirstlich Jablonowski’schen Gesellschaft zu Leipzig. Leipzig:
Verlag von B. S. Hirzel.

METSCHNIKOFF, E. ’92b. Atrophie des muscles pendant la transformation deg
Batraciens. Ann. Inst. Pasteur, T. 6, p. I.

23 (719)
Effect of phlorhizin on a dog with Eck fistula.
By N. B. Foster.

It has been stated by Rosenfeld that the administration of
phlorhizin to dogs with Eck fistula does not induce glucosuria.
This observation would have so much bearing upon our ideas of
the mode of action of phlorhizin that the subject required con-
firmatory evidence.

One gram of phlorhizin in olive oil emulsion was given to a dog
on which an Eck fistula had been done. Glucose was found in the
urine in considerable quantity for nine days subsequent to the
phlorhizin adminstration.

24 (720)
On elimination through the mucosa of the urinary bladder.
By ISRAEL S. KLEINER.

[From the Department of Physiology and Pharmacology of the
Rockefeller Institute for Medical Research.]}

In a previous communication (Journal of Exper. Medicine,
XIV, 274, 1911), I stated that after intravenous injections of large
amounts of dextrose the intestine contained on the average 1.2 per
cent. of the injected amount of destrose and in nephrectomized
animals the dextrose content of the intestine reached the average
of 2.2 per cent. The question arose then whether in the presence
of a strong hyperglycemia, and especially after double nephrec-
tomy, all mucous membranes are slightly permeable to this
substance. The presence of measurable quantities of dextrose in
the gastro-intestinal canal is no evidence for the permeability of
the mucosa as such, since the mucous membrane of this organ is
studded with numerous glandular structures which may readily

PANCREATIC TRANSPLANTATION IN THE SPLEEN. 35

be instrumental in the removal from the blood of substances
present there in excess. In the experiments I wish to communicate
here, I selected, therefore, the bladder for the study of this problem
since the mucous lining of this organ is practically the only mem-
brane which possesses no glands. The rabbit was the experimental
animal used.

After performing a double nephrectomy and tying a cannula
in the bladder, seven to ten grams of dextrose per kilogram were
injected intravenously in from twenty to fifty minutes, washing
the bladder several times during and after the injection. The
washings were then analyzed. The results are briefly as follows.
In nine experiments either no sugar or else indeterminable amounts
were found, and in four experiments only six to eleven milligrams,
1. €., 0.05 to 0.08 per cent. of the amount injected. In addition,
in some of the experiments uranine, the sodium salt of fluorescein,
was dissolved in the injection fluid. In the experiments in which
twenty milligrams were introduced, no trace of uranine, a very
diffusible dye, could be detected in the washings of the bladder.
In four experiments 100 milligrams were introduced; in two there
was not a trace and in the other two there was a barely perceptible
trace.

The conclusion may therefore be drawn that the mucosa of the
bladder is practically impermeable for diffusible substances present
in the blood in great excess, even in the absence of the kidneys,
the chief organs of elimination of the body.

25 (721)
Pancreatic transplantations in the spleen.

By JosEPpH H. PRATT, of Boston, and FRED T. Murpny, of
St. Louis.

[From the Laboratory of the Theory and Practice of Physic, Harvard
University.|

The influence of pancreatic transplants in the spleen on the

prevention of diabetes was studied. The spleen was selected

because it has been shown that transplanted bits of tissue are

especially well nourished in the pulp of this organ. In nine

animals, bits of the pancreas of various size were buried in the

36 SCIENTIFIC PROCEEDINGS (51).

pulp of the spleen. The method used by Payr in transplanting
thyroid tissue was followed. In four animals (2 dogs and 2 cats)
pancreatic cells were demonstrable in the spleen. The interval
between the operation and the death of the animal varied from
18 hours to 13 days. In five animals (all dogs) no pancreatic
remains were found. Autolysis was rapid. In one experiment
all the pancreatic cells had disappeared 21 hours after the trans-
plantation. The bit of tissue found 13 days after the transplant
measured only I mm. by 0.1 mm. _ It consisted of normal appear-
ing acini surrounded by connective tissue. No islands of Langer-
hans were demonstrable.

In one dog a large pancreatic graft was placed in the spleen
with the blood supply preserved by means of a mesenteric stalk.
Three weeks later the original blood supply was cut off and all the
pancreatic tissue except the graft extirpated. Diabetes did not
develop, but the tolerance for glucose fell within a few weeks
toalow point. At the autopsy 187 days after the second operation
a large abscess was found in the lower part of the spleen. Pro-
jecting into the spleen from the wall of the abscess was a cone-
shaped mass of fibrous tissue. In this were the remains of the
pancreatic transplant measuring less than I cm. in length. The
pancreatic tissue consisted of acini separated by connective
tissue. In some of the cells, masses of zymogen granules were
present. These were no demonstrable islands of Langerhans.
This experiment proves that pancreatic tissue implanted in the
spleen and separated from its original vascular and nervous con-
nections can live and functionate for months.

26 (722)
The production of reversed cardiac mechanism in the dog.
By ALFRED E. CoHN.

[From the Hospital of the Rockefeller Institute for Medical Research,
New York.]

In a series of experiments, which were published with Kessel
and Mason,! on the excised perfused hearts of dogs, we were able
to show that ‘excision of the sinus node results in an immedate

1 Heart, June 15, 1912, iii, 311.

PRODUCTION OF REVERSED CARDIAC MECHANISM IN THE Doe. 37

cessation of the contractions of the whole heart,’”’ that ‘after
excision of the sinus node and the ensuing stoppage, the rate of
the whole heart falls and does not again reach the original rate,”
and that ‘‘the function of the secondary pacemaker devolves on
no special portion of the heart.’’ The results of experiments per-
formed by others and differing from ours were mentioned at that
time. I have continued the experiments on dogs, anesthetized
with ether, the hearts of which were left im situ and normally
nourished. After sufficient controls were registered electro-
cardiographically, the sinus node was clamped with a specially
constructed T clamp, 5 cm. long and 1 mm. wide. In the perfused
hearts the rate fell 10 to 70 beats, usually 30 to 50 beats, after the
node was excised. In the present series the fall in rate was I0 to
90, usually about 50. Stoppage, which is a usual phenomenon in
excised hearts, occurred also in these, but very rarely. The site
of the secondary pacemaker varied in these, as it did in the per-
fused hearts. Photographs of three hearts showing the areas of
the auricular surfaces which had been included in the jaws of the
clamp, and a series of curves from each of the corresponding experi-
ments and of one in addition were demonstrated. The relation of
the disturbed function of the heart to the area clamped will be
investigated histologically and reported in detail later. The
curves indicate that in each of these cases, there was a dislocation
of the pacemaking function from the site responsible for it, and
that, after clamping had taken place, this function devolved upon
the ventricles, the junctional tissues or another portion of the
auricle. In one of the experiments, first the ventricles (more
probably the junctional tissues) and later the auricles set the pace.
One of the experiments yielded curves which bore a close re-
semblance to those obtained from a patient by Williams and
James,! in which the wave representing auricular contraction was
found between the R and T waves, and was inverted. It is
demonstrated by the experiments that after clamping an area
along the sulcus terminalis, a site other than the normal pacemaker
assumes the pacemaking function and that a ‘reversal of the
cardiac mechanism” takes place.

1 Proc. Soc. Exp. BIOL. AND MED., Oct. 16, 1912, X, 13.

38 SCIENTIFIC PROCEEDINGS (51).

27 (723)
Distribution of amino acids in the body.

By DONALD D. VAN SLYKE and GUSTAVE M. MEYER.
[From the Rockefeller Institute for Medical Research.]

In a recent paper (Journ. Biol. Chem., Sept., 1912) we have
shown by direct determinations (nitrous acid method) of the
amino acid nitrogen of the blood, that during the digestion of
protein amino acids pass from the alimentary canal into the blood.
The amino acid nitrogen reaches only a relatively low concentra-
tion in the blood (3-5 mg. per 100 c.c. after 24 hours fast, 10-12
mg. during digestion), because, as was found after intravenous
injection of alanine, the amino acids are either destroyed or
removed with great rapidity by the tissues. The object of the
present work was to obtain direct proof, by analysis of the tissues
themselves with the nitrous acid method, as to whether the amino
acids disappear from the blood as the result of destruction, of
synthesis to more complex compounds, or of mere concentration
in the tissues. The last explanation is the correct one. The
tissues (muscle, liver, kidney, spleen, pancreas) contain, stated
very roughly, about ten times the amount of amino acid nitrogen,
per 100 grams, that is found in the blood. The amount varies
in the different tissues, the brain being especially low, but about
the same order of magnitude in this respect appears to be normally
maintained among the different organs. Amino acids injected
into the blood, or absorbed during digestion, are immediately
taken up by the tissues; but the removal, although rapid, is never
complete, equilibrium being reached and maintained between
blood and tissues respectively. The means by which the amino
acids are held in the tissues appear to be physical rather than
chemical; for the acids can be extracted by even cold water or
dilute alcohol. The fact that the amino acids are many times
more concentrated in the tissues than in the blood excludes
osmosis as an explanation. The facts thus far known are con-
sistent with the assumption that the amino acids are taken up
from the blood and held in the tissues by adsorption, chemical
transformation of the adsorbed amino acids following later. This
transformation occurs much more rapidly in the liver than in the
muscles.

THE Fate oF GLYCOGEN IN DIABETES, 39

28 (724)

The fate of parenterally introduced glycogen in human and
experimental diabetes.

By RoBerT A. COOKE.

[From the Laboratory of Chemical Pathology, Cornell University
Medical School.|

It has been shown by F. Voit and P. Mayer that glycogen
parenterally introduced into the normal body of man or lower
animal, is utilized, there being no elimination of glycogen, dextrin
or glucose in the urine. Mendel repeated this work with rabbits,
cats and dogs and found, on the contrary, that 5 per cent. to 18
per cent. of the glycogen introduced was eliminated as a dextrin
over a period of two days.

In human and experimental pancreas diabetes the administra-
tion of glycogen gave the following results:

I. Human diabetes, first trial, E. P., age 16 yrs., wgt. 110 lbs.
Glycogen subcutaneously, 28.0 gm.
Glycogen in urine as gluc. 28.0 gm.
Elimination time, under 24 hours.
II. Human diabetes, second trial (same case).
Glycogen subcutaneously, 21.5 gm.
Glycogen in urine as gluc. 21.5 gm.
Elimination time under 24 hours.
III. Complete pancreas extirpation; bitch, 8.5 kilos.
Glycogen intraperitoneally, 9.5 gm.
Glycogen recovered in urine as glucose 6.0 gm. = 66 per
cent.
Glycogen recovered in urine as dextrin 3.5 gm. = 34 per
cent.
Elimination time under 24 hours.
IV. Complete pancreas extirpation; bitch, 14.5 kilos.
Glycogen intraperitoneally, 25 gm.
Glycogen recovered in urine as glucose, 22.0 gm. = 88 per
cent.
Glycogen recovered in urine as dextrin, 3.0 gm. = 12 per
cent.
Elimination time under 22 hours.

40 SCIENTIFIC PROCEEDINGS (51).

These observations show definitely that glycogen is not a utilizable
form of carbohydrate for the diabetic organism and they indicate
that there is a more rapid conversion of glycogen to glucose in
the diabetic than in the normal body.

29 (725)
Decerebration and the action of morphine in frogs.

By J. S. GITHENSs.

[From the Laboratory of Pharmacology and Physiology,
Rockefeller Institute.]

Morphin given in sufficient dose to normal frogs causes tetanus
which does not come on, however, until several hours or even days
after the injection.

I have found that decerebration hastens the onset of tetanus
and also causes a marked reduction in the amount of morphin
required to induce tetanus.

The smallest dose with which tetanus can be induced regularly
in normal frogs at room temperature is 44 of a milligram per gm.
(10 milligrams for a 30 gm. frog). This tetanus comes on in about
24 hours.

In decerebrated frogs, at room temperature, tetanus comes on
after such a dose in from 1% to 6 hours, and may be induced with
certainty after 6 to 24 hours by a dese of 1/10 milligram per gram
(3 mg. for a 30 gram frog).

When frogs are kept cold tetanus can be induced by much
smaller doses, as we have stated in an earlier paper. Thus, intact
frogs kept in the cold show tetanus after doses of 1/30 mg. per
gm. (1 milligram for a 30 gram frog). Tetanus comes on after
such a dose in from 18 to 24 hours. Decerebrated frogs show
tetanus after such a dose in from 4 to 12 hours, and it may be
induced with certainty by doses of 1/300 milligram per gm. (1/10
mg. for a 30 gm. frog) after an interval of 1 to 3 days.

Frogs with the entire brain including the medulla destroyed,
do not respond as well as frogs with the entire brain except the
medulla destroyed.

REVERSAL OF THE CARDIAC MECHANISM. 41

30 (726)
Reversal of the cardiac mechanism. An additional note.
By Horatio B. WILLIAMS and HENRY JAMES.

[From the Physiological Laboratory of Columbia University,
New York.]

Since making our previous report on this subject, electro-
cardiograms have been obtained from the same patient in which
occasionally the small downward deflection between R and T
fails. The form of the ventricular electrogram in these instances
is precisely similar to the others with the sole exception of the
absence of the small downward deflection. This we regard as
confirmatory evidence of the correctness of the view that the
downward deflection is caused by the action of the auricles and
that the usual curve is not simply an unusual ventricular complex.
In the occasional instances cited conduction seems to have failed.

When this case first came under observation it seemed desirable
to attempt to reproduce the condition experimentally. We have
performed up to the present time seven experiments with one
positive result. Rothberger and Winterberg have published
curves showing transient reversal resulting from simultaneous
depression of the sinus-region of the right auricle with cold and
stimulation of the left ganglion stellatum. The apparent perma-
nence of the condition in our patient leads us to think rather of an
anatomical lesion than of nervous influences as the cause of the
condition. Our experiments have consisted in depression (with
anode) crushing, and excision of the region of the sinus node. In
some instances nearly the entire anterior wall of the right auricle
including a large part of the walls of the vene cave where they
fuse with the auricle, have been excised.

Slowing of the heart beat was the usual result and the auricles
and ventricles generally beat nearly or quite simultaneously. In
the single positive experiment auricular fibrillation resulted from
the application of a clamp to the upper part of the sinus region
where the greater bulk of the node is usually found. This per-
sisted for twenty minutes and stopped immediately on application
of a second clamp immediately below the first. The auricles

42 SCIENTIFIC PROCEEDINGS (51).

and ventricles then beat simultaneously. The application of a
third clamp at the junction of the inferior vena cava and auricle
was followed by reversal of the mechanism which persisted until
the animal was killed.

The occurrence of reversal but once in a series of seven experi-
ments has occasioned no surprise. We had surmised that after
destruction of the usual pacemaker it would be quite fortuitous
should the next most irritable focus lie below the auricles.

Before definite conclusions can be drawn regarding any relation
which may be thought to exist between the clinical and experi-
mental conditions it will be necessary to determine whether this
is the only lesion which can give rise to a permanent reversal of
the cardiac mechanism.

31 (727)
The intercalated discs of atrophied heart muscle.
By H. E. JORDAN.
[From the Anatomical Laboratory, University of Virginia.]

In two earlier papers! I presented evidence in support of my
interpretation of intercalated discs as irreversible contraction
bands. Ina more recent paper the idea was tested by appearances
in a natural experiment, namely, extremely hypertrophied heart
muscle.2. The conditions here obtaining were in perfect accord
with, and confirmed the plausibility of, my previous interpreta-
tion. Since then I have had opportunity to study lesser degrees
of hypertrophy, as well as an excellent specimen of atrophy’
(weight of heart 180 grams). It is the purpose of this note to
complete my report of observations on intercalated discs by a
record of my findings in atrophied heart muscle, and to reémpha-
size the point that all the evidence, including ontogenetic, com-

1 Jordan, H. E., (1911) ‘‘ The Structure of Heart Muscle of the Humming Bird,
with Special Reference to the Intercalated Discs,” Anat. Rec., 5: 11; (1912; with
Mr. K. B. Steele) ‘‘A Comparative Microscopic Study of the Intercalated Discs of
Vertebrate Heart Muscle,’’ Am. Journ. Anat., 13: 2.

2 Jordan, H. E. (1912), ‘‘The Intercalated Discs of Hypertrophied Heart
Muscle,” Anat. Rec., 6: 9.

3 For this material I am indebted to Dr. W. H. F. Addison, of the University of
Pennsylvania.

INTERCALATED Discs oF ATROPHIED HEART MUSCLE. 43

parative anatomical and pathological (experimental—including the
phenomena of fragmentation and segmentation) data points to
the same conclusion, namely, that the so-called ‘‘discs’’ are
aggregations of irreversible contraction foci on the myofibrillz
in the form of bands variously modified by a variety of normal
mechanical and pathological, both chemical and mechanical,
factors.

The first step, and central fact, in the chain of observations
leading to the above conclusion respecting the origin and nature
of the discs is the close similarity, amounting practically to an
identity, between the normal contraction band in contracted
muscle fibers and the simplest type of disc. This correspondence
of appearance is very striking in the humming-bird’s heart. In
the relaxed fiber the Z-line is very conspicuous but delicate.!
The Q-disc is wide and pale. In the contracted fibers the appear-
ance is one of an alternation of robust dark (Q) and light (J) bands
of approximately equal thickness. These are the same conditions
that obtain in striped muscle generally in relaxed and contracted
states respectively. In contraction the Z-line appears to become
thickened by accumulation about it of dark (Q) substance. Of
course the changes take place in the myofibrilla. We need simply
postulate local inability (due to strain, whether an instant or
cumulative effect) on the part of a contraction band to reverse
(or relax) to pass to the first step in the formation of a disc. The
similarity between such “bands” and “‘discs,’’ as also their periph-
eral and frequently supernuclear position, renders the assump-
tion almost a certainty. Further support to this interpretation
is derived from the following ontogenetic facts: (a) the discs
begin to make their appearance coincidently with the cross stria-
tions; (d) they are at first exclusively of the simple homogeneous
band type; (c) they increase in number (and complexity) at least
to the time of full growth of the heart; (d) they are permanent
structures throughout life. Further confirmative data are these:
(a) step-forms are rare in birds, and absent in lower forms, the
prevailing disc being of a type corresponding to the earlier onto-
genetic forms in mammals; (b) in hypertrophied hearts the discs
are exactly of a form expected from a modification of a simple

1 Prepared according to Zimmermann’s technique.

44 SCIENTIFIC PROCEEDINGS (51).

“comb-type”’ under the influence of longitudinal and transverse
gross enlargement, and multiplication of myofibrille, 7. e., irregular
zig-zag form; (c) mechanical ruptures in normal hearts are not
localized along the discs, while in pathological fragmentation and
segmentation the lines of fracture are largely limited to these
levels. Of further significance is the fact that the presence of step-
forms coincides with the condition of more profuse branching,
hence absent in forms below birds. Step-forms would seem to be
due to tension at various angles to the main longitudinal axis of
the fibers.

The predominant type of disc in atrophied heart muscle is the
comb-type. Fractures again occur almost exclusively at these
levels. The discs in pathological heart muscle appear to be regions
of weakness, probably in part at least due to chemical change.
The main steps in the formation of the discs are conceived as
follows: (1) irreversible contraction band (apparently homo-
geneous); (2) comb-discs, the length of the “teeth” varying
according to the total amount, or degree, of traction on the
modified regions of the included fibrils; and (3) zig-zag discs, due
to a combination of the factors of longitudinal and transverse
tension, and increase in number of fibrils by longitudinal splitting,
e. g., in hypertrophied cardiac fibers. Under the condition of
general compaction which prevails in atrophied cardiac fibers,
we should expect to find exactly the type of disc actually present,
namely, a comb-type. Comb-discs according to the above inter-
pretation of their origin are obviously regions of relative weakness,
of relatively greater degree in pathological hearts where there is
operating very probably an additional chemical factor. Hence
in atrophied hearts fragmentation when present is limited to these
levels. The discs once formed are permanent structures under-
going various modifications according to physiological conditions.
Direct observation reveals a close similarity between the contrac-
tion band and the simplest type of disc. Hence the evidence for
the origin of the various types of discs from irreversible contraction
bands is practically complete.

STARCHES IN RELATION TO GENERA, SPECIES, ETC. 45

32 (728)

The differentiation and specificity of starches in relation to genera,
species, etc.: stereochemistry applied to protoplasmic proc-
esses and products, and as a strictly scientific basis
for the classification of plants and animals.

By EDWARD TYSON REICHERT.

[From the S. Weir Mitchell Laboratory of Physiology, University of
Pennsylvania.|

Under the foregoing title the results of an elaborate investiga-
tion will shortly appear as Publication No. 173 of the Carnegie
Institution of Washington. This research is supplementary and
complementary to, and in support of, Publication No. 116 entitled,
“The Differentiation and Specificity of Corresponding Proteins
and Other Vital Substances in Relation to Biological Classification
and Organic Evolution: The Crystallography of Hemoglobins.”
A preliminary review of the latter will be found in the PROCEED-
INGS of the Society, 1908, V, 66-68.

Studies of the histological, physical, physico-chemical and
chemical properties of over 300 starches from various plant-sources
were made. Among the most important conclusions reached are
the following:

1. Starch is not a unit substance but exists in a vast number of
stereoisomeric forms. Starch from any given plant and of every
mature starch grain is a mixture of different forms of starch-
substance.

2. Starches from different plants exhibit constant and specific
characters in relation to genera, species and varieties, by which
the plant can be identified.

3. The stereochemic peculiarities of starches and other complex
organic metabolites constitute a strictly scientific basis for the
classification of plants and animals.

46 SCIENTIFIC PROCEEDINGS (51).

33 (729)

The motor cortex and pyramid tract in the raccoon
(Procyon lotor Linn.).

By SUTHERLAND SIMPSON.

[From the Physiological Laboratory, Medical College, Cornell
University, Ithaca, N. Y.]

The raccoon is a very intelligent animal and, as one would
expect, relatively to the size of its body, it possesses a large brain
with a highly convoluted cerebral cortex. Little appears to be
known, however, about its cortical topography or the fiber tracts
of its central nervous system.

The present note refers to the experimental localization of the
motor areas and the subsequent tracing of the pyramid tract by
the method of ablation and secondary degeneration. Five full-
grown specimens were obtained. The cerebral cortex was exposed
on the left side, under ether anesthesia, the motor areas were
localized both by the bipolar and unipolar methods and then
removed. The wound was closed and the animal allowed to live
for about two weeks when the left hemisphere was exposed and
explored in the same way before the animal was finally dispatched
by an overdose of ether. The brain and spinal cord were removed
and treated by the Marchi method.

The region from which muscular responses were obtained is
relatively large and well defined. It occupies the whole free
surface of what may be termed the post-cruciate convolution,
extending from the mesial border of the hemisphere to a little
way beyond the lateral extremity of the cruciate sulcus. Unless
when the current was comparatively strong, no movements were
obtained from the cortex in front of the cruciate sulcus. From
the mesial border lateralwards the order of the responsive areas
was as follows: anus, tail, hind limb and digits, body, fore-limb
and digits, head and eyes, and face, mouth and tongue, the last
curving forwards around the lateral extremity of the sulcus. The
movements were readily elicited and the areas well defined but
there was always some overlapping at the margins. The forearm
area appeared to be the most easily excited, that is to say, it

NoTE ON THE ACTION OF TONSILLAR EXTRACT. 47

responded to a weaker current than was necessary for the stimula-
tion of the other areas.

The pyramid tract fibers were traced from their origin in the
cerebral cortex, caudalwards, to the spinal cord. They occupied
the usual positions in the crusta, pontine bundles and anterior
pyramid above the decussation. In the posterior part of the
medulla oblongata most of them crossed the median raphe and
turned caudalwards into the lateral column of the spinal cord, but
in three of the specimens examined a considerable number of
fibers remained uncrossed and formed a direct ventral pyramid
tract, extending along the margin of the ventral median fissure.
This tract could be traced to the middle of the thoracic region
where it disappeared. A few uncrossed fibers were also found in
the lateral column.

It is generally believed that this uncrossed ventral tract (direct
pyramidal tract) is limited to man and the anthropoid apes; such,
however, is not the case, for it is present in the raccoon and is more
pronounced still in the porcupine.!

The crossed lateral pyramid tract could be followed to the last
of the sacral segments. In the raccoon this is a large tract both
in relative area, in transverse sections of the spinal cord, and in
the number of fibers which it contains. In no animal below the
macaque monkey have I found it so extensive.

34 (730)
Note on the action of tonsillar extract.

By Isaac OTT and Joun C. Scott.

[From the Laboratory of Physiology, Medico-Chirurgical College,
Philadel phia.|

We used the dried powdered tonsil of the calf. When a
filtered infusion of two grains of the powdered tonsil was injected
into the cat by the jugular vein in divided doses, it produced a
great fall of blood-pressure, lasting about a minute, followed by
a rise above normal, with a much slower and stronger heart beat.
Increase of this dose suddenly arrested the heart. In the same
animal it was also a diuretic, increasing the flow of urine twenty
times the original amount.

1 Sutherland Simpson, Proc. Soc. Exper. BIOL. AND MED., 1912, Oct. 18, p. 15.

48 SCIENTIFIC PROCEEDINGS (51).

35 (731)

The diffusion of iodo-eosin from ether through rubber
membrane into ether.

By JacoB ROSENBLOOM.

[From the Laboratory of Biological Chemistry of the University of
Pittsburgh, Pittsburgh, Pa.]

Rosenbloom and Gies! have shown that many ether soluble
substances diffuse from ether through rubber membrane into ether,
but oddly various phospholipins do not possess this property.

Together with Boas,? I was able to show that various choles-
terolesters diffuse from ether through rubber membrane into ether.
This is very interesting on account of the high molecular weight
of these esters. Cholesterol-stearate with a molecular weight of
652.51 diffuses very readily.

For some time I have been trying to find an ether soluble
substance of higher molecular weight than cholesterol-stearate
and which could be easily detected in the diffusate. The free
dye-acid of iodoeosin fulfilled these requirements. This free dye-
acid has been employed by Professor Ehrlich as a very delicate
reagent for free alkali in the erythrocytes of man.*

Iodo-eosin is the potassium salt of tetraiodo-fluorescein with
the following formula,

CoH ./CHI(OK)
CO GX CHL OK)°-

As a salt it dissolves in dilute alkalies with red color, but is insoluble
in ether or any other organic solvent. The free dye-acid, however,
is obtained as a yellow precipitate from the alkaline solution of
iodo-eosin by adding hydrochloric acid in excess, and it dissolves
readily in ether, or in any other organic solvent, but is insoluble in

1 Proc. Amer. Soc. Biol. Chem., 1910, 2, p. 8; Jr. Biol. Chem., 1911, IX, p. xiv.
Biochemical Bulletin, 1912, 11, p. 64; George Crocker Special Research Fund, Vol. 3;
(In press.)

2 Proc. Soc. Exp. BIOL. AND MED., I1g11, VIII, p. 132.

3 Ehrlich-Lazarus, ‘“‘Die Anemia,” Vienna, 1898.

Curonic LEAD PoIsONING IN GUINEA Pics. 49

water. This free dye-acid has the following formula,

CeHa /CeHI2(OH)
C06 PK Ca (OH)O

with a molecular weight of 836.

The dye-acid of iodo-eosin may be made by dissolving ten
grammes of iodo-eosin (commercial dye) in one per cent. potassium
hydroxide and then adding hydrochloric acid in excess. The dye-
acid is precipitated at once, it can then be filtered off and the
precipitate washed with hot water till the washings are acid-free.
The precipitate after drying is easily soluble in ether, forming
a beautiful yellow-colored solution.

When this free dye-acid in ether solution is placed inside of an
intact rubber membrane immersed in ether, it can readily be noted
that in a few minutes diffusion currents are visible and the ether
outside of the bag becomes colored, showing that the free dye-
acid has diffused.

The bearing of our results on the question of permeability and
impermeability of membranes will be considered later.

ABSTRACTS OF THE COMMUNICATIONS, PaciFIC CoAsT BRANCH.
First meeting.
San Francisco, California, December 4, 1912.
36 (732)
Chronic lead poisoning in guinea pigs.
By W. OpHULs.
[From the Pathological Laboratory of Stanford University.]

Of the twenty-eight guinea pigs treated with sublethal doses of
carbonate of lead seven (25 per cent.) showed a peculiar condition
to which so far attention does not seem to have been directed.
There has developed a hemorrhagic, sero-fibrinous inflammation
of the pericardium, of the peritoneum in the upper part of the
peritoneal cavity and occasionally also of the pleure. In the
pericardium the lesion commences with a hemorrhagic exudate
followed by the formation of fibrinous deposits especially on the
parietal layer and ending with organization with marked thicken-

50 SCIENTIFIC PROCEEDINGS (51).

ing and occasionally adhesions. In the pleure one finds as a
rule a simple serous exudate without deposits but occasionally
heavy deposits of fibrin also. In the peritoneum also the condition
commences with fibrinous deposits and eventually leads to marked
peritoneal thickening more especially in the upper part of the
peritoneal cavity. The capsules of liver and spleen are much
thickened and the contraction of the thickened capsule often
causes marked deformity of the liver. The lesions produced in
this way are closely analogous to Curschmann’s ‘Zuckerguss-
leber.’’ The spleen is always more or less enlarged, but this is due
to the excessive destruction of erythrocytes by the lead and is
found in all animals.

It was only natural to suspect the presence of a bacterial
infection of the serous membranes in these cases. A very careful
bacteriological examination was made in several of the cases, but
it was entirely negative.

Whether the hemorrhagic exudation which ushers in the
process is due to the lead anemia appears to me a question well
worth debating. Smears of the exudate show few leucocytes and
often many nucleated red blood corpuscles, even megaloblasts.

In one case (46) adhesions had formed in the upper part of the
peritoneal cavity with partial strangulation of duodenum and
intestine and extreme dilatation of the stomach.

In the liver the periportal connective tissue is moderately but
distinctly thickened in some cases. One always finds small areas of
fatty degeneration and necrosis of the liver tissue. In one case
(46) they were quite large and show beginning liquefaction in the
center (due to apparently secondary invasion of bacteria). Whether
these degenerative lesions are due to circulatory disturbances pro-
duced by the shrinkage of the thickened capsule or whether they
are independent of this and toxic in origin, it is difficult to decide,
although in one case the lesions in the liver were well marked but
those in the capsule only slight. The bearing this may have on
the modern conception of cirrhosis as closely connected with
certain types of anemia is evident.

1It is interesting to know that Charcot and Gombault found acute pericarditis
in their guinea pig 4, hemorrhagic pericarditis in their guinea pig 9, subacute perie

carditis in their guinea pig 15, and pericardial thickening in their guinea pigs 6 and 14
(5 out of 15).

Curonic LEAD PoIsoNING IN GUINEA Pics. 51

It is interesting to observe in these animals a condition so en-
tirely analogous to what has been described in man as polysero-
sitis, or a polyorrhomenitis by Italian investigators, of which
Curschmann’s Zuckergussleber and Pick’s pericarditic pseudocir-
rhosis are only special manifestations. The relations of these con-
ditions in man tochronic lead poisoning and to chronic anemia
remains to be studied.

37 (733)

Chronic lead poisoning in guinea pigs. Its relation to chronic
nephritis.

By W. OPHULS.
[From the Pathological Laboratory of Stanford University.]

During the last years I have been repeating the experiments of
Charcot and Gombault! on chronic lead poisoning in guinea pigs.
Twenty-eight guinea pigs were given sublethal doses of carbonate
of lead for periods ranging from one month to three years and ten
months. Fourteen of these experiments lasted over one year.

The lesions in the kidneys were much less striking than one
would expect from the report of Charcot and Gombault. In all
cases there was a limited necrosis and desquamation of the epi-
thelium with marked evidences of regeneration especially in the
ascending limbs of the loops of Henle. In some cases the epi-
thelium in places was heavily pigmented. Occasionally there were
seen a few glomeruli with slightly thickened capsules. There were
only two of the twenty-eight experiments in which more advanced
lesions were discovered in the kidneys. In one case (69) in which
the guinea pig had received over thirty grams of carbonate of lead
in three years the kidneys were actually granular and the cortex
distinctly narrow. The lesions in this case consisted in collapse
of tubules over large areas with marked development of fibrous
tissue between them. There were many casts. The glomeruli
showed marked fibrous thickening of the capsules and cystic
dilatation. The other guinea pig (51) which received about four
grams of carbonate of lead in twenty months showed similar con-

1 Charcot et Gombault, ‘‘Note relative a l'étude anatomique de la nephrite
saturnine expérimentale,” Arch. de Phys., 1881, 2 s., VIII, 126.

52 SCIENTIFIC PROCEEDINGS (51).

ditions but they were not so well marked. Calcareous infarcts
were not found in any of the kidneys. No vascular lesions were
ever found in the aorta or in the branches of the renal arteries.
On the whole the condition produced in these few animals does
not resemble human nephritis, but is much more similar to the
lesions observed in experimental uranium nephritis.

38 (734)
Agglutination of encapsulated bacteria.

By J. G. FITZGERALD.

[From the Hearst Laboratory of Pathology and Bacteriology, University
of California.|

During the past year a systematic study of the group of en-
capsulated bacteria (including Bacterium pneumoniae Friedlander,
Bacterium rhinoscleromatis v. Frisch, Bacterium ozene Abel-
Lowenberg, and Bacterium capsulatus mucosus Fasching) has been
carried on, employing for the purpose biometrical methods some-
what similar in character to those suggested by Winslow! in his
work on the Coccacez.

During the course of the investigation, immunological methods
have been used. At the outset, the reaction of agglutination was
tried. Paltauf was the first (quoted by Beham?) to suggest that
the agglutination of encapsulated micro-organisms is inhibited
because the bacilli are surrounded by a slimy nucleo-protein
capsule. Porges* was able to supply experimental proof of this.
v. Eisler and Porges* then elaborated a method of removing the
capsule, after the application of which these bacteria were agglu-
tinable.

My own work done independently of Beham has given results
in harmony with his. I have found that Bacterium rhinosclero-
matis on injection into rabbits yields a potent agglutinating serum.

Using this serum, agglutination not only of the homologous
microérganism has been obtained but a positive result was found

1 Winslow, “Systematic Relationships of ,the Coccacee,” John Wiley & Sons,
New York, 1908.

2 Beham, Central. f. Bakt., Abt. 1, Orig., Bd. 66, Heft 1, p. 110.

3 Porges, Wiener klinische Wochenschrift, 1905, No. 26.
4y. Eisler and Porges, Central. f. Bakt., Abt. 1, Orig., Bd. 42, Heft 7, 660.

AGGLUTINATION OF ENCAPSULATED BACTERIA. 53

to occur also when four other strains of the same species, from
widely different sources, were tested. The bacteria agglutinated
in a dilution of the serum, ranging from 1-400 to 1-800. Two
strains of Bacterium ozene, one strain of Bacterium capsulatus
mucosus Fasching, and one strain of Bacterium pneumoniae Fried-
lander, were not agglutinated at allby thesameserum. Thestrains
of Bacterium rhinoscleromatis which agglutinated were no longer
encapsulated. All the other species still showed a capsule. One
of the strains of rhinoscleromatis when originally isolated was not
agglutinated by the serum of the patient from whom it was
isolated and at that time it was encapsulated (Thro, Proceedings
New York Pathological Society, April and May, 1910). This
micro6rganism, then, was not agglutinable when encapsulated
shortly after isolation. Later, it lost its capsule and became
agglutinable. As far as my work has gone, I have found that of
the species here considered rhinoscleromatis most easily loses its
capsule when grown on ordinary laboratory media and becomes
agglutinable.

Beham has shown that encapsulated as well as non-encapsu-
lated species of this group have antigenic properties and may
produce agglutinins. I have no data on this point as yet. The
acid agglutination reaction of Michaelis, as modified by Beniasch!
did not agglutinate Bacterium rhinoscleromatis without a capsule,
or Bacterium capsulatus mucosus possessing a capsule.

Bacteria of the species here considered are not agglutinable
when encapsulated, but may become so when they lose their capsule
in the course of months of growth on laboratory media. Having
lost their capsules, these species may be differentiated by the
reaction of agglutination. Thus Bacterium rhinoscleromatis has
been shown to differ from Bacterium ozene and from Bacterium
pneumoniae Friedlander, in that it more easily loses its capsule and
is then agglutinated by its specific serum.

1 Beniasch, Zeitschrift fir Immunitatsforschung, Vol. XII, Heft 3, 1912, 268.

54 SCIENTIFIC PROCEEDINGS (51).

39 (735)
Relative frequency of B. coli communior in contaminated water.

By J. G. FITZGERALD.

[From the Hearst Laboratory of Pathology and Bacteriology,
University of California.]}

Some recent work of my students has been interesting in con-
nection with the relative frequency of occurrence of the species of
the B. coli group in water contaminated with human feces. The
B. coli group consists of four species: B. communior (Durham),
B. communis (Escherich), B. aerogenes (Escherich), B. acidi
lactict (Hueppe). The general characteristics common to all
species are: short, Gram-negative, cocco-bacilli, with rounded
ends, non-spore bearing, fermentation of dextrose, glucose and
lactose with gas production, non-liquefaction of gelatin after 14
days, indol production, and reduction of nitrates to nitrites. In
general, also, litmus milk is acidified and coagulated. Further
differentiation of the members of the group is possible by means of
fermentation tests in the sugars dulcite, saccharose, mannite and
raffinose. B. communior and B. communis ferment dulcite; B.
aerogenes and acidi lactict do not. B. communior and B. aerogenes
ferment saccharose. B. communis and B. acidi lactici do not.
MacConkey! isolated a large number of strains of these species
from feces and found that their relative frequency was as follows:

Per Cent.
IB, COTINUNAON 5a fea Sail to aloes ole ner aueue eee ele teis Tolatehenereeea eee 23
Bi COMMUNES rah s iaie inate cotohei cl levee data raleks iol ehensdetare lous enevevalev esc teres 37
BBs: CERO SENOS HANS coca Sys Seo ails ole Va PNG aiot ake Ore te tare overcast a era avateyeeetets I5
BB OCU GHELCE « f6ta8 a aici'si& sede cs fobotoi sis aera de elase laverctcteloheteteieieretnreeiore 25

Graham Smith? in the analysis of thirty-five strains isolated from
flies found, on the other hand, a predominance of communior
(43 per cent.), communis occurring much less frequently (17 per
cent.). B. aerogenes (11 per cent.) and B. acidi lactict (29 per
cent.) did not differ greatly in their occurrence.

Winslow and Walker*® in twenty-five strains isolated from

2 Graham-Smith, Report of Local Government Board, London, 1909.
3 Winslow and Walker, Science, 1907, Vol. 26, p. 675.

PRODUCTION OF CREATININE BY BACTERIA. 55

feces found 28 per cent. of communior, 60 per cent. communis, 4
per cent. aerogenes, and 8 per cent. acidi lactici.

The results of an analysis of thirty-two strains isolated in my
laboratory from human feces were as follows:

Per Cent,
B. coli communior.........+ JOOS DISRO COORD MOR OSD CRD TOOTS 65
IBCOU COMMUNES crete relia cele ieiciote pleleiteleletel hele stticeie ics 28
ES CEVORERES EE a eretad Tetcis eraieteretetetchetotetclciertatctelstel sTeveiauerersicvers rele 3-5
BRO UBGT Bn Sra Ob OO nOSOAOe Ao OOODODOOD ODL COO OUN.COL 3.5

The saccharose fermenting species, then, may predominate in
water recently contaminated with human feces.

40 (736)
Production of creatinine by bacteria.

By J. G. FITZGERALD and Cart L. A. SCHMIDT.

[From the Hearst Laboratory of Pathology and Bacteriology,
University of California, Berkeley, California.]

Very little work has been done to determine whether creatinine
is produced by bacteria growing on media free from creatin and
creatinine. Germ4n,! in a brief review of the literature on this
subject, makes mention of only three contributions. In his paper,
GermAn gives the result of an investigation of thirty-five species
of bacteria to determine whether they were able to produce
creatinine. In summing up, he says that this characteristic might
be of value in the differentiation of closely allied species.

Our work was undertaken to ascertain: (1) Whether creatinine
production is of any value in the differentiation of groups of
closely allied species; (2) to determine the method best suited
for studying creatinine production by bacteria; (3) to ascertain
what amounts of creatinine were produced so that certain quanti-
tative studies might be undertaken.

A medium composed of 2 per cent. Witte’s peptone and .5 per
cent. salt furnishes a creatin and creatinine free medium. Twenty-
eight strains of bacteria, belonging to the mucosus capsulatus
group of microérganisms, including B. rhinoscleromatis, B. ozene
and B. lactis aerogenes, were grown on the above medium for eight

1 German, Ceniralbl. f. Bakt., I. Abt. Orig., Bd. 63, Heft 7, 1 June, 1912.

56 SCIENTIFIC PROCEEDINGS (51).

days. Ten cubic centimeters were then tested for creatinine by
Weyl’s method (adding several c.c. of a 10 per cent. NaOH solu-
tion and several drops of a freshly prepared sodium nitroprusside
solution, a positive reaction being indicated by the immediate
appearance of a dark red zone which soon turns to a greenish
color) and 50 c.c. were tested by the Folin method (adding 7.5 c.c.
saturated picric acid solution and 2.5 c.c. sodium hydrate, a
positive reaction being indicated by the production of an orange-
red color similar to the color produced by a potassium bichromate
solution). As controls, a strain of each of B. coli communis, B.
proteus, V. cholere asiatica, M. aureus, M. albus, and V. Metchni-
kovii were examined at the same time. A sterile peptone solution
itself gives with the Folin method a slight color which to a certain
extent interferes with the test; this, however, can be eliminated
by comparing the solution to be tested with the control. In only
one case, that of B. proteus, was the Folin reaction decidedly
positive, while V. cholere asiatica, and several others gave a some-
what doubtful positive result. With the Weyl reaction a much
more accurate determination of the presence of creatinine can be
made.

As regards the Mucosus capsulatus group, we found that
creatinine production is no criterion in differentiating the species,
since nearly all the strains gave negative reactions. That the
amount of creatinine which is produced as a result of bacterial
metabolism is very small is indicated in the results obtained by
the Folin method. That certain bacteria produce creatinine more
readily and in larger amounts than others is shown by the more
strongly positive reactions given by B. proteus. GermA&n’s results
tend to show the same thing. His inability to get a positive
creatinine reaction with certain organisms in twenty-four hours,
which after a longer period of time gave positive reactions, does
not show that no creatinine was produced during the first twenty-
four hours, but rather that the amounts produced were so small
that the test appeared to be negative. That there are certain
bacteria which either do not produce any creatinine when grown
on a peptone medium or produce amounts of creatinine so small
that the Weyl reaction appears negative is shown both by Ger-
m4n’s and our results.

REGENERATION OF BONE FROM PERIOSTEUM. 57

We were unable in our work to parallel GermAn’s results in the
case of all the species which he claims produce creatinine. This
we believe to be due to the fact that the ability to produce crea-
tinine is a characteristic which may easily be lost by bacteria
just as is the power of producing indol.

41 (737)

Regeneration of bone from periosteum.
By S. L. Haas.
[From the Pathological Laboratory of Stanford University.]}

In an endeavor to establish the factor that periosteum plays
in the regeneration of bone, the following experiments were per-
formed on the ribs of rabbits.

Several experiments consisting merely of a subperiosteal re-
section of the rib showed after 20 days a cartilaginous-like material
filling the entire space; while after a longer time complete filling
in with bone.

The next endeavor was to isolate the periosteum so as to
prevent bony elements from growing in to the periosteal space.

In one experiment the rib ends were capped with lead—after
12 days no evidence of regeneration. This method was discarded
as it was thought better to try and raise the rib from its periosteal
covering without severing the ends, and isolate it by sewing muscle
beneath the raised rib.

In 10 experiments of this nature the results uniformly showed
a tendency for bone to grow in at the angle where the rib was
raised from its periosteal bed. In one experiment after 59 days
the whole area was filled in by bone, although in another after
8 months there was only a small spicule of bone growing in at the
sternal side. In only one experiment after 26 days was an isolated
island of bone found, free from connection with bone elements.

Although this does not prove the point sought, it at least
emphasizes the tendency of bone to grow in the direction of existing
periosteum.

At this stage it was noticed that there was a difference at
times in bone regeneration and it was suggested to try the effect

58 SCIENTIFIC PROCEEDINGS (51).

of blood clot. The experiment was performed in the same way
as the previous one except that blood clot was placed in the peri-
osteal gutter left by the raised rib. After periods of 48, 44, 39, 23,
and even 14 days, the whole area was found to be occupied by a
calcareous material. Two ribs in the same animal were similarly
treated with the exception that in one no blood clot was used.
The one containing blood clot showed considerably more bone
formation. This showed the marked increase of bone in the
presence of blood clot.

In order to rule out ingrowth of bone from the side, the follow-
ing method was resorted to. Extensive resections of a rib from
near the vertebral junction, to and including about I cm. of costal
cartilage. Hot paraffin was poured in at the vertebral and sternal
sides, allowing a free gap between. This gap was filled with blood
clot and all sutured in place. In four experiments of this nature
there always was found calcareous material, separated grossly
from any bony tissue. A few experiments without blood clot
were used as controls to the last. These showed some calcareous
material. In one particular instance considerable bleeding was
noticed at the end of operation, and in this case a heavy bony
deposit occurred. Thus the factor of post-operative hemorrhage
might explain discrepancies of results of these experiments and
those of other experimenters.

An interesting point was noticed in cases where costal cartilage
was dissected away, in that there was an especially marked
tendency for regeneration of bone at costal cartilaginous junction.
If we consider this junction as an epiphysis it is remarkable that
we should get so much regeneration after its destruction, evidently
from the periosteum.

In these experiments the periosteum has been considered as
one usually leaves it after surgical removal of bone. In conclusion
it appears that periosteum under certain conditions has the power
of regenerating bone, especially in the presence of blood clot.
Also, that it always exerts a stimulating effect on bone formation.

INFLUENCE OF LECITHIN AND CHOLESTERIN UPON TUMORS. 59

42 (738)

Preliminary report on the influence of lecithin and cholesterin
upon the growth of tumors.

By T. BRAILSFORD ROBERTSON and THEO. C. BURNETT.

From the Rudolph Spreckels Physiological Laboratory of the
University of California.]

Sixty-one white rats, obtained from local sources and hereafter
designated ‘‘Local’’ were inoculated in the axillary region with
portions of a Flexner-Jobling carcinoma which was obtained from
the Rockefeller Institute for Medical Research through the kind-
ness of Dr. Peyton Rous. The number of successful inoculations,
determined after 19 days, was 42, or 69 per cent.

Sixty-four rats obtained from dealers in Chicago and hereafter
designated ‘‘Chicago”’ were similarly inoculated with portions of
a Flexner-Jobling carcinoma which had been obtained from the
Rockefeller Institute for Medical Research early in the year and
propagated through four generations of rats in this laboratory.
The number of successful inoculations, determined after 18 days,
was 55, or 86 per cent.

Both sets of tumors grew rapidly, attaining average diameters
of 15.0 and 11.9 millimeters after 19 and 18 days respectively.

Beginning upon the 19th day in the case of the ‘‘Local’”’ and
upon the 18th day in the case of the “‘Chicago”’ animals, each
of the two groups of animals was sorted without selection into
three batches, of which one (consisting of 12 animals in each case)
served as controls, another (10 ‘‘Local” and 13 ‘‘Chicago’’)
received I c.c. of a 3.9 per cent. suspension of cholesterin in N/1o
sodium oleate solution, which was injected hypodermically directly
into the tumors every 2d or 3d day (three times a week); the
third batch (20 “‘Local’”’ and 30 ‘“‘Chicago’’) received 1 c.c. of a 2
per cent. aqueous emulsion of lecithin similarly injected into the
tumors upon the same days.

It was found that in the pre-metastatic stage cholesterin causes
a very notable acceleration of the primary growth, the gain in
diameter between the 19th (18th) and 24th (23d) days being 11.6
mm. in the ‘“Local’”’ and 11.4 mm. in the ‘“Chicago’”’ animals as

60 SCIENTIFIC PROCEEDINGS (51).

compared with 4.8 mm. and 2.9 mm. respectively in the controls.
The acceleration of the growth of the primary tumor by cholesterin
is not very evident in the metastatic stage (31st to 38th days), but
the tendency to form metastases and the rate of metastatic growth
are very markedly increased.

Lecithin, on the contrary, diminishes the tendency to form
metastases, retards metastatic growth when it does occur and,
in some instances (the ‘‘Local” animals) also retards the growth
of the primary tumor in the post-metastatic period.

43 (739)
Note on the cock’s comb test for the activity of ergot.

By ALBERT C. CRAWFORD and JAMES P. CRAWFORD.
[Laboratory of Pharmacology, Leland Stanford Junior University.]

The cock’s comb test has become a popular method of deter-
mining the activity of ergot preparations. It was introduced
because gangrene frequently occurred in epidemics of ergot
poisoning, and because bluing of the cock’s comb was believed to
be due to arterial constriction. This view was supported by von
Recklinghausen’s interpretation of the microscopical examination
of the comb in chronic ergot poisoning of cocks. We were im-
pressed by the fact that in ergot poisoning often the only patho-
logical feature was venous dilatation and we believe that venous
dilatation is probably the real cause of the bluing of the comb.
It is admitted that the intravenous injection of epinephrin causes
a rise in blood pressure, mainly from vaso-constriction and we
have found that it will blue the cock’s comb, but the bluing only
comes on as the blood pressure falls and persists for an hour or two;
in other words it occurs at a time when arterial-constriction has
subsided. Large doses of paraldehyde given per os, or the inhala-
tion of amyl nitrite, will also cause bluing of the comb.

On subcutaneous injection neither adrenalin nor p. oxyphenyl-
ethylamine caused this bluing. Dale claims that much of the
activity of ergot preparations is due to p. oxyphenylethylamine.
Now while the subcutaneous injection of 25 mg. of p. oxyphenyl-
ethylamine caused marked symptoms in cocks, there was no
bluing, hence one would argue that the subcutaneous injection

An EXPERIMENTAL STUDY OF POISON Oak. 61

of ergot into cocks cannot be considered a quantitative method for
testing ergot. Oncometric studies of the cock’s comb during acute
ergot poisoning are now being made.

44 (740)
An experimental study of poison oak.

By EDWARD VON ADELUNG.

[From the Hearst Laboratory of Pathology and Bacteriology,
University of California.]

Experiments show that the toxic principle of Rhus diversiloba
while not volatile can poison at a distance by means of mechanical
carriers. It is not destroyed by subjection for one hour to 100° C.
and is carried, potent, by the smoke from burning Rhus plants.
The dermatitis produced by this plant is a purely local affection
and is not spread by the blood or lymph or by the serum of the
blebs. The reaction of the sweat has no relation to individual
susceptibility.

Absolute immunity in man has been claimed but was not
found to exist, on repeated attempts, in any of the individuals
tried (6 persons). In spite of the work of Ford, the conclusion is
arrived at that experimental immunity in animals to Rhus toxin
has not been proved. The author, working with pure toxin
(glucoside) produced by the method of Syme, was unable to
intoxicate animals with any reasonable amount. Ford’s work
was done with a commercial fluid extract containing various
impurities. A permanent aqueous suspension of the alcoholic
solution of the pure toxin can be prepared, and remains toxic for
the human skin. As much as 0.025 gm. of this preparation of
toxin can be given intravenously to a 2,000-gram rabbit without
fatal effect, and as much as 0.03875 gm. can be given to a 280-gm.
guinea pig subcutaneously without fatal effect. Pure alcoholic
extract when given subcutaneously produces severe necrosis and
death owing to the alcohol itself used asa menstruum, but with no
lesions attributable to the toxin. No skin lesions were produced
in monkeys, rabbits, or guinea pigs asa rule, but a slight dermatitis
was produced on the rabbit’s ear at times.

62 SCIENTIFIC PROCEEDINGS (51).

Inasmuch as animals can not be killed by the pure toxin, it is
impossible to demonstrate antitoxic effect in the serum of animals
that have received repeated doses of the toxin. The Bordet-
Gengou fixation reaction failed to demonstrate the presence of
antibodies in the sera of animals so treated.

The simplest prophylactic measure against Rhus poisoning is
to wash well with soap and hot water as soon after exposure as
possible. The protection of the skin by anointing with cotton-
seed oil before exposure and washing this off within a few hours
with soap and water, renders prophylaxis fairly certain. The
following remedies have therapeutic value: hot water, ichthyol
collodion, permanganate of potassium, magnesium sulfate, and
tincture of iodine.

45 (741)
The failure of union between antigen and precipitin

when present in the same serum.
By HANS ZINSSER and STEWART YOUNG.
[Stanford University.]

It has been frequently observed that the serum of rabbits,
immunized with foreign protein, may, at certain times, contain not
only precipitin, but also unaltered antigen remnants. Such sera
not only precipitate the antigen, but also give precipitates when
mixed with other antisera prepared with the same antigen (Linosier
et Lemoine, Eisenberg, Michaelis, and Fleischmann, Ascoli, Von
Dungern). They have been recently studied by Gay and Rusk.
It has been difficult to explain why such sera do not spontaneously
precipitate since both reacting factors are present.

In the cases of two sera recently studied by us the phenomena
observed were as follows: Sera “3” and “4,” obtained by injecting
two rabbits with horse serum on three successive days and bleeding
eight days after the last injection, were perfectly clear and showed
no spontaneous precipitation on standing several days. Serum
“2” precipitated horse serum in dilutions of I to 1,000, and serum
“4” in dilutions of 1 to 500. When mixed with equal quantities of
an antihorse serum precipitin, which contained no antigen, both
sera were precipitated, ‘‘3’’ more strongly than ‘‘4.”’ Neither of

FAILURE OF UNION BETWEEN ANTIGEN AND PRECIPITIN. 63

these sera fixed complement. When “3” and ‘4”’ were mixed,
slow and slight, but distinct precipitates occurred, unmistakable
after 12 hours in the ice chest.

On standing for one month in the ice chest sera “3” and “4”
were found to have spontaneously precipitated. The precipitate
_ showed slight complement fixation and the supernatant fluid was
found on titration to have lost about one half of its precipitating
power.

Such spontaneous precipitation of precipitin sera has been
repeatedly observed after prolonged conservation.

Von Dungern who has done much work on this question has
assumed a multiplicity of antigens and precipitins to account for
this failure of union between antibody and antigen in such sera.
He says ‘“‘Neben dem gebildeten Praezipitin bleibt aber ein
anderer Theil der Praezipitablen Substanz der keine Affinitaet
zu dem gebildeten Praezipitin besitzt, bestehen, solange bis ein
anderes Partial Praezipitin von den Kaninchen geliefert wird,
welches sich mit Gruppen der in Loesung gebliebenen Eiweiss-
koerper vereinigen kann.”’

We do not believe that this explanation is tenable in our cases
since eventual precipitation, therefore union, took place after
prolonged standing.

Another explanation (Eisenberg) accounts for the phenomenon
by assuming that antigen and precipitin unite according to the
laws of mass action, establishing an equilibrium, in which un-
united portions of each of the reacting factors are therefore found.
This explanation seems incompatible with the observation that
the two sera did not bind complement, pointing to the absence of
united antigen and precipitin in the sera.

We believe that the failure of union of the two elements may
be regarded as closely analogous to phenomena occurring under
the influence of protective colloids.

This is compatible with the original failure of the two sub-
stances to unite; with their precipitation by other sera in which
there is present one or the other of the reacting bodies; and with
their final spontaneous precipitation on prolonged standing.

It is more difficult to account, on this basis, for the mutual
precipitation of these sera which occurs when they are mixed.

64 SCIENTIFIC PROCEEDINGS (51).

For this we have sought analogy. Fresh dog serum precipitates
colloidal trisulphide of arsenic. Heated dog serum precipitates
the arsenic only when small quantities of the serum are added.
Larger quantities again disperse the precipitate and then protect
the colloidal metal from precipitation by subsequently added
fresh serum.

The addition of further arsenic trisulphide to such protected
mixture will disturb the balance and lead to precipitation.

It should be possible to make up two clear solutions, each con-
taining the three bodies, but in entirely different proportions, such
that upon mixing the two, the protective action of the heated serum
is so reduced that precipitation will occur. Slight, slowly appear-
ing precipitates have been produced in this way, but the results
thus far obtained have merely sufficed to emphasize the likelihood
of the explanation we offer. The protective action of the heated
serum is so enormous that it requires a very accurate adjustment
of relative amounts to ascertain the exact limit of protection.

SCIENTIFIC PROCEEDINGS.
ABSTRACTS OF COMMUNICATIONS.
Fifty-second meeting.

Physiological Laboratory, College of Physicians and Surgeons,
February 19, 1913. President Ewing in the chair.

46 (742)
Further light on the conjugation of Paramecium.
By Gary N. CALKINS.
[From the Depariment of Zoology, Columbia University.]

These experiments were undertaken to test the variability
after conjugation of Paramecium caudatum in respect to the power
to conjugate. An individual immediately after separating from
conjugation possesses a new fertilization micronucleus and the
old macronucleus. The new micronucleus divides into two, these
two into four, the four into eight. During these three divisions
the old macronucleus goes to pieces and the fragments are gradu-
ally absorbed in the protoplasm. Four of the eight new nuclei
then metamorphose into four macronuclei, and then for the first
time after conjugation, the cell divides. The progeny have two
macro- and two micronuclei each. These cells immediately divide
again without further nuclear division, forming four cells, each
with one macronucleus and one micronucleus. The normal con-
dition of Paramecium is then attained. The problem to be solved
by experiment was: Do these four cells give rise to progeny which
vary in respect to the power to conjugate? In order to get as
many chances as possible the products of the first three divisions
of each of the four cells were kept isolated, thus giving eight lines
of cells from each of the original four. These sets of eight,
thirty-two in all, I shall speak of as the Ist, 2d, etc., quadrants.
The thirty-two lines were kept isolated in vials and fed at intervals
of from six to eight days under conditions prohibitive of conjuga-

65

66 SCIENTIFIC PROCEEDINGS (52).

tion, from the 27th of last July to the present. From the outset
marked differences in the size of the progeny of different lines
were noted; the statistical proof, however, is not yet ready for
presentation. A marked difference in division rate was also noted;
for example, the daily division rate of representative individuals
of all the lines was carefully kept for a period of 30 days in October
and November. The average rate for individuals of the first
quadrant was 0.65 divisions per day; of the 2d, 0.91 per day; of
the 3d, 0.81, and of the 4th, 0.95 per day, a difference of from
fifteen to twenty per cent. between the rates of individuals from
the Ist quadrant and those of the other three. The weaker vitality
of progeny of the Ist quadrant is also shown by the extinction by
death of all the progeny of four lines of the eight, whereas only
one line died out from the progeny of quadrants 2 and 4, and none
from quadrant 3. (In quadrant 2, two lines failed to start by
reason of a pathological division in the third generation.) The
power to conjugate is correlated with these variations in vitality.
Conjugation tests (too complex to be described here) were started
in October and November, and many pairs were found in two
of the four remaining lines of the Ist quadrant, none in the others.
The December tests gave similar results; three of the four lines
of the first quadrant giving many pairs while none were found in
the other twenty lines. Again in January and February all four
lines of the first quadrant furnished many pairs, but not a single
pair was found in the other lines subjected to exactly the same
treatment. A specific example will show how extensive the ob-
servations were. In the January test, for example, not less than
150,000 Paramecium belonging to the non-conjugating lines passed
under my eyes, not a single conjugating pair being seen. Of the
four conjugating lines not less than 50,000 Paramecium were
observed, of which more than 2,000 pairs were in conjugation.
It is evident therefore that a very decided difference exists in the
total progeny of an ex-conjugant; some are potential germ-cells,
others apparently are not. These experiments may give a clue
to the divergent results obtained by Maupas, Woodruff and my-
self, which cannot be harmonized on the ground of abnormal
conditions or bacterial poisons in one case and not in another.
The divergence must be due to some more deeply lying cause in

THE INTRAPERITONEAL LysIs OF TUBERCLE BACILLI. 67

the organisms themselves. The race that I worked with in 1901
was a conjugating race which died out in the 742d generation.
Woodruff’s long line of over 3,500 generations is a non-conjugating
race and the two races cannot be compared in regard to vitality,
since normal conjugation was prevented in the conjugating race,
whereas in the non-conjugating race there has been no artificial
prevention of a normal process. The following conclusions may
be drawn; they must be considered provisional, however, since the
experiments are not yet concluded.

1. The traditional view that each Paramecium is a potential
germ cell is not true.

2. Some descendants of an ex-conjugant are potential germ
cells, others are not.

3. The life history of conjugating lines has shown that if
conjugation is prevented, the race dies out.

4. Weismann’s hypothesis that natural death is absent in
protozoa is not borne out by the facts.

5. Care must be exercised in arguing that one effect of con-
jugation is to bring about variations because of amphimixis until
we know that such variations are not brought about by every
individual ex-conjugant in its normal development.

47 (743)

Note on the intraperitoneal lysis of tubercle bacilli.
By WILFRED H. MANWARING and J. BRONFENBRENNER.

[From the Laboratories of the Rockefeller Institute for Medical Re-
search, New York.|

In a previous communication,! it was shown that tuberculous
guinea-pigs, rabbits and dogs react to intraperitoneal reinoculation
with tubercle bacilli by causing rapid degenerative changes in
the injected bacteria and a rapid decrease in their number, not
observed in normal animals. The question now arose as to the
mechanism of this heightened peritoneal resistance. From the
similarity between this phenomenon and the Pfeiffer reaction

1 These TRANSACTIONS, 1912, X, p. 30.

68 SCIENTIFIC PROCEEDINGS (52).

attempts were made to determine whether or not the specific
antibodies, upon which the intraperitoneal lysis may be supposed
to depend, are present in the circulating fluids.

To test this, guinea-pigs, rabbits and dogs were made tubercu-
lous by inoculating them subcutaneously with tubercle bacilli.
After an interval of from five to eight weeks, these animals were
bled and their blood tested 1m vitro and in vivo. In a number of
these experiments direct transfusion of the blood was made from
the tuberculous animals into normal animals, an amount o blood
often as great as three quarters of the total blood-volume being
thus passed into the circulating system of the normal animals, the
normal animals having been previously bled to free them as much
as possible from normal blood. The transfused animals were sub-
sequently tested by intraperitoneal injections of tubercle bacilli.

Neither in the test-tube experiments, nor in normal animals
injected subcutaneously, intravenously or intraperitoneally with
tuberculous serum, nor even in the normal animals directly trans-
fused with large quantities of the unaltered blood of tuberculous
animals, has the reaction thus far been obtained. The substances
responsible for the heightened peritoneal resistance, therefore,
apparently do not exist in appreciable quantities as circulating
antibodies, at least at the stage of the disease studied. The
heightened tuberculous resistance, therefore, is apparently due to
substances held in fixed tissue cells.

Evidences of tuberculolytic substances have, however, been
obtained in the peritoneal fluids of tuberculous guinea-pigs, soon
after the introduction of tubercle bacilli. If these fluids are with-
drawn, centrifuged free from formed elements and then introduced
into the peritoneal cavities of normal guinea-pigs, they confer
upon the normal peritoneal cavities a slight power of destroying
tubercle bacilli. It is suggested, therefore, that fixed tuberculolysins
are set free by the peritoneal cells in response to the presence of
tubercle bacilli, and that these lysins account for the heightened
resistance to intraperitoneal reinoculation with tubercle bacilli.

SPECIFIC REACTIONS EXHIBITED BY HAy FEVER CASES. 69

48 (744)

A preliminary communication on certain specific reactions ex-
hibited by hay fever cases.

By GEorRGE H. A. CLOWES.

[From the Laboratory of Biological Chemistry, State Institute for
the Study of Malignant Disease, Buffalo, New York.]|

Dunbar was the first to demonstrate conclusively that the
European or spring variety of hay fever is caused by the pollen of
timothy and other members of the graminacez family. He found,
that, even in midwinter an aqueous extract of timothy pollen
produced flushing and intense irritation when introduced into the
eye of a susceptible individual, whilst normal individuals were
entirely unaffected by such a procedure.

Sufferers from the American or autumnal form of hay fever
(caused by the pollen of ragweed, golden rod, and other members
of the Composite family) exhibit a similar sensitiveness when
tested with aqueous extracts of ragweed pollen. This reaction
is also strictly specific, and only those who suffer from both spring
and autumnal hay fever react to the extracts obtained from both
varieties of pollen. The amount of pollen extract required to
produce flushing in the eyes of sensitive individuals varies from
1/20 of a c.c. of a solution of one in five hundred thousand to 1/20
of a c.c. of a solution of one in five hundred. This test can be
utilized quantitatively and affords a fair index of the measure of
immunity or resistance possessed by sensitized individuals.

A specific cutaneous reaction capable of quantitative applica-
tion may be obtained by making a slight abrasion of the skin and
applying a drop of pollen extract of suitable concentration. A
large white welt similar to that resulting from a mosquito or bee
bite accompanied by a distinct itching sensation develops on
susceptible individuals within 15 minutes, while normal controls
are unaffected. A small dose of pollen extract (1 c.c. of a one in
a million for example) injected subcutaneously produces in highly
sensitized individuals an itching or prickly sensation followed
immediately by considerable swelling at the point of injection and
redness extending sometimes over an area of several inches. The

70 SCIENTIFIC PROCEEDINGS (52).

injection of a somewhat larger dose (1 c.c. of a one in fifty thou-
sand) caused a considerable swelling and pain, and small blisters
resembling hives developed at the point of injection. This was
followed by dizziness and a sense of general discomfort and a slight
attack of hay fever was precipitated.

Specific precipitin and complement deviation reactions using
pollen extracts as antigens were exhibited by certain cases before
the commencement of the hay fever season. These reactions dis-
appeared under treatment (see following paper) reappearing a
few weeks later. Since the reactions in question, while quite
definite, were not exhibited by all the cases, it is proposed to make
them the subject of further investigation. The leukocyte count
and body temperature were frequently above the normal in the
course of an acute attack of hay fever. All attempts to induce
hay fever symptoms in normal individuals by introducing into
their eyes pollen extracts mixed with the serum or nasal secretions
of sensitized individuals have failed. It may be concluded from
these results that specific enzymes capable of splitting the pollen
protein and liberating a toxic factor, or immune bodies capable of
combining with the specific body present in the pollen extract to
form a toxic combination are either not present or not demon-
strable by this method in the blood or secretions of hay fever
cases. Preliminary experiments which require confirmation would
indicate rather that the serum of highly immunized hay fever
cases when mixed with pollen extract may exert aslightly neutral-
izing effect on the latter, protecting the eyes of sensitive indi-
viduals to a certain extent from pollen toxin.

49 (745)

A preliminary communication on the treatment of autumnal hay
fever by vaccination with an aqueous extract of the pollen
of ragweed.

By GEorGE H. A. CLOWES.

[From the Laboratory of Biological Chemistry, State Institute for
the Study of Malignant Disease, Buffalo, N. Y.]

Dunbar was the first to attempt active immunization against
hay fever by vaccination with aqueous pollen extract. He used

VACCINATION FOR AUTUMNAL HaAy FEVER. 71

too large a dose (about ten thousand times that which we have found
most satisfactory), and the patient on whom he tried the experiment
experienced such a violent anaphylactic shock that he decided to
abandon direct vaccination, and subsequently resorted to passive
procedures. Noon and Freeman succeeded in considerably alle-
viating the condition of sufferers from European or spring hay
fever by injecting small doses of timothy pollen extract.

So far as we are aware no previous attempts have been made to
immunize against American autumnal hay fever by vaccination.
The extracts which we have employed for this purpose were pre-
pared from the pollen of ragweed by one of the two following
procedures: (1) That previously employed by Dunbar consisting
of repeated freezing and thawing in a 5 per cent. aqueous suspen-
sion, and (2) an original method, which consists of precipitating
the pollen with acetone and extracting with water.

A series of eight cases were vaccinated last summer, using
doses ranging for the most part from I c.c. of a one in five
million solution to 1 c.c. of a one in five hundred thousand. The
size of the initial dose, the increase in the amount at each injec-
tion, the number of doses administered (ranging from three to
twelve), and the time intervals (ranging from two to six days)
were regulated by making frequent ophthalmic, cutaneous and
blood tests and noting the measure of immunity indicated by
these tests and the general condition of the patient. All the
cases treated experienced a marked alleviation of general symp-
toms corresponding very closely with the physical tests. In the
following table are recorded the initial resistance, maximum
resistance, and resistance five months after completion of treat-
ment exhibited by a series of cases. The figures represent the
number of units of pollen toxin required to produce a definite
ophthalmic reaction, one unit being the soluble constituents of
one twenty-millionth of a gram of pollen. The figures included

RESISTANCE CLASSIFIED UNDER TREATMENT.

Initial. Maximum. Five Months After
Treatment.
| Dio CAA ae eIcIa DIGG 2 2,000 (40,000) 10
WU Viterricenciore ce veiora eats 40 2,000 (20,000) 20
Dr CDs cisciac anes 20 1,000 10
DD Trae Peseta yous! syst dcyn-ale.8 40 2,000 Io

INES Seis cielisllereve @ ae 2 1,000

72 SCIENTIFIC PROCEEDINGS (52).

in brackets indicate the number of units required to produce an
itching sensation in the eye which apparently runs considerably
higher than the flushing point in highly immunized individuals.
Extremely small doses (less than those recommended by Noon
and Freeman in treating European hay fever) appear to give the
best results. One case, for example, received four injections at
intervals of four days, the amounts being .12, .18, .24, and .36 c.c.
of a one in one hundred thousand extract of ragweed pollen, and
showed at the end of this period a fifty-fold increase in resistance.
Another case recently immunized against spring hay fever for
experimental purposes, received three doses of timothy pollen
extract, I c.c., 14 c.c. and 2 c.c. of a solution of one in five million
at intervals of six days, and at the end of this period exhibited a
hundred-fold increase in resistance. In another case in which
the resistance had already been raised by previous treatment a
thousand-fold, a dose of 214 c.c. of a one in one hundred thousand
solution produced urticaria, dizziness and other unpleasant symp-
toms referred to in the preceding paper and obviously constituted
an overdose. The employment of doses in excess of I c.c. of one
in one hundred thousand is unnecessary, possibly dangerous and
certainly to be avoided. That the changes noted above are not
due solely to the development of natural immunity in the course
of the disease is proved by the fact that three cases after experienc-
ing typical hay fever symptoms for two or three weeks were
immunized in the middle of the season and immediately showed a
marked improvement, and three cases treated long after the close
of the season rapidly developed a greatly increased resistance
to the pollen extract employed. The specific nature of the im-
munity developed by injecting specific pollen extracts was proved
in two cases suffering from both spring and autumnal hay fever,
one of whom, Mrs. K., was recently partially immunized against
timothy without changing the ragweed sensitiveness, the other,
Miss R., being similarly immunized against ragweed without
affecting the timothy resistance (see table).

Initial Maximum. Initial. Maximum.

R. 200 200 R 200 800
T 200 200

ABDERHALDEN’S BIOLOGICAL TEST OF PREGNANCY. 73,

50 (746)
Abderhalden’s biological test of pregnancy.
By P. F. WILLIAMS and R. M. PEARCE.

[From the John Herr Musser Department of Research Medicine,
University of Pennsylvania.|

SUMMARY.

The use of Abderhalden’s test for pregnancy, employing the
dialysis method and the ninhydrin color reaction has given
positive results with each of 28 sera from pregnant women and
with eight from women in the post-partum period, including one
abortion. The test has never been negative in a known preg-
nancy. On the other hand, the serum of pregnancy reacts with
tissues (kidney, heart, uterus) other than placenta. Also sera of
two cases of nephritis, one of tabes and one of infection (carbuncle),
and occasionally of some individuals in apparent perfect health
have given the reaction with placenta and other tissues.

In the use of Abderhalden’s dialysis method we have found
the ninhydrin reaction far superior to the biuret reaction. It is
also important that Schleicher and Schull’s smaller dialysis sacks
should be used rather than the fish skin membranes originally
recommended.

Results as satisfactory as those obtained by dialysis are ob-
tained by mixing tissue and serum in tubes and after incubating
for 24 hours testing the filtrate obtained on coagulation by heat
and acetic acid with ninhydrin.

Inactivation of the serum causes a great diminution in the
degree of reaction, but does not cause it to disappear entirely.
At zero temperature no reaction occurs. The power of a serum
to cause the reaction persists when the serum is kept under proper
conditions of temperature for at least 7 days.

As the result of our studies we feel that this test cannot be
accepted as an accurate clinical method until it has been more
thoroughly investigated and the possible sources of error corrected.
This conclusion however applies only to Abderhalden’s dialysis
method and not to his optical method with which we have had
no experience.

74 SCIENTIFIC PROCEEDINGS (52).

51 (747)

The Kernplasmarelation during the life of a pedigreed race of
Oxytricha fallax.

By LoRANDE Loss WooDRtUFF.
[Sheffield Biological Laboratory, Yale University.|

The following conclusions were reached from a study of cells
during periods of characteristically different reproductive activity
of an 860 generation pedigreed race of Oxytricha fallax:

1. A wide variation in the size of the cells and of the nuclei
occurs at all periods of the life of the race.

2. The mean size of the cell is smallest at periods of high repro-
ductive activity and becomes progressively larger as the division
rate falls.

3. The mean size of the nucleus is smallest at periods of high
reproductive activity and becomes progressively larger as the
division rate falls.

4. The Kernplasmarelation of individual cells shows a wide
variation at all periods of the life of the race.

5. The mean proportion of nuclear to cytoplasmic material is
highest during the period of greatest reproductive activity.

6. The size of the cell and the size of the nucleus as well as the
Kernplasmarelation are interpreted as an incidental result rather
than as a cause of the rate of cell division.

52 (748)
The reaction between oxygen and hemoglobin.
By E. E. BUTTERFIELD.
[From the Rockefeller Institute for Medical Research.|

There are at present four different views as to the nature of
the absorption of oxygen by blood.
The first and oldest view is expressed by the reation,
1 mol. hemoglobin + 1 mol. oxygen = 1 mol. oxyhemoglobin.
According to the law of mass action one would have

ab = ke, (1)

THE REACTION BETWEEN OXYGEN AND HEMOGLOBIN. 75

in which a represents the concentration of hemoglobin in solution,
b the oxygen concentration, ¢ the concentration of oxyhemoglobin,
and & a constant.

The second view is that of Bohr. Bohr assumed that hemo-
globin in aqueous solution is hydrolytically split into globin + Fe-
component, and that the Fe-component combines reversibly with
oxygen. Bohr’s formulation of this hypothesis leads to an equa-
tion of the fourth degree. Without going into details it will
suffice to call attention to the main points of Bohr’s work as
far as we are concerned here. They are, in addition to the
assumed hydrolysis of hemoglobin in aqueous solution, first, the
inapplicability of formula (1) to his results and secondly, the

oxygen absorbed
total hemoglobin concentration
total hemoglobin concentration! at constant pressure.

The third view is that of Wo. Ostwald who applied the older
adsorption formula,

with the

variation of the ratio

x = ke, (2)

in which x = oxygen adsorbed, ¢ = oxygen concentration, k and
nm are constants, to some of the data of Paul Bert and Loewy and
found that log x and log ¢ plotted in rectangular codrdinates gave
a straight line.

The fourth and last view is that brought forward by Manchot,
who found that the quantity of oxygen or CO absorbed by 100
c.c. blood, laked or unlaked, varied with the dilution, 7. e., with
the total hemoglobin concentration. On 10-fold dilution this
quantity approached a maximum value of 2 mol. oxygen to I mol.
hemoglobin.

The experiments which I have to report were carried out during
the last 4 years in the course of other studies. These experiments
are restricted to the influence of change in the total hemoglobin
concentration on the amount of gas bound pro gram hemoblogin
at constant gas pressure. The results also hold only for hemo-
globin in solution, 7. e., aqueous solutions of crystalline oxyhemo-
globin or centrifuged red corpuscles dissolved in H2O. It is to
be emphasized that the few experiments cited here have been

1 By total hemoglobin concentration is meant here and in the following the total
weight of blood coloring matter (hemoglobin + oxyhemoglobin) in unit volume.

76 SCIENTIFIC PROCEEDINGS (52).

selected solely with reference to the lowest and highest total
hemoglobin concentration in each series. Four entirely different
methods were employed and they all lead to the same result.

TABLE I.

Absorption of CO. Hemoglobin spectrophotometrically.

co
a ¢ Hp
654 3.59 1.29
16 658 4.86 1.30 Laked ox blood.
668 2.88 1.29
616 5.40 1.27 Laked human blood.
Reduction with palladium-hydrogen. Hemoglobin gravimetrically.
O2
D ¢ Hb
II. 156 1.23 1.09
154 II.50 1.08 Oxyhemoglobin from ox
blood.
Oxygen capacity by ferricyanide method. Hemoglobin spectrophotometrically.
O§
"4 Hb
100m 1.64 1.37
4.27 1.40 Laked rabbit blood.
Spectrophotometric.
log 4
Dilution 2
log Jai
A
2 I.12
10 1.13 Laked ox blood.
1.5 1.03 Laked ox blood after extrac-
Io 1.13 tion with ether.
IV. 200 Tests
log is
zs
7,
log 7
1.14 1.13
7.00 1.13 Oxyhemoglobin from ox

blood.

The first group of experiments was carried out with laked ox
blood and with laked human blood. The volume of CO absorbed
by oxygen-free blood ((NH2)2.H2O and vacuum pump) was deter-
mined. The quantity of CO physically absorbed was deducted
from the total volume, using the coefficient of absorption for CO
in H.O at the temperature of the experiment (20° C.) for the
calculation. Total hemoglobin concentration determined spectro-
photometrically. It will be seen from the table that for a small

THE REACTION BETWEEN OXYGEN AND HEMOGLOBIN. 77

range of concentration (2-fold) the CO absorption pro gram hemo-
globin is practically constant for laked ox blood or human blood.
(In the table, c= hemoglobin concentration, gram in 100 c.c., CO=
volume CO absorbed by hemoglobin, Hb. = total hemoglobin
quantity in grams, and p = pressure of CO in mm.) The devia-
tions are no greater than could be accounted for by the slight
decrease of the solubility of CO in hemoglobin solutions of ascend-
ing concentrations. It will be noticed that at a CO pressure of
650 mm. the limit 1 mol. CO to 1 mol. hemoglobin is almost
reached (theor. 1.34 c.c. CO pro gram hemoglobin). The differ-
ence again is in all probability due to the difference in coefficients
of absorption of CO in H.O and in blood of the concentrations
studied. The reason for the good agreement between the values
for ox blood and human blood rests in the identity of the optical
constants for human hemoglobin and ox hemoglobin.

The second series of results was obtained by the method of
Paal. Reduction of dialyzed solution of oxyhemoglobin, saturated
with air at the prevailing atmospheric pressure, in an atmosphere
of hydrogen in the presence of colloidal palladium. The quantity of
hydrogen absorbed was measured. It was assumed that under the
conditions of the experiment oxyhemoglobin + H;, = H.O + hemo-
globin. Previously the quantity of Hz absorbed by 0.1 gram Pd
in I10 c.c. H2O was determined. This was deducted from the
total H2 absorbed in the experiments with hemoglobin. In all
of these experiments 0.1 gram Pd and IIo c.c. solution were
constantly used. Hemoglobin concentration determined gravi-
metrically. It will be seen from the table that for an almost Io-
fold variation in the hemoglobin concentration the quantity of
O2 pro gram Hb. remains constant. The value is considerably
lower than for CO at high pressures. It is not necessary to go
into the various possibilities (incomplete reaction, adsorption of
hemoglobin by Pd, etc.) which may account for this difference.
It suffices here to call attention to the fact that the quantity of
O2 given off pro gram Hb. to a reducing agent is independent of
the total hemoglobin concentration if the solutions have been
previously saturated at the same partial pressure of oxygen.

The third series! was obtained by the ferricyanide method of

1 From an unpublished study on the action of pneumococcus on blood, by Dr.
F. W. Peabody and myself.

78 SCIENTIFIC PROCEEDINGS (52).

Haldane and Barcroft with laked rabbit saturated with oxygen
at the prevailing partial pressure in the air. Total hemoglobin
concentration determined spectrophotometrically, using the con-
stants of oxyhemoglobin from ox blood. Here again for a 3-fold
variation in concentration O./Hb. is constant. The apparent
slight excess over I mol. O2 to 1 mol. Hb. is probably due to a
slight difference in the optical constants of oxyhemoglobin from
rabbit blood and oxyhemoglobin from ox blood.

The fourth and last series was obtained from measurements of
the light absorption of laked ox blood and oxyhemoglobin from
ox blood at the wave-lengths 577, 579 uu (double line, mercury
arc) and 546up. Log (J1/h’) is the log of the ratio of initial
and final intensities in the solution at 577, 579 up, log (Le/Is’)
the log of the corresponding ratio at 546 wu. Between the quotient
of the logs of these ratios and the concentration of hemoglobin
(oxygen-free) in the presence of oxyhemoglobin there exists the
following relation:

log a (Bi — a) x + ay
1
Sea an toes (3)
log ZT! (Bo — az)x + ae
2
in which x = relative concentration of hemoglobin, a1, a2, 61, Be
qT,
log =
constants which can be determined experimentally. If — =
log a
2

remains constant on dilution then x must be constant. That is if

iT.
log —

remains constant on dilution at constant Oz pressure the

relative composition of the solution (oxyhemoglobin and hemo-
globin) also remains constant.

We will now apply these results to the existing views on the
nature of the absorption of oxygen by hemoglobin. Several con-
siderations not embodied in the table will also be of aid in selecting
the most probable formula.

The Bohr formula falls out for the following reasons:

THE REACTION BETWEEN OXYGEN AND HEMOGLOBIN. 79

1. When the globin is split off from the hemoglobin molecule
the iron-component can be identified as hematin (or its reduction
product, hemochromogen), which has a characteristic spectrum.
All of our spectroscopic and spectrophotometric evidence is against
the presence of hematin or hemochromogen in normal blood or in
solutions of freshly prepared oxyhemoglobin. I have recently
been able to split hemoglobin in glycerin solution by means of
(NHe)2.H2O reversibly into globin + hemochromogen. The re-
action, however, only takes place under special conditions and is
always accompanied by sharp spectral changes.

2. Granted for the moment that hemoglobin is hydrolyzed in
aqueous solution. Then it must be possible to shift the equilib-
rium by changing the concentration of H’ or OH’ ions. Following
experiments! show that such a shift of equilibrium does not
occur or at least if it does it has no influence on the combination
of oxygen with hemoglobin at constant pressure. Similar results
were obtained with KOH. On increasing the H’* or OH’ concen-
tration beyond the quantities given in the table a constitutive
change occurs in the oxyhemoglobin molecule; methemoglobin is
formed.

TABLE II.
uantity of 2/10 HCl Added
c O,-Capacity. = on 4-5 ce Washed Rabbit
b Corpuscles in c.c.

I 4.27 5.97 1.40 0.0
II 4.47 6.25 I.40 0.5
Iil 4.48 6.37 1.42 1.0
IV 4.40 6.35 I.44 2.0

3. From Bohr’s experiments and formula the quantity of Oz
absorbed pro gram hemoglobin is a function of the total hemo-
globin concentration. According to the experiments here pre-
sented the quantity of O2 bound pro gram hemoglobin is inde-
pendent of the total hemoglobin concentration at constant gas
pressure.

The arguments 2 and 3 against the Bohr formula also hold for
Manchot’s work. In addition it is to be noted that in our experi-
ments the limit value for the oxygen (or CO) absorption is 1 mol.
oxygen to I mol. hemoglobin. In no case does it approach 2 mol.

1 From the pneumococcus study by Dr. Peabody and myself.

80 SCIENTIFIC PROCEEDINGS (52).

The spectrophotometric data are conclusively against Manchot’s
views. Manchot reached the conclusion that undiluted blood
contains about 33 per cent. hemoglobin uncombined with oxygen,
while at 10-fold dilution the amount of uncombined hemoglobin
approaches zero. From the spectrophotometric data it follows that
x (or 1o0ox, the percentage of uncombined hemoglobin) is prac-
tically independent of dilution at constant oxygen pressure.

Of the four views mentioned in the beginning, this leaves the
choice between the mass action formula for a monomolecular
chemical reaction and the adsorption formula. Our experimental
results furnish little aid in this choice, except the fact that the
limit value of oxygen (or CO) absorption is 1 mol. gas for each
mol. hemoglobin. This would indicate, of course, a mono-
molecular reaction. A very strong argument against the adsorp-
tion theory is the fact that combination of hemoglobin with
oxygen is accompanied by a sharp change in the spectrum, which
as far as I know can only be interpreted as constitutive molecular
change. Not having any data of my own suitable for a test of
the adsorption formula, I have taken the data of Bohr, Hiifner,
and Loewy. Inno case is m log c + log k a straight line function
of log x. Even from Loewy’s numerical data it is impossible to
find support for the adsorption theory—it will be recalled that
Wo. Ostwald bases his claim for the adsorption of oxygen by
blood chiefly on a curve of Loewy’s. The formula x = kc” is
only of limited application to adsorption phenomena. On trying
the more general formula of Arrhenius on the same data one finds
that k is not constant. So on the whole it may be said that there
is no evidence to support the view that oxygen is adsorbed by
hemoglobin. It is possible that adsorption may play a réle in
the taking up of oxygen by intact red corpuscles, but there too
the chief phenomenon is more probably the chemical combination
of oxygen with hemoglobin.

This leaves finally the first and oldest view of reaction between
oxygen and hemoglobin as the only one of the four which is at
the same time tenable and compatible with the results here
presented. These results are entirely in accord with formula (1),
b being practically constant in each series (or varying slightly
with the solubility of the gas in solutions of different total hemo-
globin concentration). We would have then,

STIMULATION OF THE LABYRINTH OF THE EAR. 8I

conc. hemoglobin

conc. oxyhemoglobin
which is a uniform result at constant pressure and temperature,
obtained by four different methods. It is necessary in closing
to call attention to the fact that in all experiments the gas pres-
sures were relatively high and that under these conditions one is
working in the neighborhood of maximum saturation of hemo-
globin with CO or oxygen.

—CONSt

53 (749)

The effects of stimulation of the labyrinth of the ear in the living
animal. (With demonstration.)

By J. GorDON WILSON and F. H. PIKE.

It may be shown that if one sits in a moving railway train
and observes objects out of the window, they may seem to be
moving in a direction opposite to that of the train. If one notices
the movements of one’s eyes in observing these objects, it will
be seen that the eyes move slowly backward, opposite to the
direction in which the train is going, and more quickly return to
the median position, the quick movement of the eye being in the
same direction as the movement of the train. We may express
this relation between eye-movements and apparent movement of
external objects or real passive movement of the one’s own body
by the statement that the apparent movement of external objects
is in the direction of the slow deviation of the eyes, and the real
passive movement of one’s body is in the direction of the quick
movement of the eyes. The otic labyrinth is not involved in
these reactions. We may extend this relationship still further
and say that under whatever conditions of this kind, with one
possible exception,! one may be placed the movement, either real
or apparent, of external objects is in the direction of the slow
deviation of the eyes, and passive movement, either real or ap-
parent, of one’s own body is in the direction of the quick movement
of the eyes. While many of the facts have long been known, we
have not seen any general expression of the relationships in terms
of the slow and quick eye movements.

1 Holt, E. B., Psychological Review, 1909, XVI, p. 388.

82 SCIENTIFIC PROCEEDINGS (52).

Stimulation of the otic labyrinth in a number of different ways,
including rotation about a particular axis of the head, evokes
movements of the eyes, and sensations of apparent movement of
external objects or of one’s own body, or both. The movement
of the eyes is slow to the stimulated side if a weak stimulus is
used, and slow to the opposite side if a strong stimulus is applied.!
Here also the real or apparent movement of external objects is in
the direction of the slow deviation of the eyes, and the apparent
passive movement of the body, or the real passive movement, if
rotation occurs, is in the direction of the quick movement of the
eyes, if it occurs. This quick movement of the eyes is lacking in
some lower forms, such as the turtle.

When, by any means of labyrinthine stimulation, sensation of
apparent (passive) movement of the body is produced in an animal
at rest, the active compensatory movement of the animal is
opposite in direction to that of the apparent movement of the
body, and is in the direction of the slow movement of the eyes.
This is the basis of the explanation of the fact, observed clinically,
that patients with labyrinthine lesions fall or make other body
movements in the direction of the slow deviation of the eyes, a
relationship not previously sufficiently emphasized.

In electrical stimulation of the labyrinth by the direct current,
the animal (turtle) moves toward the side on which the anode
is placed. Reversing the direction of the current changes the
direction of movement of the animal.

We may give a description of the passive movements, either
real or apparent, and the active or compensatory movements of
the body, under these various conditions, in terms of eye move-
ments by the statement that the passive movements are in the
direction of the quick movement of the eyes, and the active
compensatory movements are in the direction of the slow devia-
tion of the eyes.

1 Wilson and Pike, Phil. Trans. Royal Society, London, Series B, 1912, Vol. 203,
p. 135.

PROTEIN CONCENTRATION AND ABSORPTION OF ANTIBODIES. 83

54 (750)
The pancreatic lipase of infants in acute intestinal disturbances.
By ALFRED F. HEss.
[Research Laboratory, Dept. of Health, N. Y. C.]

In the former communication which considered the pancreatic
secretions in chronic malnutrition in infants, it was found that
various ferments of the gland are normally secreted even in
advanced instances of marasmus or atrophy. In the present
study of an acute disease, of acute intestinal indigestion or ali-
mentary intoxication, which was carried out also by the direct
method, by the use of the duodenal catheter the lipase was found
deficient, although the two other pancreatic ferments were present
in considerable amount. The deficiency of lipase seemed to some
degree characteristic of this disturbance; it is not a general char-
acteristic of all febrile conditions and was not met with in pneu-
monia or empyema. It is possible that the lack of lipolytic
activity in this disease should be correlated with the clinical
manifestation of fat intolerance, and the metabolic studies showing
a deficient absorption of fat.

55 (751)

The influence of protein concentration upon the absorption of
antibodies from the subcutaneous tissues.

By W. H. Park, L. W. FAMULENER and E. J. BANZHAF.
[From the Laboratories of the Department of Health, New York City.]

The experiments so far completed indicate that the concen-
tration of protein up to double its normal amount in an antitoxic
or agglutinating serum or globulin solution has but little influence
upon the absorption of the contained antitoxin or agglutinin
from the subcutaneous tissues of man or animals unless the
increased concentration of protein together with other substances
gives rise to a greater local inflammatory reaction. The absorp-
tion of agglutinin was markedly less in a number of rabbits in
which the subcutaneous injections of the high proteid solutions
were followed by infiltration and necrosis of the adjacent tissues.

84 SCIENTIFIC PROCEEDINGS (52).

The high proteid concentration did not appreciably lessen the
amount or rapidity of absorption, when no such reaction in the
tissues took place. Contrary to the conclusion of Walbum the
results obtained in four healthy men did not show any appreciable
difference in absorption of antitoxin from an antitoxic globulin
solution, the proteid concentration of which was equal to that of
normal horse serum and one in which the concentration was
double that amount.

56 (752)

The influence of the vagus nerves on the faradized auricles in
the dog’s heart.

By G. CanBy ROBINSON.

[From the Hospital of the Rockefeller Institute for Medical Research,
New York.|

The effect of vagus stimulation on the abnormal cardiac
activity set up by faradization of the dog’s auricle was studied in
twenty-three experiments. Faradization of the auricles threw
them into a tumultuous activity which in fifteen of the experi-
ments persisted after faradization was discontinued from five
minutes to overan hour. In these experiments opportunities were
afforded for studying the nature of the abnormal auricular activity
set up by faradization and for determining what effect stimulation
of each vagus nerve had upon it. In several experiments the effect
of cutting the vagi while the abnormal activity was present was
observed. In eight experiments in which the abnormal activity
could not be established independently, the effect of vagus stimula-
tion was observed by beginning it before ending the faradization
of the auricles. When this was done the abnormal auricular
activity usually continued until after the end of vagus stimulation
and was affected in the same manner as the continuous or estab-
lished tumultuous activity.

The auricular activity resulting from auricular faradization
consisted in very rapid movements, apparently contractions of
the whole auricles, which were sufficient to produce definite move-
ments of the recording tambour attached to the auricular myo-
cardiograph. Beside this rapid auricular tachycardia, fine fibril-

INFLUENCE OF VAGUS NERVES ON AURICLES. 85

latory movements in the various fibers could be seen. When the
right vagus was stimulated with a faradic current of moderate
strength, the coarser movements ceased and the typical fine
fibrillations persisted, and when the stimulation was removed the
coarser movements could be seen definitely, gradually returning
and being coexistent with the fine fibrillation. This effect pro-
duced a change in the electrocardiogram and the undulations
representing auricular activity became more rapid, blurred and
often almost disappeared. This characteristic change in the
electrocardiograms occurred in 89.5 per cent. of the experiments.

When the left vagus was stimulated the coarser movements of
the auricles were not disturbed, but appeared sometimes perhaps
even more distinctly than before. It was difficult to determine
with certainty whether the fibrillatory movements ceased or were
influenced. There was a definite difference in the electrocardio-
grams when the right and left vagi were stimulated during the
abnormal auricular activity in 70.5 per cent. of the experiments,
the larger waves of auricular activity being much less or not at
all disturbed by stimulation of the left nerve.

Thus the difference between the action of the two vagi in the
dog, pointed out by Cohn,! is further extended.

Cutting the vagi after the establishment of the abnormal
auricular activity had little or no effect upon it, but the ventricular
rate was sometimes much increased. Vagus stimulation increased
the susceptibility of the auricles to faradization, and in four experi-
ments the abnormal auricular activity could be made to continue
after the faradization was stopped only by stimulating the vagi
synchronously with or for a few seconds after faradization. In
two experiments an auricular activity identical with that following
faradization was set up by right vagus stimulation alone.

The normal sequential beat is also often restored by vagus
stimulation. It replaces the abnormal auricular activity not
during, but a few seconds after the termination of vagus stimula-
tion. Left vagus stimulation seems somewhat more effectual in
producing this effect than right vagus stimulation.

1 Cohn, Jour. Exper. Med., 1912, XVI, 732.

86 SCIENTIFIC PROCEEDINGS (52).

57 (753)

Prolonged complete heart-block, without lesion of the bundle of
His and with frequent changes in the idio-ventricular
electrical complexes.

By B. S. OPPENHEIMER and H. B. WILLIAMS.

[From the Department of Physiology, Columbia University, New
York.|

Electrocardiograms were obtained at intervals from an old
hemiplegic patient who was known to have complete heart-block
from February 26, 1912, up to the day of his death, December 31
of the same year. During many months of this period he had
pronounced Cheyne-Stokes respiration. The interest in this case
lies in the fact that there were frequent changes in the electrical
complexes of the ventricular beat and correlated with this the
fact that histological examination revealed no organic lesion to
account for the block, in the auriculo-nodal junction, the node
of Tawara, or the main stem and its branches. The nodal artery
was sclerotic.

The variations in the ventricular complexes were seen not
only from one examination to the next, but often from beat to
beat.

The waves Q, R, S and T all showed variations; for example
with leads I and II the wave R was sometimes upright, sometimes
inverted. With lead III the wave R was always inverted.

The auricular rate was strikingly reduced during the dyspneic
period (44.49 beats per minute), as compared with its rate during
apnea (94.17 beats per minute). The ventricular rate was only
slightly reduced during dyspnea (30.46 beats per minute as com-
pared with 31.25 beats per minute). In other words the vagus
still had a marked chronotropic effect on the auricle and little if
any on the ventricle. Two atropin tests were made during which
the heartblock was not relieved; and, again, the auricular rate
was decidedly increased, the ventricular rate very slightly so.

To explain the divergent types of ventricular complexes we
may consider the possibility that the intrinsic ventricular pace-
maker was frequently shifting, or that the different impulses

PRODUCTION OF TEMPORARY VALVULAR LESIONS. 87

started at the same point and traveled either along different
routes, or at varying rates along the same route.

Complete heart-block without anatomical lesion in the auriculo-
ventricular system may possibly be of neurogenic or of circulatory
origin, or it may be ascribed to chemical agents, to asphyxia, or
to some hindrance to the passage of impulses from the terminal
arborizations of the conducting system to the ventricular muscula-
ture.

A previous example of possible functional heart-block was
reported by Dr. Alfred Cohn.! In his case of transient complete
dissociation showing constantly varying ventricular complexes,
the patient recovered, so that there was no opportunity of deter-
mining whether or not there was an organic lesion in the aur iculo-
ventricular system.

58 (754)
Methods for the production of temporary valvular lesions.

By Car. J. WIGGERS and EUGENE F. Du Bots.

[From the Physiological Laboratory, Cornell University Medical
College, New York City.]

Experimental valvular lesions have been induced by a number
of investigators—stenoses by tightening of ligatures or clamps
about the valvular orifices, insufficiencies by tearing of valves
with sounds and glass rods or by cutting with specially constructed
valvulotomes. Such experimental stenoses may, if desired, be
temporary, and normal circulatory conditions may be subsequently
reéstablished. Experimental insufficiencies such as have been
described, must, owing to the traumatic nature of the lesion, be
permanent. As no method for the production of temporary in-
sufficiencies has apparently been described, the following method,
which also permits a study of the intraventricular pressure changes,
was devised.

Method.—A curved metal catheter (22 cm. long, internal
diameter 6 mm.) having toward the tip one or two openings
(6 mm. in diameter) and three centimeters from the tip a longi-

1 Cohn, Proc, Soc. FOR ExPER. BIOL. AND MED., Vol. IX, No. 2, p. 24, December,
IQII.

88 SCIENTIFIC PROCEEDINGS (52).

tudinal slot (6 mm. wide and 25 mm. long) is fitted with a lubri-
cated rubber tube (4 mm. internal diameter) so as to occlude the
longitudinal slot. To produce aortic insufficiency, the metal
catheter with its rubber obturator is introduced, free from air
and without hemorrhage, into the left subclavian artery and
aorta. The catheter is so adjusted by palpation that the valves
close about it near the middle of the occluded slot in the catheter.
The inner rubber tube may now be connected directly with a
manometer.! Aortic pressure records with Frank’s manometer
show that a catheter of such size that it can be introduced into
the subclavian does not impede the systolic discharge so as to
cause stenosis in the physiological sense. By drawing out the
obturating rubber tube to such an extent that the slot is opened, a
valveless circuit with a minimal resistance is established between
aorta and ventricle. The intraventricular pressure may still be
recorded.

Upon entirely withdrawing the metal catheter, normal con-
ditions are restored. By inserting the catheter through the auricu-
lar appendage, the method may be used to induce mitral regurgi-
tation. Postmortem examination of six hearts showed no damage
of the valves.

The advantages of being able to produce temporary valvular
insufficiencies are several:

1. It permits the consecutive demonstration of all valvular
lesions and their circulatory effects on the same animal. In such
a series of demonstrations we produce mitral and tricuspid stenosis
by simple invagination of the auricular wall into the auricular
ventricular openings with a finger.

2. By subsequently reéstablishing normal conditions, it per-
mits a control of the reflex circulatory and respiratory disturbance
incidental to the production of experimental valvular lesions. It
is quite possible for such reflex effects to entirely overshadow or
complicate the mechanical effect of the lesion and, unless so
controlled, the results cannot be considered comparable to clinical
lesions.

1In the case of Frank’s optical manometer, the connection can be made rigid
and without elastic connection.

REGENERATION OF AN ANNELID Worm. 89

59 (755)

The influence of the central nervous system in regeneration of
an annelid worm.

By A. J. GOLDFARB.
[From the Biologic Laboratories of the College of the City of New York.]

In a previous investigation, it was found that the Morgan and
Nussbaum methods of operating worms resulted in a large mor-
tality, excessive injury, retarded healing and little regeneration.
The same results were obtained with the marine annelid worm
Amphinoma pacifica.

By the method used successfully with Lumbricus 90 Amphi-
nomas were operated in such a manner that the nerve cord was
completely removed from 2 to 6 or more segments next the ampu-
tated level and with little or no injury to adjoining tissues. Over
151 worms served as controls.

Regeneration was limited to definite levels. Within these
regions the head or the tail was regenerated in most of the worms,
even though the nerve cord had been removed. On subsequent
examination of serial sections it was found that in one group of
worms, the nerve cord had regenerated from the broken end as
far as the amputated level. Some of these worms had begun to
proliferate new tissues, others showed no sign of proliferation.
In a second large group, the regenerated nerve cord of the new
head was continuous with the old nerve cord.

In a third group, the old nerve cord had grown a few ganglia
towards the amputated level, the remaining segments were entirely
devoid of nerve cord. Nevertheless the head had been regenerated
with its typical supraesophageal ganglia or ‘“‘brain,’’ its commis-
sures and even ventral nerve ganglion which grew posteriorly
towards the old cord. This group demonstrated that Amphinoma
as well as Lumbricus, and other adult organisms could regenerate
the missing organ without the contact of or stimulation from the
central nervous system.

90 SCIENTIFIC PROCEEDINGS (52).

60 (756)

Changes in concentration of sea water and their influence upon
regeneration.

By A. J. GOLDFARB.
[From the Biologic Laboratories of the College of the City of New York.|

The regeneration under changed densities of sea water was
observed under conditions that ensured the elimination or uni-
formity of associated factors such as size of medusz, volume,
surface and depth of solutions, extent of injury, level of amputa-
tion, temperature, crowding, aeration, etc. Furthermore, the
dilute solutions were made with water containing a known quan-
tity of sea salts, and the concentrated solutions were made by
slow evaporation in the sun’s heat, which corrected certain errors
in previous experiments of this nature. The results were checked
by repeated observations, plotted for intervals of 14, 24 and 30
days, after amputation of the arms of Cassiopea xamanacha.

These medusz lived in solutions ranging from 40 to 153 per
centum of the salts of sea water. They regenerated however in
50 to 133 per cent. solutions. The regeneration of normal arms
and supernumerary arms was limited further to 75 to 105 per
cent. solutions. Beyond these limits regeneration was atypic.
Optimum regeneration occurred not in sea water but in sea water
diluted 95 to 90 per cent. With increasing dilution the amount
regenerated was diminished very slowly, with increasing con-
centration very rapidly. The subsequent examinations revealed
the fact that in the sub-optimum solutions the arms regenerated
absolutely and relatively faster than in any of the other solu-
tions, correspondingly changing the character of the curve.

When the results were compared with those of Loeb it was
found that both the hydroid Endendrium of Woods Hole, Mass.,
as well as Cassiopea of Dry Tortugas, Fla., differed radically from
the classic experiments of Loeb on Tubularia of Naples, in respect
to the range of solutions in which animals lived or regenerated,
the optimum solutions, the normality of the regenerated parts
and the character of the curve. It is altogether probable that
Loeb’s curve is limited to Tubularia of Naples and does not

BACTERIA OF THE DUODENUM. gI

represent the behavior of organisms to changes of density of sea
water, and that the differences in the behavior of these three
organisms can hardly be correlated with the differences in con-
centration of the sea water in which they normally live.

61 (757)

Variations in the amount of transformed atoxyl (trypanotoxyl)
produced by varying the strength of atoxyl incubated with
blood.

By B. T. TERRY.

[From the Laboratories of the Rockefeller Institute for Medical Re-
search, New York City.]

A Io per cent. solution of atoxyl in blood incubated for I to 3
hours at 37° C. is more than Io times as toxic for Trypanosoma
brucei as a I per cent. solution in blood similarly incubated. To
avoid misleading results due to the transforming action of red
blood corpuscles upon unaltered atoxyl continuing after the dilu-
tions are made, immediately after incubation all red blood cor-
puscles should be removed by centrifugalization from the fluid
containing the transformed atoxyl.

62 (758)

Some observations on bacteria of the duodenum.
By W. J. MAcNEAtL and A. F. CHACE.

[From the Laboratories of the New York Post-Graduate Medical
School and Hospital.|

The duodenal tube was sterilized by boiling in water for ten
minutes and the lower end was covered with a tightly-fitting
gelatin capsule which had been soaked in alcohol for several days.
The gelatin capsule was finally coated with shellac and dried.
The tube was ordinarily given late at night and the sample of
fluid aspirated on the following morning, usually without any
food being taken in the interval. In a few cases the fluid was
obtained an hour after giving an Ewald test meal.

92 SCIENTIFIC PROCEEDINGS (52).

The study of the fluid included inspection, direct microscopic
count of the bacterial cells, plating of measured amounts on
litmus lactose agar and ascitic-fluid agar, separation cultures of
measured quantities in tall tubes of ascitic-fluid agar and fer-
mentation-tube cultures of measured quantities in dextrose broth
and lactose broth. In addition, a portion of the fluid was heated
to 80° C. for ten minutes and inoculated in measured quantities
into tall tubes of glucose ascitic-fluid agar and into fermentation
tubes of glucose broth and lactose broth.

Thirty-five samples were studied, of which the first nine were
unsatisfactory because of defects in technic. The results obtained
on the remaining twenty-six serve as a basis for this report.

The results can be presented here only in summary form.
In general, the number of bacterial cells seen microscopically
varied from 600,000 to 960,000,000 per c.c. and their number
bore no evident relation to the number of colonies obtained in
cultures nor to the clinical condition of the patient. On the
other hand, the results of the culture work indicate that the normal
duodenal fluid is practically free from living bacteria when food
is absent, and that the number of cultivable bacteria obtained
in a given case is a rough index of the digestive derangement.
Gas-forming microbes developed in the fermentation tubes in
ten of the twenty-six fluids examined, all from cases with gastro-
intestinal disturbance. Bacterial spores capable of resisting a
temperature of 80° C. for twenty minutes were not found in any
examination.

One hundred ten subcultures were isolated and studied. Forty-
three of these were Gram-positive, non-liquefying cocci. These
were isolated from a majority of the samples and doubtless are
the organisms most frequently present in health and in slight dis-
turbances. Gram-negative, gas-forming bacilli which liquefy gela-
tin were isolated from five of the twenty-six fluids, all derived
from cases of severe gastro-intestinal disorder. In some cases these
bacilli were very abundant. Liquefying cocci, yeasts and various
kinds of bacilli were isolated in less abundance. Only three of the
110 strains belonged in the B. coli group. Four of the twelve
strains isolated from the one case of typhoid relapse proved to be
B. typhosus.

EFFECTS OF MORPHINE ON Doc’s HEART. 93

63 (759)

The effect of morphine on the mechanism of the dog’s heart
after removal of one vagus nerve.

By ALFRED E. CoHN.

[From the Hospital of the Rockefeller Institute for Medical Research,
New York.]|

When injected intravenously, morphine was shown by v.
Egmond to cause cardiac arhythmia in dogs. Ejinthoven and
Meek and Eyster studied the results of such injections electro-
cardiographically. Einthoven concluded that these were due to
stimulation of the vagus centers. He found complete lack of
uniformity in the results. Meek and Eyster believed the effects
to be due to disturbances in conduction between sinus and auricle
and between auricles and ventricles.

The similarity of some of Einthoven’s curves to those resulting
from faradic stimulation of the right vagus nerve and of others
from stimulation of the left nerve rendered it probable that
morphine sometimes had a preponderating influence on the right
and at others on the left vagus nerve and center. Experiments
were accordingly carried out in twelve dogs, in six of which the
right and in six of which the left vagus nerves were removed
aseptically. In 3 morphine was injected before operation. Reg-
istration was galvanometric. In three dogs (one right during
three attempts and two left during two and three attempts)
arhythmia characteristic of morphine poisoning was not obtained.!
In three it was obtained on the first attempt, in five on the second
and in one on the third. In five right vagus and in four left vagus
dogs injection succeeded and the resulting arhythmias were directly
comparable to those obtained on faradic stimulation. In the right
vagus dogs the auricles, except for occasional contractions which
escaped at long intervals, and also the ventricles ceased to beat.
The circulation was carried on by ectopic ventricular contractions.
That there was no defect in conduction was shown in a number of
ways. In the left vagus dogs the auricles were merely slowed.
There was depression in A-V conduction in all of them,—in three

1 The dogs are named according to the nerve retained.

94 SCIENTIFIC PROCEEDINGS (52).

it was of moderate degree and resulted in an As-Vs ratio of 2:1
or 3:1. In another it was severe; except occasionally, the ven-
tricles received no impulses from auricular contractions, but main-
tained the circulation by idioventricular contractions in complete
dissociation with those of the auricles. All the dogs recovered.
The conclusions are warranted: that morphine stimulation of
the vagus center in dogs having one vagus nerve only has the same
effect as faradic stimulation of the same nerve in other dogs;
that the results of the morphine method substantiates a former
conclusion based on faradic stimulation that characteristic differ-
ences between the two vagi exist; and that the lack of uniformity
found by Einthoven in the arhythmia of the heart obtained on
morphine injections is only apparent and is capable of analysis.

64 (760)

The influence of temperature on the minimal dose of strychnin
in frogs and on the time of onset of tetanus.

By T. S. GITHENS.

[From the Department of Physiology and Pharmacology of the
Rockefeller Institute.]

Rana pipiens were used exclusively and all injections were
given in the dorsal lymph sac. Temperatures from 40° F. to
80° F. were studied. With doses of 0.0006 mg. per gm. (about
0.02 mg. for a 30 gm. frog) tetanus was constantly obtained at all
temperatures. With doses of 0.0005 mg. per gm. (about 0.015
mg. per frog) strong tetanus was obtained constantly at tempera-
tures about 40° F. and about 80° F. It was occasionally seen at
55° F. and never at temperatures from 65° to 75° F. With doses
of 0.0003 mg. per gm. strong tetanus was constant about 40° F.
and occurred frequently at 80° F., but was never seen at tempera-
tures from 55° F. to 70° F.

In regard to the time elapsing before tetanus; with minimal
doses it is very variable but on the whole is less the higher the
temperature. Thus with a dose of 0.0006 mg. per gm. tetanus
came on at 40° after 14 to 4 hours; at 55° after 1 to 214 hours; at
70° after 34 to 2 hours; and at 85° after 4% to 1 hour. Witha

GREHANT’s ANESTHETIC AND MORPHINE-ETHER. 95

dose of 0.001 mg. per gm. tetanus came on at 40° after 14 to 4
hours; at 55° after 34 to 114 hours; at 70° after 14 to 34 hours.

These results show that tetanus may be induced by strychnin
at low and at high temperatures by doses which will not cause
tetanus at temperatures between. They also show that the inter-
val between injection and the onset of tetanus grows less with
higher temperatures, although there is no constant ratio.

65 (761)

On the difference in the effect of Gréhant’s! anesthetic and of
morphine-ether on the total output of urine and the
composition of the urine in normal dogs.

By WM. DEB. MACNIDER.
[From the Laboratory of Pharmacology, University of North Carolina.]

In a recent study of the action of various diuretics in uranium
nephritis,” it was shown that the anesthetic employed in the experi-
ments not only influenced the output of urine, but that following
the anesthetic, diuretic substances such as caffeine, theobromine,
and 0.9 per cent. sodium chloride lost their diuretic value.

The following experiments are being conducted to ascertain
if the previously mentioned anesthetics have any effect in reducing
the output and in changing the composition of the urine in the
normal dog and if the anesthetics differ from one another in their
action.

When full grown dogs are given Gréhant’s anesthetic in the
strength usually employed there is a marked reduction in the
output of urine and the animal becomes glycosuric, the percentage
of glucose varying between 0.165—3.33 per cent. Acetone has so
far been constantly present in the urine of these full-grown
animals.

An albuminuria is induced and is accompanied by the appear-
ance of casts, hyaline, or hyaline and granular. In one of the

1 Gréhant’s anesthetic. The animal is given 14 c.c. per kilogram of a 4 per cent.
solution of morphine. This is followed in half an hour by 10 c.c. per kilogram of the
following mixture: chloroform, 50 c.c.; alcohol and water, each 500 c.c.

2 The Journal of Pharmacology and Experimental Therapeutics, Vol. III, No. 4,
March, 1912.

96 SCIENTIFIC PROCEEDINGS (52).

animals bile appeared in the urine on the second day after re-
covery from the anesthetic and persisted for two days.

The anesthesia is complete for from ten to eighteen hours.
Three animals have failed to recover.

When Gréhant’s anesthetic is given to full-grown dogs in half
the strength usually employed the output of urine is usually but
slightly diminished, the animals however become glycosuric, the
percentage of glucose having varied between .10I-—.301 per cent.
The urine does not contain acetone.

Albumen is present with hyaline or hyaline and granular casts.

The anesthesia is imperfect. Recovery is usually complete
within twelve hours.

In a final series of animals, puppies were used. The age of
these animals ranged from six weeks to four and a half months.
Gréhant’s anesthetic was given in full strength. The animals
were completely anesthetized for two and a half to nine hours.
The output of urine was decidedly decreased and showed both
glucose and albumen. The percentage of glucose varied from
.O70I-.202 per cent. Acetone was present in the urine of the
animal four and a half months old. All of the animals recovered.

In the experiments conducted with morphine-ether, only full-
grown animals have so far been employed. The anesthetic was
given in sufficient quantity to keep the animal completely anes-
thetized for three hours. All of the animals recovered. The
recovery has been usually complete within six to eight hours.

The output of urine in the twenty-four hours following the
anesthetic has been but slightly reduced, excepting in one animal
that was very old. In this animal the urine was reduced from
515 c.c. on the day prior to the anesthetic to 320 c.c. in the twenty-
four hour period following the anesthetic. The urine of this dog
showed a fairly heavy precipitate of albumen and .104 per cent.
of glucose. Acetone was not present.

The remaining animals of this series have not developed a
glycosuria, and the urine has been free from acetone, albumen, and
casts.

Experiments will be continued to ascertain if there exists any
relation between the age of the animal and the duration of the
anesthesia, and between the duration of the anesthesia and the
appearance of various abnormal constituents in the urine.

[From the Physiological Laboratory, Medical College, Cornell
University, Ithaca, N. Y.]

Observations have been made on twenty-one litters.

RATE OF GROWTH IN Doe.

The rate of growth in the dog.

By SUTHERLAND SIMPSON.

66 (762)

97

As soon

after birth as possible, when the amniotic fluid had been dried
off, each individual was weighed and at the same time it was ear-
marked and the sex determined. For the first three or four weeks
the weights were taken daily, at the same hour (4 to 5 p.m.), and
later every third day. From the figures obtained curves were
plotted showing graphically the absolute weight increments and
the rate of growth. The figures for the number of young, the
proportion of the sexes, the birth mortality, and the body weights
when born, are given in the following table.

pee Eo Average Weight at
Birth.

Sey cres | DP UWwW | DSS ASA SS

emer Mel te lee cle eth AS Ske le (ela |

Dead.

Ee ey

No. of
Litter. oe

rot

I 4
2 4
3 —
4 4
5 6
6 4
7 3
8 I
9 I
10 4
It 2
12 3
13 3
14 4
15 3
16 I
17 4
18 I
19 —
20 5
21 I
58

Total.

Lal

Leal
AMWNHHNHADWAAUAW OI DON OH~1 ©

Lal
w
P=

292

279

The number of young in a litter varies from 2 to 12, families of
8 occurring most frequently. The proportion of the sexes is in
favor of the female, the ratio being 100 males to 119 females.

98 SCIENTIFIC PROCEEDINGS (52).

This does not agree with the statistics for the human subject nor
with Minot’s results in the guinea-pig, but, of course, the number
of individuals so far examined by me is too small to draw any
general conclusion from. The birth mortality is highest in the
males, as is also the average body weight when born.

Minot found that in male guinea-pigs, as in newborn children,
there is an actual loss of weight for the first 3 or 4 days after birth.
Such, however, is not the case in the dog. In almost all the litters
there is some gain in 24 hours, and this is very decided at the end
of the second day. There is a post-natal retardation of growth
but it is of relatively short duration.

67 (763)

The influence of experimental cretinism upon nitrogenous
metabolism in the sheep.

By ANDREW HUNTER.

[From the Department of Physiology and Biochemistry, Cornell
University Medical College, Ithaca, N. Y.]

The object of the investigation was to determine whether
athyroidism in sheep is associated with any striking abnormality
of intermediary metabolism, such as might be revealed by the
nitrogen partition of the urine. The subjects were three sheep
which have already been described before this Society by Simpson.!
At the age of two months they had suffered the loss of the thyroid
and internal parathyroid glands, and had subsequently developed
into typical cretins. One year after the first operation the external
parathyroids also had been removed. While the later condition
was of course not that of uncomplicated athyroidism, symptoms
referable to the loss of the parathyroids were but slightly marked.
Tetany, in particular, was never observed. When the animals
came under my care they were one and a half to two years old.

To furnish a basis of comparison two normal sheep, nearly
four years of age, were included in the investigation. As the
most convenient way of avoiding the difficulties caused by variable
appetite, etc., all five animals were starved. The urine was
collected as voided.

1 Simpson, Proc. Soc. For Exp. BIOL. AND MEDICINE, 1911, IX, 2.

EXPERIMENTAL CRETINISM IN SHEEP. 99

The normal animals endured fasts of five days’ duration with
little apparent distress or enfeeblement. Of the others two rapidly
weakened, and succumbed within six days; the third was in
excellent condition after seven days without food. Below is shown
the ascertained distribution of nitrogen in the urine of one normal
and one operated animal. The records of the others were of
corresponding character.

NORMAL SHEEP—WEIGHT 42 KILOGRAMS.

|

Per Cent. Nitrogen as

Day of Starva- Total

ton. Nitrogen. Wrea Am- | Creati- | Cre- | Allan- | Uric | Purine | Undeter-
“| monia. | nine. atine. toin. Acid. | Bases. | mined.

2-3 8.04 84.4 2.3 3.8 2.0 1.8 0.3 0.4 5.0

3-4 7.64 83.6 3.2 4.2 1.8 rity / 0.4 0.4 4.7

4-5 6.40 80.3 4.3 4.2 2.5 m5 0.5 0.5 6.2

5-6 5-78 79.8 6.3 4.9 2.5 1.7 0.4 0.5 3-9

OPERATED SHEEP—WEIGHT 15 KILOGRAMS.

I-2 3.83 84.4 1.8 3-5 1.2 2.1 0.4 0.7 5-9

2-3 6.82 85.8 1.6 2.4 2.1 I.I 0.2 0.4 6.4

3-5 4.98 84.5 2.4 2.5 Spi 1.4 0.3 0.4 5.4

5-6 2.31 76.6 2.2 2.5

5:9 TAS 0.5 0.5 10.3

Nitrogen partition in the urine of the normal sheep is seen to
follow the general mammalian type, the only point worthy of
special note being the comparatively small proportion of purine
nitrogen excreted as allantoin. In the urine of the thyroidec-
tomized animal the only constituents apparently affected are
ammonia and creatine. The percentage of the first is lower than
the normal at corresponding stages of starvation. (One only of
the three operated sheep—the one least affected by fasting—
showed figures similar to the controls.) The creatine rises early
in the fast to a higher level than the creatinine. The significance
of this ‘‘creatine crossing’”’ is not clear. It might be supposed to
be related to the susceptibility of the animals to withdrawal of
food; but, curiously, in the one subject that survived a week,
creatine exceeded creatinine from the very first day of observation.

Complete data will be published shortly.

100

SCIENTIFIC PROCEEDINGS (52).

68 (764)

infantilism of the Lorain type.

Metabolism studies in a case of hypopituitarism, with

By DEWITT STETTEN and JACOB ROSENBLOOM.
[From the German Hospital, New York, and the Laboratory of Bio-

chemistry of the University of Pittsburgh, Pittsburgh, Pa.|

This report contains the results obtained on studying certain
phases of metabolism in a case of a man aged 22, who presented
the classical symptoms of infantilism of the Lorain type, with
symptoms definitely pointing to a benign tumor, probably a cyst

of the hypophysis.!

The arrest of the body growth and of sexual

development with the moderate adiposity stamped this case as a

Date.

Date.

Total Nitrogen.

TABLE I. (NITROGEN PARTITION.)
Urine.
Urea- Ammonia- Creatinine- Uric Acid- Undeter-
Nitrogen. Nitrogen. Nitrogen. Nitrogen. mari bs
ois a Th Sine ee
sso eas) ee) sa 0 et)
r=] as bp es bo aS bo = ase] g] 28 wo
5 |de8| & |d88| & |d8e| & 1588/5) 532
ee TAs = Es = 2
ile INAS (A Aol MeN peu|Le
13.5 | 83.8 | 0.36 2.3 0.60 3.8 0.20 TA) i t-4ly Oe7
13.3 | 81.6 | 0.41 2.5 0.58 a5 0.24 I.5 |1.8] I1.0
12.6 | 80.3 | 0.40 2.5 0.56 3.5 0.26 big f qatkal) ae yat
12.9 | 81.7 | 0.44 BT 0.54 | 3-4 0.23 15) ht-7|| .10:8
12.3 | 80.5 | 0.40 2.6 0.50 3.3 0.27 Te 70|| 8 || eke
12.6 | 80.2 | 0.48 Byit 0.55 3-5 0.29 1.9 |1.8] 11.5
TABLE II. (SULPHUR PARTITION.)
Urine.
Total |Ethereal| Ior-
Sul- Sul- anic | Neutral} Sulphate-S Hees imaee, Sun ewe:
phate | phate | Sul- Sul- |—Totals Total S.
Sul- Sul- es phur otal S Total S Total S 4
ul- er
vbur, ; Pane ; Bee Gm. | Per Cent. | PerCent. | PerCent.| Cent.
m.
0.45 | 0.04 | 0.41 | 0.12 78.6 6.8 71.8 21.3
0:30) )1/10:03'5|00:27,5)| (0.15 72.5 72 65.3 27.4
0.66 | 0.06 | 0.60 | 0.14 82.4 G Gist) 17.5
0.69 | 0.04 | 0.65 | 0.21 76.7 4.3 72.4 23-3
0.69 | 0.05 | 0.64 | 0.21 76.7 5.5 7 fifo 23.3
0.53 | 0.05 | 0.48 | 0.34 61.7 6.4 55-3 38.3
0.66 | 0.04 |! 0.63 ! 0.30 69.2 4.0 65.2 30.8

1 At operation a cyst of the hypophysis was found.

RELATION OF PANCREATIC EXTRACT TO SUGAR. IOI

typical example of the Froehlich typus or the so-called dystrophia
adiposo-genitalis of Bartels.

The patient was placed on the high protein diet of Folin and
the following tables contain the data obtained in this study.
The high undetermined nitrogen and neutral sulphur of the urine,
can readily be noted, while the absorption of fat and protein

was normal,
TABLE III.

Summary of data pertaining to total nitrogen and fat metabolism.

Total Nitrogen. Feces. Fat.
Total Nitrogen.

Ae See a 2 Percent-

Date. Ingested, Excreted, Balance, Fat, palGunt hey age of
Grams. Grams. Grams. Grams. poand: of Total ‘ny | Grams. ate

“| Ingested.

6/26 18.2 17.9 +0.3 2.2 1.8 9.9 140 98.4

6/27 18.6 18.1 +0.5 2.2 1.8 9.7 140 98.4

6/28 18.3 17.5 +0.8 2.2 1.8 9.9 140 98.4

6/29 18.1 17.9 +0.2 1.8 2.1 11.6 146 98.8

6/30 17.6 17.4 +0.2 1.8 2.1 II.9 146 98.8

9(1 17.8 17.8 oO 1.8 2.1 II.9 146 98.8

Total 108.6 106.6 +2.0 I2.00 | II.70 _ 858 —
Average | 18.2 | 177+ | +033 | 2.00] 1-951 10.8 ! 143 | 98.6_
69 (765)

The relation of pancreatic extract to the sugar of the blood.
By ERNEsT L. SCOTT.
[From the Department of Physiology of Columbia University]

In January, 1912, I reported a preparation of pancreas which
when injected intravenously into dogs that had been rendered
diabetic by pancreatectomy, lowered both the output of sugar
and the D/N ratio. This preparation seemed to offer a ready
means of attack for several of the problems bearing on the relation
of the pancreas to sugar metabolism. Possibly the simplest of
these is the relation between the amount of sugar present in the
blood and the abundance of the pancreatic hormone present, and
this is a preliminary report of my work on this subject.

Cats were killed and their blood was collected. The protein
was removed and the blood was decolorized by a modification of
the phosphotungstic acid method reported by Oppler. In deter-

102 SCIENTIFIC PROCEEDINGS (52).

mining the amount of sugar Munson and Walker’s ‘“ Uniform
method of sugar determination’? was used and gave consistent
results. In order to test its efficiency the method was controlled
by division of a sample of blood into two portions. In one of
these the glucose was estimated directly. To the other a known
amount of glucose was added and then this amount was subtracted
from the total recovered, leaving a remainder which should equal
the amount found in the portion estimated directly. The results
of one such control are shown in Table I.

TABLE I.
Blood sample 6 (two cats). Total blood used 190.05 gm.

a b c d
Gm bloodiinisamples, wereis lene el etotatele 47.51 47-51 47.51 47-51
Gm. glucose recovered...........--+- 0.0314 0.0328 0.0501 0.0497
Gmglucose added't.-- eile lieretelovsieisie ats 0.0178 0.0178
Gmyelucoseni blood nr-iieiasieatete sete el 0.0314 0.0328 0.0323 0.0319
Gm. glucose per 100 gm. blood........ 0.0661 0.0690 0.0680 0.0671
Average for ab and cd...........+.+:- 0.0676 0.0676

It was necessary next to determine the amount of sugar
in the blood of normal cats. The results for a series of ten cats
are given in Table II. It will be noticed that with the exception
of the cat which gave 76 mgm.—obviously an anomalous case—
the greatest variation from the average is 4 mgms. The starred
numbers indicate controls such as are reported in Table I. The
results are calculated to grams of glucose per hundred grams of
blood.

TABLE II. . TABLE III.
0.0676 . 0.1026
0.0676 0.0783
0.0618 0.0719
0.0760 0.0811
0.0653 0.0692
0.0628 Average... .0.0806
0.0627
0.0646 =
0.0646
0.0691 *

Average... .0.0662

The extracts of the pancreas that were used in the following
experiments were prepared as reported previously, except that the

BLoop PRESSOR SUBSTANCES IN IMMUNITY. 103

temperature was at all times kept below 50° C. instead of being
allowed to go to 65° as before. So far five cats have been injected
with the extract, the first three with two injections each and the
others with one each. All injections were made beneath the skin
of the back. The results are shown in Table III and are calculated
to grams of glucose per hundred grams of blood as in Table I.
The average for the injected cats is over 21 per cent. above that
for the normal ones, and moveover the amount of sugar is greater
in each injected cat than for any normal animal except the one
that gave 76mgm. This increase is very surprising and of peculiar
interest. At present I am not willing to venture any explanation.
There are however several possibilities which are amenable to
experiment and I hope that further work will throw some light
on it. In any case it would seem to put in grave doubt the idea
that the pancreatic hormone always tends to increase the storage
of glycogen in the liver at the expense of sugar in the blood.

There are a number of factors entering into the experiments
so far performed that might cause the individual variations in the
experimental results. Some of the more probable are: the length
of time intervening between the injection and the death of the
animal; the amount of extract injected; the age of the extract;
the number of injections and the time intervening between them,
etc. At present I am trying to find some of the optimum condi-
tions.

70 (766)

On some blood pressor substances and adrenal separations
in experimental immunity.

By J. P. ATKINSON and C. B. FITZPATRICK.

One of us in a work! on “The Preparation of Diphtheria
Antitoxin”’ endeavored to demonstrate by charts of the systemic
reaction following injections of cultures of the bacillus of diphtheria
and its toxin that the real crux of the process of immunization was
to determine when to re-inject. This question is still unsettled;
in short, of two animals treated the same, upon being re-injected
the one, which may apparently be the better prepared, dies and
the other recovers.

1 Fitzpatrick, N. Y. Medical Journal, April 27, 1895.

104 SCIENTIFIC PROCEEDINGS (52).

This aspect of immunization is very well brought out in the
following example.

R6émer and Joseph! re-infected two tuberculous sheep with
a culture of which I mg. per Io kg. killed healthy sheep in one
month. One of these sheep was re-infected with I mg. per Io kg.
ten months after a previous inoculation with a .2 mg. per Io kg.
and 15 months after a first injection of .1 mg. per 10 kg. This
animal died in 48 hours. The other one of these sheep was re-
infected with the same dose (I mg. per 10 kg.) ten months after
previous injection of .2 mg. per 10 kg. This animal responded
with an intense reaction. The reaction was followed by a return
to health.

We have shown in studies read before this society that the
blood of tuberculous animals especially when about to die of
tuberculosis contains a depressor substance. The use of this
blood in conjunction with tuberculin was likewise shown to give
an effective immunity against fatal infection with the B. tubercu-
losis in one case, and in some others the fatal ending was delayed
beyond the controls. It was also demonstrated that the injection
of this serum shortly after the injection with the tuberculin
rapidly caused death. We have also found depressor substances
to be present in the blood serum of other diseases.?

The first part of our present report consists of results of the
injection of blood serum obtained from animals recovering from
inoculations of B. tuberculosis and the possible application of these
observations to the practical therapy of infection and intoxication.

Protocol’ April 19, 1912.—A 14 lb. female dog, sensitized
April 18, 1912, with 34 c.c. of crude tuberculin.

8 c.c. of the serum of a calf recovering from repeated doses of
culture of B. tuberculosis (human type), when injected into the
femoral vein of this dog gave a decided rise in blood pressure.
This rise was preceded by a very slight depression. This dose
was repeated in six minutes with a like result. Five minutes later
the same dose was injected and gave no response.

1“‘Die Tuberculése Re-infection,” Beitrége zur Klinik der Tuberculose, 1910,
XVII, pages 330-333.

2 Atkinson & Fitzpatrick, Proc. Soc. Exp. BIoLoGy AND MEDICINE, I9g1o, VII,
PP. 77-79; ibid., pp. 104-107; zbid., VIII, pp. 24-28; ibid., pp. 49-51, 1912.

33 c.c. normal calf serum gave no response when injected into the femoral vein
of dog sensitized with tuberculin.

BLoopD PRESSOR SUBSTANCES IN IMMUNITY. 105

Protocol July 13, 1912.—7 c.c. of serum of same calf, bled
June 21, 1913, gave a fair rise. The extract in saline solution of
the adrenal gland of this calf gave a very high rise, I /16 c.c.
being sufficient to cause a very marked straight rise.’ 7 c.c. of
tuberculous (bovine type) dog serum gave no response. This dog
was killed and was probably dying according to the autopsy
findings.

Protocol August 28, 1912.—8 c.c. of serum from a dog, recover-
ing according to autopsy findings from tuberculosis (bovine type)
gave a fine rise. Repeated with same results three times.

Protocol January 5, 1910.—2 c.c. of the serum from a rabbit
with tuberculosis, bovine type, gave a fair, sustained rise.

Protocol April 6, 1912.—8 c.c. of serum from a dog recovering
from tuberculosis, bovine type, gave a marked rise. Repeated
three times.

8 c.c. of this serum, injected into a dog sensitized with tuber-
culin, gave a marked rise, when added to 10 drops of tuberculin
No. 5, which ordinarily caused a fall.

Further observations of the occurrence of pressor substances
were made as follows in animals about to die from hydrophobia.?

Protocol 16.—7 c.c. of serum obtained from a rabbit (June 21,
1912), sick with hydrophobia, gave a marked rise. Femoral
injection.

7 c.c. of serum from a goat with hydrophobia gave a slight rise.
Femoral injection.

Protocol March 17, 1911.—7 c.c. of serum from a dog with
hydrophobia, gave a slight depression.

Protocol April 8, 1911.—5 c.c. of serum of rabid dog, bled
April 8, 1911, gave marked rise. Repeated once.

4 c.c. of serum from collie with hydrophobia also gave a marked
rise.

7 c.c. from a dog with dumb rabies gave a rise, bled April 5,
IQII.

7 c.c. from a dog with rabies (March 17, I9II) gave a rise.

1 The extract of the adrenal glands of rabbits which died of tuberculosis showed
little or no pressor substance.

2 We found the pressor substance present in the serum of animals (dog and goat)

suffering from street rabies, bled just before their death. These animals with the
exception of the rabbit were naturally infected.

106 SCIENTIFIC PROCEEDINGS (52).

6 c.c. from a collie with rabies gave a marked rise, bled March
24, IQII.

Protocol of March 31, 1911.—3)4 c.c. of serum from collie
with hydrophobia gave a very marked rise. Repeated three times.

6 c.c. of serum from fox terrier, with hydrophobia, tested
March 17, I91I, gave a very marked rise. Repeated twice.

Protocol November 9, 1910.—1)4 c.c. of serum from a dog with
hydrophobia, bled November 3, I910, gave a slight rise; 5 c.c.
gave a fair rise.

Protocol January 6, 1912.—7 c.c. of serum from a hydrophobia
dog, of January 2, 1912, gave a fair rise. ;

The apparent contradiction between the recovering tuberculous
animal and the dying hydrophobia animal may be explained by our
previously reported observations,! where the continued injection
of a depressor substance instead of causing death led to an actual
increase in blood pressure.’

It would appear that animals possessing these pressor sub-
stances do not die as readily from infection or intoxication as those
which do not have these pressor substances. It appears to us
that we may have here a general law for many, if not all infections
and intoxications.

We have found two substances in adrenal gland preparations.

A substance causing increased pressure and a depressor sub-
stance.

In commercial preparations we found these two substances to
be present.

Separations were made by alcohol, ether, chloroform and normal
saline solution.

In saline extracts of fresh glands, the substance causing increased
pressure was present alone or so greatly in excess as to dominate
the reaction.

In the adrenals of certain diseased animals (tuberculosis,

1 Proc. Soc. Exp. BIOLOGY AND MEDICINE, 1912, IX, pp. 49-51.

2 It would also be of interest to study the origin of these pressor substances in
normal and diseased tissues. Protocol of January 16, 1913, for example shows
that the injection of 5 c.c. normal dog brain extract, gave no response, while the
same doses of tuberculous and hydrophobia brain extracts gave a slight rise. These
extracts were made with saline solution.

COMPLEMENT DEVIATION IN PREGNANCY. 107

vaccinia, septic pneumonia, etc.) the substance causing increased
pressure and the depressor substance were found.

The organs which possess pressor substances are apparently
important organs of immunization. The organs with the depressor
substances are possibly eliminating or fixing the split-products or
poisons arising from the disease. The adrenals are probably
simply one of a number of organs, which possess pressor producing
tissues, and which when acting altogether in the whole living
organism produce the general pressor substances or immuno-
pressor substances. In vivo these pressor substances are probably
present in every tissue and form part of the factors which over-
come infection. They may be the first step or a very important
step in overcoming infection.

These results apparently furnish an indicator as to when and
how to re-inject animals already afflicted artificially or naturally
with an infection or intoxication, with the purpose of immunizing
and healing the diseased animals. That the rdle of these pressor
substances, present in experimental immunity is important, we do
not doubt. The use of an immunizing dose which is just sufficient
to cause their production may be what these observations indicate.

71 (767)

Preliminary communication on a complement deviation reaction
exhibited in pregnancy.

By G. H. A. CLowEs, Francis C. GOLDSBOROUGH,
and F. WEsT.

[From the Laboratory of Biological Chemistry, State Institute for the
Study of Malignant Disease, Buffalo, N. Y.]

In a series of twenty-five normal pregnant women at term in
which syphilis could be excluded the blood of the mother was taken
from the vein during labor and the blood of the infant from the
cord at the time of delivery. The sera after separation from the
clots were frozen and allowed to remain in the icebox 48 hours
before being employed. A series of complement deviation tests
were carried out, using both unheated and heated sera of mothers

108 SCIENTIFIC PROCEEDINGS (52).

and infants in conjunction with a large variety of antigens with
the following results. The unheated mothers’ sera invariably
contained antibodies capable of causing a well-marked deviation
of the complement when used in conjunction with an antigen
obtained by extracting human blood clots with alcohol. The
unheated infants’ sera tested under the same conditions invariably
showed completely negative results. A large variety of alcoholic
extracts of tissues used as antigens gave similar although some-
what less marked deviation with unheated mothers’ sera and
absolutely negative results with unheated infants’ sera. The
deviating body concerned in this reaction is destroyed by heating
for 14 hour at 58° C. The mother’s sera tested after heating were
negative to the antigens enumerated above, and those of the
infants either negative or very slightly positive, but exhibiting
on the whole a somewhat greater capacity to deviate than that
possessed by the heated mothers’ sera. The deviating capacity
of unheated mothers’ serum varies greatly, certain cases exhibiting
a complete deviation only when employed in concentrations as
high as .05 to .075 c.c. of serum, others giving a complete deviation
when amounts as small as .ooI c.c. of serum were employed. It
is important to note that the antibodies in question occasionally
fail to make their appearance until after the serum has been
frozen for two or three days as indicated above. Similar non-
specific immune bodies destroyed by heating at 58° C. have been
observed in cancer and other pathological conditions,! and to a
certain extent in supposedly normal individuals. The entire
absence of these bodies in the blood of newborn infants and their
invariable occurrence in the blood of pregnant women at term
indicates that in this case at least they probably bear some relation
to the reaction of the body against detached fetal cells or proteid
or enzymatic bodies of fetal origin. The occurrence of this re-
action to a marked extent in cancer, particularly in those cases
in which tumors are absorbing under treatment, lends further
support to this point of view.

1 Clowes, Zeitschrift f. Krebsforschung, 1912, Vol. XII, page 421.

ANTITRYPTIC REACTION IN PREGNANCY. 109

72 (768)
On the antitryptic reaction exhibited in pregnancy.
By G. H. A. CLowEs and FRANcIS C. GOLDSBOROUGH.

[From the Laboratory of Biological Chemistry, State Institute for the
Study of Malignant Disease, Buffalo, N. Y.]

The antitryptic index has been determined in a series of twenty-
five pregnant women and their infants. The blood was taken
during labor from the vein of the mother, and from the cord of
the infant at birth, was allowed to clot and the separated serum
employed for tests which were carried out by means of the Oswald
viscosimeter, making use of a method previously described.1. The
antitryptic index of the mother’s serum was found to range from
1.5 to 2.5, averaging about 2, whilst that of the infants was found
to range from .9 to I.2, averaging slightly over 1. The antitryptic
index of a series of cancer cases! previously reported shows a
range of variation from 1.2 to 3.5 and gives an average over 2.
It will thus be seen that the blood of this series of twenty-five
infants appears to be practically normal. On the other hand the
blood of the mothers contains an extremely high percentage of
antibodies to trypsin, averaging over twice the normal, a charac-
teristic also exhibited by cancer blood. This antitryptic reaction
is destroyed by heating the serum to 60° for 144 hour in which
respect it resembles the complement deviating reaction referred
to in the previous paper from which it may be concluded that
whilst these two reactions do not follow absolutely parallel lines
in a quantitative sense, they probably have a common origin.
The fact that reactions of this type invariably occur in preg-
nant women and in cancer cases and are entirely absent in
infants lends support to the theory that they result from a re-
action of the body against enzymes or other products of fetal
origin in pregnancy, and similar products derived from the tumor
in cancer.

1 Clowes, Zeitschrift f. Krebsforschung, Vol. XI, page 146.

110 SCIENTIFIC PROCEEDINGS (52).

73 (769)

On a new factor in passive anaphylaxis.
By RICHARD WEIL.

[From the Department of Experimental Therapeutics, Cornell Uni-
versity Medical School, New York City.]

After active sensitization guinea-pigs remain hyper-sensitive
for at least three years—probably for life. After passive sensitiza-
tion with homologous serum, 7. e., with the blood of an immune
guinea-pig, they retain the sensitive condition so conferred for a
period of 60 or 70 days at least. After passive sensitization with
heterologous serum, 7. é€., with the blood of immune rabbits, they
lose the sensitive condition in 10 days or less. This rapid loss has
never been explained. It might theoretically be due to the devel-
opment by the injected guinea-pig of immune substances directed
against the introduced rabbit serum, which would then neutralize
or destroy the rabbit anti-bodies, on the presence of which sen-
sitization depends. This theory would explain the fact that the
injected guinea-pigs retain their sensitiveness for about ten days,
which would correspond to the time necessary to develop anti-
bodies.

In order to test this idea, normal guinea-pigs were given a
subcutaneous injection of normal rabbit serum. After eight days
they were sensitized with a very large dose of the serum of a
rabbit immunized against horse serum, given intra-peritoneally.
Two days later, when tested by an intravenous injection of horse
serum, they failed to manifest any symptom of anaphylaxis.
The controls, although sensitized with a much smaller dose of
immune rabbit serum, and intoxicated with about one fiftieth
of the amount of horse serum used in the previous series, suc-
cumbed without exception.

The refractory state towards passive sensitization, as thus
induced, may occasionally be demonstrated within three days of
the first injection.

Partial immunization of guinea-pigs against rabbit antibodies
can sometimes be obtained by the previous injection -of sheep
serum. It is, therefore, not strictly specific.

HypERTROPHY OF THYROID GLAND. III

74 (770)

Hypertrophy of the thryoid gland. Revision of experiments
made 25 years ago.

By WILLIAM STEWART HALSTED.

For some years I have thought that the hyperplasia of the
remaining thryoid tissue which has followed excision of a portion
of the thyroid gland might be due to infection of the wound, and
was, in most cases, not a compensatory hypertrophy; and for the
following reasons:

1. In 1888 I found that hyperplasia of the thyroid glands of
dogs occurred after the injection of several c.c. of a bouillon
culture of Staphylococcus aureus into the peritoneal cavity, and
also when a mild form of peritonitis had been produced in these
animals—a peritonitis which was not rapidly fatal.

2. Experiments conducted in 1906 and 7 in the Hunterian
laboratory seemed to prove that for the successful transplantation
of a parathyroid glandule, a considerable deficiency must be
created.

3. Have observed that symptoms of hyperthyroidism and even
exophthalmic goiter may develop promptly after tonsillitis, ap-
pendicitis, pneumonia, typhoid fever and other infections.

4. Twice in the course of the past five years I have had the
opportunity to examine the remaining lobe of the thyroid gland
after excision of the other in dogs whose wounds had healed
throughout without suppuration and have noted that there was
no hyperplasia of the former.

5. The inconstant results obtained by other experimenters.

Last October I proposed to Dr. Hunnicutt, my assistant the
Hunterian laboratory, that he undertake a series of experiments
with view to determining the matter definitely. Observing
aseptic precautions in the strictest manner Dr. Hunnicutt has
made a large number of experiments and we are able to report
that in the nine dogs whose thyroids thus far have been examined
there has been not the slightest evidence of hyperplasia in a
single instance. The average time allowed to elapse between the
removal of the first and second lobes was 55 days, the shortest

II2 SCIENTIFIC PROCEEDINGS (52).

interval being 30 and the longest 81 days. In one of the dogs
there was a superficial stitch infection for a few days, otherwise
there was in no case the slightest evidence of suppuration.

From a restudy of the report of my experiments on extirpation
of the thyroid gland made in 1888, I find that, for the major part
of the experiments, the wounds of the dogs were left open, and
that after 22 days, with 3 exceptions (Nos. 105, 126 and 127)
there was hypertrophy, macroscopic and microscopic, of the remain-
ing gland in the animals whose wounds were permitted to heal
by granulation, whereas when the wound healed absolutely per
primam the hyperplasia of the remaining thyroid tissue did not
develop except in dogs which died from some intercurrent disease
such as pneumonia or distemper.

Of the three exceptions referred to the tip only of one thyroid
gland was ligated in two (nos. 126 and 127) and no tissue removed
at the operation. 132 days later, examination of the thyroid lobe
revealed no hyperplasia, except perhaps to a very slight and
questionable degree in one of the sections of the lobe of one of
these dogs. The absence of hypertrophy in dogs 126 and 127
after 132 days may possibly have been due to the long interval
between the first and second operations. The hyperplasic picture
may, possibly, have been present and vanished, although from
observations of 25 years ago we know that extreme hyperplasia
may persist for 104 days and we have no absolute proof from
them that having been once established it disappears. From a
study of the elaborate and important work of David Marine
and Marine and Lenhart I am quite convinced that the return
to normal is to be expected. In one of Dr. Hunnicutt’s dogs
the lobe first removed showed marked hyperplasia, whereas
87 days later the second lobe presented the normal histological
appearance.

In the remaining case (No. 105) the veins at the upper poles of
both lobes were ligated and unnecessary manipulation carefully
avoided. On the 51st day there was no evidence of hyperplasia.

That there is such a thing as true compensatory hyperplasia
is proved, I think, by my experiments in transplantation of the
parathyroid glandules. Thus when both thyroid lobes have been

1 Johns Hopkins Hospital Reports, Vol. I, Table IV.

OccLuUSION OF AORTA BY LIVING TISSUE. 113

removed and only a film of thyroid transplanted with one para-
thyroid gland, this film hypertrophies enormously and on micro-
scopical examination displays the typical picture of hyperplasia.
In one instance the graft was examined 15 months after operation.!
It would be interesting to determine the amount of deficiency
which it is necessary to create for the successful transplantation
of the thyroid gland; also whether hyperplasia necessarily ensues
when the transplant lives. In other words will a thyroid graft
always fail to take unless the deficiency created is so great that
hyperplasia must develop?

Should it become a definitely established fact that hyperplasia
may be produced by infection, not only are a number of things
explained which otherwise seemed inexplicable, but new lines of
investigation suggested.

75 (771)
Partial occlusion of the thoracic and abdominal aortas by bands
of fresh aorta and of fascia lata

By W. S. HatsTED, M.D.

Ligation of the human abdominal aorta has been made 1g or
20 times and always with fatal result.

Dubois, Assalini, Bujalsky, Pirigoff, Cooper, Keen and perhaps
others attempted to occlude the abdominal aorta gradually by
means of cleverly devised instruments which, carrying snares of
silk, metal or catgut might be tightened or loosened at will.
The instruments traversed the abdominal wall and hence infection
was a complication common to all of the methods and defeated
the plans of the operators.

In 1904 assisted by Dr. W. F. M. Sowers, I began a series of
experiments on dogs in the hope of finding a safe method of oc-
cluding the aorta and curing aortic aneurysm. Bands of silver
and aluminum curled about the aorta by an instrument con-
structed for this purpose were rolled tighter by the fingers until
the desired degree of occlusion of this vessel was obtained. The
abdominal wounds were closed with the expectation that they
would have to be reopened one or more times for the purpose of

1 Journal of Experimental Medicine, 1912, Vol. XV, Plate 30, Fig. 2.

114 SCIENTIFIC PROCEEDINGS (52).

progressively occluding the lumen of the artery. But in the
course of our experiments we had opportunities to make trial in
the human subject of partially occluding bands on other arteries
(innominate, subclavian, carotid, femoral, popliteal) whose blood
streams in some instances it seemed unsafe to cut off suddenly
and completely, and found that incomplete occlusion of an
artery sufficed to cure the aneurysm, possibly quite as surely as
might have been expected of total occlusion. Hence, tentatively,
I abandoned the idea of progressive closure of the aorta, deter-
mining, instead, to obliterate the lumen of this vessel, in the
attempt to cure its aneurysm, to an extent which we had found
quite safe in the dog.

I have applied an aluminum band to the human aorta four
times—twice in one subject—and twice with promising results
so far as the cure of the aneurysm is concerned. But the experi-
mental work on animals had led me to expect that ultimately the
metal bands must cut through the artery, because in cases observed
for 7 months or less the wall of the aorta had become atrophied to
an extreme degree and there was no adhesion between the
infolded, attenuated surfaces. That my fears were well founded
was proved by an experience in Europe, about 18 months ago.
The patient was an aged woman with dilated and badly functioning
heart. The large aortic aneurysm was well located for the placing
of a band which was applied just below the renal vessels. Within
a few days the aneurysm which before operation was distinctly
visible from the seats of the operating amphitheater was barely
discernible at the bedside, and at the end of six weeks had dis-
appeared so completely that the patient was discharged apparently
cured. But, walking out of the door of the hospital she was
seized with a pain and returned to her bed. The following morn-
ing she died from hemorrhage. The aorta had ruptured at the
side of the band, but the aneurysm was found to be nearly cured.

Stimulated by the results in this case to further experimenta-
tion it occurred to me to test the behavior of cuffs and spiral
strips of the fresh aorta of one dog when wound about the aorta
of another. So on the 29th of April, 1912, I operated upon two
dogs, partially occluding the aorta of.one of them with a spiral
aortic band and of the other with a cuff cut from the same vessel.

OccLusIon OF AoRTA BY LIVING TISSUE. 115

Strips of aorta were employed rather than of fascia lata, for ex-
ample, because I hoped that the elastic tissue, in case it did not
live, might, at least, serve its purpose for a time sufficient to cure
an aneurysm.

At the end of two months one of the dogs was killed and I was
pleased to find that the cuff which had been used in this experi-
ment was apparently organized and had not stretched to any
appreciable extent. Above the cuff the aortic pulse was forcible,
but below the constriction it was very feeble, though countable
and accompanied by a thrill.

The other dog operated upon at the same time and in the
same manner, except that a spiral band of aorta instead of a cuff
had been employed died (cause of death unascertained) about
three weeks after the operation. In this instance the aorta had
been almost completely occluded by the spiral aortic strip. The
weldlike band had not stretched and seemed to be organized.
The aorta on being split longitudinally was seen to be greatly
and characteristically infolded and almost occluded at the site
of the band. Sections of the specimens indicate that the elastic
coats of the bands as well of the included artery are intact. Dur-
ing the present winter I have made many similar experiments em-
ploying various fresh tissues for the bands with most encouraging
results. We have learned, however, that whereas the spiral bands
seem to be perfectly safe there is danger in the employment of the
cuffs. In two instances of twelve or more experiments one of the
mattrass sutures taken to hold the flaps of the cuffs together cut
through one side and thus being brought in contact with the aortic
wall cut a minute hole in the vessel through which the animal
bled to death. Such an accident can hardly happen with the
employment of the spiral strip, for not only is the strain on the
stitches very slight when this form of band is used, but even if
it were so great that a thread might cut through the spiral at any
point, it could hardly be brought to bear upon the aorta in such
way as to wear into its wall.

The spiral strip is wound twice about the aorta, and when one
or perhaps two stitches have been taken at one end to hold the
contiguous edges of the spiral together at this point, the other end
of the strip is pulled upon until the aorta is occluded to the desired

116 SCIENTIFIC PROCEEDINGS (52).

amount, and then two additional stitches are taken to maintain
this degree of constriction.’

ABSTRACTS OF THE COMMUNICATIONS, PACIFIC Coast BRANCH.
Second meeting.

San Francisco, California, February 1, 1913.
76 (772)
Studies on the nature of biological specificity.
By FREDERICK P. Gay and T. BRAILSFORD ROBERTSON.

[From the Hearst Laboratory of Pathology and Bacteriology and the
Rudolph Spreckels Physiological Laboratory of the University
of California.]

Our previous immunological studies? with the split products
of casein and a compound of casein with protamin, which were
undertaken for the purpose of gaining some insight as to the nature
of biological specificity have apparently been fruitless in so far
as the main point at issue was concerned, with one exception,
In our comparative studies of the antigenic properties of split
paranuclein with paranuclein synthesized by the reversible action
of pepsin from the products of peptic digestion, we were apparently
able to demonstrate the genesis of an antigenic property.

The present study deals with the investigation of the antigenic

1 About 3 weeks ago I received from Dr. Francesco Nassetti a reprint of a
paper by him entitled “‘Avvolgimento di vasi Sanguigni con lembi liberi di aponeurosi,”’
and published April 26, 1912, in the Atti della R. Academia dei Fisiocritici in Siena.
Dr. Nassetti’s experiments were made in the Istituto di Patalogia Speciale Chir-
urgica della R. Universita di Siena, which is under the direction of Prof. A. Salomoni.
His first experiment (a band of fascia about the carotid artery) antedates mine by
56 days, and his article appeared about three months before the publication by me
of a brief account of my first experiments with spiral strips of aorta (Johns Hopkins
Hospital Bulletin, July, 1912, p. 217). My first experiment (Apr. 29, 1912) was
made 3 days after the publication of Nassetti’s report. Hence the credit for the
idea of wrapping blood vessels with bands of fresh tissue belongs, I am happy to
say, to Italy, the country of the famous surgeon, Luigi Porta, who was, I think, the
first to attempt the partial occlusion of an artery (the aorta). Ihave the impression
that Porta used for this purpose a strip of diachylon plaster.

2 Gay, F. P., and Robertson, T. B., Jour. Exper. Med., 1912, XVI, 470; Jour.
Biol. Chem., 1912, XII, 233; Jour. Exper. Med., 1912, XVI, 479.

O6cyYTIN. 117

properties of our second compound of casein with a non-antigenic
protein, globin caseinate. We find that globin is non-antigenic
and highly toxic, producing in guinea-pigs the typical symptoms
and lesions of anaphylaxis. When compounded with casein, it
still remains slightly toxic. An anti-serum derived by repeated
injections of rabbits with globin caseinate contains fixation bodies
for casein, globin caseinate, and, curiously enough, for globin,
although globin alone does not produce such antibodies. By
absorption experiments it may be shown that the antibodies in
anti-globin caseinate serum are two in number, one for casein and
one for globin. Thus it appears that the change in globin brought
about by this combination with casein renders it antigenic.

A further study of similar and of more complex compounded
proteins should give further insight as to the nature of specificity.

77 (773)

On the nature of odcytin; the fertilizing and cytolyzing substance
in mammalian blood-sera. (Preliminary communication.)

By T. BRAILSFORD ROBERTSON.

[From the Rudolph Spreckels Physiological Laboratory of the Uni-
versity of California.]

I have elsewhere shown! that the agent in ox-serum which
brings about the formation of fertilization-membranes in sea-
urchin eggs? can be isolated in an impure condition by a process
consisting, essentially, in precipitating the substance by barium
chloride, re-solution of this precipitate in dilute acid, removal
of the excess of barium by excess of sodium sulphate, and re-
precipitation by acetone.

The preparations thus obtained were found to be contaminated
by a considerable proportion of sodium sulphate, precipitated
together with the fertilizing agent by the acetone. They also

1T. Brailsford Robertson, Journal of Biol. Chem., XI, 1912, p. 339; XII, 1912,
p. 163; Arch. f. Entwicklungsmech., XXXV, 1912, p. 64.

2 J. Loeb, Arch. f. d. ges. Physiol., 118 (1907), p. 36; 122 (1908), p. 96; 124 (1908),
p. 37; ‘““Die chemische Entwicklungserregung des tierischen Eies,’’ Berlin, 1909,
p. 185.

118 SCIENTIFIC PROCEEDINGS (52).

contained appreciable traces of phosphates. I have obtained a
much purer preparation by a process consisting, essentially, in
precipitation from the serum by acetone, extraction of the precipi-
tate with hot N/1o HCl, exactly neutralizing the extract with
Ba(OH)., re-dissolving the precipitate in N/1o HsSO, and re-
precipitating it with acetone. The yield from a liter of ox-serum
lies between 10 and 4o milligrams.

The substance which is thus obtained is soluble in dilute acids,
alkalies and salt-solutions. It is much more rapidly dissolved
when these solvents are hot. Its solutions are not coagulated
by boiling. It is thrown out of solution by chlorides of the alkaline
earths. It yields the Millon, Acree-Rosenheim and xanthoproteic
tests for protein. One part of the substance rubbed up in 512,000
parts of sea-water caused membrane-formation in 80 per cent. of
Strongylocentrotus purpuratus eggs which had previously been
sensitized by 4 minute’s immersion in 3/8 m SrClo.

The active substance, therefore, is either a protein or a peptone,
or else, by the above methods of preparation, is precipitated to-
gether with a protein or peptone.

I find that Witte’s ‘‘peptone’’ contains the membrane-forming
substance, since one part of Witte’s ‘‘peptone’”’ dissolved in
16,000 parts of sea-water caused membrane-formation in 32 per
cent. of sensitized purpuratus eggs. Hence the membrane-form-
agent is digested either with difficulty or not at all by pepsin.

I find that the addition of 0.08 per cent. of lecithin or cholesterin
to rabbit serum does not affect, either qualitatively or quantita-
tively, the membrane-forming and cytolyzing action of the serum.
It would appear very unlikely, therefore, that the active substance
is a lipoid.

78 (774)
On the influence of lecithin upon the development of sea-urchin
embryos. (Preliminary communication.)

By T. BRAILSFORD ROBERTSON.
[From the Rudolph Spreckels Physiological Laboratory of the Uni-
versity of California.]

When the eggs of Strongylocentrotus purpuratus are fertilized
by sperm in a mixture of 50 c.c. of sea-water and 5 c.c. of a 1.7

INFLUENCE OF LECITHIN ON EMBRYOS. 119

per cent. suspension of lecithin in m/2 NaCl, the inner and outer
fertilization-membranes are slowly dissolved by the lecithin, with
the result that in the course of some six hours the cleavage-cells
which have formed fall apart and ultimately disintegrate.

The eggs of Strongylocentrotus purpuratus female were divided
into two portions. Both portions were placed in sea-water and
fertilized with sperm. After 24 hours both lots of eggs had de-
veloped into free-swimming blastule. One portion was now trans-
ferred to a mixture of 50 c.c. of sea-water and 5 c.c. of a 1.7 per
cent. suspension of lecithin in m/2 NaCl for a period of 24 hours
and then returned to normal sea-water. The other portion was
left in normal sea-water. The following table shows the relative
development of the two portions:

Time After

Fertilization. Portion x (Controls). Portion 2.

TAY ierepcieis ster Blastule. Blastule. (These were now trans-
ferred to the lecithin mixture for 24
hours.)

PAY Sine <yeie ets Gastrule. Blastule. (These were now trans-
ferred to normal sea-water.)

BidayS: <5 cise. Gastrule. Blastule.

ANGAY Si = creletsicrc Gastrule and early plutei. Blastule.

SiMAYSis eicisis vers Fully developed plutei. Blastule and 25 per cent. gastrule.

OGEN Sy Gandane Advanced plutei. Early gastrule with narrow, un-
branched intestine and large, clear body-
cavity.

PAYS tie ravave eve Advanced plutei. Unchanged.

SIGAY Sine acs, vc Advanced plutei. The gastrule are now retrograding;
the intestine has almost disappeared.

QO days. sa. Unchanged. Unchanged.

Bacterial invasion terminated the experiment after the 9th
day. It is evident that the immersion of blastule for 24 hours
in a 0.15 per cent solution of lecithin enormously retards their
development.

If purpuratus eggs are fertilized by sperm in more dilute
solutions of lecithin in sea-water (0.003 per cent. to 0.015 per cent.)
the fertilization-membranes are not dissolved sufficiently rapidly
to affect the development. In these solutions development is not
appreciably retarded until the blastula stage is reached. There-
after development is very markedly retarded, and the retardation
is greater the greater the concentration of the lecithin. The

120 SCIENTIFIC PROCEEDINGS (52).

embryos are not injured by the lecithin, however, as they will
ultimately develop to normal plutei if left in these solutions for a
sufficient time.

If cholesterin, suspended in a mixture of m/100 sodium oleate
and m/2 NaCl be mixed with the lecithin in equal proportions
the retarding action of the lecithin upon the development of
purpuratus eggs is almost completely neutralized. The slight
retardation which is observed in these mixtures may be due to
the sodium oleate which is employed to keep the cholesterin in
suspension, since sodium oleate is very toxic for sea-urchin eggs
and embryos.

Cholesterin itself, when added to sea-water, has no influence
upon the rate of development of the eggs. The emulsions of
cholesterin are, however, coagulated by the salts in sea-water
and the cholesterin is completely thrown out of suspension in the
form of coarse flocculli.

79 (775)
On acid agglutination as a method of differentiation of bacteria.

By H. J. SEArRs.

[Division of Bacteriology, Department of Medicine, Stanford
University.|

Michaelis and several other workers following him have claimed
for the phenomenon of acid agglutination a specificity comparable
with that of specific serum agglutination. The reaction is specific,
they say, in that optimum agglutination is produced in suspensions
of bacteria of a single species by a definite concentration of
hydrogen ions, irrespective of the acid used, and in that this
concentration is, in general, different for different species. Their
method has been to prepare solutions of definite hydrogen ion
concentrations by using mixtures of a weak acid with its sodium
salt, the concentrations being calculated from the formula
Coat ”

where & represents the dissociation constant of the acid used.
The differences between the constants obtained in this way for

Cy =k:

DIFFERENTIATION OF BACTERIA. I2I

the different species are especially marked in the case of the
typhoid-colon group. Schidorsky and Reim claim to have had
considerable success in the practical diagnosis of typhoid using
this method.

Jaffé, working with a number of strains in the typhoid-colon
group, obtained constants differing somewhat from those of
Michaelis, and observed also several strains of each species which
showed markedly different behavior toward the reagents from
that of the general average.

In my investigations I made use of six strains of B. typhosus
and four of B. coli. In two sets of experiments, using in one case,
acetic acid and sodium acetate, and in the other, lactic acid and
sodium lactate, the behavior of five strains of B. typhosus was shown
to be the same toward both of these reagents. Three of the strains
regularly failed to agglutinate at all even when concentrations of
hydrogen ion varying between .7 and 900 X 10— were used.
The other two strains agglutinated about equally in all the con-
centrations between 14 and 200 X 10—°. Variations in the con-
centrations of salt present between N/40 and N/200 made no
noticeable difference.

Using acetic and lactic acids without the presence of the
salt, the same three strains failed to agglutinate, the other two,
however, showing agglutination between the values of 60 and
300 X 10—. The hydrogen ion concentrations in these cases were
calculated from the formula

Cy ae V B- Cacia:

Comparing these acids with the strong acid HCl, which can be
assumed to be completely dissociated in dilutions above N/t1oo,
it was found that the three strains which showed resistance
toward the weaker acids were partially agglutinated by values of
Cy of from 50 to 500 X 10— while the other two were completely
agglutinated by these concentrations. A sixth strain also showed
partial agglutination by the same values.

The conclusions to be drawn from these results would seem to
be, first, that the hydrogen ion concentrations necessary to bring
about agglutination in suspensions of typhoid bacilli may differ
considerably with the way in which these concentrations are ob-

122 SCIENTIFIC PROCEEDINGS (52).

tained; second, that equal agglutination is obtained in wide
ranges of concentrations; and third, that these facts together
with the existence of atypical strains of both the B. typhosus and
the B. coli make the differentiation of these two species by this

method extremely uncertain.

SCIENTIFIC PROCEEDINGS.

ABSTRACTS OF COMMUNICATIONS.
Fifty-third meeting.

Physiological Laboratory, University and Bellevue Hospital Medical
College, April 16, 1913. President Ewing in the chair.

80 (776)
Heliotropism and galvanotropism in Euglena.

By FRANK W. BANCROFT.

[From the Department of Experimental Biology of the Rockefeller
Institute for Medical Research.]

Hitherto positively heliotropic Euglene have always been
found to give the motor reaction when suddenly shaded, and not
when suddenly illuminated. Conversely negatively heliotropic
individuals were found to react only to sudden illumination and
not to sudden shading. Upon this association Jennings has based
his theory according to which heliotropic orientation in Euglena
is by ‘‘trial and error.”

It has been found, however, that it is possible to obtain posi-
tively heliotropic Euglene which give the motor reaction when
suddenly illuminated and not when suddenly shaded. It is also
possible to obtain at will negatively heliotropic organisms which
give motor reactions when suddenly shaded, and not when sud-
denly illuminated. Under certain conditions the motor reaction
to shading is given by Euglene in which no heliotropism can be
demonstrated at that light intensity. Under other conditions
distinct negative heliotropism is obtained with a light intensity
which does not bring about any motor reactions when allowed to
shine suddenly on the organisms, or when they are suddenly
shaded. These facts show that heliotropic orientation in Euglena
does not depend upon the motor reactions, but upon a separate

mechanism.
123

124 SCIENTIFIC PROCEEDINGS (53).

Galvanotropism, which has so far eluded observation in Euglena
was obtained by using culture media containing citric acid. With
this material it was found that the method of orientation is
identical in both galvanotropism and heliotropism. Consequently
the orientation to light is as direct as the locomotor mechanism
of Euglena permits, and does not take place by ‘“‘trial and error.”

81 (777)

The fat content, morphology and length of life of cells growing in
diluted blood plasma.

By ROBERT A. LAMBERT.

[From the Department of Pathology of the College of Physicians and
Surgeons, Columbia University.]

Cells growing in unmodified blood plasma (Harrison’s
method) exhibit regularly an accumulation of fat droplets in their
cytoplasm. In the case of the cells of the chick embryo this fat
accumulation is quite marked after 24 to 48 hours, and reaches a
maximum after five to seven days, at which time the cells are
distended with fat droplets.

The experiments herewith reported were planned to determine
the effect of a reduction in the fat content of the culture medium,
brought about by dilution of the plasma, on the accumulation of
fat by the cells. In the course of the experiments the influence
of dilution on the length of life and morphology of the cells was
also observed.

One part of plasma added to twenty or twenty-five parts of
Ringer’s solution forms a medium which coagulates satisfactorily
in hanging drops. Studies were made with dilutions of I : 2,
I:5, 1:10, 1:15, and 1:20 of pigeon plasma in Ringer’s
solution, containing 0.9 per cent. NaCl. Pieces of chick embryo
heart were used for cultivation. The tissue was finely divided
into pieces of suitable size for cultures, which were washed in
Ringer’s solution for a half hour before using. Cultures from the
various series were fixed in formalin at the end of two, three and
four days and stained with hematoxylin and Sudan III.

The results may be briefly summarized:

ABSORPTION OF WATER BY THE SKIN OF THE FROG. 125

In plasma diluted 1 : 2 the cells live practically as long (5 to 10
days without transfer), as in pure plasma, and show a similar
accumulation of fat.

In a1: 5 dilution the fat content is slightly diminished; there
is little or no effect on the length of life or on the morphology of
the cells.

In a 1: 10 dilution the fat content of the cells is definitely
reduced; the cells appear smaller and are stained more deeply,
and the duration of life is shortened (3 to 5 days).

In higher dilutions (1 : 15 and 1 : 20) the accumulation of fat
is reduced to a minimum, a majority of the cells showing at the
end of two days a complete absence of fat granules. The cells
which do contain fat show, as a rule, a single rather large droplet
instead of a number of small droplets, as in the controls in un-
diluted plasma. Two to three days represents, as a rule, the
limit of activity of the preparation. When stained the cells in
cultures of high dilution exhibit a rather striking contrast to those
in pure plasma cultures: they are smaller, more irregular in
shape, and take a deeper stain in both nucleus and cytoplasm.

In all experiments observations upon the living cells were con-
firmed by a study of stained and fixed preparations. Diluted prep-
arations and controls were, of course, fixed atthe same moment.

The results of these studies which show, in brief, that the
amount of fat accumulated by cells in cultures varies directly
with the fat content of the plasma medium, afford further evi-
dence in favor of the view that these fatty accumulations are not
degenerative in origin, but are the result of some disturbance in
the metabolism of the cells.

82 (778)
The rate of absorption of water by the skin of the frog, in relation
M. H. Fischer’s theory of edema.
By J. F. McCLenpon.

[From the Department of Anatomy, Cornell University Medical
College, New York City.]
Fischer observed swelling of amputated frog’s legs, in water.
The question arises: is this a phenomenon of osmosis due to the
osmotic pressure of cellular and intercellular fluids?

126 SCIENTIFIC PROCEEDINGS (53).

A frog’s leg was tied tightly and amputated above the ligature.
The second leg of the same frog was skinned and the skin filled
with a 0.7 per cent. NaCl solution and tied at the same level as the
first leg. The two legs were weighed, placed in water, and weighed
at intervals to determine the water absorbed. The leg filled with
NaCl solution absorbed water more rapidly than the other leg.

An amputated and ligatured leg was placed in 0.7 per cent.
NaCl. Its weight remained constant.

The ratio of the skin areas of a whole frog except the head,
to the hind legs below the knees, was found to be about 3.5. Two
frogs of the same size were selected. The hind legs of one were
tied just above the knees and amputated above the ligatures and
placed in water. The other frog was put in a harness that kept
the head out of water, and a canula with rubber bag attached
was inserted into the cloaca. This experiment was repeated a
large number of times. The water absorbed by the whole frog
within 6 hours was always more than 3.5 times as much as that
absorbed by the two hind legs. The water absorption for longer
periods of time is being studied.

Conclusion.—The swelling of frog’s legs, in which the circula-
tion of the blood is stopped, may be accounted for by osmotic
pressure.

83 (779)
The dynamics of a model of cell division.

By J. F. McCLenpon.

[From the Department of Anatomy, Cornell Unwwersity Medical
College, New York City.]}

A low beaker is half filled with distilled water and a funnel
inserted so that the stem extends to the bottom. A saturated
solution of NaCl is slowly poured into the funnel and forms a
layer beneath the pure water. About I c.c. of a mixture of 2
parts chloroform and 3 parts rancid olive oil is sucked up into a
pipette and injected into the beaker so that it forms a drop
suspended between the NaCl solution and the pure water. Two
pipettes with capillary openings are filled with 1/10 normal NaOH
solution and inserted into the beaker. The NaOH solution is

THE Dynamics oF A MopDEL oF CELL DIVISION. 127

allowed to flow onto opposite poles of the drop at the same time
and rate. The drop quickly elongates toward the pipettes, 7. e¢.,
toward the poles, and constricts along the equator, and some-
times divides into two. The smaller the drop, the more certain
the division, provided the operator has sufficient skill.

The alkali forms soap which reduces the surface tension on the
polar areas, and the hydrostatic pressure within the drop causes
these areas to bulge, whereas the relatively higher surface tension
of the equatorial region causes it to constrict until a barrel-shaped
figure is formed, which rapidly becomes hour-glass shaped. The
equatorial surface film contracts and the polar surface film spreads,
causing vortex movements. The enlargement of the polar fields
spreads the soap over larger areas, and the area of unaltered
surface tension is reduced to a narrow equatorial band. This
band, being partially released by reduction of tension at its edges,
acts as a sphincter and constricts until it cuts the oil drop into
two. This constriction of the oil drop may be considered as a
rough model of cell division.

T. B. Robertson in a recent paper! claims that exactly the
opposite changes take place in cell division. He divided the oil
dropp by lacing on ita linen thread 0.4 mm. in diameter, previously
soaked in the alkali. If the drop is not more than 1/10 c.c. in
volume the thread cuts it in two. This is due to gravitation of
the thread, the alkali merely lessening the resistance to the cutting.
I found that better results were obtained by adding a little alkali
to the water instead of soaking the thread in it.

The various points in Robertson’s argument cannot be con-
sidered here, and the reader is referred to his paper. The most
striking fallacy is that in Fig. 2, p. 699, M, and M, are not the same
distance below the curved line alsmd.

1 Arch. f. Entwicklungsmech., 1913, XXXV, p. 692.

128 SCIENTIFIC PROCEEDINGS (53).

84 (780)
Antitoxic action of sodium iodid on morphin.

By T. S. GITHENs and S. J. MELTZER.

[From the Department of Physiology and Pharmacology of the
Rockefeller Institute.]

A Preliminary Communication.

The relation of iodids to morphin was studied by Reid Hunt.
He found that feeding mice, rats and guinea-pigs with potassium
iodid increases their susceptibility to morphin poisoning. This
may be considered as the chronic influence of iodids. In our
experiments we studied their acute effects. Our experiments were
made, in the first place, on rabbits and sodium iodid in 5 per cent.
solution was the salt employed. Morphin was administered intra-
venously. The iodid solution was given intravenously, and sub-
cutaneously, ten to thirty minutes before the morphin injection.
The quantity of sodium iodid administered to each rabbit was
quite large; 15 cubic centimeters intravenously and 30 cubic
centimeters subcutaneously. Such injections of sodium iodid
alone seemed to cause no ill-effects in rabbits.

Morphin, if not rapidly fatal, causes narcosis, paresis, tremors,
convulsions and finally death. The fatal dose of morphin for
rabbits is somewhat variable, it is therefore difficult to study the
influence which other substances may exert upon the toxic action
of morphin. The most definite results we obtained have been
with doses of 300 and 250 milligrams of morphin per kilo body-
weight. Eighteen rabbits received morphin alone; 11 of these
animals received the drug (300 milligrams in each case) through
the ear vein, while in 7 animals the morphin (300 or 250 milli-
grams) was injected through the jugular vein. Sixteen rabbits
received sodium iodid besides morphin. In ten of these animals
the injection (300 milligrams) was given through the ear vein and
in six through the jugular vein. The difference in the results was
quite striking. Of the eighteen animals which received morphin
alone, ten died immediately after the injection, five lived less than
2 hours, one lived four days and two survived. Of the sixteen
rabbits which received sodium iodid besides morphin, only one

NATURE OF THE SEMI-PERMEABLE MEMBRANES. 129

died immediately after the injection, three lived longer than three
hours, 7. e., between 3 and 20 hours (died in night), one lived
eighteen, one twenty-three, and one thirty-six hours; one lived
two days, one nine and one 16 days, and six rabbits survived.
The meaning is quite unmistakable; the injection of sodium
iodid undoubtedly reduced the mortality or postponed death in a
palpable manner. We may add that the favorable effect of the
iodid seemed to be more manifest in white than in gray rabbits.

The experiments seem to demonstrate also that sodium iodid
antagonizes essentially the tetanic effects of morphin, while the
depression is perhaps even more manifest in the iodid animals.
However, we shall not discuss these particulars for the present.

We experimented also with mice. For mice we can only say
for the present that iodid seems to retard perceptibly the onset
of convulsions and the fatal outcome of morphin poisoning.

85 (781)

On the nature of the semi-permeable membranes which surround
the fibers of striated muscle.

By EDWARD B. MEIGs.

[From the Wistar Institute of Anatomy and Biology.|

The view that the fibers of striated muscle are surrounded by
semi-permeable membranes has received a wide acceptance among
physiologists, and there has been a good deal of speculation re-
garding the nature of these membranes. The hypothesis that
they are composed of lipoids has received much attention. Arti-
ficial lipoid membranes, however, have been found to be either
impermeable both to water and to dissolved substances or else
nearly equally permeable to water and to dissolved substances.
It is a general rule that artificial membranes composed of pure
colloids are either impermeable to both water and dissolved salts;
or else nearly equally permeable to water and salts, and imper-
meable only to colloids. The best known artificial membranes
which are semi-permeable with regard to salts dissolved in water
are composed of precipitates of insoluble salts such as copper
ferrocyanide and calcium phosphate.

130 SCIENTIFIC PROCEEDINGS (53).

The animal body can present the conditions necessary for the
precipitation of calcium phosphate, as, for instance, in the case of
bone formation. The striated muscle fibers contain considerable
amounts of dipotassium phosphate, and are surrounded by lymph
which contains calcium chloride, so that it is far from incon-
ceivable that thin layers of calcium phosphate might be precipi-
tated at the surfaces of the muscle fibers.

I have examined some of the properties of celloidin membranes
impregnated with calcium phosphate. Celloidin membranes free
from precipitate are quite permeable both to water and to dis-
solved salts. Such membranes were filled with a dipotassium
phosphate solution and immersed in a calcium chloride solution.
Under these circumstances they become impregnated with calcium
phosphate, and at the same time they become markedly semi-
permeable with regard to salts dissolved in water. That is to
say that if they separate salt solutions of different osmotic pres-
sures, water passes rapidly from the less concentrated to the more
concentrated solution.

Celloidin membranes impregnated with calcium phosphate
were filled with a 1.3 per cent. solution of dipotassium phosphate
and immersed in isotonic solutions of various substances: it was
then determined whether or not water passed through the mem-
brane from the outer solution to the inner one against a moderate
hydrostatic pressure. Such experiments were tried with solutions
of NaCl, KCI, CaCls, cane sugar, alanin (an amino acid), glycerine,
urea, andethylalcohol. The results indicated that the membranes
were highly impermeable to salt, sugar, and amino-acid, somewhat
permeable to glycerine and urea, and highly permeable to ethyl
alcohol. In all these respects they resemble the muscle membranes
with the possible exception of the case of KCl. There is some
reason to believe that the muscle membranes are more or less
permeable to KCl.

The supposition that calcium phosphate plays a part in giving
to the muscle membranes their semi-permeable properties would
explain two great classes of facts—those facts, namely, which
show that muscles rapidly lose their semi-permeable properties
in acid solutions; and those which show that calcium may play
an important part in maintaining the semi-permeable properties

FAcTorsS OF EXCESSIVE ARTIFICIAL RESPIRATION. 131

of muscle. Precipitates of calcium phosphate are rapidly dis-
solved by acids, and would dissolve slowly in neutral calcium-
free solutions. In view of these considerations I propose to
devote some time to the study of the osmotic properties of calcium
phosphate precipitates, as well as to those of precipitates of
calcium carbonate and of magnesium phosphate and magnesium
carbonate.

86 (782)

The mechanical factors of excessive artificial respiration and a
consideration of their relation to the acapnial theory of shock.

By H. H. JANEway and E. M. EwINe.

[From the Department of Physiology, Unweristy and Bellevue
Hospital Medical College.|

It has been claimed that the most important factor in the
causation of shock is diminution of CO, within the blood, and
that this diminution is a regular consequence of all influences
resulting in shock. That CO, possesses important physiological
functions cannot be denied. An investigation therefore of the
true significance of a diminution of its normal amount within the
blood is important and bears a special relation to various methods
of artificial respiration utilized in thoracic surgery. The present
experiments were undertaken for the purpose of investigating the
effects of acapnia and the relation of some factors concerned in its
production to shock. In all of them dogs were used. The first
series was performed for the purpose of studying the effect
of variations in intrapulmonic air pressure upon the blood
pressure. The thorax was opened laterally, a T-tube connected
with a water manometer was tied in a small bronchus, and the
heart enclosed in a Henderson cardiometer in series with a record-
ing tambour. The blood pressure was recorded from the
carotid artery. The thorax was closed and the animal was
subjected to intratracheal insufflation from an apparatus pro-
vided with an exhaust valve which reduced the pressure to
approximately zero about four (4) to twelve (12) times per
minute. The blood pressure averaged 150 mm. Hg. when the
intrapulmonic air pressure was not allowed to exceed 6 mm. Hg.

132 SCIENTIFIC PROCEEDINGS (53).

In one experiment, with an increase of intrabronchial pressure
from 8 mm. to 30 mm. Hg, blood pressure fell from 122 mm. to
55 mm. Hg and the volumetric tracing indicated that the output
from the heart had diminished about 44 per cent. These varia-
tions in blood pressure were completed within a few seconds
after the change in intrabronchial pressure, and could be dupli-
cated at will.

A rise of intrabronchial pressure above 8 to 10 mm. Hg always
caused a fall in blood pressure, and it was concluded that the varia-
tion in pressure was the result of a diminished venous return to
the heart, resulting from compression of both the pulmonary and
systemic veins in the thorax.

In view of the marked changes in heart output and blood
pressure resulting from small variations in intrapulmonic pressure,
it is evident that, in any experiments attempting to estimate the
part played in the production of shock by a diminution of CO,
which is caused by hyperartificial respiration, the effect of the
increase of intrapulmonic pressure upon the return flow of blood
to the heart must be considered.

In the second series of animals, Henderson’s experiments were
duplicated, the dogs being artificially respired by means of a force
and suction pump, working about 70 times per minute. The
animals were given morphine, and ether was administered only
when necessary. In these experiments, blood pressure fell about
40 per cent. within one minute after artificial respiration was begun,
and then decreased more slowly throughout the experiment to
between 40 and 50 mm. Hg. At the end of the experiment when
the artificial respiration was stopped, blood pressure increased 60
to 90 per cent. within a few seconds. In all experiments the blood
gas analysis showed the CO, content at the end to be only 40 to
50 per cent. of the original volume. These animals, at the end of
2 to 3 hours of artificial respiration, were all in a condition of deep
shock. This degree of shock was indicated by a rapid pulse, a
marked degree of coma, and insensibility to a sensory stimulation.
Three of the animals so treated lived three days (dying of sec-
ondary effects of the experiment), and one lived 24 hours. None
of them died from the immediate effects of the experiment.
During these experiments when the artificial respiration was

Factors OF EXCESSIVE ARTIFICIAL RESPIRATION. 133

temporarily stopped or permanently stopped at the end of the
experiment the period of apnea lasted only one or two minutes,
so that no death resulted directly from acapnia. The absence
of a prolonged period of apnea is explained by the fact that the
effect of ether was not added to that of morphine.

With a third series of animals, the experiments just described
were duplicated with the exception that the CO, content of the
blood was maintained at its normal level, or slightly above it.
The conservation of CO. was accomplished by inserting a large
rubber bag, to act as a reservoir, between the suction pump and
the force pump, thus creating an almost perfectly closed circuit;
the dog thus rebreathed expired air. To replace a small amount
of air lost from the animal’s trachea, CO, was fed from a tank into
the rubber bag where the latter was diluted with air drawn in from
the trachea in smaller quantities than that amount lost through
this route. In these experiments the animals went into the same
degree of shock in 2 or 3 hours as those of Series II in which the CO,
content of the blood was diminished to 40 per cent. of the original
volume. One animal died on the table just before the experiment
closed, the others lived one to three days. Blood pressure changes
in the two series were similar, but a characteristic of the experiment
when the CO, was kept high, was the slow, strong heart beat, in
place of the rapid, weak pulse observed when the CO, was di-
minished. Conclusions drawn from Series II and III can only
be that the reduction in the CO, content of the blood is not the
important factor in the production of shock by hyper-respiration,
but that in the shock so produced, the essential factor is an in-
terference with the venous return to the heart.

In the fourth series of experiments the effects of aeration and
handling of the intestines were studied. A celluloid window was
placed in the abdominal wall, and a stream of warm, moistened
air was passed over the intestines for a period of three hours.
During this procedure the animals breathed normally, the blood
pressure was 163 mm. Hg, CO, content was normal, and there was
no evidence of shock. The celluloid was then removed, the
intestines spread out, and the aeration continued. After 45
minutes the CO, determination showed 38.8 volume per cent.,
and blood pressure was 153 mm. Hg. The intestines were then

134 SCIENTIFIC PROCEEDINGS (53).

handled, and in ten minutes blood pressure had fallen to 98 mm.,
and in 20 minutes to 56 mm. Hg. In 40 minutes there was still
31.6 vol. per cent. CO» in the arterial blood.

In another experiment the intestines were exposed and aerated
(not handled). The CO: content of the blood was maintained by
connecting a long tube to the trachea. After one hour and a half,
blood pressure had changed but one mm. Hg and the animal was
in good condition. The intestines were then handled, and in ten
minutes the blood pressure fell from 122 to60 mm. Hg. The CO,
content was 45.I per cent. In 25 minutes the blood pressure was
46 mm. Hg, the CO, content normal, and the dog in shock to such
a degree that the sciatic nerve could be dissected out without the
administration of ether, the animal making no movement what-
ever. In these experiments on the abdominal cavity the primary
factor concerned is unquestionably the manipulation of the intes-
tines, and not any diminution of CO, caused thereby. It will
be remembered that in the similar experiments of aeration of the
intestines, reported by Henderson, the intestines were ‘‘ handled
gently.’’ We have been unable to find that he notes any mention
of aerating the abdominal cavity beneath a celluloid membrane
with air as a control experiment.

His control experiment, in which he did not secure shock, was
merely aerating the abdominal cavity beneath a celluloid window
placed in the abdominal wall with a stream of air plus CO».

The present experiment shows that aeration of the intestines
without the addition of CO, does not produce shock.

87 (783)
A note on the transmission of spiroch etes.
By Joun L. Topp.

Leishman and other authors have shown that the anal and
coxal excretions voided by ticks (Ornithodoros moubata), capable of
transmitting spirochetes (Spirocheta duttoni), are infective and
that susceptible animals inoculated with these fluids develop a
spirochetal infection. It has been asserted that animals upon
which infected ticks have fed will not become infected, unless

EXPERIMENTALLY FusED LARV&. 135

these fluids are voided while the ticks feed, and it has been sug-
gested that the infection is transmitted by the flowing of the fluids
into the wounds made by the mouth-parts of the ticks in feeding.
Spirochzetes had not been seen in these fluids and it was suggested
that they existed there in a coccoid form.

On several occasions, coxal and anal fluids, excreted by infected
ticks, coming from Uganda and British Central Africa, have been
examined. In every instance the fluid was taken while the ticks
fed upon an uninfected animal. The fluid collected was free
from blood and, in two instances, coxal fluid was collected ap-
parently free from anal excretion. On six occasions, after the
fluid had been centrifugalized, spirochetes were found in it;
their morphology is not distinct from that of Spirocheta duttont.
Spirochetes were found in the fluid that apparently contained no
anal excretion.

88 (784)
Experimentally fused larve.

By A. J. GOLDFARB.

[From the Department of Natural History, The College of the City
of New York.|

When the eggs of Toxopneustes variegatus were subjected to a
5/8 molecular NaCl, after the removal of the fertilization mem-
brane, considerable numbers were subsequently fused together.
I have counted as many as forty percentum, in the optimum
solutions, of agglutinated and fused pairs, triplets, etc. Few of
these reached the pluteus stage of development due to the early
death of all fusions of more than three eggs, and to the large
mortality of even the double embryos.

The plutei contain at least three characteristic tissues, namely,
body wall,archenteron, skeleton. The first two of these behaved
essentially as described by Driesch in various European species,
and by the writer in the American species Arbacia punctulata;
1. é., the body walls or the archentera of plutei derived from
separate eggs were united either incompletely or so completely
as to give little or no evidence of the original dual character of the
larve.

136 SCIENTIFIC PROCEEDINGS (53).

It was supposed that the skeletal structures united in the same
manner as the archentera. The evidence furnished by Toxo-
pneustes however clearly shows that in this species at least no
fusion of the skeletal parts occurred. Instead some very interest-
ing changes took place which may be stated briefly as follows:
One of the pair of fused larve developed normally in every detail,
the other developed in nearly every instance, incompletely. An
almost perfect series of fused larvee were obtained in which the
incomplete pluteus lacked more and more of the characteristic
parts that constitute the perfect larval skeleton; and the order
of their disappearance was in the reverse order of their appear-
ance in ontogeny.

The union of the two larve involved the approximation of their
branched and complex skeletons, whose parts frequently over-
lapped but never fused.

89 (785)
Metabolism studies in a case of myotonia atrophica.

By JAcoB ROSENBLOOM and BENSON A. COHOE.

[From the St. Francis Hospital, and the Laboratory of Biochemistry
of the Unwersity of Pittsburgh, Pittsburgh, Pa.]

In a thirteen day metabolism study on an individual suffering
from myotonia atrophica, we have studied the nitrogen metab-
olism, and urinary nitrogen partition, the sulphur metabolism
and urinary sulphur partition, and the calcium, magnesium,
phosphorous, chlorine and fat metabolism. The creatinine excre-
tion was normal. The only striking metabolic anomaly noted in
this study was the marked loss of calcium.

90 (786)
Sugar from lactic acid in human diabetes,

By NELus B. Foster, M.D.

[From the Medical Service of The New York Hospfital.]
The evidence presented by experimental diabetes seems to
indicate that in the transformation of amino-acids into glucose
lactic acid is an intermediary step. It has also been suggested

VACCINIA BLOooD PRESSOR SUBSTANCES IN RABBITS. 137

that in the combustion of glucose lactic acid may arise from the
cleavage of the sugar molecule. On account of this significant
place held by lactic acid it appears of interest to investigate the
relation it may hold to sugar production in human diabetes.
Only cases of diabetes of considerable severity are suitable for
the experiment, since in the milder degrees of the human disorder
relatively large amounts of carbohydrate may be burned or stored
and the sugar output is apt to fluctuate. In a patient it was
ascertained that with a uniform diet for three days a fairly constant
glucose excretion could be depended upon and lactic acid as the
sodium salt was given on the second day of the experimental
period with the result that there was a rise in sugar excretion
shown by the following abstract from the protocol.

Amount Sugar, Total N NH.N. |Acetone and} B-oxy

urine. grams. : ao diacetic. butyric.
Vegetable diet.... 1,810 2.89 3.62 stasiy/ Aris’ 0.79
Oatmeal diet..... 1,450 2.80 4.80 1.22 +59 1.41
Meat diets... cas. 2,300 6.56 11.46 1.42 -76 ater/7/
15 gm. Na lactate. 2,010 19.90 14.06 1.56 .82 1.49
2,050 2.91 14.92 0.71 .86 1.97

gt (787)

On some vaccinia blood pressor substances in rabbits.
By J. P. ATKINSON and Cuas, B. Fitzpatrick, M.D.

[From the Chemical and Research Laboratories, Department of
Health, City of New York.|

These notes constitute a continuation of our observations on
some blood pressor substances in experimental immunity, reported
at the last meeting of this society, and apparently tend to confirm
the same. We have endeavored to ascertain when the pressor
and depressor substances appeared in the blood serum of living
rabbits after they had been successfully infected with vaccinia
virus. The height of the production of the vesicles from which
the virus was collected was reached on the fifth day. They then
rapidly healed and the rabbits appeared outwardly to be in good
health.

Observations were also made to ascertain the effect of aging
on the presence of the depressor substance.

SCIENTIFIC PROCEEDINGS (53).

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139

VACCINIA BLooD PRESSOR SUBSTANCES IN RABBITS.

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140 SCIENTIFIC PROCEEDINGS (53).

Summary.—t. Depressor and pressor substances arise after
vaccinia infection in the blood-serum of rabbits. 2. Aging tends
to eliminate the depressor substance and a pressor substance then
comes in evidence.

Note.—A serum obtained from a rabbit after streptococcus infec-
tion, which had 12 months previously given a profound depressor
reaction was also tested (3-18-13) and was found to give no reac-
tion. 8 c.c. of a saline extraction of the adrenals of a 30 day
vaccinia rabbit, gave (3-28-13) no reaction. This extraction was
made in 20 c.c. of physiological saline solution and was kept 6
days in the ice-box. 7 c.c. of a saline extraction of the adrenals
of a two day vaccinia rabbit gave (3-8-13) a fine rise followed by
a marked fall. This extraction was made in 20 c.c. and was kept
24 hrs. in the ice-box, 7. e., since immediately after removal.

ABSTRACTS OF THE COMMUNICATIONS, PaciFIc CoAstT BRANCH.
Third meeting.
San Francisco, California, April 2, 1913.

92 (788)
Preliminary communication on the part played by cholesterol in
determining the incidence of carcinoma.

By T. BRAILSFORD ROBERTSON and THEODORE C, BURNETT.

[From the Rudolph Spreckels Physiological Laboratory of the Uni-
versity of California.]

We have elsewhere shown! that cholesterol, when injected
directly into rat carcinomas, causes a marked acceleration both
of the primary and of the metastatic growth of the tumors.

This led us to form the opinion that cholesterol is probably a
factor of importance in determining the incidence of carcinoma.

It has been shown by Dorée and Gardner, Ellis and Gardner,
and others? that cholesterol is not synthesized by animals, the

1T. Brailsford Robertson and Theodore C. Burnett, Proc. Soc. ExpEer. BIOL,
AND MED., I0 (1912), p. 59; Journal of Exper. Med., 17 (1913), p. 344.

2C. Dorée and J. A. Gardner, Proc. Roy. Soc. London, 80 B (1908), p. 227; 81 B
(1909), G. W. Ellis and J. A. Gardner, ibid., 81 B (1909), p. 129.

DETERMINING THE INCIDENCE OF CARCINOMA. 141

cholesterol in animal tissues being derived from their diet. This
fact suggested the possibility that the incidence of carcinoma in
inoculated animals might be diminished by feeding them for a
considerable period prior to the inoculation upon a diet poor in
cholesterol.

Accordingly twenty-two white rats, about two months old,
were divided without exercising any selection into two lots, One
lot of 15 were fed upon a diet composed exclusively of milk; the
remainder were fed upon a mixed diet of oats and meat.

The content of cholesterol in milk, while by no means negligible,
is extremely small in comparision with the content of cholesterol
in other foodstuffs. Thus Tolmatscheff finds that human milk
contains from 0.025 per cent. to 0.039 per cent. of cholesterol,}
while Bémer and Kirsten find that the fats in cow’s milk contain
0.5 per cent. cholesterol, corresponding to a content of less than
0.02 per cent. in the whole milk.2 Meat, on the other hand, con-
tains from 0.07 to 0.08 per cent. of cholesterol,? while the content
of phytosterols in seeds is considerable.*

Both lots of animals thrived well, the milk-fed animals present-
ing an especially well-nourished appearance.

At the end of two months both lots of rats were inoculated in
the axillary region with portions of a Flexner-Jobling carcinoma.
The diet of each lot of rats was maintained unaltered. At the
end of twenty days the proportion of successful inoculations in
each batch of animals was determined, with the following results:

Milk diet: 10 out of I5 = 67 per cent.
Mixed diet: 7 out of 7 = 100 per cent,

Another batch of half-grown animals obtained at the same time
from the same dealer, which were fed upon a mixed diet and
inoculated with Flexner-Jobling carcinoma yielded the following
results:

Mixed diet: 55 out of 64 = 86 per cent.

While the milk-fed animals yielded the lowest percentage of

successful inoculations it is evident that the difference between

1 Tolmatscheff, Hoppe-Seyler’s Med.-Chem. Unters., 2 (1867), p. 272.

2R. W. Raudnitz, Ergeb. d. Physiol., 2, I (1903), p. 264.

3 C. Dormeyer, Arch. f. d. ges. Physiol., 65 (1896), p. 99; G. W. Ellis and J. A.
Gardner, Proc. Roy. Soc. London, 81 B (1909), p. 507.

4F. Czapek, ‘‘ Biochemie der Pflanzen,”’ Jena, 1905, Vol. I.

142 SCIENTIFIC PROCEEDINGS (53).

the milk-fed and the normal animals in this respect was not so
marked as one would be inclined to anticipate if cholesterol were
really a prime factor in determining the incidence of carcinoma,
and if feeding for two months upon a diet unusually low in choles-
terol really brought about any appreciable diminution in the choles-
terol-content of the tissues.

That cholesterol is an important factor in determining the inci-
dence of carcinoma can hardly be doubted in view of our previous
results, cited above, and of the fact, recently discovered by
Wacker, that the cholesterol content of the fatty deposits in the
subcutaneous tissues and mesenteries of persons who have car-
cinoma is no less than 66 per cent. greater than the cholesterol
content of the fatty deposits in normal persons.!_ An increase in
the cholesterol content of reserve-fats was also observed by Wacker
in aged persons and in persons afflicted with tuberculosis or
diabetes.

Our failure to observe a more striking difference between the
incidence of carcinoma in milk-fed and in normal rats we are
inclined to attribute to the fact which has been demonstrated by
Ellis and Gardner? that cholesterol is strictly conserved in the
animal economy, since the cholesterol which is excreted in the bile
is reabsorbed in the intestine and does not appear in the feces,
while, on the other hand, cholesterol contained in the food is in
large part absorbed. Hence removing cholesterol from the diet
of an animal does not lead to an appreciable removal of cholesterol
from the tissues. More conclusive results might be anticipated
if animals were fed from the time of birth upon a cholesterol-free
diet, since the tissues of newly-born animals contain relatively
little cholesterol. Experiments in this direction are being under-
taken.

In conclusion we wish to point out that since the cholesterol
in the diet is partly absorbed, while the cholesterol content of the
body is strictly conserved, it follows that animals must tend to
accumulate cholesterol. We believe that this explains the exces-
sive cholesterol content of the reserve fats in aged persons which

1L. Wacker, Zeit. f. physiol. Chem , 80 (1912), p. 383-

2G. W. Ellis and J. A. Gardner, Proc. Roy. Soc. London, 81 B (1909), p. 505.
3G. W.Ellis and J. A. Gardner, Proc. Roy. Soc. London, 84 B (1912), p. 461.

INFLUENCE OF DIGITONIN UPON GROWTH OF CARCINOMA. 143

has been observed by Wacker, and that this in its turn is the reason
for the well-known increase in the incidence of carcinoma with
advancing age.

93 (789)
The influence of digitonin upon the growth of carcinoma.

By T. BRAILSFORD ROBERTSON and THEODORE C. BURNETT.

[From the Rudolph Spreckels Physiological Laboratory of the
University of California.]

It has been shown by Windaus! that digitonin combines with
cholesterol to form a very insoluble and pharmacologically in-
active compound. In view of the marked influence of cholesterol
in accelerating the growth of carcinoma we have thought it of
importance to ascertain the influence of digitonin upon the growth
of Flexner-Jobling carcinoma in rats.

The digitonin employed was Merck’s, stated to have no physio-
logical action upon the heart. By heating the preparation to
boiling in m/6 NaCl solution a soapy-looking fine suspension is
formed which settles out in the course of several hours. We
injected the digitonin, suspended either in m/6 NaCl, or in m/6
NaCl containing 1 per cent. of lecithin, directly into the tumors.

One hundred and sixty-six white rats were inoculated with
Flexner-Jobling carcinoma in the axillary region. The number of
successful inoculations, determined after 20 days, was 64, or 39
per cent.

On the 20th day after inoculation these animals were sorted,
without selection, into three batches, of which one (consisting of
12 animals) served as controls, another (12 animals) received
injections of digitonin, and the third (40 animals) received injec-
tions of digitonin together with lecithin.

We began by administering I c.c. of a I per cent. suspension of
digitonin, suspended in m/6 NaCl and in m/6 NaCl + 1 per cent.
lecithin respectively. The animals which received digitonin with-
out lecithin evinced symptoms of severe local irritation, and one

1A. Windaus, Ber. d. d. Chem. Ges., 42, I (1909), p. 238; Zeit. f. physiol. Chem.,
65 (1910), p. 110; M. T. Fraser and J. A. Gardner, Proc. Roy. Soc. London, 82 B
(1910), p. 559.

144 SCIENTIFIC PROCEEDINGS (53).

of the animals which received digitonin alone and two of those
which received digitonin and lecithin died within a few hours
after the treatment. Post-mortems showed that the heart had in
each case stopped in extreme systole, the auricles being engorged.
In subsequent treatments the dose of digitonin was reduced to
I c.c. of a 0.5 per cent. suspension, and in the case of the animals
which received lecithin, the concentration of the lecithin emulsion
was increased to 2 per cent. In the course of these treatments
two more of the animals which received digitonin and lecithin
died, in each case post-mortem examination showed that the
heart had stopped in extreme systole.

Treatments were given on the 20th, 22d, 25th, 27th, and 29th
days after the inoculation.

It was found that twenty-four hours after the first treatment
the tumors were soft and pulpy and also presented a discolored
appearance. The tumors rapidly hardened again, however, and
those which were treated with digitonin alone grew much more
rapidly than the controls or the tumors which were treated with
digitonin and lecithin together. This is shown by the following
figures, the diameter of the tumors at the beginning of treatment
being taken as 100 in each case.

Diameter of the tumors.

20 days after inoculation. 25 days after inoculation.
Controls: oc spas ce se oe 100 II4
Digitonin. . 3.6 ee ee eee 100 146
Digitonin + lecithin............ 100 II3

It is evident that lecithin antagonizes the action of digitonin
in accelerating the growth of carcinoma. This we have confirmed
by further experiments which will be reported in a later com-
munication.

Thirty per cent. of the controls, thirty per cent. of the animals
which had received digitonin alone, and twenty-five per cent. of
the animals which received digitonin together with lecithin had
undergone ‘‘spontaneous cure”’ 32 days after inoculation.

In order to ascertain whether or not the above effects were
due to digitonin itself or to some impurity contained in Merck’s
preparation, we prepared pure digitonin by dissolving Merck’s
digitonin in alcohol, pouring this solution into a large volume of

IEFLUENCE OF DIGITONIN UPON GROWTH OF CARCINOMA. 145

water, collecting the precipitate, washing it in water and drying
over H:SO, at 36° C. The substance which was thus obtained
was pure white. It is soluble in alcohol and forms unstable sus-
pensions in water. One c.c. of a I per cent. suspension injected
subcutaneously in rats produced no symptoms whatever, either
local or general. A solution in alcohol precipitated cholesterol
from alcoholic solution, and a suspension in water coagulated a
suspension of cholesterol in N/100 sodium oleate.

Fourteen rats with carcinoma, which had been inoculated 20
days previously, were divided without selection into two lots,
the one lot of 5 served as controls, the other of 9 received I c.c.
of a I per cent. suspension of the above preparation of digitonin
in m/6 NaCl, injected directly into the tumors on the 2oth, 21st,
22d, 23d, 25th and 26th days after inoculation. Asin the previous
experiment, the earlier injections caused softening and discolora-
tion of the tumors. The tumors soon hardened, however, and
thereafter grew much more rapidly than the tumors in the controls,
as the following figures reveal:

20 days after 25 days after 29 days after

inoculation inoculation. inoculation,
GControlsier races aeons 100 I10 126
Digitonink mc acsoe se een 100 I47 168

We interpret these results in the following way: When digitonin
is injected into the tumor it first of all renders inactive the cho-
lesterol then within the tumor and this results in softening and
incipient degeneration of the tumor. The precipitated cholesterol
is, however, replaced by cholesterol from the surrounding tissues
and body-fluids, and the cholesterol bound by the digitonin is
retained within the tumor and gradually liberated from its com-
bination with the digitonin, with the result that cholesterol is
concentrated within the tumor, and the growth of the tumor is
thereby accelerated. The fact that lecithin prevents the accelera-
tion of the growth of the tumor by digitonin is in harmony with
the above interpretation and with our previous results showing
that lecithin and cholesterol have opposite actions upon the
growth of carcinoma.!

1T. Brailsford Robertson and Theodore C. Burnett, Proc. Soc. Exper. BIOL.
AND MED., I0 (1912), p. 59; Journal of Exper. Med., 17 (1913), p. 344.

146 SCIENTIFIC PROCEEDINGS (53).

94 (790)
Note on the relation of alexin to thrombin.

By THEO. C. BurNeETT (by invitation).

[From the Rudolph Spreckels Physiological Laboratory of the
University of California.]

The publishing of negative results is, as a rule, a questionable
proceeding. Occasionally however, a negative result may have
a positive bearing upon a subject, of more or less value, in which
case the objection does not hold. It is for this reason I present
the following note.

In reviewing the subject of coagulation some time ago, I was
struck by a certain similarity between thrombin and alexin.
Both are inactivated by a temperature of 56 degrees; both are
derived from leucocytes;! thrombin consists of thrombokinase and
calcium, while alexin consists of two parts, as is well known. It
occurred to me that there might be a closer relation between the
two; that they might in fact, be identical. It has probably
occurred to many another, but in a hasty glance over the literature,
I can find no mention of it. At first sight the idea seems absurd,
for alexin is contained in serum that has been collected over a
clot, and hence contains no thrombin. The alexic potency of a
serum, however, increases by standing for some hours in contact
with the clot, and it is conceivable that during that time thrombin
is excreted by the leucocytes, but is not apparent because of the
absence of fibrinogen wherewith to combine. Having occasion to
prepare some thrombin for another purpose, I determined to test
the matter.

The immune serum used was rabbit serum immunized against
ox corpuscles. Tested with guinea-pig serum it had a potency
of 1-800. (Ox blood was used on account of the ease with which
material could be obtained at any time.) Most of the experiments
were carried out with a 1-100 dilution, although similar results
were obtained with higher dilutions.

The thrombin was prepared by Howell’s method? and gave a

1 It is generally conceded now that thrombin is derived from the platelets.
2? Howell, W. H., Am. Jour. Physiol., Vol. 26, 1910, p. 453.

UNION OF ALEXIN WITH SPECIFIC PRECIPITATES. 147

coagulum with fibrinogen in a few minutes. As thrombin is made
with 8 per cent. NaCl solution, it was diluted to approximate
isotonicity, m/6. In order to still further control any possible
osmotic effects from hypotonicity, solutions of m/5 and m/4 con-
centrations were also used to dilute the serum.

The system consisted of I c.c. of I-100 immune serum, plus
0.I c.c. washed ox corpuscles (5 per cent. emulsion), plus thrombin
in amounts varying from 0.05 c.c. to 0.3. c.c. As a control one
tube was always prepared with normal serum (alexin) instead of
thrombin. The tubes were kept at a temperature of 36-37° C.

The results can be stated in a few words. The control tubes
showed complete hemolysis in from fifteen to thirty minutes. In
no instance was there a trace of hemolysis even after several
hours, in the tubes containing thrombin. The corpuscles settled
out of the solution and left a clear supernatant fluid, not even
tinged with hemoglobin. Having obtained this negative result
with thrombin, I then tried solutions of fibrinogen and of serum-
globulin, in order to make the matter complete. Exactly the
same results were obtained, and I think we may conclude that
whatever alexin may be, it is certainly mot identical with thrombin
or the globulins.

95 (791)

On the nature of the union of alexin with specific precipitates.
By HANs ZINSSER, M.D.

[Division of Bacteriology, Medical Department, Stanford University.]

In continuing studies on the nature of alexin fixation by
mixtures of unformed proteins and their antisera, it occurred to
the writer to examine whether the alexin fixation which is exerted
by specific precipitates was subject to the same conditions that
prevail in the case of sensitized cell complexes in their relations
to the alexin fractions as first obtained by Ferrata. It is well-
known, of course, that by dialysis, by dilution with weak acid in
distilled water and by a number of other methods of globulin
precipitation, the alexin or complement can be divided into two
functional parts, one which comes down with the globulins, the

148 SCIENTIFIC PROCEEDINGS (53).

so-called “midpiece’’ and the other which remains in the albumin
fraction, the so-called ‘‘endpiece.’”’ Neither of these can produce
hemolysis of sensitized cells alone. Together they functionate.
The globulin fraction can be bound to sensitized cells, forming the
so-called ‘‘persensitized cells’? which are now hemolyzable by the
endpiece alone. The albumen fraction does not become attached
or fixed to the sensitized cells except in the presence of the globulin
fraction. (The terms midpiece and endpiece are used for the sake
of clearness since they are terms which have become etablished in
the German literature. Owing to studies which are being made
by Mr. Maltaner in this laboratory we feel that a definite nomen-
clature which assumes an intermediate function of the globulin,
fraction, is premature.)

In experiments in which alexin fractions, produced by both
the method of Ferrata and by that of Sachs and Altmann, were
exposed to union with precipitates, formed in mixtures of beef
serum and its antiserum, we have found that the conditions which
prevail are entirely analogous to those which govern the attach-
ment of the alexin fractions to the sensitized cells. A specific
precipitate may fix the globulin fraction (midpiece) alone. It
may fix the albumin fraction (endpiece) in the presence of the
globulin fraction. It does not however, fix the albumin fraction
(endpiece) by itself. The experiments were in every case con-
trolled by titrations of the alexin fractions and the whole alexin
in tubes set up parallel with the main experiment.

The writer believes that these results have considerable
theoretical importance in bearing out his previously expressed
view that the “‘precipitin’’ may be regarded as a protein sensitizer,
the fact of visible precipitation being a merely secondary occur-
rence due to the union of two colloids under conditions of quantita-
tive relations and environment which favor precipitation.

ACTION OF GENTIAN VIOLET ON MUuUCOSUS CAPSULATUS. 149

96 (792)
Action of gentian violet on Mucosus capsulatus group.

By J. G. FITZERGERALD and GERTRUDE MACKINTOSH.

[From the Hearst Laboratory of Pathology and Bacteriology,
University of California.]

It has recently been shown by Churchman! in two interesting
communications bearing on the subject that the aniline dye,
gentian violet, has a selective bactericidal action on certain
bacteria. The action of the dye has been spoken of as bacterio-
static, indicating that the growth of some species of bacteria is
inhibited. In addition to this inhibiting influence, Churchman
believes the substance has also a very definite bactericidal action.
It was shown in the publications referred to that the action of
gentian violet as a bacteriostatic or bactericidal agent presented
in a general way a parallelism with the Gram stain. The majority
of gram positive bacteria are inhibited by gentian violet, while
the majority of gram negative bacteria are not. The action of
gentian violet can be observed on the divided plates or by staining
with gentian violet and determining subsequently whether the
microérganisms so treated will grow on culture media. On divided
plates, one half of an ordinary petri dish is covered with plain
nutrient agar, the other half with nutrient agar to which has been
added an aqueous solution of gentian violet; such a plate when
streaked with gram positive bacteria will show a growth only on
the side of the plate where there is no gentian violet, while gram
negative bacteria usually grow equally well on the plain agar and
on gentian violet agar.

Since it seemed possible that the differentiation of closely
related species might be accomplished by the use of divided
gentian violet plates, the method has been used in a further study
of the Mucosus capsulatus group. In all, thirty-six strains of
bacteria have been investigated; they include B. ozene, B. lactis
aerogenes, B. enteriditis, B. pneumonie Friedlander, B. rhino-
scleromatis, B. capsulatus, and B. Mucosus capsulatus. All of the

1 Churchman, Journal Experimental Medicine, Vol. XVI, No. 2, August, 1912,
pp. 221-247, and Vol. XVI, No. 6, December, 1912, pp. 822-830.

150 SCIENTIFIC PROCEEDINGS (53).

thirty-six were found to be gram negative. This is in harmony
with most of the observations heretofore recorded, although a
contrary statement is made with reference to B. rhinoscleromatis
by McFarland! and by Page.2 When studied by means of the
gentian violet agar plates, twenty-nine of the thirty-six cultures
were found to grow equally well on plain agar and on gentian
violet agar. Seven others, however, behaved differently. No
growth could be obtained on the gentian violet agar side of the
divided plates. Churchman has mentioned the fact that certain
bacteria when stained will not grow on one culture medium but
might grow on a more selective medium. The seven strains that
refused to grow on ordinary gentian violet agar were then planted
on serum agar violet, and three of them grew on serum agar violet,
although they had refused to grow on ordinary violet agar. The
other four did not grow even on the more favorable medium and
behaved in the same way as most of the gram positive bacteria.
Thinking the substitution of a selective culture medium for
ordinary medium in the case of certain of the gram positive bacteria
might result in a change in the action of gentian violet on their
growth, B. diphtherie and streptococcus were planted on serum
agar violet plates, and on agar violet plates. Both plates failed
to show a growth on the violet side.

Of the three microdrganisms that grew on serum agar violet,
two have remained constant, growing well on serum agar violet
but not at all on ordinary agar violet; the third one, however,
has shown a very interesting adaptation. The culture is one of
B. pneumonie Friedlander from the Kral collection. This bacillus
when grown on a plate containing serum violet and agar violet
can be induced to grow on agar violet once it has started to grow
on serum violet; if, however, the growth is started from the
opposite side of the plate on plain agar, it cannot be made to grow
on violet agar. The experiment was made by putting in one
plate culture, serum agar, serum agar violet, agar violet, and plain
agar, the different media merging into one another in a petri dish.
The behavior of the microérganism is thus shown to be inconstant
toward gentian violet, depending on the culture media on which

1 McFarland, ‘‘Pathogenic Bacteria and rotozoa,”’ 7th edition, 1912, p. 785.
2 Page, Journal Medical Research, Vol. XXVI, No. 3, July, 1912, pp. 489-499.

ELECTRIC POTENTIAL OF BACTERIAL SUSPENSIONS. I5I1

it is grown, that is, a microdérganism incapable of growing in the
presence of gentian violet readily does so on a more suitable
medium, and it can adapt itself to a less suitable medium if
started on a selective medium. Gentian violet has not been
found to be of any value in the differentiation of the Mucosus
capsulatus group. Further, the microérganisms of this group
which refuse to grow on agar violet are representatives of what
we at present regard as three distinct species.

It would seem from the result of the experiment here recorded
that while the action of gentian violet on bacteria is usually
constant, it is sometimes susceptible of modification and may not
be as fundamental a characteristic as originally supposed.

97 (793)

On the influence of light on the electric potential of bacterial and
other suspensions.

By S. W. Younce (by invitation).

[Laboratory of Physical Chemistry, Stanford University, Cal.]

Experiments in this laboratory have shown that light and other
forms of radiant energy exert a marked influence on the rate of
wandering in the electric current. Thus the rate of wandering
of arsenic sulphide suspensions is about twice as great in the dark
as under strong illumination in the sun, or in the carbon arc or in
the Nernst lamp. On the other hand, the rate of wandering of
mastic emulsions is increased under strong illumination, and that
to the extent of about forty per cent. of the value in the dark.
Ferric hydroxide is retarded in the light to the extent of about six
per cent., while chlorophyll suspensions are accelerated in the
light to the extent of about forty per cent. The effect of the
radiant energy is not in general instantaneous, but requires a
few minutes exposure to reach its greatest value. In general also
the effect is reversible, that is colloids whose rates of wandering
have been influenced by radiant energy, return to their original
values if kept in the dark for some minutes.

These phenomena are very interesting in connection with
photochemical reactions in general. Arsenic sulphide suspensions

152 SCIENTIFIC PROCEEDINGS (53).

may be kept almost indefinitely in the dark. On exposure to light
they oxidize rapidly. Knowing now that light also reduces the
negative charge on the colloidal particles of arsenic sulphide, one
can arrive at a comparatively simple electrical theory of photo-
oxidation. If one assumes that oxidation takes place when two
hydroxyl ions can give up their charges, forming water and
nascent oxygen available for oxidation, and that subsequently
the negative charges thus set free may react with free hydrogen
ions and free oxygen to form new hydroxyl ions which may again
discharge and oxidize, it will be readily seen that the rate of
oxidation will primarily depend on the value of the negative charge
which the oxidized particle itself carries. If this is large the
hydroxyl ions will be repelled, and oxidation will be retarded.
If any force intervenes to reduce the negative potential of the
colloid particle, the hydroxy] ions will be less repelled and oxidation
favored. The above results show that such a result is produced
by radiant energy. Since it is well known that practically all
organic matter, living or dead, carries a negative charge, and since
the oxidation of all organic matter is greatly accelerated by radiant
energy, it would seem that this theory might find quite wide
application.

It was thought interesting in this connection to determine what
might be the effect of illumination upon the rate of wandering of
bacteria in the electric current. Sarcina flava, Sarcina rosea and
Bacillus prodigiosus were investigated. The rates of wandering
were first determined in the dark, and then in the light. The
results were in all cases positive. In every case the rate of wander-
ing was less in light than the dark. The difference was in general
about twenty per cent. We have here a possible electrical expla-
nation of the toxic effect of sunlight on bacteria. Bacteria live
best in the dark. If we assume that they take on such a charge
as regulates the rate of oxidation to such a value as is demanded
by the normal metabolism, then anything which reduced this
charge would tend toward destructive oxidation. Radiant energy
is capable of reducing the negative potential of the bacteria, and
the result is clear.

Another class of phenomena which receives ready explanation
from this point of view is that of the coagulation of many proteins

ELECTRIC POTENTIAL OF BACTERIAL SUSPENSIONS. 153

under the influence of ultra-violet rays. It is generally considered
that the coagulation of colloids is favored by the reduction of the
electric field between the colloid particle and the medium in
which it exists. As has been shown radiant energy in general
affects the field and very commonly reduces it, which would explain
the coagulation in such cases.

In a purely tentative and speculative way, it may perhaps be
justifiable to raise the question as to what extent the above prin-
ciple may be applied to the explanation of any or all of the life
processes. From the physical-chemical point of view it is not
unreasonable to assume that all metabolic processes which take
place between a colloid particle and the medium in which it is
found are determined and regulated by the establishment of
certain potential differences between colloid and medium. There
will be in any particular case, such a potential developed as is
most favorable to the particular work in hand. So long as such
favorable conditions of potential are maintained metabolism will
proceed smoothly. Such influences as disturb the potential system
will disturb the metabolism, and such disturbance may be either
favorable or unfavorable. It may be going too far, but is it not
possible there is some suggestive value in the assumption that
living and dead matter are to be distinguished by the ability or
inability to maintain such potential systems as properly regulate
the metabolic processes? Certainly there would seem to be here
a possible explanation of the fact that the living stomach does not
digest itself, while the dead one does.

It is also interesting to note that bacteria, whose metabolism
is essentially one of oxidation, and which are adjusted to the dark,
have their potentials so affected by light as to lead to destruction.
On the other hand, chlorophyll, whose function is that of an agent
in a metabolism which is essentially reducing, and which is active
only in the light, has its potential so affected by light as to favor
its type of metabolic reaction. In other words the charge on
bacteria is reduced by light, that on chlorophyll is increased.

The author desires that the above be considered not as the
exposition of a theory, but merely as the suggestion of one.

SCIENTIFIC PROCEEDINGS.

ABSTRACTS OF COMMUNICATIONS.
Fifty-fourth meeting.

Zoological Laboratory, Columbia University, May 21, 1913.
President Ewing in the chair.

98 (794)

The color index and color of the red blood corpuscles.
By E. E. BUTTERFIELD.

[From the Rockefeller Institute for Medical Research.]

The present study is based largely on material observed several
years ago in Munich in the clinic of Prof. Friedrich Miiller.
The object of the study was to determine by means of exact
methods, (1) the existence of a high color index in pernicious
anemia, (2) the magnitude of the elevation of the color index,
and (3) the explanation of the phenomenon.

The calibration of the pipette and the dimensions of the count-
ing chamber used for the erythrocyte counts were checked by
special methods. The hemoglobin determinations were made
spectrophotometrically (spectrophotometer of Kénig, Martens
and Griinbaum, Nernst filament as light source). The manner
in which the hemoglobin concentration may be calculated from a
measurement of the light absorption follows from the equation for
the diminution in the intensity of homogeneous light on traversing
a planparallel layer of a colored solution:

Tl! = Te*, (1)

in which J = initial intensity, J’ = final intensity, k = a constant,

1 = linear thickness of the absorbing layer, and ¢ = the concentra-

tion of the colored substance. J’/I can be measured with spectro-
155

156 SCIENTIFIC PROCEEDINGS (54).

photometer. When this has been done with a solution of pure
oxyhemoglobin of known concentration in an absorption tube of
known length k can then be calculated. After the value of k has
been determined for pure oxyhemoglobin from human blood the
concentration of hemoglobin in laked human blood can be derived
from a measurement of J’/J._ Oxyhemoglobin from ox blood may
be conveniently substituted for human hemoblogin in the deter-
mination of k since it was shown in Tiibingen that the constant has
the same value for both hemoglobins. In the present study
homogeneous light was not employed, but the measurements were
made in a very narrow interval in the spectrum (about 4 uy).
This is not accompanied by an appreciable error as long as the
concentration and linear thickness are kept within narrow limits.
The measurements were always made in two regions in the spec-
trum, (1) at the maximum in green of the second absorption band
of oxyhemoglobin, and (2) at the minimum in yellowish green
between the two absorption bands of oxyhemoglobin. The reason
for this follows on combining the two equations for these measure-
ments. If J,’ = I,e-“!° represent the conditions in the first
region and J,’ = I,e-** those in the second region, one would
have after logarithmating and dividing,

log fi

dl ks
lon coy b i
Tf.
Therefore

I,
log I!
Ty
logrs

remains constant independent of concentration and linear thick-
ness. If

varies then some other substance with constants different from k;

COLOR OF THE RED BLOOD CORPUSCLES. 157

and k, must also be present in the solution. For pure oxyhemo-
globin k; = 19.36 and k, = 11.66. As concentration grams in
100 c.c. is used. The blood was taken in all cases from an arm
vein and immediately defibrinated. Table I gives the result of
determinations made chiefly on individuals in apparent good
health and on patients with pernicious anemia.

TABLE I.
NORMAL BLOOD.

Ery- uantity of

chins log i, Hemoglo- Aeaoerbte
Specific} Sabli | cytes. |" 71 MT Blood. [ein average | Color

gravity.|reading. aed ise Ze Grams per Para & index.
cu.mm. t, Too C.C. 10711,
Se |
Men, 1 1.057 95 4.29 | 1.66 15.4 3.6 I.I
2 1.055 77 4.19 1.65 14.3 3-4 1.0
3 1.062 | 100 5.19 1.67 17.4 3.2 0.9
4 1.061 97 5.46 | 1.68 17.8 3-3 1.0
5 1.056 87 4.57 1.64 15.3 3-4 I.0
6 1.066 | 105 5-79 1.65 19.3 2:3 1.0
7 1.060 93 4.74 | 1.65 16.9 3-6 Tek
IA VEVARES soo. .ys.0siere steels T.059 93 4.92 1.66 16.6 3.4 1.0
Women, I 1.057 83 4.42 | 1.64 15.8 3.6 U0
2 1.058 88 atgLs 1.64 15.6 300 1.0
3 1.053 Ghz] 4.44 | 1.67 1337 35 1.0
4 1.061 07 SelE |) Od. 17.0 Bes 1.0
5 I.055 85 4.46 | 1.64 13.7 ial! 1.0
6 1.056 90 4.95 1.65 15.2 Bak 1.0
IAVETA ROS cis oy eeete hehe ciate 1.057 87 4.75 | 1.65 15.2 a2 1.0
PATHOLOGICAL CASES.

Pernicious anemia I....] 1.040 21 0.74 | 1.65 3.47 4.7 | 1.5
Pernicious anemia IT... .| 1.035 23 0.87 | 1.65 3-79 4.4 | I
Secondary anemia...... | —_ | = 2.43 | 1.63 5.59 2.3 | 0.7

Sa Se BT LA ee = Sal See ee OS eT )
Polycythemia......... | 1.075) || — — | 1.66 | 23.9 a

It will be seen first that the average hemoglobin content of
defibrinated normal blood is considerably higher than the orthodox
14 grams in 100 c.c. Secondly, the value for the amount of
hemoglobin per corpuscle in normal blood is remarkably constant.
This may be taken as a measure of the color index, and the color
index may be conveniently defined as the quantity of hemoglobin
per corpuscle multiplied by that factor which renders the quantity
of hemoglobin per normal corpuscle = 1. In pernicious anemia

158 SCIENTIFIC PROCEEDINGS (54).

there is a marked increase in the quantity of hemoglobin per
corpuscle. That this is an actual increase and not a simulated
effect due to the presence of some substance with a greater light
absorption than oxyhemoglobin is shown by the constant value for

in normal blood and in the blood of pernicious anemia.

Closely related to the color index is the actual color of
the red blood corpuscles. It is well known that the color of
erythrocytes in single layer viewed microscopically in transmitted
light is yellowish green. The blood itself is deep red in color,
and the reddish tinge becomes noticeable microscopically when
several superposed corpuscles are viewed in transmitted light.
The explanation of this phenomenon becomes apparent, I think,
when one studies the spectrum of a single layer of corpuscles as
compared with several superposed layers. The absorption curve
of oxyhemoglobin presents a minimum in yellowish green (560 up)
and a region of least absorption in red (650 ypyu-660 up). In the
oxyhemoglobin spectrum k,, is much greater than k,, con-
sequently on increasing the thickness of the absorbing layer the
intensity of the transmitted light diminishes much more rapidly
in yellowish green thaninred. If we regard only these two regions
in the spectrum we would have for the intensity of the transmitted
light in yellowish green J,,’ = I,,e~*' and in red I,’ = I,e-*".
For J,,e—-*!>I,e-*" a color change would be expected. This is
known as the principle of dichromatism. Whether a color change
occurs or not with the same light source would depend on the values
of k,, and k;andon 1. The necessary conditions Ij, > I,, Rgr > Rr
for oxyhemoglobin and / sufficiently small are all realized in the
case of red blood corpuscles viewed indaylight. This formulation
isonly avery rough approximation. Theexact formulation would
require integration of the intensities over the whole spectrum, and
this cannot be done at present as long asit is not known what func-
tions J and k are of \ (the wave-length). However, several ob-
servations furnish strong support for the view that the color

CoMBINED ACTION OF MAGNESIA AND ETHER. 159

change of erythrocytes in layers of varying thickness is in accord-
ance with the principle of dichromatism. It is possible to con-
struct a thin wedge of solid oxyhemoglobin and observe the thick-
ness at which the color change occurs. In such a wedge at a
thickness of 1.3 u and less the color is identical with that of a single
layer of red blood corpuscles. Above 1.3 » adistinct reddish tinge
is noticeable, increasing with the thickness of the wedge to a deep
pure red. In this experiment the color is the same in parallel or
convergent light. This rules out the influence of the stroma and
the surface curvature on the color of the red blood corpuscles.
Finally, small (microscopic) crystals of oxyhemoglobin (second
crystallization) are of the same color as the red blood corpuscles,
while larger (i. e., thicker) crystals are bright red.

99 (795)

Combined action of magnesium and ether; evidence of a central
effect of magnesium.

By S. J. MELTZER and JOHN AUER.

[From the Department of Physiology and Pharmacology of the
Rockefeller Institute.]

We have shown about eight years ago that magnesium sulphate
is capable of causing a profound depression in animals. After an
injection of a proper dose of a solution of a magnesium salt the
animal loses for some time, all reflexes and signs of sensibility,
while the respiration remains intact. Several years before it was
found (M.) that a condition similar to this can be produced by an
intra-cerebral injection of two or three drops of a 5 per cent.
solution of magnesium sulphate, while the injection of hypertonic
solutions of other salts caused convulsions. On the basis of both
experiences we assumed as a working hypothesis that magnesium
favors an inhibition of the entire nervous system. We designated
the depressed condition of the animals as anesthesia, which implied
that the central nervous system was also affected. This interpre-
tation has not been accepted by Wiki. He called attention to
experiments of Binet and of his own to the effect that magnesium
salts paralyze the motor nerve endings, and he assumed that in our

160 SCIENTIFIC PROCEEDINGS (54).

experiments the animals were merely paralyzed and had not lost
any sensation; in short, magnesium acts, according to Wiki, like
curare, although he admits the significant difference that curare
paralyzes the respiratory motor nerves before the motor nerves of
the other parts of the body, while magnesium paralyzes all other
motor nerves before it attacks the motor nerves concerned in the
respiration. The statement that magnesium paralyzes motor
nerve endings is perfectly correct; we have seen it ourselves
numerous times. While it is true that many other inorganic salts
have also a curare-like action upon the motor nerve endings, it
has to be admitted that the effect of magnesium salts upon the
motor nerve endings exceeds that of any of the other salts. This
fact, however, is rather in harmony with our assumption that
magnesium depresses all parts of the nervous system. The ques-
tion is only whether it affects also the central part of the nervous
system. Wiki and two or three others deny it; we assume it,
and have many good reasons for this assumption. We shall,
however, not enter here upon a discussion of the entire subject.
Our sole purpose in the present communication is to report the
results of a series of experiments which make it probable that
magnesium affects also the central nervous system. In these
experiments rabbits and dogs received one half, or less, of the effect-
ive dose of magnesium sulphate. It was found that such animals
are readily deeply narcotized by inhalations of small doses of ether
which are insufficient to narcotize normal animals. You see here
a picture of three rabbits. The dose of magnesium sulphate
necessary to narcotize a rabbit is about 1.2 gm. per kilo body
weight. Rabbits No. 1 and 3 each received intramuscularly 0.6
gm. MgSO, per kilo. Rabbits No. 2 and 3 inhaled through trache-
otomy tubes, connected by means of a T-tube with the tube of a
bottle containing ether. Each rabbit received exactly the same
amount of ether which was insufficient to cause complete narcosis.
The animals were photographed soon after the discontinuation of
the etherization. Rabbit No. 1 which had only magnesium, and
No. 2 which had only ether are sitting up. Rabbit No. 3 which
had magnesium and ether is deeply narcotized and is limp. You
see here a similar picture of three dogs treated in the same manner.
If magnesium would have had only a peripheral effect there could

PRODUCTION OF AcID BY TISSUES. 161

have been no summation with the subminimum central effect of
the ether.

100 (796)
Note on the production of acid by tissues growing in vitro.
By PEytTon Rows, M.D.

[From the Laboratories of the Rockefeller Institute for Medical
Research.|

Connective-tissue cells of the chicken, growing im witro in
chicken plasma to which a little blue litmus has been added,
produce rapidly a focal, pink coloration of the medium. If a
number of small fragments of one tissue (heart muscle or the aorta
of young chicks, or chicken sarcoma) be plated out with the
plasma medium in a petrie dish, it will be found that all the tissue
bits are at first stained blue, but that those from which growth
occurs become pink, while the growing tissue itself is unstained.
The fragments remaining permanently inert keep the blue color.

Often a pink coloration of tissue bits can be observed at a time
when growth is found, microscopically, to have barely started.
The acid change is in general sharply localized to the neighborhood
of the growing tissue. When growth is checked by placing the
preparation in the ice-box, neutralization in the acid foci is often
incomplete at the end of forty-eight hours, and this even when the
bulk of alkaline plasma is relatively large and its plasma network
thinned by dilution. Diffusion in the plasma medium as thus
indicated is very slow. Under the ordinary circumstance of
im vitro life without artificial provision for a circulation of fluid,
tissue proliferation must take place almost from its beginning,
inanacid medium. This constitutes a serious fault in the method
of cultivation.

The nature of the acids produced by the growing tissue has not
been determined. Carbonic and lactic acids are presumably
present in greatest quantity. That the amount of acid formed
may be very considerable has been shown by titrating out the
as yet unclotted blue plasma to the tint acquired by the tissue
cultures. The acid does not affect methyl orange, but very
occasionally it changes congo red toward violet, a change best seen

162 SCIENTIFIC PROCEEDINGS (54).

in the interior of degenerating cells which have taken up the indi-
cator. The violet change is not due to free carbon dioxide, for
bubbling the gas through unclotted plasma containing congo red
fails to bring about an alteration in color.

The observations show that tissue cells can withstand much
more considerable changes in the reaction of the medium round
about than has been supposed; and that their growth in vitro
may be very active in an acid medium.

101 (797)

The location of the optic anlage in Amblystoma and the interpreta-
tion of certain eye defects.

By CHARLES R. STOCKARD.

[From the Department of Anatomy, Cornell Medical College, New
York City.]

The early embryos of Amblystoma were operated upon so as
to remove definite areas from the open medullary plate in order
to determine the position of the materials which would give origin
to the future eyes.

Preliminary experiments of sticking and disturbing the cells
in the anterior end of the medullary plate without actually remov-
ing these cells did not prevent the subsequent development of the
optic vesicles in an apparently normal manner. Cutting out
rectangular pieces of the anterior part of the medullary plate,
reversing the pieces and transplanting them merely caused the
eyes to develop in misplaced positions. These two experiments
demonstrate the fact that unless the future eye material is well
removed by the operation the optic vesicles may form. This
fact is important in considering the results of the following experi-
ments.

A general statement of the results of the experiments may be
expressed as follows: Thirty embryos studied after various opera-
tions in which lateral portions of the medullary plate were removed
at slightly different developmental stages showed in twenty-four
individuals, or eighty per cent. of the cases, subsequent develop-
ment of both eyes, while only six specimens, or twenty per cent.

LOCATION OF THE OpTIC ANALAGE IN AMBLYSTOMA. 163

of the cases, showed absence of the eye. In one case the presence
of the eye was questionable, in five cases one eye and in one case
both eyes were absent. The absence of the eyes in the latter
cases was possibly due to the cut having been made in a more
median position than was intended.

Nine embryos studied after having been operated upon so as
to remove a narrow median strip of cells from the anterior portion
of the medullary plate showed in four cases, or about forty-five per
cent. of the specimens, entire absence of eyes. In four other
individuals the eyes were highly defective, one specimen having
one poorly formed eye while the other was questionably present.
In only one of the nine embryos did the eyes approach the normal
condition, from this specimen an extremely narrow median piece
had been cut out of the medullary plate. The optic anlage might
have been sufficiently wide at the time of the operation to allow
its median portion to be removed and yet enough material remain
on either side of the cut to give origin to the two eyes. According
to the views of several investigators the removal of this median
material should have caused the cyclopean defect, yet it did not.
In a more extended report of these experiments I shall show that
cyclopia is not due to a coming together of lateral materials in the
median plane, but to a failure of median material to spread
laterally.

Contrasting the results obtained after the lateral and median
cuts mentioned above one must conclude that: The eye anlage in
the medullary plate occupies an antero-median position as shown by
the various abnormalities incurred when this region 1s cut away.
The failure to injure the development of the eyes in the great majority
of cases when the lateral portions of the medullary plate are removed
by operation indicates further that the eye anlagen do not occupy
lateral positions during this stage of development.

Based upon these experiments and a study of a large number of
eye abnormalities it is concluded that the cyclopean defect is a
developmental arrest. The eye anlage fails to widen laterally
so that only a single median growth center arises from which
develops the ventro-median cyclopean eye. In normal cases
the anlage widens and two more or less lateral growth centers
become established and give rise to the ventro-lateral optic

164 SCIENTIFIC PROCEEDINGS (54).

vesicles. The optic stalks, however, and later the optic nerves
following the stalks as paths always lead back to the point of
their median origin and the optic cross or chiasma is in the median
plane, below and outside the brain tissue. The attainment of this
position of the optic cross would seem mechanically impossible
if the eyes arose from lateral medullary tissues since the optic
fibers following the stalks would enter the brain laterally and would
necessarily cross within the brain tissue, not below and outside as
the nerves actually do.

There is no medullary tissue other than future eye tissue be-
tween the eye anlagen, therefore, Spemann and others are incorrect
in assuming that cyclopia is due to a failure to develop of tissues
between the eyes thus permitting the eye anlagen to slump towards
the median plane and fuse. The defect is due to a failure or
arrest in development of the eye material itself.

102 (798)
The occurrence of betaine in the muscles of invertebrates.

By D. WRIGHT WILSON.

[From the Sheffield Laboratory of Physiological Chemistry, Yale
University.]

Betaine, or trimethylglycocoll, was isolated from the muscle
tissue of two varieties of mollusc, Pecten irradians, the common
scallop, and Sycotypus canaliculatus, the periwinkle.

The tissues used were the adductor muscle of the Pecten and
the large pedal muscle of the Sycotypus. The manner of treat-
ment was the same in both cases. The muscles were finely
ground, extracted with several changes of water and the concen-
trated extract freed of colloidal material by precipitation with
alcohol and by the regular Kutscher manipulation with tannin.
The portion precipitated by phosphotungstic acid was fractioned
by precipitation with silver nitrate and barium hydroxide and
from the resulting filtrate, betaine was crystallized as the free
base and hydrochloride. In both cases, the compound was
identified by the melting points of the hydrochloride, picrate and
chloroplatinate and by the analyses of the hydrochloride and
chloroplatinate.

STUDIES IN THyroiD ACTIVITY. 165

103 (799)
Studies in thyroid activity.

I. THe CHEMICAL CONSTITUENTS OF THE THYROID GLAND.
By E. C. KENDALL.

[From the Pathological Department of St. Luke’s Hospital, New
York City.|

Interest in the chemical examination of the thyroid gland
centers around the iodine-bearing compound. The first attempt
made in this laboratory to separate and purify the iodine-bearing
compound consisted in dialyzing under varying conditions. The
entire gland was dissolved in sodium hydroxide and dialyzed from
collodion sacs. With running tap water only ten per cent. of the
total iodine would pass through the sac. At a temperature of 60
to 70° C, and under proper conditions of acidity, eighty per cent.
of the iodine would pass through the sac. These experiments led
to a study of the diffusibility of certain cleavage products of the
thyroid. As this study of the chemical constituents of the thyroid
is still being carried on, the details will be published later. The
results, however, may be briefly described as follows:

A new method of treatment has been found by which the com-
plex proteins of the thyroid gland may be broken down into simpler
constituents which have not been described by previous investi-
gators. Among the products obtained uric acid and tryptophan
in large amount are found. There are several non-iodine-bearing
compounds. The iodine has been found to exist in two distinctly
different compounds. These have been partially purified and a
compound containing 23.3 per cent. of iodine in organic combina-
tion has been separated. Of the non-iodine-bearing compounds
two are of especial interest. One reduces silver, gold, and mercury
in alkaline solution, and the other absorbs free iodine very readily.
In all, twelve distinct chemical compounds have been separated.
The properties of three of these show them to be still of high
molecular weight and they may, by further treatment, be divided
into still simpler compounds.

166 SCIENTIFIC PROCEEDINGS (54).

104 (800)
Studies in thyroid activity.

II. THe Speciric PHysioLoGicAL ACTIVITY OF CERTAIN CoN-
STITUENTS OF THE THYROID GLAND.

By E. C. KENDALL.

[From the Pathological Department of St. Luke’s Hospital, New
York City.]

Investigations of the physiological activity of the thyroid
gland have shown that the internal secretion of the gland serves
many different functions. One or more of a series of symptoms
accompany cases of thyroid deficiency. These symptoms are
relieved by the administration of the thyroid gland of certain
animals, as the sheep, hog, and ox. When this treatment is
stopped the symptoms return. It would thus appear that the
functions of the internal secretion of the thyroid may be fulfilled
by furnishing the body with the constituents of the thyroid gland
from another animal.

The separation of the various chemical constituents briefly
described above suggested the possibility of determining which
constituents controlled the various symptoms occurring in cases of
thyroid deficiency.

The first step in the method of separation of the constituents
results in two solutions. One of these contains about 60 per cent.
of the total iodine and 9 per cent. of the nitrogen. This is desig-
nated ‘“ Solution A.’”’ The other, called ‘“‘ Solution B,’’ contains
40 per cent. of the total iodine and 91 per cent. of the nitrogen.
In order to establish the physiological activity of these solutions,
experiments were carried out, first upon dogs, and then with cases
of thyroid deficiency. The number of cases treated is insufficient
to establish completely the physiological properties of these
solutions, but the results based upon a series of experiments with
two dogs extending over four months, with two typical cases of
myxedema, and with three cretins, are as follows:

Solution A was found: (1) to affect the nitrogen metabolism
and hence the body weight and temperature; (2) to produce
tachycardia; (3) to cause nervousness and tremor; (4) to relieve

STUDIES IN THYROID ACTIVITY. 167

pain and great weakness felt in the back and certain sensations of
cold felt on the head.

Solution B was found: (1) to change the dry, scaly, rough skin
to a soft and entirely normal condition, which allowed perspiration
to take place; (2) to remove the soreness of bones and joints; (3)
to prevent cramps and twitching of the muscles; (4) to have a
marked effect on the mental activity, especially noticeable in
cretins; (5) to relieve the hard, firm condition of the flesh, allowing
it to become soft and pliable; (6) to prevent burning sensations
which flash over the skin from one part of the body to another.

Solutions A and B act independently of each other. Further-
more, each of these solutions has been subdivided and the various
constituents separated have continued to act independently. It
would thus appear that each function of the thyroid gland is due
to the specific physiological activity of the separate and distinct
chemical constituents of the gland. Baumann claimed that
iodothyrin is the active principle of the thyroid gland. It is
evident that there is no one active principle of the thyroid gland,
but that there are many, each with its own specific action. The
separation of the various constituents makes it possible to treat
cases of thyroid deficiency with those portions of the gland whose
deficiency is indicated by the symptoms. Furthermore, it will
be possible to standardize the active principles of the thyroid
gland by a method based on chemical analysis.

I wish to thank Dr. F. C. Wood and the attending physicians of
St. Luke’s Hospital for the opportunity of carrying on this in-
vestigation, and Mr. A. W. Thomas for assistance in the chemical
laboratory.

Further work is being carried on, both with the chemical separa-
tion and identification of the various constituents and with the
physiological activity of the same.

168 SCIENTIFIC PROCEEDINGS (54).

105 (801)

Further studies on muscle creatine.

By M. S. FINE and V. C. Myers.
[From the Laboratory of Pathological Chemistry, New York Post-
Graduate Medical School and Hospital.)

In an earlier communication,! attention was called to the
constancy in the content of muscle creatine for normal animals of
a given species, though distinctive for different animals. It was
further pointed out that during starvation in the rabbit, the per-
centage content of muscle creatine may show either an increase or
a decrease, the latter depending in considerable part upon the
rate and amount of creatine eliminated in the urine.

These observations have been extended to rabbits which have
been fed for varying periods upon carbohydrate—without fat
or protein. Carbohydrate feeding greatly reduces the elimination
of creatine in the urine, as previously observed, though the creatine
content of the muscle does not materially differ from that found
during a similar length of starvation. In other words, it may be
markedly decreased during a long period of carbohydrate feeding.
It seems probable that the action of the carbohydrate is simply
one phase of the sparing action of carbohydrate on protein meta-
bolism, in this case allowing sufficient time for the body to handle
the creatine, 7. e., to oxidize it, or change it to creatinine.

That creatine when fed or injected does not reappear in the
urine in the form of creatinine, except in traces, or inlarge amount
unchanged, unless given in considerable quantity, has been as-
certained by a number of investigators. The possibility that this
creatine, which remains unaccounted for, is stored up in the muscle
has not been adequately studied. In four experiments on rabbits,
the creatine content of the muscle, after repeated subcutaneous
injections of creatine, has been found to be uniformly slightly
above (4-7 per cent.) the normal amount. This would appear to
indicate that a small amount of the injected creatine was deposited
in the muscles, though insufficient to account for the creatine not
eliminated in the urine either unchanged or in the form of creati-

1 Myers and Fine, Proc. Soc. Exp. BIOL. AND MED., October, 1912, X, pp. 10
and 12.

NON-PROTEIN NITROGEN OF THE BLOOD IN NEPHRITIS. 169

nine. In two similar experiments with creatinine, there was
apparently a slight increase in the creatine (total creatinine) con-
centration of the muscle, which we are not prepared to discuss at
this time.

The possible influence of autolysis upon the content of muscle
creatine and added creatine and creatinine is being investigated.

106 (802)

The total non-protein nitrogen of the blood in nephritis and allied
conditions.

By CuFForD B. Farr, M.D., and J. Haroitp Austin, M.D.

[From the John Herr Musser Department of Research Medicine,
University of Pennsylvania, Philadelphia.|

The following summary covers the results of our study of the
non-protein nitrogen of the blood by Folin’s methods in a series
of fifty-nine hospital patients. Our main concern has been with
nephritis but we have examined the blood in many other conditions
as opportunity offered.

The patients group themselves into four divisions:

I. Those showing no disturbance of renal function (17 cases).

II. Those with marked cardio-vascular disease of some type,
most of which showed urinary changes the result of renal
congestion (II cases).

III. Those showing nephritis (23 cases).

IV. Those in which certain features would lead one to suspect
nephritis, but in which the existence of nephritis is not
borne out by other findings (8 cases).

Our patients of Group I, suffering from a variety of acute
and chronic diseases, but without evidence of disturbance of renal
function, showed a total non-protein nitrogen in the blood varying
from 16 to 43 milligrams per 100 c.c. From 50 to 60 per cent. of
this was in the ammonia-urea fraction. In the patients with
cardio-vascular disease with renal congestion, but without evidence
of other renal lesion there was no increase of the non-protein
nitrogen in the blood, nor alteration of the ammonia-urea per-
centage, although albuminuria, casts and some impairment of the
phenolsulphonephthalein elimination were usually present.

170 SCIENTIFIC PROCEEDINGS (54).

In that type of chronic nephritis characterized by marked
albuminuria, cylindruria and edema, there were similar findings.
In that type of chronic nephritis associated with hypertension, the
non-protein nitrogen was increased, ranging from 40 to 181 milli-
grams per 100 c.c., and the percentage of the ammonia-urea
fraction was usually higher than in non-nephritic cases. The
nitrogen values in these patients were subject to rapid fluctuations
in the course of a few days and clinical improvement was associated
with a fall in the non-protein nitrogen content. Uremia was
almost always accompanied by some increase of the non-protein
nitrogen in the blood but no constant relation could be established
between the degree of the increase and the tendency to uremia.

We believe that this method of estimating the total non-protein
nitrogen in the blood is a valuable aid in the clinical study of
nephritis and that it can be carried out in any thoroughly equipped
clinical laboratory. The error of the method is indicated by the
duplicate analyses which were done in almost all cases and which
showed an average discrepancy between duplicates of 1.6 milli-
grams per 100 c.c. of blood. The urea method was in our hands
less reliable, and large and inexplicable discrepancies occurred at
times in our urea duplicates rendering repetition necessary and
causing us to attach less importance to the urea figures.

107 (803)

The toxicity of sodium tartrate with special reference to diet and
tolerance.

By W. SALANT and C. S. SMITH.

[From the U. S. Department of Agriculture, Bureau of Animal
Industry, Washington, D. C.]

The toxicity of the sodium salts of dextro and levo tartaric acid
was tested in experiments on frogsand rabbits. Both isomers were
found equally toxic in these animals thus contradicting the earlier
work of Chabrié! on the subject, who claimed that levo was more
than twice as toxic as dextro tartaric acid. In experiments on rab-
bits, diet proved to be an important factor in the determination of
resistance to this substance. Animals which were fed oats or oats

1 Compt. Rend. Acad. Sc., 1893, Vol. 116, p. 1410.

INFLUENCE OF PANCREATIC AND DUODENAL EXTRACTS. I7I

and cabbage succumbed to a dose of 0.4 gm. of the salt per kilo
when given by subcutaneous injection. Suppression of urine was
usually observed on the first day and death occurred in six to
seven days. In starvation, slightly smaller doses were fatal to
some rabbits. The resistance was increased considerably when
the diet was changed to carrots. Such animals stood 1.0 gm. per
kilo by subcutaneous injection, while 1.2-1.5 gm. per kilo were
toxic. A moderate degree of tolerance for tartrates was in-
duced in animals which were fed oats and cabbage. By grad-
ually increasing the dose, a large proportion (6 out of 9) of rabbits
survived 0.8 gm. per kilo which is twice the fatal dose. Rabbits
which were receiving carrots did not acquire tolerance for tar-
trates. Sodium tartrate was much less toxic when given by
mouth. 5 gm. per kilo was found to be the minimum fatal dose.

EXPERIMENTS ON CATs.

Amounts which have been found to be fatal for rabbits did
not produce any symptoms in cats. Subcutaneous injection of
one gm. per kilo produced a slight diarrhea in some individuals,
and had no effect whatever in others. 13 gm. per kilo proved
fatal to one cat but was without action in another. Out of four
cats which received 2 gm. per kilo three died, one survived. When
sodium tartrate was given by mouth vomiting frequently occurred.
In one case, however, when ten gm. per kilo were fed diarrhea was
the only effect observed.

108 (804)

The influence of pancreatic and duodenal extracts on the gly-
cosuria and the respiratory metabolism of depancreatized dogs.

By J. R. MuRLIN and B. KRAMER.

[From the Physiological Laboratory of Cornell University Medical
College, New York City.]

Several dogs completely depancreatized by Hedon’s method
and eliminating glucose and nitrogen in Minkowski’s ratio were
treated by intravenous injection of pancreatic extract prepared
by Knowlton and Starling’s method.! The urines collected in
twenty-four hour periods exhibited an increase in the D: N ratio

1 Knowlton and Starling, Journ. of Physiol., 1912, XLV, p. 146.

E72 SCIENTIFIC PROCEEDINGS (54).

on the days immediately following. When the urine was collected
in short periods after injection, a marked fall in the sugar output
was witnessed lasting from four to ten hours but this was followed
by a compensating rise which, in some instances, augmented the
total for the twenty-four hour period, in others raised it only to
the previous level.

Much greater effects were obtained with a double extract of
dog’s pancreas and duodenal mucosa. The following experiment
(No. II) is typical. The effect, however, cannot be ascribed to
the organic extract from either the pancreas or the duodenum for
the same Ringer’s solution in which the tissues were extracted
when made alkaline to about the same extent with Na,CO; gave
an identical effect.

Doc “Q.’’ OPERATED APRIL 17.

Exp. II.
Vol. | Glu- | Nitro- Ni Blood
Date. Time. urine, | cose, gen, | D:N. | Glucose per trogen | sugar,
cic; gm. gm. hour. per hour. per cent.
4/21/13 I2.15- 2.15 30

2.41 | 0.95 2.52 | 1.20 | 0.47 0.128
2.15— 4.15 50 2.40 | 1.09 | 2.20 1.20 0.54
5.00— 5.30 | 150 c.c. of pancreas and duodenum from 2 normal dogs inj.
intravenously

4-15-— 6.15 ] 30 1.30 | 0.67 | 193° (0:65 0.35
6.15— 7.15 25 0.28 | 0.33 0.87 | 0.28 0.33
7-15— 8.15 26 0.41 | 0.45 | 0.89 0.41 0.45 0.148
8.15- 9.15 26 0.59 | 0.48 | 1.21 0.59 0.48
9.15-10.15 22 0.51 | 0.39 | 1.28 0.51 0.39
P.M.
4/22/13 | 10.15— 6.00 18,66 | 6.99 | 2.68 0.93 0.35
Exp. III.
4/24/13 2.19-4.19 22 | 0.54 | 3.08 0.82 0.27 |
4.26-5.15 150 c.c. Ringer’s Sol. +1%Naz2COs injected intravenously
4.19-6.19 60 1.65 | 0.64 | 2.55 | 0.82 0.32
| 6.19-8.19 24 0.46 | 0.52 | 0.88 0.23 0.26
| A.M.
4/24-25 | 8.19-1.19 90 | 2.41 | 0.59 1.62 0.48 0.29

The similarity in these two experiments on the same dog
indicates that it is not a hormone which is responsible for the
reduction in the sugar, and the increase in the percentage of sugar
in the blood indicates that the reduced sugar elimination is in
reality due to a change in the permeability of the kidney, as
Wohlgemuth? has found it after ligation of the pancreatic ducts.

That there is no effect on the combustion of sugar attending

2 Wohlgemuth, Berl. klin. Woch., February 24, 1913, p. 339.

INFLUENCE OF PANCREATIC AND DUODENAL EXTRACTS. 173

the reduced elimination of that substance is proved by the following
experiment on another dog place in the respiration calorimeter.

Doc “S.”” OPERATED May 6.

Glucose Nice: H

Date. Time. in urine fare D:N. | COz O2, R.Q. oe Temp.
per per gm. mg. duced. | of dog.
HOUKS atranr!

5/9/13 9.45-10.45 8.62 | 9.22] 0.68 | 21.73 | 38.6
10.45-11.45 9.97 | 10.55 | 0.69 | 27.12 | 38.6
8.50-11.55 | 1.74 | 0.56 | 3.08

P.M.

12.35- 1.05 |150c.c. exts. of pancreas and duodenum from 2 normal dogs
injected intravenously. 20 gms. glucose given per os.
IT.55— 1.05 | 1.07 | 0.39 | 2.70

I.55- 2.55 13.60 | 14.13 | 0.70 39-6
2.55—- 3-55 11.84 | 11.78 | 0.73

3-55- 4-55 10.16 | 10.71 | 0.69 39-9
I.05- 5.05 0.45 1

Already in the first hour’s urine, including the period of
injection, a marked decline in the sugar elimination is seen, showing
that the typical effect on the excretion of sugar would have been
produced, if glucose had not been fed. With 20 grams of glucose
available, however, none, or an extremely small quantity at the
most, was burned. The increase in respiratory metabolism the
first two hours was due to restlessness of the dog.

Other experiments after injection of pancreatic extract alone
and after injection of normal dog’s blood likewise showed no
effect on the respiratory quotient.

Incidentally it has been found in the single experiment in the
respiration calorimeter that the heat production in the depan-
creatized dog was from 30 to 50 per cent. higher than the normal
on the same dog determined one month earlier. This confirms
the observations of Benedict and Joslin in severe cases of human
diabetes.1

1 Benedict and Joslin, Carnegie Institution of Washington Publication No. 176.

174 SCIENTIFIC PROCEEDINGS (54).

109 (805)

The carbon dioxide and oxygen content of the blood after ligation
of the abdominal aorta and the inferior vena cava.

By J. R. Muruin, L. EDELMANN and R. C. GILEs.

[From the Physiological Laboratory of Cornell University Medical
College, New York City.]

Respiration experiments on normal and depancreatized anaes-
thetized animals after ligation of the abdominal aorta and the
inferior vena cava! and on normal animals after exclusion of the
liver by joining the portal vein to the inferior vena cava? show an
increase in the respiratory quotient, which is interpreted by the v.
Noorden school to demonstrate the combustion of sugar in the
depancreatized animal and the dependence of the normal animal
upon the liver for its ability to burn protein and fat. These
experiments, however, were not accompanied by analyses of the
blood gases. It is possible that the higher respiratory quotient
after shortening of the circulating stream might be due to an
interference with the oxygen absorption (passive congestion of
the lungs) or to increased elimination of carbon dioxide by more
rapid circulation of the blood through the lungs.

Preliminary to some respiration experiments on depancreatized
dogs in which we are seeking the explanation of the altered res-
piratory quotient, we have made a number of experiments on
normal dogs analyzing the carotid blood before and after simul-
taneous clamping of the abdominal aorta and the inferior vena
cava. The results follow:

: : Oz per cent. COz per cent.
Pulse per min. | Resp. per min. auniggae 2 lood.
Dog | J E : Clamp:
Date. | No. |S2| a 3 3 3 4 (4 3 z on.
§ Bs | o 5 = & =
+>) & +2) i= I i=] 2) r=]
3 a 3 a 3 a 3 a
a 5) a is) Z D a oD
I9t3

3/22 I} 8] 108 I40 36 30 14.73 | 14.83 | 43.16 | 22.53 | 30 min.
4/10 II | 12 66 102 35 I2 16.40 | 16.45 | 38.35 | 37-47 | 23 min.
4/19 | III} 9} 132 I2I 66 77 18.88 | 16.39 | 39.42 | 24.28 | 1 hr.
5/10 | IVir1o! 108 120 33 15 I2.62 | 10.90 | 57.06 | 34.16! 1 hr.

1 Porjes, O., Biochem. Zeitschr., 1910, XXVII, p. 131; Porjes and Salomon, zdem,
p- 143.
2 Verzar, idem, 1912, XXXIV, p. 52.

INFLUENCE OF CERTAIN DiETS UPON TUMORS. 175

Chloretone anaesthesia was used. In the case of the last dog
which eliminated about 80 c.c. of CO, per minute, it may be
estimated that a reduction from 57 to 34 per cent. would remove
from the body in the course of one hour, about 800 c.c. of CO: or
enough to raise the R. Q. from 0.75 to 0.88. It cannot be assumed
as Porges has done that the blood gases, under the circumstances,
would reach an equilibrium within 10-15 minutes, for with the
circulation diminished to one half and the heart beating at its
normal rate, or higher, the blood would pour through the lungs
twice as often and would continue to lose carbon dioxide until the
tension in all the tissues became very much reduced.

In dog II the compensation in the rate of respiration was almost
sufficient to prevent loss of CO:.

110 (806)

On the influence of certain diets upon the growth of experimental
tumors.

By J. E. Sweet, E. P. CorSON-WHITE and G. J. SAxon.
[From the Laboratory of the American Oncologic Hospital,
Philadelphia, Pa.]

The study of the experimental tumors of animals has brought
forward numerous interesting observations upon the variation in
susceptibility of animals of the same species obtained from different
sources, to a given tumor strain, as well as variation in the rate
of growth of the transplanted tumors. We have undertaken the
study of the relation of certain diets to tumor growth and wish to
briefly report the results obtained with a diet based upon the
work of Mendel and Osborne. In their studies of the effect of
feeding with pure vegetable proteins they encountered numerous
combinations which effectively prevented growth, the animal
meanwhile appearing in good health. This seemed to us to offer
a most interesting opportunity to study the behavior of the tumor
cell under these conditions; in other words, regardless of whatever
the cause of cancer may be, can an inoculable tumor grow in a
host which is apparently incapable of normal cell growth?

This report, while based on a small series as tumor experi-
ments go, shows a result so uniform and striking that its con-

176 SCIENTIFIC PROCEEDINGS (54).

sideration would seem justified. In these series we have made
use of white mice, having by preliminary observations determined
that a diet made up on the basis of Mendel and Osborne’s work, of
a combination of glutenin and gliadin, would effectively retard the
growth of young white mice. One series of fifty mice inoculated
with the tumor obtained through the courtesy of Dr. Rous, of the
Rockefeller Institute, gave twenty-three tumors out of twenty-five
mice on a normal control diet, but only four out of twenty-five
on a vegetable protein diet, of which three tumors later dis-
appeared. In another series of fifty males, all again inoculated
with the same tumor, eighteen out of twenty-five on normal diet
developed tumors, with three out of twenty-five on a vegetable
protein diet; a third series of fifty females gave fifteen tumors out
of twenty-five on normal diet with seven out of twenty-five on a
vegetable protein diet. Expressed in percentage, 75 per cent. of
seventy-five mice developed a tumor under normal conditions;
19 per cent. of seventy-five mice developed a tumor when fed on
a vegetable protein diet, and further the tumors in the latter
series at thirty days were hardly larger than the tumors in the
normal fed mice at ten days.

By referring to the work of Mendel and Osborne it will be
seen that it is not a question of starvation in the ordinary sense
of the word nor of anemia, but that the most probable conclusion
is that the tumor cell is subject to the same laws of growth as is
the normal somatic cell.

I1I (807)

On the inhibitory action cf certain anilin dyes upon bacterial
development.

By CHARLES E. Simon, B.A., M.D., assisted by MartHa A.
Woop, M.D.

The triamino triphenyl methanes possess a well-marked in-
hibitory power over the development of certain pathogenic
organisms, notably staphylococci, streptococci, pneumococci and
meningococci, besides the anthrax bacillus and actinomyces.
This is quite pronounced, even in a dilution of 1 : 100,000. The
common pathogenic bacilli are not affected by the dyes in question,
in this concentration.

ANILIN DyEs AND BACTERIAL DEVELOPMENT. 177

The inhibitory effect is referable to the underlying chromo-
phoric group, and the presence of the basic auxochromic groups.
If the basicity of the dye is diminished by the replacement of the
basic by acid auxochromic groups, or if such groups are introduced
in addition to the basic groups, the inhibitory effect is destroyed.

Of the other classes of anilin dyes an inhibitory effect is pro-
duced only by those which contain the chromophonic radicles:

R R i
Nee Cre Se and i
MN —F5

\p7 NS
and here also, only by those which contain basic auxochromic
radicles. Acid dyes possess no inhibitory properties. The most
active inhibitory dyes are the triamino triphenyl methanes, while
the indamins, the oxazins, the thiazins and azins are on the whole
less active in this respect.

The inhibitory effect does not depend upon the color of the dye,
as there are violet acid dyes which are non-inhibitory, and red
basic dyes which are markedly inhibitory; similarly there are
green dyes which are active and others which are inactive in this
respect. The essential common factor evidently is the absence
of acid and the presence of basic auxochromic groups. Not all
representatives of the sensitive groups of organisms are equally
influenced by the dyes in question, nor even by a single dye, nor
are the different groups affected to the same extent. Certain
dyes will inhibit the growth of all the different sensitive groups,
while the effect of others is less extensive.

The inhibitory action of the dyes in question is most readily
explained on the assumption that the susceptible organisms
combine with the dyes by means of corresponding nutriceptors,
but are unable to cause the cleavage of the anchored molecules,
the death of the organism resulting from interference with its
normal nutritional (sc. reproductive) functions. Upon this basis
a directly toxic effect on the part of the dye need not be assumed.
It isin accord with the validity of this hypothesis that dye-resistant
strains of susceptable groups exist in nature as such, or may be
produced artificially. This was accomplished in the case of
certain hay bacilli, staphylococci and streptococci. Upon this

178 SCIENTIFIC PROCEEDINGS (54).

basis adaptation of this order would be due to the production of
receptors different from those found in dye susceptible strains,
by which the nutrition (sc. reproduction) of the organism could
be carried on, and which either possesses no affinity for the dye
in question or through which the organism can bring about its
cleavage. A limited number of observations would suggest that
in the treatment of certain infections, with staphylococci and
streptococci more especially, certain dyes might be used to ad-
vantage. In two cases of erysipelas the repeated local application
of the concentrated solution of dahlia seemed to restrict the exten-
sion of the infection.

112 (808)

Waxy degeneration of muscle in venom intoxication.
By RICHARD M. PEaRcE, M.D.

[From the John Herr Musser Department of Research Medicine
University of Pennsylvania, Philadelphia.]

In connection with the discussion by Beneke! and Wells? of
waxy degeneration of muscle occurring in anaphylactic poisoning,
and especially in view of Beneke’s reference to the action of
‘‘ brasilianischen Schlangengift ”’ (Crotalus terrificus) the following
notes on waxy degeneration in the rabbit following the intravenous
injection of the venom of Crotalus adamanteus may be of interest.

In the course of three series’? of experiments on a total of
about 45 rabbits it was noted that the intravenous injection of
venom was followed in three instances by waxy degeneration so
well-marked as to be evident macroscopically. In two of the

1 Beneke, R., ‘‘Ueber den Kernikterus der Neugeborenen,’’ Miinch. med.
Wochenschr., 1912, LIX, 387; ‘‘Ueber Muskelveranderungen bei akuten Ver-
giftungen mit Klapperschlangengift sowie bei Anaphylaxie,’’ Minch. med. Woch.,
1912, LIX, 729; Beneke, R., and Steinschneider, E., ‘‘Zur Kenntnis der anaphylak-
tischen Giftwirkungen,’’ Centralb. f. allg. Path. u. path. Anat., 1912, XXIII, 529.

2 Wells, H. Gideon, ‘‘Anaphylaxie und Wachsartige Degeneration der Muskeln,”
Centralb. f. allg. Path. u. path. Anat., 1912, XIII, 945.

3 Pearce, R. M., ‘‘An Experimental Study of the Relative Importance of Renal
Injury, Vascular Injury and Plethoric Hydremia in the Production of Edema,”
Arch. f. Int. Med., 1909, III, 422; ‘‘An Experimental Glomerular Lesion Caused by
Venom (Crotalus Adamanteus), Jour. Exper. Med., 1909, XI, 532; ‘“Experimental
Studies of the Influence of Crotalus Venom on the Kidney,’’ Jour. Exper. Med., in
press.

EFFECT OF ANIMAL ExTRACTS UPON THYROID GLAND. 179

animals, attention was called to the lesion by the presence of pale
opaque focal area in the psoas mucsle. Microscopical examination
of these areas showed that the muscle fibers were irregular in
shape, had lost their nuclei and striation and presented the typical
hyaline appearance of Zenker’s degeneration as it occurs in man.
In these animals the adjacent tissue showed no hemorrhage,
exudate or evidence of connective tissue reaction. The animals
died after 18 and 35 days respectively, the first having received
8 injections and the second 7 injections of venom in doses varying
from 0.5 to 2 milligrams.

A third rabbit died on the 38th day after the first injection,
having received nine injections, the last on the 23d day. At
autopsy mottled hemorrhages were seen in the rectus and psoas
muscles and about these hemorrhages, the peculiar opaque, whitish
appearance of hyaline degeneration. Upon microscopical ex-
amination, the picture was identical with that of true Zenker’s
degeneration. Irregular, swollen, vacuolated and varicose, hyaline
fibers, more or less fractured, without nuclei and invaded by
leucocytes occupied large irregular areas. Inthe midst of these
fibers were foci of hemorrhage and throughout an infiltration of
polymorphonuclear leucocytes, while about the necrotic areas
were wide bands of granulation tissue which sent prolongations
between the bundles of muscle fibers. In such areas the surviving
fibers frequently showed multiple nuclei.

Whether or not these lesions have fundamentally a common
relation with those caused by anaphylactic poisons is of course a
matter of doubt. It seems wise, however, to add, in support of
Beneke’s experience with the venom of Crotalus terrificus, these
observations on the effect of the venom of Crotalus adamanteus.

113 (809)
Note on the effect of animal extracts upon the volume of the
thyroid gland.
By Isaac OTT, M.D., and Joun C. Scott, M.D.
The volume of the thyroid was registered by an oncometer and

a modified piston recorder. The arterial tension was also noted.
The animals used were dogs, etherized and with a small dose of

180 SCIENTIFIC PROCEEDINGS (54).

morphia per jugular. Infundibulin per jugular had the most
powerful action in reducing the volume of the thyroid, although at
times there was a preliminary momentary increase. Adrenalin,
after a temporary increase, produced a decrement in the volume.
An infusion of the fresh ovary of a pregnant cat augmented the
size of the thyroid, a fact noted by Hallion. Mammary, corpus
luteum, thyroid, placenta, iodine and parathyroid extracts also
increased the volume. The anterior part of the pituitary decreased
the volume.

114 (810)
The relation of external temperature to hibernation.
By SUTHERLAND SIMPSON.

[From the Physiological Laboratory, Medical College, Cornell
University, Ithaca, N. Y.]

In a former communication! to this society it was shown that
the absence of food is an important factor in determining the
onset of hibernation in the woodchuck (Marmotta monax). In the
present note attention is drawn to the fact that the cause of the
awakening of these animals from their torpid condition in the early
spring is not a rise in the temperature of their surroundings.

A colony of woodchucks was kept in artificial burrows a little
over four feet? below the surface of the ground, as already de-
scribed.? At the bottom of one of these burrows, the oil bulb of a
Friez thermograph was placed, and connected with the recording
clock-drum contained in a box at the top. All the burrows were
packed with dry straw, while the one containing the bulb was
shut off from the central court, to prevent the woodchucks having
access to it.

A continuous record of the temperature at this depth has been
kept from January I, 1912, till the present time. It shows that the
lowest temperature is reached late in March or early in April—
just about the time when the hibernating woodchucks are begin-
ning to wake up. There is no appreciable rise in temperature

1The Food Factor in Hibernation, Proc. Soc. EXPER. BIOL. AND MED., Vol.
9 (April 17, 1912), p. 92.

2 Accurate measurement shows the burrows to be a few inches over four feet
below the surface and not five feet as formerly stated.

3 Loc. cit.

ANAPHYLAXIS IN IMMUNE ANIMALS. 181

till well on in April. The diurnal variation, so marked at the
surface, is almost completely abolished at this level, all the year
round, and this is a circumstance which greatly favors animals
with an imperfectly developed heat regulating mechanism, such
as the woodchuck possesses.

Records of the air temperature taken at the Ithaca station
of the U. S. Weather Bureau, situated about half a mile from the
burrows, show that in 1912 the coldest month of the year was
January, and in 1913, February, the average mean temperatures
for the first four months being as follows:

Jan. Feb. March. April.
VONCIA Sit oe Kanto ays T63c) he 21.4 28.8 44.5
TODS ene: cccteee suse AVG PY 1B, 2302 36.8 48.1

Notwithstanding the fact that the weather in these four months
was much milder in 1913 than in 1912, the temperature at the
depth of four feet, in March and April 1913, as indicated by the
thermograph, was about 2° F. lower than in the corresponding
months of 1912. The snowfall, however, was greater in I912 than
in 1913 and this will probably explain the apparent anomaly.

Jan. Feb. March. April.
Snowfall in inches, 1912... .9.0 6.7 13.3 0.5
Snowfall in inches, 1913... .2.1 5.0 2.9 0.8

In 1912, although the air was intensely cold, the comparatively
thick layer of snow effectively retarded the radiation of heat from
the surface of the ground.

It is interesting to note, then, that the woodchuck awakes from
the hibernating state and becomes active just about the time the
temperature of its surroundings has reached the lowest point for
the year, and it would appear that some cause other than the
temperature or carbon dioxide factor is at work to bring this result
about.

I15 (811)
Anaphylaxis in immune animals.
By RICHARD WEIL, M.D.
[From Cornell University Medical School.|

In his general review of anaphylaxis, Besredka! stated that an
injection of horse serum into guinea pigs produced hypersensitiza-

1 Kraus and Levaditi’s ‘‘Handbook,”’ Ig1t, p. 248.

182 SCIENTIFIC PROCEEDINGS (54).

tion, only if minute doses were employed. ‘‘ The preliminary
injection of more than .02 cubic centimeter is attended by an
uncertain result.’’ Similar statements abound in the literature.
The great majority of the experiments on which these conclusions
are founded have employed either the intracerebral or the intra-
peritoneal route for the intoxicating injections. If, however, the
second injection is made intravenously, the results are entirely
different. With this method, it can be shown that a preliminary
injection of as much as five cubic centimeters of horse, or other
serum, is invariably followed by a typical anaphylactic state, in
which death is produced by the second injection of an amount of
the same serum, which, for normal animals, is absolutely in-
nocuous. If guinea pigs are given repeated large injections, e. g.,
3 c.c. of horse serum, on three or four successive days, the total
amounting to nine or twelve cubic centimeters, exactly the same
result follows. Finally, if spaced injections are employed, as in
the procedure followed for purposes of immunization, again the
animals become typically anaphylactic. There are, however,
certain differences in the behavior of animals sensitized, on the one
hand, by means of minute, and, on the other, by very massive
injections of serum. In the former, the minimal lethal dose is
often no greater than .005 cubic centimeter of horse serum. In
the latter, it is often one hundred times this amount. Again,
after a small injection, anaphylaxis frequently does not develop
for three weeks; after massive injections, it may generally be
demonstrated in ten or twelve days. These differences are trace-
able simply to the effects exercised upon antibody production by
small, as compared with large doses of antigen. With the former,
the antibodies are produced slowly and in small number; with
the latter, rapidly and in large number. Thus, in the case of
guinea pigs sensitized by small doses, it is generally necessary to
use the entire amount of blood obtainable by exsanguination in
order to sensitize another normal pig. After large injections,
however, one tenth of the total blood may suffice for this result.
It is probably on account of the larger amount of circulating
antibody that guinea pigs sensitized by large doses require a
larger toxic injection.

THE PARATHYROIDS AND PREGNANCY. 183

116 (812)
The parathyroids and pregnancy.

By A. J. CARLSON.

[From the Hull Physiological Laboratory, University of Chicago.]

A number of investigators have found, especially in dogs, that
pregnancy seems to augment the symptoms of parathyroid tetany,
and may transform latent tetany into acute tetany in cases of
partial thyroidectomy.! The parathyroids must become func-
tional in the fetus a considerable time before birth, because there
is no record of parathyroid tetany (acute or latent) in prematurely
born infants. In view of the fact that in late pregnancy the pan-
creas of the fetus seems to be able to compensate for the total
extirpation of the pancreas of the mother to such an extent that this
operation is not followed by the usual diabetes,? an investigation
of the other organs of internal secretion with reference to the
passage of the hormones from the blood of the fetus to the blood
of the mother seemed desirable. Accordingly, in the spring and
summer of 1912, thyroid-parathyroidectomy was made in 16 dogs
and 11 cats in late pregnancy, that is, within one to three weeks of

term.
RESULTS.

1. Dogs.—Eight out of the fifteen dogs died in acute tetany
within 12 to 24 hours after the parathyroidectomy. In no case
was there evidence of labor. Three dogs developed tetany after
36 hours, one dying in tetany on the third day, and the other two
on the fourth day without any signs of labor. Two dogs gave
birth to the young on the second day and developed tetany symp-
toms on the third and fourth days respectively. These two died
in tetany and depression on the eighth day.

One dog had moderately strong tetany on the second and third
days after the operation. On the 4th and 5th days there was no
tetany, but one pup was born on each of these days. The dog
died in depression on the 6th day. Five pups were found in the
uterus. One of the pups was engaged, but the uterine contractions

1 The literature is reviewed by Biedl, ‘‘Innere Sekretion,”’ 1913, I, p. 83.
2 Carlson and Drennan, Am. Jour. of Physiol., 1911, XXVIII, p. 391.

184 SCIENTIFIC PROCEEDINGS (54).

had evidently been too feeble or incoordinated for the delivery.
Two dogs showed no signs of tetany at any time, one of the dogs
giving birth to the young on the 6th day, and the other on the 9th
day following the parathyroidectomy. Three weeks later the
dogs were killed and a search made for accessory parathyroids;
none were found.

The development of acute and fatal tetany within 24 hours
after parathyroidectomy is very exceptional in male and non-
pregnant female dogs. It does occur, however, especially after
hemorrhage or prolonged anaesthesia. It would therefore seem
that late pregnancy in dogs accelerates and intensifies the para-
thyroid tetany in the majority of cases. This may, however,
be only an addition effect (latent eclampsia + parathyroid tetany).

2. Cats.—Ten out of the eleven pregnant cats developed fatal
tetany. The slight difference between pregnant and non-pregnant
cats is shown by the following comparison:

Parathyroid tetany ; average figures for:

20 males and non-pregnant females. 10 females in late pregnancy.
Maximum...... 72 hours 48 hours
First tetany symptoms4 Minimum....... 5 hours 24 hours
ii {sii = cea Sheree 28 hours 30 hours
Maximum...... 168 hours 175 hours
Deathy Minimum....... 24 hours 75 hours
| sine CA eye ae 78 hours 120 hours

It is obvious that advanced pregnancy in cats does not accelerate
or intensify the symptoms of parathyroid tetany. On the contrary
the pregnant cats live a little longer and the excitation symptoms
are less severe than in the case of the non-pregnant controls. But
the depression seemed greater in the pregnant animals. If the
fetal parathyroids are in any way capable of functioning for the
mother, this action is so slight that it cannot be demonstrated
on the present short series of experimental animals.

It is premature to speculate on the cause of the above difference
in the relation of pregnancy to parathyroids in cats and dogs
before more data are at hand on other species.

PHYSIOLOGICAL STATE OF THYROID OF MOTHER AND FETUS. 185

117 (813)

The correlation between the physiological state of the thyroid of
the mother and of the fetus.

(PRELIMINARY NOTE.)
By A. J. CARLSON.

[From the Hull Physiological Laboratory of the University of
Chicago.|

The starting point of this investigation was some observations
(incidental to other work on the thyroids) that pups born of
mothers having active hyperplasia of the thyroids seemed to have
much larger thyroids than the pups born of mothers with normal
thyroids or with colloid goiters. The size of the thyroid in pups
from mothers with marked thyroid hyperplasia is in many cases
so great that they produce the distortion of the neck similar to
goiter in adults. These pups are apparently born with goiter.

The work was begun in the spring of 1912, and so far data have
been obtained on mother and offsprings in the case of 16 cats and
14 dogs; the work is being continued and extended to other species.

It is well known that goiter (active hyperplasia and colloid)
is prevalent in dogs in the Great Lakes region of United States,
while in cats in the same region goiter is practically unknown.
The goiter of the newborn of mothers with thyroid hyperplasia
may be (1) primarily hereditary, that is, due to defects in the
germ plasm, or (2) it may be due to some temporary metabolic
disturbance in the mother,—toxins or abnormal concentration of
normal products of metabolism, acting alike both on the maternal
and on the fetal thyroid. If the fetal goiter is due primarily to
the constitution of the ovum rather than to the maternal environ-
ment during intrauterine life, we would expect the goitre to persist
in varying degrees after birth. We would also expect to meet with
fetal goiter in the case of mothers with colloid goiters, because the
colloid state appears to be preceded by active hyperplasia. On
the other hand, if the fetal goiter is due primarily to some intoxica-
tion or temporarily altered metabolism of the mother, acting
alike on the fetal and on the maternal thyroid so as to produce

186 SCIENTIFIC PROCEEDINGS (54).

hyperplasia, there ought to be no fetal goiters in the case of mothers
with normal thyroid or with colloid goiters; and the fetal goiters
of mothers with active hyperplasia ought to diminish after birth.

1. Results in Dogs.—During intrauterine life the body in-
creases in weight faster than the thyroid gland so that the ratio of
the weight of the thyroid to the body weight becomes gradually
larger. But in the case of mothers with normal (or nearly normal)
thyroids, the ratio of thyroid to body weight is always greater in
the mother than in the pups; in the case of mothers with colloid
glands the ratio may be greater in the pups than in the mother
while in the case of mothers with thyroid hyperplasia the ratio
may be the same, or it may be higher or lower, depending on the
degree of hyperplasia of the maternal thyroid.

The newborn of mothers with active hyperplasia have invari-
ably much larger thyroids than the pups of normal mothers, but
the thyroids of the former do not exhibit any greater degree of
differentiation toward adult structure. It would thus seem that
the conditions causing thyroid hyperplasia in the mother lead in
the fetus to excessive growth of the thyroid rather than to specific
thyroid differentiation.

In the case of mothers with colloid goiter the thyroids of the
pups are on the whole of the same size as in the pups born of
normal mothers. A few thyroid ratios may be cited for illustra-

tion.
Ratio of thyroid to body weight.
Mother. Pups.
WV, Normal! thyroid\rt.y 11a rie eicreniele I-6,600 I-—3,000
XX. |Colloidigoiter: <tepeeie «ian clexcielcistereiene I-I,300 I-4,200
III. Thyroid hyperplasia.............. I-I,100 I- 480

On the whole, the relative weight of thyroids in dogs, both
adult and newborn, is large in comparison with that in cats.
There may be considerable variation in the thyroid: body weight
ratio in pups of the same litter. We have not been able to deter-
mine whether this is due to primary difference in the ova or to the
varying factor of accessory thyroids.

2. Results in Cats.—Active hyperplasia or colloid goiter has
not yet been found in our pregnant cats. But there is some vari-
ations in relative bulk of thyroid tissue. Thus the extremes of
thyroid-body weight ratio are 1-4,680, and 1-16,000 respectively.

PARATHYROID TETANY AND ACTIVE IMMUNITY. 187

In general the adult pregnant female weighing 3-3.5 K. has
thyroids weighing (fresh) 0.20 g. to 0.30 g. While thyroid hyper-
plasia has not yet been found in pregnant cats, it is a striking fact
that mothers with relatively large thyroids give birth to kittens
with relatively large thyroids and vice versa. The following

figures may be cited as typical:
Ratio of Ehyroid to body weight.

other. Kittens.
IN Os ENGarsia vane hs Poteet a cueheraca nena sisi ersintshere I-16,250 I-7,326
INOS LW ire stots sretspotetene at aen shad ah gant te she lanesaye I— 4,680 I-2,000

The results so far point to the following conclusions:

1. Active thyroid hyperplasia is not associated with hyper-
secretion of the thyroids, because hypersecretion of the thyroids
in the mother would retard rather than augment thyroid growth
in the fetus.

2. Since fetal goiter is always present in the offsprings of
mothers with active thyroid hyperplasia, and never present in the
offsprings of mothers with normal thyroids or colloid goiter, it
would seem that the fetal goiter is not due primarily to the condi-
tion of the germ cells, but to some intoxication of the mother or
altered condition of the maternal metabolism. Since the maternal
environment acts on the fetus only by the way of the blood, the
goiter must be due to substances in the blood acting alike on the
fetal and the maternal thyroid to produce cell division and growth,
rather than specific thyroid differentiation and secretion. This
hyperplasia is therefore not compensatory.

118 (814)
Parathyroid tetany and active immunity.

By A. J. CaRLSON.

[From the Hull Physiological Laboratory of the University of
Chicago.]

Parathyroid tetany in dogs seems to be associated with dimin-
ished resistance to bacterial invasion of the mucous membranes,
as shown by the frequent infection of the eyes, the nose, and the
respiratory passages. This diminished resistance may be due to
(1) depression of the processes of active immunity; (2) local de-

188 SCIENTIFIC PROCEEDINGS (54).

pression of the cells of the mucous membranes; (3) disturbance of
the body heat regulating mechanism. It is also possible that all
three factors are involved.

In order to determine whether the first possibility is a factor,
dogs were immunized at varying periods before and after para-
thyroidectomy, and the influence of the tetany condition on the
immunity reaction noted. The particular antibody studied was
the lysin developed by the injection of goat erythrocytes into dogs.
The immunizing dose consisted of a single intravenous injection of
I c.c. of a 10 per cent. suspension of goat corpuscles per kilo body
weight of dog.

The results so far obtained show that the active immunity is
decreased by the condition of parathyroid tetany. None of our dogs
developed lysin to goat corpuscles to the degree observed in the
series of normal dogs reported by Hektoen and Carlson, and by
Luckhardt and Becht.!. The greatest concentration of the specific
lysin in our tetany dogs was represented by a serum dilution of
1-8,000; the lowest concentration by a serum dilution of 1-400.
Two typical experiments may be cited:

Dog V. Nov. 1. No lysis in 1-50. Complete thyroidectomy.
Nov. 2. No lysis in 1-50. 13 c.c. goat corp. susp. inj.
Nov. 3. No lysis in 1-50. No tetany.
Nov. 4. No lysis in 1-50. Strong tremors.
Nov. 5. Lysis in 1-50. No tetany.
Nov. 6. Lysis in 1-400. Tetany.
Nov. 7. Lysis in 1-1,600. Violent tetany.
Nov. 8. Lysis in I-2,000. Strong tetany.
Nov. 9g. Lysis in I—2,000. Tetany and depression.
Nov. 10. Lysis in 1-2,000. Tremors and depression (died).
Dog VI. Nov. 2. No lysis in I-50. II c.c. goat corp. susp. inj.
Nov. 3. No lysis in 1-50.
Nov. 4. No lysis in I-50. Complete thyroidectomy.
Nov. 5. Lysis in I-50. Dog seems normal.
Nov. 6. Lysis in 1-400. Tremors.
Nov. 7. Lysis in I-1,000. Mild tetany.
Nov. 8. Lysis in 1-1,600. Violent tetany.
Nov. 9. Lysis in I-2,000. Violent tetany, depression.
Nov. 10. Lysis in 1-—4,000. Tetany and depression.
Nov. 11. Lysis in 1-4,000. Tremors and depression (died).

1Hektoen and Carlson, Jour. of Inf. Dis., 1910, VII, p. 319; Luckhardt and
Becht, Am. Jour. of Physiol., 1911, XXVIII, p. 257.

SOLUBILITY OF LEAD SALTs IN HuMAN GASTRIC JUICE. 189

119 (815)

The solubility of certain lead salts in human gastric juice, and its
bearing on the hygiene of the lead industries.

By A. J. CARLSON and A. WOELFEL.
[From the Hull Physiological Laboratory, University of Chicago.]

I. SOLUBILITY OF WHITE LEADS IN HUMAN GASTRIC JUICE.

WHITE LEAD PAINT DUSTS.

Solubility in pure gastric juice (25 c.c. G.J. to 0.5 gr. lead):

Basicilead: carbonate: paintiGust) rs cse)eusicin seeds 8 oie iaie) sXe) ofelinr sy) salle 'o cers 46 per cent.

Basicileadistlphateypaint Custer. erevsoer alors: clolelssereicie sis) sie/creislans) s+ 5.7 per cent.
Solubility in gastric juice+peptone (25 c.c. G.J., 0.1 gr. peptone, 0.5 gr. lead):

Basicsdead: carbonate) pains Gust aa. iccnect st u-iciss oi oles alarelies cel sie) cleseael ace’: 46 per cent.

Basiqleadsusulphateypain trust. rye circuit cree ele eile etclets telat) aials. viehe 7.3 per cent.
Solubility in gastric juice+milk (G.J. 1-milk 1):

Basicileadicarbonate paintidust. ste (ev< 21 ce niece tee eres e es none =O per cent.

Basiculead sulphates pain tudust-iera nis ciocele lee Glee etal) sierelerene none =0 per cent.
Solubility in 0.5 per cent. HCI (25 c.c. HCl to 0.5 gr. lead):

Basicleadicarbonate: paint GuUSt ie sy eis tale yell sie lee eyeiedar seis a) © 66 per cent.

Basic lead*sulphate paint) dust)... «ali. toreictercrcveteconeicie® 6.0 heuer) ersaiere 6.7 per cent-
Solubility in 0.5 per cent. HCl+milk (HCl 1-milk 1):

Basic leadicarbonate paintidusty, cists). cialeiciensiolel<eicte1a Aoveiciele none =0 per cent.

Basicveadiculphateipaing Gusti.) lero cis aeiciiaeiasiesteiienseere css none =O per cent.
Solubility 0.5 per cent. HCl+milk (HCl 2-milk 1):

Basictleadicarbonate) paint7dust = eters chet crcis.e: oko oo) sidishorehete svete/atarars 25.4 per cent.

Basic leadisulphate) paint Gust acisrattelersieleue ver sleiee = ate clio a eiste mia 1.5 per cent.
Solubility in 0.5 per cent. HCl+milk (HCI 4-milk 1):

Basic leadicarbonatespaint dusts ey. riser cyclers ors oie: nie iahelsrs cceiel eo) sae 83.5 per cent.

Basic lead sulphates paint Gusts si iciterts) ners chels ates = levels sie le ores oe 6.9 per cent.

WHITE LEADS.

Solubility in pure gastric juice (25 ¢.c. G.J. to 0.5 gr. lead):

Weadicarbonate.(@ Old DutcheProcessia) ire ereraensscerses cles sieve suctelens = 53 per cent.

Basicvleadvsl phates ctysscrs ao aya crcterstole toketeneca cock sne! sacienele arate Otererctchs 25 per cent.
Solubility in gastric juice +peptone (25 c.c. G.J. to 0.5 gr. lead):

Meadicarbonate:( Olds utche Process.) arn cicdeneie os ers cteleiete cicneie + 57 per cent.

Basictleadssulpbaterr-trccrs nce ole nie entiasedorsi ol otevehaver dave otuciescrencders 27 per cent.
Solubility in pure gastric juice (50 ¢.c. G.J. to 0.5 gr. lead):

Weadseanponaten(a Old sD utcheProcess=)iejenrsrerere cites eel eine ciaice: 69 per cent.

Basic lead stl phate racy perry cress scote wievetersasvera eels over sesmverene ti wvoyacaver sus 30 per cent.
Solubility in gastric juice+milk (G.J. 4—milk 1):

WeAadhGarDponate syne circverciy a avseie ee ckevere elation -ceie's) Siete oro) oie) creiereie ouayeks 90 per cent.

Basicvlead: sul phateryaict scicsvanie a osits ewer sho seayaye vs.» yee a yereleaa ns 34.8 per cent.

190 SCIENTIFIC PROCEEDINGS (54).

2. TOXICITY OF WHITE LEADS WHEN FED To Docs AND Cats.

The lead carbonate is much more toxic than the lead sulphate.
But both salts produce acute lead poisoning when given in quan-
tities of 0.1 gr. per kilo body weight daily.

3. THE INFLUENCE OF MILK.

When milk and gastric juice is mixed in the proportion of
I-1, the hydrochloric acid of the gastric juice is so completely
fixed by the milk proteins or neutralized by the carbonates in the
milk that the mixture has virtually no solvent action on the lead
salts, but when gastric juice is present in excess the lead salts g.
into solution in proportion to the excess of the gastric juice
When milk is taken into the stomach there occurs, of course, a
similar fixation of the hydrochloric acid, and, in addition, the total
quantity of gastric juice is diminished owing to the inhibitory
action of the fats in the milk on the processes of secretion.

On the basis of our work we venture to offer these two practical
suggestions: (1) The lead carbonate is so much more toxic than the
lead sulphate that lead workers as well as the state should aim at
the elimination of the use of the carbonate in all industries where
this is possible. (2) In addition to taking other important pro-
phylactic measures, the lead workers should drink a glass of milk
between meals (say 10 A.M. and 4 P.M.) in order to diminish the
chances for any swallowed lead to be dissolved by free hydrochloric
acid of the gastric juice, as in some persons there is considerable
secretion of gastric juice in the empty stomach.!

1 Hornborg, Skand. Arch. f. Physiologie, 1904, XV, p. 209; Carlson, Am. Jour.
of Physiology, 1912, XXXI, p. 151.

TYROSINASE IN AMPHIBIAN EGG JELL. 191

120 (816)

Certain observations on the occurrence of tyrosinase in amphibian
egg jell.

By A. M. Banta and Ross AIKEN GORTNER.

[From the Station for Experimental Evolution, The Carnegie
Institution of Washington.|

In experiments having for their object the modification of
pigment in amphibian larvae fresh eggs and young embryos of the
following species of amphibians were placed in tyrosin solutions
of 0.025 per cent. concentration: Ambystoma punctatum (L.),
Spelerpes bilineatus (Green), Bufo lentiginosus Shaw, Hyla picker-
ingit Holbrook, Rana clamitans Lat. and Rana sylvatica Le Conte.

No coloration of the egg membranes or jell appeared in either
Ambystoma or Spelerpes. There were some indications of colora-
tion in Bufo jell but the coloration was so slight as to be inconclu-
sive. Hyla egg jell showed a pink to violet coloration within 24
hours, after being placed in the solution, and the solution itself
took on much the same color. Within 2 or 3 days the jell was
much blackened, and the solution was fairly dusky with suspended
humin. The egg jell of Rana clamitans showed some darkening
within a day and after forty-eight hours was conspicuously black-
ened while the ‘solution was likewise very perceptibly darkened.
Rana sylvatica egg jell took on the rose coloration within one to
three hours after being placed in the solution, and the colora-
tion rapidly proceeded through violet to an intense black so that
the eggs were invisible at a depth of 3-4 cm.

These results indicate the presence of tyrosinase in the egg
jell of Hyla pickeringit, Rana clamitans and Rana sylvatica, and
possibly in Bufo, but not in Ambystoma punctatum or Spelerpes
bilineatus.

If the slight evidence of tyrosinase in Bufo is taken to indicate
its actual occurrence there, tyrosinase is found in the egg jell of
four species of Anura but is absent in the two species of Urodela
examined. This may possibly prove to be an additional dis-
tinguishing character between these two orders of amphibians.

192 SCIENTIFIC PROCEEDINGS (54).

121 (817)

Experiments on the light and tactile reactions of a cave variety
and an open water variety of an amphipod species.

By A. M. BAnNTa.

[From the Station for Experimental Evolution, The Carnegie
Institution of Washington.|

The amphipod, Eucrangonyx gracilis (Smith), occurs in the
waters of many caves in southern Indiana and is very generally
and abundantly distributed in small surface streams in the same
region. The individuals occurring in caves almost without excep-
tion have very little or no body pigment though possessed of the
normal pigment in the eyes. Those living in surface streams
have the usual amount of pigment for a crustacean. In other
structural characters the forms are apparently alike.

In an attempt to determine whether there was a physiological
difference between the two forms they were tested for light and
tactile reactions. These amphipods are negative to all intensities
of light to which they respond. Extended series of light experi-
ments using horizontal illumination were conducted during which
every effort was made to maintain as precise conditions as possible.
The cave form was found to be somewhat less responsive to light
than the form living outside caves, but this difference was not
large and was distinctly observable only with relatively small
intensities of light.

In order to test the tactile reactions a delicate camel’s hair
fastened to the end of a glass rod was used to stimulate various
parts of the body of several individuals of the cave form and the
same number (of equal size) of the surface form. Complete tabu-
lations of the nature and vigor of each response, when a response
occurred, were kept and failures to respond were likewise recorded.
This data showed a greater number of failures to respond and on
the average a slightly less vigorous response on the part of the
surface form as compared with the cave form.

Hence the cave form appears to be less responsive to light and
more responsive to tactile stimulation than its outside relative.

RECAPITUESHON, Ob wine NAMES »OF
THE AUTHORS AND OF THE-TITLES
OF THE COMMUNICATIONS:

VOLUME X.
Atkinson, J. P.
766. [with C. B. Fitzpatrick.| On the presence of pressor
substances in experimental immunity.
787. [with C, B. Fitzpatrick.] On some vaccinia blood
pressor substances in rabbits.
Auer, J.
795. See Meltzer, S. J.
Austin, J. H.
802. [with C. B. Farr.| The total non-protein nitrogen
of the blood in nephritis and allied conditions.
Bancroft, F. W.
776. Heliotropism and galvanotropism in Euglena.
Banta, A. M.
816. [with R. A. Gortner.| Certain observations on the
occurrence of tyrosinase in amphibian egg jell.
817. Experiments on the light and tactile reactions of a
cave variety and open water variety of an amphipod species.
Banzhaf, E. J.
751. See Park, W. H.
Bronfenbrenner, J.
717. See Manwaring, W. H.
743. See Manwaring, W. H.
Burnett, T. C.
788. See Robertson, T. B.
789. See Robertson, T. B.
790. Note on the relation of alexin to thrombin.
Butterfield, E. E.
748. The reaction between oxygen and hemoglobin.
794. The color index and color of the red blood corpuscles,
193

194 SCIENTIFIC PROCEEDINGS (54).

Calkins, G. N.
742. Further light on the conjugation of paramecium.
Carlson, A. J.

812. The parathyroids and pregnancy.

813. The correlation between the physiological state of
the thyroid of the mother and of the foetus.

814. Parathyroid tetany and active immunity.

815. [with A. Woelfel.] The solubility of various lead
salts in the human gastric juice and its bearing on the hygiene
of the lead industries.

Chase, A. F.

758. See MacNeal, W. J.
Clark, P. F.

697. See Flexner, S.
Clowes, G. H. A.

744. Certain specific reactions in hay fever.

745. Treatment of autumnal hay fever by vaccination.

767. [with F. C. Goldsborough and F. West.] On a
complement deviation reaction exhibited in pregnancy.

768. [with F. C. Goldsborough.] On the antitryptic re-
action exhibited in pregnancy.

Cohn, A. E.

702. On the differences in the effects of stimulation of the
two vagus nerves in the dog’s heart.

722. The production of reversed cardiac mechanism in
the dog.

759. The effects of morphine on the mechanism of the

dog’s heart after removal of one vagus nerve.
Cohoe, B. A.

785. See Rosenbloom, J.
Cooke, R. A.
724. The fate of parenterally introduced glycogen in
human and experimental diabetes.
Corson-White, E. P.
806. See Sweet, J. E.
Crawford, A. C.
739. [with J. P. Crawford.] Note on the cock’s comb
test for the activity of ergot.

NAMES OF AUTHORS. 195

Crawford, J. P.
739. See Crawford, A. C.
Dresbach, Melvin.
701. [with S. A. Munford.] Interpolated extra-systoles,
of frequent occurrence, in an otherwise normal heart.
Du Bois, E. F.
754. See Wiggers, C. J.
Edelmann, L.
805. See Murlin, J. R.
Ewing, E. M.
782. See Janeway, H. H.
Famulener, L. W.
751. See Park, W. H.
Farr, C. B.
802. See Austin, J. H.
Fine, M. S.
703. See Myers, V. C.
704. See Myers, V. C.
801. [with V. C. Myers.] Further studies on muscle
creatine.
Fitzgerald, J. G.
734. Agglutination of encapsulated bacteria.
735. Relative frequency of B. coli communior in con-
taminated water.
736. [with C. L. A. Schmidt.] Production of creatinine
by bacteria.
792. [with Gertrude Macintosh.] Action of gentian
violet on Mucosus capsulatus group.
Fitzpatrick, C. B.
766. See Atkinson, J. P.
787. See Atkinson, J. P.
Flexner, S.
697 [with P. F. Clark.] The mode of infection in experi-
mental poliomyelitis.
Foster, N. B.
719. The effect of phlorhizin on dogs with Eck fistula.
786. Sugar from lactic acid in human diabetes.

196 SCIENTIFIC PROCEEDINGS (54).

Gay, F. P.
772. [with T. Brailsford Robertson.] Studies on the
nature of biological specificity.
Giles, R. C.
805. See Murlin, J. R.
Githens, T. S.

715. [with S. J. Meltzer.] Differences in the toxic effects
of ether and chloroform as observed under intratracheal
insufflation.

725. Decerebration and the action of morphine in frogs.

760. The influence of temperature on the minimal dose of
strychnine in frogs and the time of onset of tetanus.

780. [with S. J. Meltzer.] Antitoxic action of sodium
iodide on morphine.

Goldfarb, A. J.
755. The influence of the central nervous system in re-
generation of the annelid worm.
756. Changes in concentration of sea water and their
influence upon regeneration.
784. Experimentally fused larvae.
Goldsborough, F. C.

767. See Clowes, G. H. A.

768. See Clowes, G. H. A.
Haas, S. L.

737. Regeneration of bone from periosteum.
Halstead, W. S.

770. Hypertrophy of the thyroid gland. Revision of
experiments made 25 years ago.

771. Partial occlusion of the aorta by bands of living
tissue.

Hess, A. F.

750. The pancreatic lipase of infants in acute intestinal

disturbances.
Hunter, A.
763. The influence of experimental cretinism upon nitrog-
enous metabolism in the sheep.
James, Henry.
705. See Williams, H. B.
726. See Williams, H. B.

NAMES OF AUTHORS. 197

Janeway, H. H.

782. [with E. M. Ewing.] The mechanical factors of
excessive artificial respiration and a consideration of their
relation to the acapnial theory of shock.

Jordan, H. E.

710. Heterochromosomes in mammals.

727. The intercalated discs of atrophied heart muscle.
Kendall, E. C.

799. The chemical constituents of the thyroid gland.

800. The specific physiological activity of certain con-
stituents of the thyroid gland.

Kleiner, I. S.

720. On elimination through the mucosa of the urinary

bladder.
Kramer, B.

706. The rdle of lipoids, and particularly lecithin in
narcosis.

804. See Murlin, J. R.

Lambert, R. A.
777. The fat content, morphology and length of life of
cells growing in diluted blood plasma.
Macintosh, Gertrude.
792. See Fitzgerald, J. G.
MacNeal, W. J.

758. [with A. F. Chase.] Some observations on bacteria

of the duodenum.
MacNider, W. deB.

761. The difference in the effect of Gréhant’s anesthetic
and of morphine-ether on the total output and composition of
the urine in normal dogs.

Manwaring, W. H.

698. [with J. Bronfenbrenner.] Effects on meningeal
tuberculosis of the local injection of foreign leucocytes.

716. On the relation of leucocytic extract to body fluids.

717. [with J. Bronfenbrenner.] The Kraus-Hofer phe-
nomenon in tuberculosis.

743. On lysis of tubercle bacilli.

198 SCIENTIFIC PROCEEDINGS (54).

McClendon, J. F.

778. The rate of absorption of water by the skin of the
frog, in relation to M. H. Fischer’s theory of edema.

779. The dynamics of a model of cell division.

McCrudden, F. H.
699. The importance of calcium for growth.
Meigs, E. B.

781. On the nature of the semipermeable membranes

which surround the fibers of striated muscle.
Meltzer, S. J.

712. The effects of strychnine in cardiectomized frogs
with destroyed anterior lymph sacs; a demonstration.

713. See Wollstein, Martha.

714. See Wollstein, Martha.

715. See Githens, T. S.

780. See Githens, T. S.

795. [with J. Auer.] Combined action of magnesium
and ether; evidence for a central effect of magnesium.

Meyer, G. M.
723. See Van Slyke, D.
Morse, Max.
718. The réle of phagocytosis in involution.
Munford, S. A.
701. See Dresbach, M.
Murlin, J. R.

804. [with B. Kramer.] The influence of pancreatic and
duodenal extracts on the glycosuria and the respiratory
metabolism of depancreatized dogs.

805. [with L. Edelmann and R. C. Giles.] The carbon
dioxide and oxygen content of the blood after ligation of the
abdominal aorta and the inferior vena cava.

Murphy, F. T.
y721. See Pratt; J. H:
Myers, V. C.

703. [with M.S. Fine.] The creatine content of muscle
under normal conditions. Its relation to urinary creatinine.

704. The creatine content of muscle during starvation
and its relation to urinary creatine.

801. See Fine, M. S.

NAMES OF AUTHORS. 199

Ophuls, W.

732. Chronic lead poisoning in guinea pigs. Its relation
to polyserositis.

733. Chronic lead poisoning in guinea pigs. Its relation
to chronic nephritis.

Oppenheimer, B. S.

753. [with H. B. Williams.] Prolonged complete heart
block without lesion of the bundle of His with frequent
changes in the idio-ventricular complexes.

Ott, Isaac.

708. [with J. C. Scott.] The action of glucose upon the
amount of epinephrin in the blood.

709. [with J. C. Scott.] The inhibitory and synergistic
hormones of the secretion of milk.

730. [with J. C. Scott.] Note on the action of tonsillar
extract.

809. [with J. C. Scott.] Note on the effect of animal
extracts upon the volume of the thyroid gland.

Park, W. H.

751. [with L. W. Famulener and E. J. Banzhaf.] Influ-
ence of protein concentration upon the absorption of anti-
bodies from subcutaneous tissues.

Pearce, R. M.

746. See Williams, P. F.

808. Waxy degeneration of muscle in venom intoxication.
Pike, F. H.

749. See Wilson, J. G.
Pratt, J. H.

721. [with F. T. Murphy.] Pancreatic transplantations
in the spleen.

Reichert, E, T.

728. The differentiation and specificity of starches in rela-

tion to genera, species, etc.
Robertson, T. B.

738. [with T. C. Burnett.] The influence of lecithin and
cholesterin upon the growth of tumors.

772. See Gay, F. H.

773. On the nature of Oécytin; the fertilizing and cytolyz-
ing substance in mammalian blood-sera.

200 SCIENTIFIC PROCEEDINGS (54).

774. On the influence of lecithin upon the development
of sea-urchin embryos.

788. [with T. C. Burnett.] Preliminary communication
on the part played by cholesterol in determining the incidence
of carcinoma.

789. [with T. C. Burnett.] The influence of digitonin
upon the growth of carcinoma.

Robinson, G. Canby.
752. The influence of the vagus nerves on the faradized
auricles in the dog’s heart.

Rosenbloom, J.

731. The diffusion of iodo-eosin from ether through
rubber into ether.

764. See Stetten, DeWitt.

785. Metabolism studies in a case of myotonia atrophica.

Rous, Peyton.

796. Note on the production of acid by tissues growing

in vitro.
Rusk, G. Y.

711. A comparison of chemical with micro-chemical
methods for the determination of varying amounts of glycogen
in the liver.

Salant, W.
803. [with C.S. Smith.] The toxicity of sodium tartrate
with special reference to diet and tolerance.
Saxon, G. M.
806. See Sweet, J. E.
Schmidt, C. L. A.
736. See Fitzgerald, J. G.
Scott, E. L.

765. The relation of pancreatic extract to the sugar of

the blood.
Scott, J. C.

708. See Ott, I.

709. See Ott, I.

730. See Ott, I.

809. See Ott, I.

NAMES OF AUTHORS. 201

Sears, H. J.

775. On acid agglutination as a method of differentiation

of bacteria.
Simon, C. E.

807. [with M. A. Wood.] On the inhibitory action of

certain anilin dyes upon bacterial development.
Simpson, Sutherland.

700. The pyramid tract in the porcupine.

729. The motor cortex and pyramid tract in the raccoon.

762. The rate of growth in the dog.

810. The relation of external temperature to hibernation.

Smith, C.S.
803. See Salant, W.
Stetten, DeWitt.

764. [with J. Rosenbloom.] Metabolism studies in a

case of hypopituitarism, with infantilism of the Lorain type.
Stockard, C. R.

797. The location of the optic anlage in Amblystoma and

the interpretation of certain eye defects.
Sweet, J. E.

806. [with E. P. Corson-White and G. M. Saxon.] The
influence of certain diets upon the growth of experimental
tumors.

Terry, B. F.

757. Variations in the amount of transformed atoxyl
(trypanotoxyl) produced by varying the strength of atoxyl
incubated with blood.

Todd, J. L.
783. A note on the transmission of spirochaetes.
Van Slyke, D.
723. [with G. M. Meyer.] The distribution of amino
acids in the body.

von Adeling, E.

740. An experimental study of poison oak,
Weil, R.

769. On a new factor in anaphylaxis.

811. Anaphylaxis in immune animals.
West, F.

767. See Clowes, G. H. A.

202 SCIENTIFIC PROCEEDINGS (54).

Wiggers, C. J.
754. [with E. F. DuBois.] Methods for the production
of temporary valvular lesions.
Williams, H. B.
705. [with Henry James.] Reversal of the cardiac
mechanism.
726. [with Henry James.] Reversal of the cardiac
mechanism; an additional note.
753. See Oppenheimer, B. S.
Williams, P. F.
746. [with R. M. Pearce.] Experience with Abderhal-
den’s test for pregnancy.
Wilson, D. Wright.
798. The occurrence of betaine in the muscles of inverte-
brates.
Wilson, J. G.
749. [with F. H, Pike.] A demonstration of the effects
of electrical stimulation of the labyrinth of the ear.
Wollstein, Martha.
713. [with S. J. Meltzer.] Pulmonary lesions produced
by intrabronchial insufflation of cultures of B. megatherium.
714. [with S. J. Meltzer.] The development of experi-
mental pneumonia; a demonstration.
Wood, M. A.
807. See Simon, C. E.
Woodruff, L. L.
747. The kernplasma relation during the life of a pedi-
greed race of Oxytricha fallax.
Yerkes, R. M.
707. Habit and its relations to the nervous system in the
earth worm.
Young, Stewart W.
741. See Zinsser, H.
793. On the influence of light on the electrical potential
of bacterial and other suspensions.
Zinsser, H.
741. [with Stewart Young.] The failure of union between
antigen and precipitin when present in the same serum.
791. On the nature of the union of alexin with specific
precipitates.

EXECUTIVE, PROCEEDINGS.

Fiftieth meeting.

Cornell University Medical College, October 16,1912. President
Ewing in the chair.

Members present: Auer, Benedict, Brooks, Burrows, Butter-
field, Cohn, Eisenbrey, Field, Githens, Goldfarb, Kleiner, Lee,
Longcope, McClendon, McCrudden, Manwaring, Murlin, Norris,
Pike, Robinson, Senior, Simpson, Swift, Terry, Wadsworth,
Wallace, Wiggers, H. B. Williams.

Members elected: E. A. Park, M. Sittenfeld.

Fifty-first meeting.

The Rockefeller Institute for Medical Research, December 18,
Io12. President Ewing in the chair.

Members present: Atkinson, Auer, Bancroft, Beebe, Berg,
Brooks, Cohn, E. M. Ewing, James Ewing, Field, Fitzpatrick,
Gies, Githens, Goldfarb, Hatcher, Kleiner, Lee, Lusk, McCrudden,
MacNeal, J. A. Mandel, Manwaring, Meltzer, Norris, Oppen-
heimer, Pratt, Rous, Senior, Sittenfeld, Swift, Terry, Van Slyke,
Wadsworth, Wallace, H. B. Williams.

Members elected: Jacob Bronfenbrenner, P. F. Clark, Melvin
Dresbach, Paul J. Hanzlik, Arthur D. Hirschfelder, J. B. Murphy,
V. C. Myers, E. L. Scott.

Fifty-second meeting.

College of Physicians and Surgeons, February 19, 1913. Prest-
dent Ewing in the chair.

Members present: Atkinson, Auer, Austin, Bancroft, Beebe,
Bronfenbrenner, Butterfield, Calkins, Clowes, Cohn, Cole, Draper,
J. W. DuBois, Dunham, Eisenbrey, Elsberg, Emerson, Ewing,
J., Famulener, Fitzpatrick, Flexner, Gies, Githens, Goldfarb,
Halstead, Harris, Hess, Jackson, Jacobs, Joseph, Kast, Kleiner,
Lambert, Lee, Levin, I., Levene, P. A., Longcope, Lusk, Mac-

203

204 SCIENTIFIC PROCEEDINGS (54).

Callum, W. J., MacNeal, McCrudden, Mandel, J. A., Manwaring,
Meltzer, Murphy, Myers, Norris, Oppenheimer, Pappenheimer,
Park, Pearce, Pike, Ringer, Robinson, Rous, Scott, Senior, Sitten-
feld, Stewart, H. A., Steinhart, Terry, Wadsworth, Wallace, Weil,
Wiggins, Williams, Anna, Williams, H. B., Wood.

Members elected: R. A. Cecil, C. Eggleston, G. K. Falk,
Davenport Hooker, P. E. Howe, C. J. West.

Officers elected: President, James Ewing; Vice-President, Cyrus
W. Field; Treasurer, Charles Norris; Secretary, Holmes C.
Jackson.

This was the tenth annual meeting of the Society, permanent
organization of which was effected on February 25, 1903. At
the conclusion of the regular meeting, the tenth anniversary was
celebrated by an informal dinner at the Hofbraii Haus at which an
unusually large number of non-resident members was present.
The dinner closed with speeches congratulatory in character.
The following attended: Auer, Bancroft, Butterfield, Bronfen-
brenner, Clowes, Cohn, Draper, DuBois, Dunham, Eisenbrey,
Ewing, J., Field, Fitzpatrick, Foster, Gies, Githens, Harris, Hess,
Jackson, Jacobs, Janeway, H. H., Joseph, Kast, Kleiner, Larhbert,
Lee, Levene, Levin, Lieb, Loeb, J., Longcope, Lusk, MacNeal,
McCrudden, Mandel, A. R., Mandel, J. A., Manwaring, Meltzer,
Meyer, Mosenthal, Murlin, Myers, Norris, Noguchi, Oppenheimer,
Pappenheimer, Park, W. H., Pearce, Pike, Ringer, Robinson,
Rous, Scott, G. G., Scott, E. L., Senior, Sittenfeld, Stewart, Swift,
Terry, Van Slyke, Wadsworth, Wallace, West, Wiggers, Williams,
H. B., Wood.

Fifty-third meeting.

University and Bellevue Hospital Medical College, April 16,
1913. President Ewing in the chair.

Members present: Bancroft, Beebe, Bronfenbrenner, Brooks,
Butterfield, Eisenbrey, Ewing, E. M., Ewing, J., Field, Fitzpatrick,
Githens, Goldfarb, Jackson, Kleiner, Lambert, McClendon, Meigs,
Meltzer, Murphy, Norris, Pike, Stockard, Wastenays, Wiggers.

Members elected: A. O. Gettler, J. A. Hartwell, G. G. Scott,
Martha Wollstein.

EXECUTIVE PROCEEDINGS. 205

Fifty-fourth meeting.

Zoological Department, Columbia University, May 21, 1091}.
President Ewing in the chatr.

Members present: Atkinson, Auer, Austin, Bronfenbrenner,
Butterfield, Calkins, Eisenbrey, Ewing, J., Field, Fitzpatrick,
Jackson, Kleiner, Meigs, Meltzer, Morgan, Murlin, Myers, Norris,
Oppenheimer, Rous, Salant, Scott, E. L., Stockard, Terry,
Wiggers, Wilson, Wood.

Members elected: A. H. Caulfeild, J. W. Churchman, H. R.
Gaylord, H. H. Janeway, J. R. Mohler, A. R. Moore, R. Pember-
ton, G. B. White.

REGISTER OF NAMES AND ADDRESSES
OP TEE MEMBERS OF THE SOCIETY
FOR EXPERIMENTAL BIOLOGY
AND MEDICINE.

ABBOTT; ALEXANDER) Gere on ce accenirid dae mas University of Pennsylvania.
ABREU, JOHN ME socctertace de ars cisieietuaud wavs ster ena ai oeters Johns Hopkins University.
ADAMPNI hy GEORGE a yeti acieranhae iis rao ee aieeee! McGill University, Montreal.
ADLER, HERMAN M......... Psychopathic Hospital, 74 Fenwood Road, Boston.
ADLER ISAAC H Haters ehaycieiets toustenshes seus artis New York Polyclinic Medical School.
ALSBERG, CARL...... U. S. Department of Agriculture, Washington, D. C.
ANDERSON, JOHN F......... U. S. Public Health and Marine-Hospital Service,
Hygienic Laboratory, Washington, D. C.
ATKINSON, JAMES) Peer nctcs sie Department of Health, New York City.
PAUIERTDIOHING ope te sie esterday ears alee Rockefeller Institute for Medical Research.
PATISTING ae) cde wecttccs Nayatenrie meet ayes oc was. suniebetey eit aleve University of Pennsylvania.
BANCROET We Wasenrpuneccasies Rockefeller Institute for Medical Research.

Banta, A. M...Carnegie Institution, Station for Experimental Evolution, Cold
Spring Harbor, Long Island, N. Y.

BANZHAES (EDWIN) Jemcnioe sce es clo. Department of Health, New York City.
BARDEEN? CHARLES Ririnte iste sete skoiers 9S eile ntile sok wre University of Wisconsin
BEEBE OUGAGH bsceve cial ores seks eysrcterers ts) aera Cornell University Medical College.
BENEDICT, FRANcIs G...Nutrition Laboratory, Carnegie Institution, Boston.
BENEDICT STANLEY, Ravine aa ceieliscee rete Cornell University Medical College.
BerG, WILLIAM N.....U. S. Department of Agriculture, Washington, D. C.
BERGEV DAVID IH sclshiz, fae o gore cselere snc jeienca traveisl « University of Pennsylvania.
BIRCHARD Heenan: U. S. Department of Agriculture, Washington, D. C.
BRONFENBRENNER, JACOB........ Rockefeller Institute for Medical Research.
BROOKS PELARU OW pratcntiacrts aot teh a cle ee ea et cne aise PIN ohare New York University.
BUNTING WG lato hetacs ce he nae fone OR ROR eee University of Wisconsin,
BURROWS; Ma donee acne eee Cornell University Medical College.
BURTON-OPIIZH RUSSELL sever jek tou tolers ciceerenetiotovehs fevenerevecie Columbia University.
BUSTEREIED DY Ea Beenie se cela Rockefeller Institute for Medical Research.
BUXTON BER ees ed tes eae Cornell University Medical College.
CALKINS a GARVaE Nines nominee cierto eee see nines Columbia University.
CANNONS IWAUTEREB ris sror ncitaeinlrevulnie ek wreltner ee teoieiicme Harvard University.
CARLSON PAG Mitriie a icketcicueig eed fete nie bonne kis Seite University of Chicago.
GWARREES AUEXISH tots cs cera Rockefeller Institute for Medical Research.
GRCTIN IR, FAL ne nie crate teers ei neers Presbyterian Hospital, New York City.

207

208 SOCIETY FOR EXPERIMENTAL BioLoGy AND MEDICINE.

CHITTENDEN, ROH caexs fasinates Yale University.
CURR KAUR EEC. ren ran eke (a eee Rockefeller Institute for Medical Research.
CLOWES, "Gs AA ee en he ere eras Gratwick Laboratory, Buffalo, N. Y.
COCA PACH raat ay Meee ae See Cornell University Medical College.
COHN; PAUEREDSE eee eee nee Rockefeller Institute for Medical Research.
Oo) 515 Dn Pape Oe a eI Re A University of Wisconsin.
Core RUFUSTI Ceo eee Rockefeller Institute for Medical Research.
COLLINS, KATHARINE R:..........5... State Board of Health, Atlanta, Ga.
CONKLIN SEDWINIG.. 7 epee ete eee Princeton University.
COOKE Jeo Vise iacietant Gee ee ee eee University of California.
COUNCILMAN A WILLYAMMI Eee eee eee EEE Ene Harvard University.
CrAmpTon, ©. WARD) 25.-520 0000 Department of Education, New York City.
CRAMPTON; “HENRY. E. bic seo nee ee eee Columbia University.
CRAWFORD VAL BERT (Ga. ons aa eee Leland Stanford University.
CRILE,} GEORGE Ws: n.4e4 ce eee Western Reserve University, Cleveland.
CUNNINGHAM, (RICHARD) Hee. ee ee eee ne er eee Columbia University.
CUSHING, HARVEY Wi.334 4k Sone ee eee Harvard University.
DAKIN VHSDe ee nae eee ee ee 819 Madison Avenue, New York City.
DAVENPORT, CHARLES B............ Carnegie Institution, Station for Experi-
mental Evolution, Cold Spring Harbor, Long Island, N. Y.
DICKSON; ENC. siecaumsthaeks ce eee nor Stanford University Medical School.
IDOCHEZ) ACURA omc aoe eee Rockefeller Institute for Medical Research.
IDONALDSON; Ec IE eae eerie Wistar Institute of Anatomy, Philadelphia.
DRAPER; (GEORGE cece eae Presbyterian Hospital, New York City.
DRAPER; «J. Wi. 3c 5a veisrercee oma orem eee Pee New York University.
DRESBACH, OM. 05 ra etek cvs GPA OR A ea Ie Cornell University.
Du Bois (ERs peice Coon eee ee Cornell University Medical College.
DUNHAM, EDWARD) Kec cise en cet earner eras New York University.
DUVAL; 'CHAREES Wissacceic nee Ee De ene Tulane University.
EDMUNDS, GC. Wii seats h.cc eee ee eee De eRe University of Michigan,
EDSALL, DAVID) Liccian cscs oe care chee Totes ee DOeEe Harvard University.
EISENBREY,; Al IB. yaaniee nee ener St. Luke’s Hospital, New York City.
ISGGLESTON; (\CARV accep eine ee eee Cornell University Medical College.
ELSBERG; CHARLES) JA®. on canine eee eee Mount Sinai Hospital.
EUseR; WiIGHIAM Waters fone cheno Cornell University Medical College.
IEMERSON;: JHAVEN 22.6 c'ciGiens ste Chee enya eter eiere eee IE Columbia University.
ERLANGER) AJOSEPH =. 2 chic cine ee one Washington University, St. Louis.
EWING (BM it ang Se eae OE ee eee New York University.
EWING, JAMES):.cleccstahicose chain sa ae Cornell University Medical College.
PAUK G. aKa anohen no eee eee Roosevelt Hospital, New York City.
PAMULENER sls Veiner interac ee Department of Health, New York City.
FIELD, (CVRUSIW sy. tack Se ee COR Bellevue Hospital, New York City.

FISCHER) | MARTIN HH iia sak otto onan hese University of Cincinnati.

Ro__ oF MEMBERSHIP. 209

PRTZGHRADD Jet Gen chiar octet ci Board of Health, Toronto, Canada.
BITZPATRICK, CaB es aeoeacn ee eae Department of Health, New York City.
NEEXNER | SIMON 5 cn enone Rockefeller Institute for Medical Research.
BYOURNOY, UBOMAS Wachee ieee Mercy Hospital, Pittsfield, Mass.
FROLING OLTOP ede tic tes Paes eae oan ee tiene eee eA aera s Harvard University.
BORD WAVWILDEAM YS Wiis tater eter ncietss titrate aleuct esses Johns Hopkins University.
ROSTERS INELEIS Bessa t etree oar Cornell University Medical College.
BROSTAIW. i Heese ae U. S. Public Health and Marine-Hospital Service,
Hygienic Laboratory, Washington, D. C.
GAGER) (GUSTUARTA tc occ eee sane ceselneneld eine Brooklyn Botanic Garden.
GAxcBREDERICKO Pes get eine Wee ee Gane ee aaa ee ee University of California.
GRETUER WAR Oe ele cate erreice acter eee ares New York University.
GIBSON ROBERTI BY 550 5 enero Philippine Medical School, Manila, P. I.
GIES WUE TAME Ree yrs. 5 tracts cin, olen aiefocstaie. don aie cere Os Columbia University.
GITHENS Hi pote oie faeces Rockefeller Institute for Medical Research.
GiASERH ORTO!Caatiey seers incsssiewa tiers. ool ican aaoay one voy hay ove sbelee University of Michigan.
GOLDRARB Ata) ec, cians cncustiaire Siete ceharecsh oust College of the City of New York.
(GORDNER RG WAGs ci jcichee Carnegie Institution, Station for Experimental
Evolution, Cold Spring Harbor, Long Island, N. Y.
(GUPNTHER WAS EA cer ciccoce acc University of Nebraska, Lincoln, Nebraska.
EU RHRTEN Cal Gr ee stats ea chrsieecrnra nt aie ercktia ia eereron cope soe University of Pittsburgh.
JRUATD NWA MG oe ares otic U. S. Public Health and Marine-Hospital Service,
Hygienic Laboratory, Washington, D. C.
EVAR STED a WALETAMy Serie eta scslleud a clntaenic aietee sree Johns Hopkins University.
FUANZETK erp iatencraieroe aise Western Reserve Medical School, Cleveland, Ohio.
ARR IS SEL SAA Gir rc crerueoereueeis esc Arlington Chemical Co., Yonkers, N. Y.
EVARRISON ROSS Ws Grins 2 cies rire Seer yee aR Son A clei eS Yale University.
EVAR TWEDL DT [eeepc tats Gos the rea conser, Soe evel Cornell University Medical College.
IAT CHER ROBERT Ale aye ecateieicre or Cornell University Medical College.
EL ATAT. SHINKISHI Ai os /seicl oie teen Wistar Institute of Anatomy, Philadelphia.
TA WK BHIRTPSB 35 1 cievcrectarie nes Jefferson Medical College, Philadelphia, Pa.
FIEKRORN ME IEUD VIG reas orci etme a eee IS University of Chicago.
HESS PALRREDEE pr cttn ye calc atenhis Department of Health, New York City.
PUR WIE ETS Atm Wietrais te ccsroeacticls «s2ene otter caro Giottlzue ieiatenss University of Michigan.
HIGRSCHRELDER VAs etre fag eesebania cid ssiticpe. 6 eieacasere Johns Hopkins University.
HOOKER ED )AVENPORD io 5 aciom icles sore wise es erciatelerectuuehs olerscsvetecas Yale University,
LOWE aE Bente fale carey ais Maa avee aia @ cceeawe Seer s Columbia University.
FIGWELD ve V AID UAMIGED 915 5 stetOn ale ee macteleen dueynis ars Johns Hopkins University.
FLOWMAND a OHN =a) teesat cect vasterepsta <5) colors Gitavels Johns Hopkins University.
FLUBER G GalGARi poy ar sfecoaretsccis ecokssee sicker atesenssarsreuaien er University of Michigan.
HUNT EREID Cenc U. S. Public Health and Marine-Hospital Service,

Hygienic Laboratory, Washington, D. C.
HIGNTER) ANDREWieccciiacsicenhiacice cee: Cornell University, Ithaca, N. Y.

210 SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE.

JACKSON, (HOLMES) ©ih oc nictes on oe ana ceeon eras New York University.
JACOBS SWALTER Atria sn eee Rockefeller Institute for Medical Research.
JANEWAY, SPHEODORE"Gsi7 7... -C crete eae soe ore Columbia University.
JENNINGS; JHU Sine oe anc aieni ik meet eee Johns Hopkins University.
JOBLING, JAMES WirrcoArcomenenace. Michael Reese Hospital, Chicago, IIl.
JONES) WALTER hie acias on lenee aehincan ee enion Johns Hopkins University.
JORDAN, EDWIN Ohio et oh ncn eie ane tone oer University of Chicago.
JORDAN; "HABE wae calet ete ae hee oe Pelco hear University of Virginia.
JOSEPH; DONURGaacose seis pore amie ane Tee Dee rian Bryn Mawr, Pa.
IEARSNER, He SD eicasd ny eb eerie eee eee ee Harvard University.
IKAST EUDWIGH =a nae New York Postgraduate Medical School.
KASTUP, “JOSEPHUM aa tase cer Kentucky Agricultural Experiment Station,
Lexington, Ky.
IKERINERG Siac panier Rockefeller Institute for Medical Research.
KEOnTz; OSRART AG pac: biG Denk eae Mee Cee University of Pittsburgh.
IVAMAR! URIGHARD eV.ccroLe mnie Rockefeller Institute for Medical Research.
LAMBERT, RAG. 5 csdican doc eicas Rao eae Columbia University.
LEATHES) J. Bosciacckuiwe peeec le Gee eee eee University of Toronto.
LEE, SF REDERICISW32 ic Gh Sa nee Eee Columbia University.
LEVENE} (Po fAVe herp aclnneneten Rockefeller Institute for Medical Research.
IGEVIN; ISAAC) Lia gih, cc eaeivad ce tle erOn een OTe teens Columbia University.
EWIS;) PAUL Ayo act ous ang nee tae Phipps Institute, Philadelphia.
eB Gai b cote eee 6 es ee ne eee Columbia University.
LILLIE, SRA Risegcters perch creo terete eee ener ae COO University of Chicago.
LILLIE RALPH: Ss. tcc meek OC eerie. University of Pennsylvania.
LOEB, AACOQUES ners Rockefeller Institute for Medical Research.
TLLOEB) (LEOb2. 2 hosts Gt Cee Barnard Cancer Hospital, St. Louis.
LOEVENHART, “ARTHUR Slik aetoastonineecie nh ren etn University of Wisconsin.
LOMBARD}; WARREN Ps. 3020 cnc ctentisoen hereon: University of Michigan.
ILONGCOPE, ‘WilBn csc 38 Gite aa skeet ceteris Columbia University.
LUSK; (GRAHAM 5 Seg ic ech shoe oe Cornell University Medical College.
LYON, | Bs Be ace hiot ave Since le ie oe ec oreo bee ee St. Louis University.
MACCALLUM: Al Bist 2422... 6 coanoatioose eee University of Toronto.
MacGavpum: WeiGtin, £72 ee etait ane teen eee Columbia University.
MACDOUGAL, IDISIt chee eer Carnegie Institution, Washington, D. C.
MACLEOD, JicyMRi 22 ek pao noe Western Reserve University, Cleveland.
MACNEAtA WARD! .7.2h24 0s soar New York Post-Graduate Medical School.
IMMACGNIDER Wee DEN BS. ace baat eee University of North Carolina.
MCCEENDON SR e2.2 eu wis see een Cornell University Medical College.
McCruppben, F. M............. Rockefeller Institute for Medical Research.
MALL, PRANKVIN Pao. ch acide Genet aes Sea? Johns Hopkins University.
MANDEL; “ARTHUR Ris i) loaone se chteid sb Oe eine New York University.

MANDEL} JOHNWASa a5 Lact erect ccm tenner New York University.

RoLL OF MEMBERSHIP. 211

MANWARING, W. H............. Rockefeller Institute for Medical Research.
MAYER, ALFREDG............. Marine Laboratory of the Carnegie Institution,
Tortugas, Fla.
MEIGS EDWARD) Ben. saamracaceraebiceriaes 2 cl: Wistar Institute of Anatomy.
IMMEIZER Su Vdave mister aceon Rockefeller Institute for Medical Research.
IMMENDET SUA PAV ETT yD tericin ccc icrevacaian tdeiatohars) sieve leilvohcds ote cuetels Yale University.
IMEVERS ADOLPH ALE escricioosie. 1 neice tateha eerie cleiees Johns Hopkins University.
MEYER, GUSTAVE M........... Rockefeller Institute for Medical Research.
MORGAN; DHOMASHEem eo ae er gercisaiereie tras iuc errata) Columbia University.
MORSE SRNUAKG eo Oe ET ce onetetoiee ae aie echo eiet iol sscsnonesal nue eave Trinity College.
IMOSENTHAT: JEIERMAN) O\e eee ie sitisierevei cites rattle «ernst Columbia University.
MURDINA JOHNURS amnion ies ncrs wrecked Cornell University Medical College.
MURPHY, JAMESOB.. ceprtec ete- Rockefeller Institute for Medical Research.
Murpny, Joun B......... Northwestern University Medical School, Chicago.
MVERS Haig Cir jer heise chacet New York Post-Graduate Medical School.
INOGUGHIWEIDEYON snc ieee oes Rockefeller Institute for Medical Research.
INORRIS= | CHARLESH) /cjcisic s sie oreietereG arsine Bellevue Hospital, New York City.
INOVVGIHREDERTCRIGS ¢ ci neiie-ievisg se ecstie les sptlcis.ece les ntl University of Michigan.
OERDEE, UH ORSD uj aie ccoterovee 8 o suouheka the esge eee 30 West 44th Street, New York.
OPHULS.) WILLTAMG cloiici) ose baie oe Cooper Medical College, San Francisco.
OPIEVISUGENEM Ea 3 cl oaeciicks het e bieisieg hae en Washington University, St. Louis.
OPPENHEIMER: Bs S25 0.5 ccm, 6) see aevevede ct ete acee wie aerate nares Columbia University.
OSBORNE, THOMAS B........... Connecticut Agricultural Experiment Station.
New Haven, Conn.
(One ISU NC Saas 2 ahd ns Gee Teo Medico-Chirurgical College, Philadelphia.
OTTENBURG! Ras scree iaicrsicieiinelsh oko nl auc pers eee ener a Mount Sinai Hospital.
IPAPPENHEIMERS AL VIN IMIG cue si aielers choleiehe. c's latesreeo tie mucre Columbia University.
PAR Km a WAt piers tartar ad Ae arate claes sia sieves ainlers aes Johns Hopkins University.
PARK VIED TAM dees aorta cle pnisieva ache 5 terse eva tie sears New York University.
IPARKERH GEORGE (lay tor cr ie Sic ciate acshe alesis keri Heelan reais Harvard University.
PEAR CEWRICHARD Mi traciae Sach cio terec ates entrances University of Pennsylvania.
PEARL, RAYMOND..... Maine Agricultural Experiment Station, Orono, Maine.
PARES EIRIAIN Zarate bkece ce diane oa aisteteuenceel olons tolster a eee aot Harvard University.
124 04 on a Bee Ae oto G COTE OCS Critics orp ECan ae Columbia University.
PORTER; | WRU TA Mi prunes ial acvenic oisjevcrychernenelstel 3 nhs Harvard University.
PRATT TOSEPHUEL se eps oilers codon olesiviais eo ecale eboalore teh nieyarecs,s Harvard University.
AVENEL Te WUAZAICK Mb 2 etree crn rer eteciaie te ee University of Wisconsin.
REICHERT) EDWARD) dicticiacis sei sil ae enless basvieun iene University of Pennsylvania.
IRIGHARDS; -ALERED Neeecinchisicsioc cen asar ene University of Pennsylvania.
IRINGER EAN eer errata ett ota cetenen eras t be University of Pennsylvania.
ROBERTSON; Di. BRAILSRORD)y 15 -)./012 s)evooneietelelare sis clei University of California.

ROBINSON; G, CANBY. 0). 2c. ne 1 Rockefeller Institute for Medical Research.

212 SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE.

ROSENAWV, (MILTON Jinn (oes ae eet e eee enor Harvard University.
ROSENBLOOM: JACOB Aes nacheeen con oe eee University of Pittsburgh.
ROUS; {PEYIOND.ceceee eee Rockefeller Institute for Medical Research.
SACHS; |ERNSDI.Oe). craeescen cee abt tere Washington University, St. Louis.
SALANT, WILLIAM....... U. S. Department of Agriculture, Washington, D. C.
SCHL UTZ Hit Winis chee nce ts ce cee enioe eEE eee University of Minnesota.
SCHULTZ,“W-His- eee bore aciee Gee U. S. Public Health and Marine-Hospital
Service, Hygienic Laboratory, Washington, D. C.
SCHWYZER, RITZ .00 3.4 Weds erro ae ee One eee Switzerland.
Secor; BiG assis eae Oe ee EOE Pine Columbia University.
SCOPT) (Gs 'Gue oe icone ee, RE eRe ee College of the City of New York.
SENIOR? Hi) (Die hab dee SOE Oe New York University.
SHAFFER: (PHInip iA: oe cew el eee ae Washington University, St. Louis.
SHAKER) AWwO).ee be oher oe Sree Philippine Medical School, Manila, P. I.
SHERMAN, HENRY<G. ..ciai.adureciacinte ot pee CE Ee Columbia University.
SIMON; \CHARERSJE Soe. eee ce ee eae ee Baltimore Medical College.
SIMPSON;) SUTHERLAND) «ee eee eee Cornell University, Ithaca, N. Y.
SITTENFELD; IM icone el rrcce sti eee eee Columbia University.
SMITH, SHHEROBADD. yea sonin eee coe oe eee Harvard University.
SOLEMAN “LORALD eee tr hire ners Western Reserve University, Cleveland.
SOUTHARD sae oe as Con eee REE oe Ee eee Harvard University.
STEINHARDT: SEDNAG aa eile Department of Health, New York City.
STEWART, (GEORGE UN:. 2 ce here eee Western Reserve University, Cleveland.
STILES, JPERCVAGo woe eee Massachusetts Institute of Technology.
STOCKARD; (CHAS. (Rasen nan eee ree Cornell University Medical College.
STOOKEY, LYMAN B........... University of Southern California, Los Angeles.
STOREY; THOMAS) Avi. see ec eee College of the City of New York.
STRONG, RICHARD! Po cto aieoeee ee eee eee Harvard University.
SWEET, Js DWING:. coc ccceinee shhine ae ere seer University of Pennsylvania.
Swart; Hicker se eee eee Oe Rockefeller Institute for Medical Research.
SYMMERS DOUGLAS). cho Peek Oe eee Gee New York University.
‘TAYLOR; ALONZO) Epis tcc eceseee cee eee University of Pennsylvania.
TEAGUE, OSCARS acc maidens eee oe ee Cornell University Medical College.
TERRY; Bx Wipe t enacts cliche crore King’s County Hospital, Brooklyn, N. Y.
Topp, JOHN IG.. sesencuses Bees oes eee McGill University, Montreal.
SORREY,| JOHN\Giscosr eee ee ee Cornell University Medical College.
DY2ZZERS ES Bes eae Be ee ee Oe Harvard University.
UNDERRBINL) "FRANK (PHee ioe ee ree Oe Yale University.
VAN SLYKE, DONALD D......... Rockefeller Institute for Medical Research.
WADSWORTH, JAUGUSTUSE Beene ee ee Ree Columbia University.

WALLACE, GEORGE SBi aa oen ee ee oEe New York University.

RoL_t or MEMBERSHIP. 213

WWARTHING ‘ALDRED) | Sy.fc ans + c.camisiisrtiomaciecia scum University of Michigan.
NWWASTIENAW Sekdtn ik, tm, one. cticu searclnencroras suiiaisinre at eatauewale Pacific Grove, California.
WIRTH | RICHARD. soaker enemies Cornell University Medical College.
NVIED CH, SWALETAMs Els. a-yousiscetave cicresiontersets enelsiereiss Johns Hopkins University.
\VEEES© es GIDRON oc os corte seo oiccrenlc. Ce asim okenenes University of Chicago.
WEST GAS iinehinscs nea ae os Rockefeller Institute for Medical Research.
IWIGGERS s Can Jicecies cushace o ouerctteueceyererecsimusi Cornell University Medical College.
NVIETTARIS WAININAU Warners rset Department of Health, New York City.
WAT TAMS SEL Bi sertrctettrc tales: biecd Stole! arcutercior ia aacyers. easels Columbia University.
WIRE TAMS WEDERIBER TMU cote mrcycle feyerdsserecctereisprictckvarers isi ate University of Buffalo.
WIESON EDMUND! Daseichyec cosine etd oclet namics acne Columbia University.
NVIINSEO We Garb Atay aserstocstentene: «custo ettaencts College of the City of New York.
WOEBACH ASS BURT varnsstas sy taceeeeie solr ielckoetelicieca’asies Harvard University.
Worm CHARD mst Gr le cic one oi iaette napchecnaneus eee oie Cambridge, England.
WOLLSTEIN, MaRTHA.......... Rockefeller Institute for Medical Research.
MOODS ERANICIS] Cs co ctasievernas mio sas eiciuan eraaliec meamiel on Columbia University.
NVOODRUEELORANDEMILOSS# an svat ciierteyarcseratic sn are = Yale University.
WIA TSU INIA OHEDE = cers A misieresla wierd oieke Oreysin cyeycuels cue ise University of Japan.
Wanangy IRGnoy tN GL Ge jopedbosae seldom ben anca capes Harvard University.
TARRY, IBUIRNSno dbonboosod daveoes poEaO CUT 6.00 6 cole Columbia University.

Total number of members at the close of the academic year, 1912~’13: 255.

on
pl S

ey on
mae

: bp.)

vo ar ‘Wu

Sa ee ae
| Mee ae

OFFICERS.’

1903-1914.
1903-04 1904—'05 1905-'06
Presidentiayes 4 -tsrt-teye' Meltzer Meltzer Wilson
Vice-President...... Park Ewing Dunham
Wibratianyetter stir Lusk Lusk Lusk
PIT CASLILED,.Nevoletslslcisiehe Calkins Calkins Calkins
Secretaryis cree ce Gies Gies Gies

I909Q—"I0 IQIO—'II IQ1I—'I2

President.......... Lee Morgan Morgan
Vice-President...... Gies Gies Levene
MY CASULET aia) -teleieiets ere Lusk Lusk Lusk
Secretary sicneiocse sess Opie Opie Wallace

1 Council—The Past Presidents and the Officers.

215

1906-07

Flexner
Dunham

Calkins
Gies

1912-13

Ewing
Levene
Norris
Wallace

1907-08

Flexner
Morgan

Calkins
Gies

I9I3—14
Ewing
Field
Norris
Jackson

1908-09

Lee
Morgan

Lusk
Gies

a eine tL ee

CEASSIFIED: LISt OF-MEMBERS OF THE
SOCIETY FOR) EXPERIMENTAL
BIOLOGY AND MEDICINE.

Resident (Greater New York).

College of the City of New York.—A. J. Goldfarb, G. G. Scott, Thomas A.
Storey, C. E. A. Winslow.

Columbia University.—Russell Burton-Opitz, Gary N. Calkins, Henry E.
Crampton, Haven Emerson, William J. Gies, P. E. Howe, Theodore C. Jane-
way, R. A. Lambert, Frederic S. Lee, Isaac Levin, C. C. Lieb, W. F. Long-
cope, W. G. MacCallum, Thomas H. Morgan, H. O. Mosenthal, B. S. Oppen-
heimer, Alwin M. Pappenheimer, F. H. Pike, E. L. Scott, Henry C. Sherman,
M. Sittenfeld, H. B. Williams, Augustus B. Wadsworth, Edmund B. Wilson,
Francis C. Wood, Hans Zinsser.

Cornell University Medical College-—S. P. Beebe, Stanley R. Benedict,
M. J. Burrows, B. H. Buxton, A. F. Coca, E. F. DuBois, C. Eggleston, William
J. Elser, James Ewing, Nellis B. Foster, J. A. Hartwell, Robert A. Hatcher,
Graham Lusk, J. F. McClendon, John R. Murlin, Chas. R. Stockard, Oscar
Teague, John C. Torrey, Richard Weil, C. J. Wiggers.

Hospitals. Bellevue—Cyrus W. Field, Charles Norris. King’s County.
—B. T. Terry. Mt. Sinai.—Charles A. Elsberg, R. Ottenberg. Roosevelt.
—G. K. Falk. Presbyterian—R. A. Cecil, George Draper. St. Luke’s.—
A. D. Ejisenbrey.

New York City Departments. Education—C. Ward Crampton. Health.
—James P. Atkinson, Edwin J. Banzhaf, L. W. Famulener, C. B. Fitzpatrick,
Alfred F. Hess, Edna Steinhardt, Anna W. Williams.

New York Polyclinic Medical School.—Isaac Adler.

New York Post-Graduate Medical School.—Ludwig Kast, W. J. MacNeal,
V. C. Myers.

New York University—Harlow Brooks, J. W. Draper, Edward K. Dun-
ham, E. M. Ewing, A. O. Gettler, Holmes C. Jackson, Arthur R. Mandel,
John A. Mandel, William H. Park, H. D. Senior, Douglas Symmers, George B.
Wallace.

Rockefeller Institute for Medical Research.John Auer, F. W. Bancroft,
Jacob Bronfenbrenner, E. E. Butterfield, Alexis Carrel, P. F. Clark, A. E.
Cohn, Rufus Cole, A. R. Dochez, Simon Flexner, T. H. Githens, Walter A.
Jacobs, I. S. Kleiner, Richard V. Lamar. P. A. Levene, Jacques Loeb, W.
H. Manwaring, S. J. Meltzer, Gustave M. Meyer, F. M. McCrudden, James
B. Murphy, Hideyo Noguchi, G. Canby Robinson, Peyton Rous, H. F. Swift,
Donald D. Van Slyke, C. J. West, Martha Wollstein.

207

218 SCIENTIFIC PROCEEDINGS (53).

Brooklyn Botanic Garden.—C. Stuart Gager.
819 Madison Avenue.—H. D. Dakin.
30 West 44th Street.—Horst Oertel.

Non-Resident.

Agricultural Experiment Stations. Connecticut (New Haven).—Thomas
B.Osborne. Kentucky (Lexington).—J.H. Kastle. Maine (Orono).—Raymond
Pearl.

Carnegie Institution of Washington—Francis G. Benedict ( Nutrition
Laboratory, Boston), A. M. Banta, Charles B. Davenport, R. A. Gortner
(Station for Experimental Evolution, Cold Spring Harbor, N. Y.), D. T.
MacDougal (Washington), Alfred G. Mayer (Marine Laboratory, Tortugas,
Fla.)

Colleges. Bryn Mawr.—Don R. Joseph. Medico-Chirurgical (Phila-
delphia).—Isaac Ott. Jefferson Medical ( Philadelphia).—P. B. Hawk. Philip-
pine Medical (Manila).—R. B. Gibson, A. O. Shaklee. Trinity (Hartford).
—Max W. Morse.

Georgia State Board of Health (Atlanta). (Katharine R. Collins.

Hospitals. Barnard Skin and Cancer (St. Louis)—Leo Loeb. Mercy
(Pittsfield, Mass.)—Thomas Flournoy. Michael Reese (Chicago).—James
W. Jobling.

Institutes. Massachusetts, of Technology—Percy G. Stiles. Phipps
(Philadelphia).—Paul A. Lewis. Wistar (Philadelphia) —H. H. Donaldson,
Shinkishi Hatai, E. B. Meigs.

U. S. Departments. Agriculture (Washington, D. C.).—Carl L. Alsberg,
William N. Berg, F. J. Birchard, William Salant. Treasury (Public Health
and Marine- Hospital Service).—John F. Anderson, W. H. Frost, W. H. Hale,
Reid Hunt and W. H. Schultz, Washington, D. C.

Universities. Buffalo—G. H. A. Clowes, Herbert U. Williams. Cal-
fornia.—J. V. Cooke, F. J. Fitzgerald, F. P. Gay, T. Brailsford Robertson,
H. Wastenays. Chicago.—A. J. Carlson, Ludvig Hektoen, Edwin O. Jordan,
Frank R. Lillie, H. Gideon Wells. Cincinnati—Martin H. Fischer. Cornell.
—Melvin Dresbach, Andrew Hunter, Sutherland Simpson. Harvard.—Her-
man M. Adler, Walter B. Cannon, W. T. Councilman, Harvey Cushing,
David L. Edsall, Otto Folin, H. T. Karsner, G. H. Parker, Franz Pfaff, W. T.
Porter, Joseph H. Pratt, M. J. Rosenau, Theobald Smith, E. E. Southard,
Richard P. Strong, E. E. Tyzzer, S. Burt Wolbach, Robert M. Yerkes. Japan.
—Naohidé Yatsu. Johns Hopkins.—John J. Abel, W. W. Ford, W.S. Halsted,
A. D. Hirschfelder, William H. Howell, John Howland, H. S. Jennings, Walter
Jones, Adolph Meyer, E. A. Park, William H. Welch. Leland Stanford.—A. C.
Crawford, E. C. Dickson, W. Ophiils. McGill (Montreal).—J. George Adami,
John L. Todd. Michigan.—C. W. Edmunds, Otto C. Glaser, A. W. Hewlett,
Carl G. Huber, Warren P. Lombard, Frederick G. Novy, Aldred S. Warthin.
Minnesota.—F. W. Schlutz. Nebraska.—A. E. Guenther. North Carolina.—
W. de B. MacNider. NorthwesternJohn B. Murphy. Pennsylvania.—J. H.

CLASSIFIED List OF MEMBERS. 219

Austin, Alexander C. Abbott, D. H. Bergey, Ralph S. Lillie, Richard M. Pearce,
Edward T. Reichert, Alfred N. Richards, A. I. Ringer, J. Edwin Sweet, A. E.
Taylor. Pittsburgh.—C. C. Guthrie, Oskar Klotz, Jacob Rosenbloom. Prince-
ton.—Edwin G. Conklin. Southern California (Los Angeles)—Lyman B.
Stookey. St. Louis —E.P. Lyon. Toronto.—J. B. Leathes, A. B. Macallum.
Tulane.—Charles W. Duval. Virginia —H. E. Jordan. Washington (St.
Louis).—Joseph Erlanger, Eugene L. Opie, Ernst Sachs, Philip A. Shaffer.
Wisconsin.—Charles R. Bardeen, C. H. Bunting, L. J. Cole, Arthur S. Leoven-
hart, Mazyck P. Ravenel. Western Reserve (Cleveland).—George W. Crile,
Paul J. Hanzlik, J. J. R. Macleod, Torald Sollmann, G. N. Stewart. Yale.—
R. H. Chittenden, Ross G. Harrison, Davenport Hooker, Lafayette B. Mendel,
Frank P. Underhill, Lorande Loss Woodruff.

Cambridge, England.—C. G. L. Wolf.

Switzerland (Lucern).—Fritz Schwyzer.

Yonkers, N. Y.—Isaac F. Harris.

INDEX

OF THE

SCIE NMFIC PROGCBEDINGS:

[THE NUMERALS IN THIS INDEX CORRESPOND WITH THE NUMERALS IN

PARENTHESIS ABOVE THE TITLES OF THE ABSTRACTS.

PAGES

ARE NOT INDICATED.]

Abderhalden’s test for pregnancy, 746.

Absorption, rate of, of water by skin of
frog, 778.

Acid produced by tissues growing in
vitro, 796.

Agglutination of bacteria, 734, 775.

Amino acids, distribution of, in body,
723.

Anaphylaxis, new factor in, 769; in
immune animals, 811.

Annelid worm, regeneration of, 755.

Anilin dyes, effect upon _ bacterial
growth, 807.

Animal extracts, effect upon thyroid,
809.

Antibodies, protein concentration upon
absorption of, 751.

Antigen and precipitin, failure of union
between, 741.

Antitoxic action of sodium iodide on
morphin, 780.

Antitryptic reaction, in pregnancy, 768.

Aorta, occlusion by living tissue, 771;
ligation, effect upon carbon dioxide
and oxygen in blood, 805.

Artificial respiration, in relation to
shock, mechanical factors of, 782

Atoxyl, 757.

Auricles, vagus nerves on faradized,
752.

B. coli communior, frequency in water,
735-

Bacteria, acid agglutination to differ-
entiate, 775; growth of, effect of
anilin dyes on, 807; of the duodenum,
758; production of creatinine by,
736.

Betaine, in invertebrate muscle, 798.

Blood, color of red corpuscles, 794.

Body fluids, relation of leucocytic
extract to, 716.

Bone, regeneration of, 737.

Calcium, importance for growth, 699.

Carbon dioxide, in blood after ligation
of aorta, 805.

Cardiac mechanism, reversed, 705, 722,
726.

Cardiectomized frogs, effects of strych-
nine in, 712.

Cell division, dynamics of, 779.

Cholesterin and lecithin, influence upon
tumor growth, 738; relation to
chronic nephritis, 733.

Color index of red blood corpuscles,
794.

Complement deviation in pregnancy,
767.

Conjugation of paramoecium, 742.

Creatin in muscle, relation to urinary
creatinine, 703; during starvation,
704; 801.

Creatinine, production of, by bacteria,
736.

Cretinism, effect upon sheep’s nitrog-
enous metabolism, 763.

Decerebration and action of morphine
in frogs, 725.

| Diabetes, fate of parenterally intro-
duced glycogen in, 724.

Diffusion of iodo-eosin through rubber,
731.

Dog, growth rate in, 762.

Duodenum, bacteria of, 758.

Eck fistula, phlorhizin on dogs with,
719.

Elimination through the mucosa of
urinary bladder, 720.

Embryos, sea-urchin, influence of leci-
thin upon, 774.

Epinephrin in blood, action of glucose
upon, 708.

221

222

Ergot, cock’s comb test for activity of,

39.
Ether and chloroform, toxic effects, 715.
Extra systoles, in otherwise normal
heart, 701.
Eye defects, interpretation of, 797.

Fat content, morphology and length |

of life of cells in plasma, 777.

Galvanotropism and heliotropism in,

Euglena, 776.

Gastric juice, solubility of lead salts in, |

815.

Glucose and amount of epinephrin in|

blood, 708.

Glycogen, fate of parenterally intro- |

duced, in diabetes, 724; determina-
tion of, in liver, 711.

Gréhant’s anesthetic and morphine-
ether upon dog’s urine, 761.

Growth, importance of calcium for, 699;
in the dog, 762.

Glycosuria, effect of pancreatic and
duodenal extracts upon, in de-
pancreatized dogs, 804.

Habit, relation to nervous system in|

worm, 707.

Hemoglobin, relation between oxygen
and, 748.

Hay fever, specific reactions in, 744;
autumnal, treatment by vaccination,

745-

Heart block, complete, without lesion |

of bundle of His, 753.

Heliotropism and galvanotropism in|

Euglena, 776.

Heterochromosomes in mammals, 710. |

Hibernation, effect of external tem-
perature upon, 810.

Hormones of secretion of milk, 709.

Hypopituitarism, metabolism in, Lor-
ain’s type, 764.

Immunity, experimental, pressor sub-
stances in, 766; and parathyroid
tetany, 814.

Intercalated disks of atrophied heart |

muscle, 727.

Intestinal disturbances, lipase of in-
fants in, 750. .

Intrabronchial insufflation, pulmonary
lesions produced by, 713.

Intratracheal insufflation, toxic effects
of ether and chloroform under, 715.

SCIENTIFIC PROCEEDINGS (53).

Involution, rdle of phagocytosis in, 718.
Iodo-eosin, diffusion of, through rub-
ber, 731.

Kernplasma relation during life, 747.
Kraus-Hofer phenomenon in tubercu-
losis, 717.

\Labyzinth, electrical stimulation of,

| 749.

Lactic acid, sugar from, in diabetes,
786.

Larve, production of grafted, 784.

Lead poisoning, chronic, in guinea pigs,
polyserositis in, 732; relation to
chronic nephritis, 733; salts, solu-
bility in gastric juice, 815.

Lecithin and cholesterin on tumor
growth, 738; influence upon develop-
ment of embryos, 774.

| Length of life, fat content, morphology,
of cells in plasma, 777.

Leucocytes, foreign, injection on men-
ingeal tuberculosis, 698.

Leucocytic extracts, relation to body
fluids, 716.

Light and tactile reactions in amphi-
pods, 817.

Lipase, pancreatic, in infants in in-
testinal disturbances, 750.

Lipoids, in narcosis, 706.

Lysis of tubercle bacillus, 743.

Magnesium, central effect of, 795.

Metabolism, cretinism effect upon
nitrogenous, in sheep, 763; in case of
hypopituitarism—Lorain type, 764;
in myotonia atrophica, 785.

Morphin, decerebration and action of,
in frogs, 725; -ether and Gréhant’s
anesthetic upon dog’s urine, 761;
on dog’s heart after removal of one
vagus, 759.

Motor cortex and pyramidal tract in
raccoon, 729.

Muscle, striated, semi-permeable mem-
branes surrounding, 781; inverte-
brate, betaine in, 798.

Myotonia atrophica, metabolism in, 785.

Narcosis, lipoids in, 706.
Nitrogen, non-protein,
blood, 802.

in nephritic

Oocytin, nature of, 773.
Optic anlage, location of, 797.

INDEX

Oxygen and hemoglobin, relation be-
tween, 748; in blood after ligation
of aorta, 805.

Oxytricha fallax, kernplasma, relation |
in life of, 747.

Pancreas, transplantation in spleen,
721.

Pancreatic extract, relation to blood |

sugar, 725; effect on glycosuria and
respiratory metabolism in depan-
creatized dogs, 804.
Paramecium, conjugation of, 742.
Parathyroids and _ pregnancy,
tetany and immunity, 814.
Phagocytosis, réle of, in involution, 718.

812;

Phlorhizin on dogs with Eck’s fistula, |

719.

Pneumonia, development of experi-|

mental, 714.

Poison oak, study of, 740.

Poliomyelitis, infection in experimental,
697.

Precipitin and antigen, failure of union
between, 741.

Pregnancy, Abderhalden’s test for, 746;
antitryptic reaction exhibited in, !

768; complement deviation in, 767; |

parathyroids and, 812.

Pressor substances in experimental im- |

munity, 766.

Protein concentration and absorption |

of antibodies, 751.
Pulmonary lesions produced by intra- |
bronchial insufflation, 713.
Pyramidal tract and motor cortex in |
raccoon, 729; in porcupine, 700.

Regeneration of bone from periosteum, |
737; of annelid worm, 755; concen-
tration of sea water upon, 756.

Respiratory metabolism in depancre-
atic dogs, 804.

Secretion of milk, hormones of, 709.

Semipermeable membranes surround- |

ing striated muscle, 781.

Shock, artificial respiration in relation |

to acapnial theory of, 782.

Sodium iodide, antitoxic action of, on.

morphin, 780.
Specificity, biological, studies of, 772.
Spirochetes, transmission of, 783.
Spleen, pancreatic transplantations of,
721.

223

| Starches in relation to genera, species,
728.
Starvation, muscle creatine and urinary
creatinine during, 704.
| Strychnine, effects of, in cardiectomized
| frogs, 712; temperature effect upon
minimum dose of, 760.
Sugar from lactic acid in diabetes, 786;
of blood, relation of pancreatic
extracts to, 765.

Tactile reaction, light and, in amphi-
pods, 817.

| Tartrate, sodium, toxicity of, 803.

| Temperature, effect on dose of strych-
nine, 760.

Tetanus, temperature effect on onset of,
after strychnine, 760.

Thyroid gland, hypertrophy, 770; con-
stituents of, 799; physiological ac-
tivity of, 800; volume of, effect of
animal extracts upon, 809; of mother
and of fetus, 813.

| Tolerance to sodium tartrate, 803.

| Tonsillar extract, action of, 730.
| Tubercle bacilli, lysis of, 743.

| Tuberculosis, injection of leucocytes
on meningeal, 698; Kraus-Hofer phe-
nomenon in, 717.

| Tumor growth, lecithin and cholesterin
upon, 738; effect of diets upon, 806.

Tyrosinase, in amphibian egg jell, 816.

|

| Urinary bladder, elimination through
mucosa of, 720.

| Vaccination, treatment of hay fever by,

| 745.

Vagi, differences in the dog’s heart, 702.

| Vagus, morphine effects after removal
of one, 759; nerves on faradized
auricles, 752.

Valvular lesions, production of tem-
porary, 754.

Venom intoxication, waxy muscle de-
generation in, 808.

Water absorption, rate of, by skin of
frog, 778.

| Worm, earth, habit and relation to

| nervous system, 707.

| Waxy degeneration in venom intoxica-
tion, 808.

PROCEEDINGS

OF THE

SOCIETY FOR

EXPERIMENTAL BIOLOGY AND MEDICINE

FIFTIETH MEETING

CORNELL UNIVERSITY MEDICAL COLLEGE

NEW YORK CITY

OCTOBER 16, Ig12

VOLUME X

No. 1

NEW YORK

IgI2

CONTENTS.

S. J. MeLTzer: The effects of strychnine in cardiectomized frogs with destroyed
anterior lymph sacs; a demonstration. 16(712).

MARTHA WOLLSTEIN and S. J. MELTZER: Pulmonary lesions produced by intrabron-
chial insufflation of cultures of B. megatherium. 17 (713).

MARTHA WOLISTEIN and S, J. MELTZER: The development of experimental pneu-
monia ; a demonstration. 18 (714).

T. S GITHENs and S. J. MELTZER: Differences in the toxic effects of ether and chloro-
form as observed under intratracheal insufflation. 19 (715).

W. H. MANwarinc: On the relation of leucocytic extract to body fluids. 20(716).

W. H. MANWARING and J. BRONFENBRENNER : The Kraus-Hofer phenomenon in tuber-
culosis. 21 (717).

MAX Morse: The role of phagocytosis in involution. 22(718).

N. B, Foster: The effect of phlorhizin on dogs with Eck fistula. 23 (719).

I. S. KLEINER : On elimination through the mucosa of the urinary bladder. 24 (720).

J. H. Pratrand F. T. MurpHy: Pancreatic transplantationsin the spleen. 25 (721).

A. E. Coun: The production of reversed cardiac mechanism in the dog. 26(722).

D. VAN SLYKE and G. M, MeEyver: The distribution of amino acids in the body.
27 (723).

R. A. ees (by invitation): The fate of parenterally introduced glycogen in human
and experimental diabetes, 28 (724).

T. S. GirHEeNs: Decerebration and the action of morphine in frogs. 29 (725).

H. B. WILiiAMs and Henry JAMES: Reversal of the cardiac mechanism ; an addi-
tional note. 30 (726).

H. E. JoRDAN: The intercalated discs of atrophied heart muscle. 31 (727).

E. T. REICHERT: The differentiation and specificity of starches in relation to genera,
species, etc. 32 (728).

SUTHERLAND SIMPSON: The motor cortex and pyramid tractinthe raccoon. 33 (729).

Isaac OTT and J. C. Scorr: Note on the action of tonsillar extract. 34(730).

JAcoB RosENBLOOM: The diffusion of iodo-eosin from ether through rubber into ether.
35 (731).

W. ee Chronic lead poisoning in guinea pigs. Its relation to polyserositis.
36 (732).

W. Grane { Chronic lead poisoning in guinea pigs. Its relation to chronic nephritis.
37 (733).

HES AM Se tg Agglutination of encapsulated bacteria. 38 (734).

J. G. FitzGERALD: Relative frequency of B. coli communior in contaminated water.
39 (735).

J. G. Ais 2a and Cari L. A. ScHMIDT: Production of creatinine by bacteria.
40 (736).

S. L. Haas (by invitation) : Regeneration of bone from periosteum. 41 (737).

T. BRAILSFORD ROBERTSON and T. C, BURNETT: The influence of lecithin and choles-
terin upon the growth of tumors. 42 (738).

ALBERT C. CRAWFORD and JAMES P. CRAWFORD: Note on the cock’s comb test for
the activity of ergot. 43 (739).

EDWARD VON ADELUNG (by invitation): An experimental study of poison oak. 44

740).

Hie Prwisih and STEWART YOUNG: The failure of union between antigen and pre-

cipitin when present in the same serum, 45 (741).

The proceedings of the Society for Experimental Biology and Medicine are pub-
lished as soon as possible after each meeting. Regular meetings of the Society are held
in New York on the third Wednesdays of October, December, February, April and
May. A volume of the proceedings consists of the numbers issued during an
academic year. _

The price per volume, sent postage prepaid, is one dollar. The price of copies
of the proceedings of any meeting is twenty cents each, postage prepaid. Subscriptions
are payable in advance.

PRESIDENT — James Ewing, Cornell University Medical College.

VICE-PRESIDENT — Phoebus A. Levene, Rockefeller Institute for Medical
Research. .

SECRETARY — George B. Wallace, University and Bellevue Hospital Medical
College. r

TREASURER — Charles Norris, Bellevue Hospital.

MANAGING EpirorR —The Secretary, 338 E. 26th St., New York City.

CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR
EXPERIMENTAL BIOLOGY AND MEDICINE.

Resident (Greater New York).

Bellevue Hospital.—Cyrus W. Field, Charles Norris.

Columbia University.—Russell Burton-Opitz, Gary N. Calkins, Heyrn
E. Crampton, Richard H. Cunningham, Haven Emerson, Nellis B. Foster,
William J. Gies, A. E. Guenther, Philip H. Hiss, Theodore C. Janeway, R.
A. Lambert, Frederic S. Lee, Isaac Levin, C. C. Lieb, W. F. Longcope, W.
G. MacCallum, Thomas H. Morgan, H. O. Mosenthal, Alwin M. Pappen-
heimer, B. S. Oppenheimer, F. H. Pike, E. L. Scott, Henry C. Sherman,
M. Sittenfeld, Hugh A. Stewart, Augustus B. Wadsworth, Edmund B. Wilson,
Francis C. Wood.

Cornell University Medical College.—S. P. Beebe, Stanley R. Benedict,
M. J. Burrows, B. H. Buxton, A. F. Coca, E. F. DuBois, William J.
Elser, James Ewing, Robert A. Hatcher, Graham Lusk, J. F. McClendon,
John R. Murlin, Chas. R. Stockard, John C. Torrey, Richard Weil, C. J.
Wiggers.

Mé. Sinat Hospital.—Charles A. Elsberg, R. Ottenberg.

New York City College.—A. J. Goldfarb, Thomas A. Storey, C. E. A.
Winslow.

New York City Departments. Education.—C. Ward Crampton.
Flealth.—James P. Atkinson, Edwin J. Banzhaf, L. W. Famulener, C. B.
Fitzpatrick, Alfred F. Hess, Edna Steinhardt, Anna W. Williams.

New York Polyclinic Medical School.—Isaac Adler.

New York Post-Graduate Medical School.—Ludwig Kast, W. J. Mac-
Neal, V. C. Myers.

New York University. — Harlow Brooks, J. W. Draper, Edward K. Dun-
ham, E. M. Ewing, Holmes C. Jackson, Arthur R. Mandel, John A. Mandel,
William H. Park, H. D. Senior, Douglas Symmers, George B. Wallace.

Rockefeller Institute for Medical Research.—John Auer, F. W. Bancroft,
Jacob Bronfebrenner, E. E. Butterfield, Alexis Carrel, P. F. Clark, A. E. C
Cohn, Rufus Cole, A. R. Dochez, George Draper, Simon Flexner, T. H.
Githens, Walter A. Jacobs, Don R. Joseph, I. S. Kleiner, Richard V. Lamar,
P. A. Levene, Jacques Loeb, W. H. Manwaring, S. J. Meltzer, Gustave M.
Meyer, F. M. McCrudden, J. B. Murrphy, Hideyo Noguchi, G. Canby
Robinson, Peyton Rous, H. F. Swift, B. T. Terry, Donald D. Van Slyke,
H. Wastenays.

Sage Institute of Pathology.— Horst Oertel.

St. Francis Hospital.—F¥ritz Schwyzer.

St. Luke's Hospital—A. B. Eisenbrey.

Brooklyn Botanic Garden.—C, Stuart Gager.

819 Madison Avenue.—H. D. Dakin.

Non-Resident.

Baltimore Medical College. —Charles E. Simon.

Carnegie Institution of Washington.—Francis G. Benedict (Vutrition
Laboratory, Boston), A. M. Banta, Charles B. Davenport, R. A. Gortner
(Station for Experimental Evolution, Cold Spring Harbor, N. Y.), D. T.
MacDougal (Washington), Alfred G. Mayer (Marine Laboratory, Tortugas,
¥la.).

Connecticut Agricultural Experiment Station (New Haven).—Thomas
B. Osborne.

Geergia State Board of Health (Atlanta).—Katharine R. Collins.

Kentucky Agricultural Experiment Station (Lexington).—J. H. Kastle.

Maine Agricultural Experiment Station (Orono).— Raymond Fearl.

Massachusetts Institute of Technology.—Percy G. Stiles.

Medico-Chirurgical College (Philadelphia).—Isaac Ott.

Mercy Hospital (Pittsfield, Mass.).—Thomas Flournoy.

Michael Reese Hospital (Chicago).—James W., Jobling.

Jefferson Medical College.—P. B. Hawk.

Philippine Medical School (Manila).—A. O. Shaklee.
Trinity College (Hartford ).—Max W. Morse.
Yonkers, N. Y.—Isaac F. Harris.

U.S. Departments. Agriculture (Washington, D.C.).—CarlL. Alsberg,
William N. Berg, F. J. Birchard, William Salant; Juterior (Philippine
Islands, Bureau of Science, Manila).—Richard P. Strong, Oscar Teague.
Treasury (Public Health and Marine-Fospital Service).—John F. Anderson,
W. H. Frost, Wm. H. Hale, Reid Hunt and W. H. Schultz, Washington,
DAC,

Universitics. Buffalo.—G. H. A. Clowes, Herbert U. Williams. Cad-
fornia. —¥. P. Gay, J. V. Cooke, F. J. Fitzgerald, T. Brailsford Robertson.
Chicago. —R. R. Bensley, A. J. Carlson, Ludvig Hektoen, Edwin O. Jordan,
Frank R. Lillie, H. Gideon Wells. Czxctnattz.—Martin H. Fischer. Corned.
Melvin Dresbach, Andrew Hunter, Sutherland Simpson. Harvard.—Her-
man M. Adler, Walter B. Cannon, W. T. Councilman, David L. Edsall,
Otto Folin; H. T. Karsner, G. H. Parker, Franz Pfaff, W. T. Porter, Joseph
H. Pratt, M. J. Rosenau, Theobald Smith, E. E, Southard, E. E. Tyzzer, S.
Burt Wolbach, Robert M. Yerkes. /afan.—Naohidé Yatsu. johns Hop-
kins. —John J. Abel, Harvey W. Cushing, W. W. Ford,W. S. Halsted, A. D.
Hirschfelder, William H. Howell, John Howland, H. S. Jennings, Walter
Jones, F. P. Mall, Adolph Meyer, E. A. Park, William H. Welch. Leland
Stanford. — A. C. Crawford, E. C. Dickson, W. Ophiils, Hans Zinsser. Mc-
Gill (Montreal ).—J. George Adami, John L. Todd. A&ichigan.—C. W. Ed-
munds, Otto C. Glaser, A. W. Hewlett, Carl G. Huber, Warren P. Lombard,
Frederick G. Novy, Aldred S. Warthin. Mznxnesota.—Robert B. Gibson, F.
W. Schlutz. orth Carolina.—W. de B. MacNider. orthwestern. —J.
B. Murphy. Pennsylvania.—J. H. Austin, Alexander C. Abbott, D. H.
Bergey, Ralph S. Lillie, Richard M. Pearce, Edward T. Reichert, Alfred N.
Richards, A. I. Ringer, J. Edwin Sweet, A. E. Taylor. Pittsburgh. —C.
C, Guthrie, Oskar Klotz, Jacob Rosenbloom. Pyincefon.—Edwin G. Conk-
lin. Southern California (Los Angeles).—Lyman B. Stookey. S?. Louis.—
E. P. Lyon. YZoronto.—T.G. Brodie, J. B. Leathes, A. B. Macallum.
Tulane. —Charles W. Duval. Virginia. —H. E. Jordan. Washington
(St. Louzs).—Joseph Erlanger, Eugene L. Opie, Ernst Sachs, Philip A.
Shaffer. Wisconsin.—Charles R. Bardene, C. H. Bunting, Arthur S. Loeven-
hart, Mazyck P. Ravenel. Western Reserve (Cleveland).—George W. Crile,
Paul J. Hanzlik, J. J. R. Macleod, Torald Sollmann, G. N. Stewart. Ya/e.
—R. H. Chittenden, Ross G. Harrison, Lafayette B. Mendel, Frank P.
Underhill, Lorande Loss Woodruff.

Cambridge, England.—C. G. L. Wolf.

University College (London).—Arthur R. Cushny.

Phipps Institute (Philadelphia).—Paul A. Lewis.

Wistar Institute of Anatomy (Philadelphia).—H. H. Donaldson, Shin-
kishi Hatai, E. B. Meigs.

Barnard Skin and Cancer Hospital (St. Louis).—Leo Loeb.

Members present at the fifty-first meeting:

Atkinson, Auer, Bancroft, Beebe, Berg, Brooks, Cohn, E. M. Ewing,
James Ewing, Field, Fitzpatrick, Gies, Githens, Goldfarb, Hatcher, Kleiner,
Lee, Lusk, McCrudden, McNeal, J. A. Mandel, Manwaring, Meltzer,
Norris, Oppenheimer, Pratt, Rous, Senior, Sittenfeld, Swift, Terry, Van
Slyke, Wadsworth, Wallace, H. B. Williams.

Jacob Bronfebrenner, P. F. Clark, Melvin Dresbach, Paul J. Hanzlik,
Arthur D. Hirschfelder, J. B. Murphy, V. C. Myers, E. L. Scott.

Dates of the next two regular meetings:
February 17, 1913—April 16, 1913.

CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR
EXPERIMENTAL BIOLOGY AND MEDICINE.

Resident (Greater New York).

Bellevue Hospital.—Cyrus W. Field, Charles Norris.

Columbia University.—Russell Burton-Opitz, Gary N. Calkins, Henry
E. Crampton, Richard H. Cunningham, Haven Emerson, Nellis B. Foster,
William J. Gies, A. E. Guenther, Philip H. Hiss, Theodore C. Janeway, R
A. Lambert, Frederic S. Lee, Isaac Levin, C. C. Lieb, W. F. Longcope, W.
G. MacCallum, Thomas H. Morgan, H. O. Mosenthal, Alwin M. Pappen-
heimer, B. S. Oppenheimer, F. H. Pike, Henry C. Sherman, Hugh A.
Stewart, Augustus B. Wadsworth, Edmund B. Wilson, Francis C. Wood.

Cornell University Medical College.—S. P. Beebe, Stanley R. Benedict,
M J. Burrows, B. H. Buxton, A. F. Coca, E. F. DuBois, William J.
Elser, James Ewing, Robert A. Hatcher, Graham Lusk, J. F. McClendon,
John R. Murlin, Chas. R. Stockard, John C. Torrey, Richard Weil, C. J.
Wiggers.

Mt. Sinai Hospital.—Charles A. Elsberg, R. Ottenberg.

New York City College.—A. J. Goldfarb, Thomas A. Storey, C. E. A.
Winslow.

New York City Departments. Education.—C. Ward Crampton.
flealth.—James P. Atkinson, Edwin J. Banzhaf, L. W. Famulener, C. B.
Fitzpatrick, Alfred F. Hess, Edna Steinhardt, Anna W. Williams.

New York Polyclinic Medical School.—Isaac Adler.

New York Post-Graduate Medical School.—Ludwig Kast, W. J. Mac-
Neal.

New York University. — Harlow Brooks, J. W. Draper, Edward K. Dun-
ham, E. M. Ewing, Holmes C. Jackson, Arthur R. Mandel, John A. Mandel,
William H. Park, H. D. Senior, Douglas Symmers, George B. Wallace.

Rockefeller Institute for Medical Research.—John Auer, F. W. Bancroft,
F. J. Birchard, E. E. Butterfield, Alexis Carrel, A. E. Cohn, Rufus Cole,
A. R. Dochez, George Draper, Simon Flexner, T. H. Githens, Walter A.
Jacobs, Don R. Joseph, I. S. Kleiner, Richard V. Lamar, P. A. Levene,
Jacques Loeb, W. H. Manwaring, S. ). Meltzer, Gustave M. Meyer, F. M.
McCrudden, Hideyo Noguchi, G. Canby Robinson, Peyton Rous, H. F.
Swift, B. T. Terry, Donald D. Van Slyke, H. Wastenays.

Sage Institute of Pathology.— Horst Oertel

St. Francis Hospital.—Fritz Schwyzer.

St. Luke's Hospital—A. B. Eisenbrey.

Brooklyn Botanic Garden.—C. Stuart Gager.

$19 Madison Avenue.—H. D. Dakin.

Non-Resident.

Baltimore Medical College.—Charles E. Simon.

Carnegie Institution of Washington.—Francis G. Benedict (/Vuirition
Laboratory, Boston), A. M. Banta, Charles B. Davenport, R. A. Gortner
(Station for Experimental Evolution, Cold Spring Harbor, N. Y.), D. T.
MacDougal (Washington), Alfred G. Mayer (Marine Laboratory, Tortugas,
fila.).

Connecticut Agricultural Experiment Station (New Haven).—Thomas
B. Osborne.

Georgia State Board of Health (Atlanta).—Katharine R. Collins.

Kentucky Agricultural Experiment Station (Lexington).—J. H. Kastle.

Maine Agricultural Experiment Station (Orono).— Raymond Fearl.

Massachusetts Institute of Technology.—Percy G. Stiles.

Medico-Chirurgical College (Philadelphia).—Isaac Ott.

Mercy Hospital (Pittsfield, Mass.).—Thomas Flournoy.

Michael Reese Hospital (Chicago).—James W. Jobling.

Northwestern University Medical School (Chicago).—]. B. Murphy.
Philippine Medical School (Manila),—A. O. Shaklee.

Trinity College (Hartford ).—Max W, Morse.

Yonkers, IV. Y.—Isaac F. Harris.

U.S. Departments. Agriculture (Washington, D.C.).—Carl L. Alsberg,
William N. Berg, William Salant ; /terior (Philippine Islands, Bureau of
Science, Manila).—Richard P. Strong, Oscar Teague. Treasury (Public
flealth and Marine-Hospital Service).—John F. Anderson, W. H. Frost,
Wm. H. Hale, Reid Hunt and W. H. Schultz, Washington, D. C.

Universities. Buffalo.—G. H. A. Clowes, Herbert U. Williams. Caiz-
fornia. —F. P. Gay, F. J. Fitzgerald, T. Brailsford Robertson. Chicago. —
R. R. Bensley, A. J. Carlson, Ludvig Hektoen, Edwin O, Jordan, Frank R.
Lillie, H. Gideon Wells. Cizucznatté.—Martin H. Fischer. Corne//.—Andrew
Hunter, Sutherland Simpson. Harvard.cHerman M. Adler, Walter B.
Cannon, W. T. Councilman, David L. Edsall, Otto Folin, H. T. Karsner, G.
H. Parker, Franz Pfaff, W. T. Porter, Joseph H. Pratt, M. J. Rosenau, Theo-
bald Smith, E. E. Southard, E. E. Tyzzer, S. Burt Wolbach, Robert M.
Yerkes. ///inots.—Philip B. Hawk. /Jafan.—Naohidé Yatsu. /Johus Hopkins.
—John J. Abel, Harvey W. Cushing, W. W. Ford, W. S. Halsted, William
H. Howell, John Howland, S. H. Jennings, Walter Jones, F. P. Mall, Adolph
Meyer, William H. Welch. Leland Stanford.—J. V. Cooke, A. C. Craw-
ford, E. C. Dickson, W. Ophiils, Hans Zinsser. McGill (Montreal ).—J.
George Adami, John L. Todd. Michigan.—C. W. Edmunds, Otto C. Glaser,
A. W. Hewlett, Carl G. Huber, Warren P. Lombard, Frederick G. Novy,
Aldred S. Warthin. J/innesota.—Robert B. Gibson, F. W. Schlutz. Vorth
Carolina.—W.de B. MacNider. Pennsylvania.—J. H. Austin, Alexander C.
Abbott, D. H. Bergey, Ralph S. Lillie, Richard M. Pearce, Edward T.
Reichert, Alfred N. Richards, A. I. Ringer, J. Edwin Sweet, A. E. Taylor.
Pittsburgh. —C. C. Guthrie, Oskar Klotz, Jacob Rosenbloom. Princeton.
—Edwin G. Conklin. Southern California (Los Angeles).—Lyman B. Stookey.
St. Louts.—E. P. Lyon. TZoronto.—T. G. Brodie, J. B. Leathes, A. B.
Macallum. Zulane.—Charles W. Duval. Virginia. —H. E. Jordan.
Washington (St. Louzs).—Joseph Erlanger, Eugene L. Opie, Ernst Sachs,
Philip A. Shaffer. Wésconsin.—Charles R. Bardene, C. H. Bunting, Ar-
thur S. Loevenhart, Mazyck P. Ravenel. Western Reserve (Cleveland).
—George W. Crile, J. J. R. Macleod, Torald Sollmann, G. N. Stewart.
Ya/e.—R. H. Chittenden, Ross G. Harrison, Lafayette B. Mendel, Frank
P. Underhill, Lorande Loss Woodruff.

Cambridge, England.—C. G. L. Wolf.

University College (London).—Arthur R. Cushny.

Phipps Institute (Philadelphia).—Paul A. Lewis.

Wistar Institute of Anatomy (Philadelphia).—H. H. Donaldson, Shin-
kishi Hatai, E. B. Meigs.

Barnard Skin and Cancer Hospital (St. Louis).—Leo Loeb.

Members present at the fiftieth meeting:

Auer, Benedict, Brooks, Burrows, Butterfield, Cohn, Eisenbrey, Field,
Githens, Goldfarb, Kleiner, Lee, Longcope, McClendon, McCrudden, Man-
waring, Murlin, Norris, Pike, Robinson, Senior, Simpson, Swift, Terry,
Wadsworth, Wallace, Wiggers, H. B. Williams.

Members elected at the fiftieth meeting :
E. A. Park, M. Sittenfeld.

Dates of the next two regular meetings:
December 18, 1912 — February 17, 1913.

PROCEEDINGS

SOCIETY FOR

EXPERIMENTAL BIOLOGY AND MEDICINE

FIFTY-SECOND MEETING

PHYSIOLOGICAL LABORATORY
COLLEGE OF PHYSICIANS AND: SURGEONS

NEW YORK CITY
FEBRUARY 1g, I913

AND

SECOND MEETING
PACIFIC COAST BRANCH, SAN FRANCISCO, CALIFORNIA

FEBRUARY 1, 1913

VOLUME X

No. 3

NEW YORK
1913

CONTENTS.

Gary N. CALKINS: Further light on the conjugation of paramecium. 46 (742).

W. H. MANwarinc and J. BRONFENBRENNER : On lysis of tubercle bacilli. 47 (743),

G. H, A. CLowes: Certain specific reactions in hay fever. 48 (744).

G, H. A. CLowss: Treatment of autumnal hay fever by vaccination, 49 (745).

P. F, WILLIAMS and R. M. Pearce: Experience with Abderhalden’s test for preg-

- mancy. 50(746).

L. L. Wooprurr: The kernplasma relation during the life of a pedigreed race of
Oxytricha fallax. 51 (747).

E. E, BuTTERFIELD: The reaction between oxygen and hemoglobin. 52 (748).

J. GORDON WILson and F, H, Pike: A demonstration of the effects of electrical
stimulation of the labyrinth of the ear. 53 (749). :

A. F. Hess: The pancreatic lipase of infants in acute intestinal disturbances, 54 (750).

W. H. Park, L. W, FAMULENER and E, J. BANZHAF: Influence of protein concen-
tration upon the absorption of antibodies from subcutaneous tissues. 55 (751).

G. CANBY ROBINSON: The influence of the vagus nerves on the faradized auricles in
the dog’s heart. 56 (752).

B. S. OPPENHEIMER and H. B, WILLIAMS: Prolonged complete heart block without
lesion of the bundle of His with frequent changes in the idio-ventricular complexes.
57 (753).

Cry. Wieoks and E. F. DuBois: Methods for the production of temporary val-
vular lesions. 58 (754).

A, J, GOLDFARB: The influence of the central nervous system in regeneration of the
annelid worm. 59 (755).

A. J. GOLDFARB: Changes in concentration of sea water and their influence upon re-
generation. 60 (756).

B. T. TeRRy: Variations in the amount of transformed atoxyl (trypanotoxyl) pro-
duced by varying the strength of atoxyl incubated with blood. 61 (757).

W. J. MAcNEAL and A, F. CHAcE: Some observations on bacteria of the duodenum.
62 (758).

A. E. Pde The effects of morphine on the mechanism of the dog’s heart after re-
moval of one vagus nerve. 63 (759).

T. S, GirHEns: The influence of temperature on the minimal dose of strychnine in
frogs and the time of onset of tetanus. 64 (760).

Wo. DE B. MACNIpDER: The difference in the effect of Gréhant’s anesthetic and of
morphine-ether on the total output and composition of the urine in normal dogs.
65 (761).

SER Gh) SIMPSON : The rate of growth in the dog. 66 (762).

ANDREW HunTER: The influence of experimental cretinism upon nitrogenous meta-
bolism in the sheep. 67 (763).

DEWITT STETTEN and JACOB ROSENBLOOM: Metabolism studies in a case of hypo-
pituitarism, with infantilism of the Lorain type. 68 (764).

E. L. Scotr: The relation of pancreatic extract to the sugar of the blood. 69 (765).

J. P. ATKINSON and C. B. Fitzparrick: On the presence of pressor substances in
experimental immunity. 70 (766).

G. H. A. CLowEs, FRANCIS C. GOLDSBOROUGH and F. WeEsT: On a complement
deviation reaction exhibited in pregnancy. 71 (767).

G. H. A. CLowes and Francis C, GoLDsBoROUGH: On the antitryptic reaction
exhibited in pregnancy. 72 (768).

RICHARD WelL: On a new factor in anaphylaxis. 73 (769).

W. S. HAtsTEAD: Hypertrophy of the thyroid gland. Revision of experiments made
25 years ago. 74(770).

W. S. HatstTreaD: Partial occlusion of the aorta by bands of living tissue. 75 (771).

F, P. Gay and T. BRAILSFORD ROBERTSON: Studies on the nature of biological
specificity. 76 (772).

T. BRAILSFORD ROBERTSON: On the nature of Odcytin; the fertilizing and cytoly-
zing substance in mammalian blood-sera. 77 (773).

T. BRAILSFORD ROBERTSON: On the influence of lecithin upon the development of
sea-urchin embryos. 78 (774).

H. J. SEARs (by invitation): On acid agglutination as a method of differentiation of

bacteria. 79 (775).

The proceedings of the Society for Experimental Biology and Medicine are pub-
lished as soon as possible after each meeting. Regular meetings of the Society are held
in New York on the third Wednesdays of October, December, February, April and
May. A volume of the proceedings consists of the numbers issued during en
academic year. ‘

The price per volume, sent postage prepaid, is one dollar. The price of copies
of the proceedings of any meeting is twenty cents each, postage prepaid. Subscriptions
are payable in advance.

PRESIDENT — James Ewing, Cornell University Medical College.

VicE-PRESIDENT — Cyrus W. Field, Bellevue Hospital.

SECRETARY — Holmes C. Jackson, University and Bellevue Hospital Medical
College.

TREASURER — Charles Norris, Bellevue Hospital.

MANAGING EpiIToR — The Secretary, 338 E. 26th St., New York City.

PROCEEDINGS

OF THE

SOCIETY FOR
EXPERIMENTAL BIOLOGY AND MEDICINE

FIFTY-THIRD MEETING

PHYSIOLOGICAL LABORATORY
UNIVERSITY AND BELEEVUE HOSPITAL
MEDICAL COLLEGE

NEW YORK CITY

APRIL) 16, 51g F3

AND

THIRD MEETING
PACIFIC COAST BRANCH, SAN FRANCISCO, CALIFORNIA
APRIL 2, 1913

VOLUME X

No. 4

NEW YORK
1913

CONTENTS.

Frank W. BANncroFT: Heliotropism and galvanotropism in Euglena. 80 (776).

R. A. LAMBerT: The fat content, morphology and length of life of cells growing in
diluted blood plasma. 81 (777).

J. F. McCLenpon: The rate of absorption of water by the skin of the frog, in relation
to M. H. Fischer’s theory of edema. 82 (778).

J. F. McCiLenpon : The dynamics of a model of cell division. 83 (779) .

T. S. GiTHeEns and S. J. MELTZER: Antitoxic action of sodium iodide on morphin.
84 (780).

E. B. Meics: On the nature of the semipermeable membranes which surround the
fibres of striated muscle. 85 (781).

H. H. JANeway and E. M. Ewinc: The mechanical factors of excessive artificial
respiration and a consideration of their relation to the acapnial theory of shock.
86 (782).

J. L. Topp: A note on the transmission of spirochaetes. 87 (783).

A. J. GoLpFArB: Experimentally fused larvae. 88 (784).

J. RosENBLOOM and B. A, CoHoz: Metabolism studies in a case of myotonia atrophica.
89 (785).

N. B. Fosrer: Sugar from lactic acid in human diabetes. 90 (786).

J. A. ATKINSON and C, B, FITZPATRICK : On some vaccinia blood pressor substances
in rabbits. 91 (787).

T. BRAILSFORD ROBERTSON and THEODORE C, BURNETT: Preliminary communication
on the part played by cholesterol in determining the incidence of carcinoma. 92
(788).

T. BRAILSFORD ROBERTSON and THEODORE C, BURNETT: The influence of digitonin
upon the growth of carcinoma. 93 (789).

THEODORE C. BuRNETT: Note on the relation of alexin to thrombin. 94 (790).

HAns ZINSSER: On the nature of the union of alexin with specific precipitates. 95
(791)

J. G. FitzGERALD and GERTRUDE MACKINTOSH : Action of gentian violet on Mucosus
capsulatus group. 96 (792).

S. W. Younc (by invitation): On the influence of light on the electrical potential of
bacterial and other suspensions. 97 (793).

The proceedings of the Society for Experimental Biology and Medicine are pub_
lished as soon as possible after each meeting. Regular meetings of the Society are held
in New York on the third Wednesdays of October, December, February, April and
May. A volume of the proceedings consists of the numbers issued during an
academic year.

The price per volume, sent postage prepaid, is one dollar. The price of copies
of the proceedings of any meeting is twenty cents each, postage prepaid. Subscriptions
are payable in advance. ;

PRESIDENT — James Ewing, Cornell University Medical College.

VICE-PRESIDENT — Cyrus W. Field, Bellevue Hospital.

SECRETARY — Holmes C. Jackson, University and Bellevue Hospital Medical
College.

TREASURER — Charles Norris, Bellevue Hospital.

MANAGING Epiror —The Secretary, 333 E. 26th St., New York City.

PROCEEDINGS

OF THE

SOCIETY FOR

EXPERIMENTAL BIOLOGY AND MEDICINE

FIFTY-FOURTH MEETING

ZOOLOGICAL LABORATORY

COLUMBIA UNIVERSITY

NEW YORK CITY,

MAY 21, I913

VOLUME X

No. 5

NEW YORK

IgI3

CONTENTS.

E. E BurrerFIELD: The color index and color of the red blood corpuscles. 98
(794).

S. J. MELTZER and J. AuER : Combined action of magnesium and ether ; evidence for
a central effect of magnesium. 99 (795).

PiyTon Rous: Note on the production of acid by tissues growing in vitro, 100 (796).

C. R SrockarpD: The location of the optic anlage in Amblystoma and the interpreta-
tion of certain eye defects. 101 (797).

D. WriGHT WILSON: The occurrence of betaine in the muscles of invertebrates. 102
(798).

E. C. KENDALL: The chemical constituents of the thyroid gland. 103 (799).

E. C. KENDALL: The specific physiological activity of certain constituents of the thy-
roid gland. 104 (800).

M. S, FINE and V. C. Myers: Further studies on muscle creatine. 105 (801).

J. H. Austin and C. B. Farr: The total non-protein nitrogen of the blood in nephri-
tis and allied conditions. 106 (802).

W. SALANT and C, S. SmitH: The toxicity of sodium tartrate with special reference
to diet and tolerance. 107 (803).

J. R. Mur In and B, KRAMER: The influence of pancreatic and duodenal extracts on
the glycosuria and the respiratory metabolism of depancreatized dogs. 108
(804).

J. R. Muriin, L. EDELMANN and C. W. GiLEs: The carbon dioxide and oxygen
content of the blood after ligation of the abdominal aorta and the inferior vena
cava. 1c¢9 (805).

J. E. Sweet, E, P. Corson- WHITE and G. M. SAxon: The influence of certain diets
upon the growth of experimental tumors. 110 (806).

CHARLES E. SIMON and MArTHA A. Woop: On the inhibitory action of certain anilin
dyes upon bacterial development. 111 (807).

R, M. PEARCE: Waxy degeneration of muscle in venom intoxication, 112 (808).

Isaac Ort and J. C. Scorr: Note on the effect of animal extracts upon the volume of
the thyroid gland. 113 (809).

SUTHERLAND Simpson: The relation of external temperature to hibernation. 114
(810).

RICHARD WEIL: Anaphylaxis in immune animals. 115 (811).

A. J. CARLSON: The parathyroids and pregnancy, 116 (812).

A. J. CARLSON: The correlation between the physiological state of the thyroid of the
mother and of the fetus. 117 (813).

A. J. CARLSON: Parathyroid tetany and active immunity. 118 (814).

A, J. CARLSON and A. WOELFEL: The solubility of various lead salts in the human
gastric juice and its bearing on the hygiene of the lead industries. 119 (815).

A. M. BANTA and R. A. GORTNER : Certain observations on the occurrence of tyrosin-
ase in amphibian egg jell. 120 (816).

A. M. BANTA: Experiments on the light and tactile reactions of a cave variety and
open water variety of an amphipod species. 121 (817).

The proceedings of the Society for Experimental Biology and Medicine are pub-
lished as soon as possible after each meeting. Regular meetings of the Society are held
in New York on the third Wednesdays of October, December, February, April and
May. A volume of the proceedings consists of the numbers issued during an
academic year.

The price per volume, sent postage prepaid, is one dollar. The price of copies
of the proceedings of any meeting is twenty cents each, postage prepaid. Subscriptions
are payable in advance.

PRESIDENT — James Ewing, Cornell University Medical College.

VicE-PRESIDENT — Cyrus W. Field, Bellevue Hospital.

SECRETARY — Holmes C. Jackson, University and Bellevue Hospital Medical
College.

TREASURER — Charles Norris, Bellevue Hospital.

MANAGING EDITOR — The Secretary, 338 E. 26th St., New York City.

CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR
EXPERIMENTAL BIOLOGY AND MEDICINE.

Resident (Greater New York).

College of the City of New York.— A. J. Goldfarb, G. G. Scott, Thomas
A. Storey, C. E. A. Winslow.

Columbia University.—Russell Burton-Opitz, Gary N. Calkins, Henry
E. Crampton, Haven Emerson, William J. Gies, P. E. Howe, Theodore C.
Janeway, R. A. Lambert, Frederic S. Lee, Isaac Levin, C. C. Lieb, W. F.
Longcope, W. G. MacCallum, Thomas H. Morgan, H. O. Mosenthal, B.
S. Oppenheimer, Alwin M. Pappenheimer, F. H. Pike, E. L. Scott, Henry C.
Sherman, M. Sittenfeld, H. B. Williams, Augustus CB. Wadsworth, Edmund
B. Wilson, Francis C. Wood, Hans Zinsser.

Cornell University Medical College.—S. P. Beebe, Stanley R. Benedict,
M. J. Burrows, B. H. Buxton, A. F. Coca, E. F. DuBois, C. Eggleston,
William J. Elser, James Ewing, Nellis B. Foster, J. A. Hartwell, Robert
A. Hatcher, Graham Lusk, J. F. McClendon, John R. Murlin, Chas. R.
Stockard, Oscar Teague, John C. Torrey, Richard Weil, C. J. Wiggers.

FHlospitals. Bellevue.—Cyrus W. Field, Charles Norris. <Aznug’s County.
—B. T. Terry. Mt. Sinmaz.—Charles A. Elsberg, R. Ottenberg. Rooseveld.
—G. K. Falk. Presbyterian.—R. A. Cecil, George Draper. Sz. Lukes.—
A, B. Eisenbrey.

New Vork City Departments. Education.—C. Ward Crampton.
Health.—James P. Atkinson, Edwin J. Banzhaf, L. W. Famulener, C. B.
Fitzpatrick, Alfred F. Hess, Edna Steinhardt, Anna W. Williams,

New York Polyclinic Medical School.—Isaac Adler.

New York Post-Graduate Medical School.—Ludwig Kast, W. J. Mac-
Neal, V. C. Myers.

New York University. — Harlow Brooks, J. W. Draper, Edward K. Dun-
ham, E. M. Ewing, A. O. Gettler, Holmes C. Jackson, Arthur R. Mandel,
John A. Mandel, William H. Park, H. D. Senior, Douglas Symmers, George
B. Wallace.

Rockefeller Institute for Medical Research.—John Auer, F. W. Bancrott,
Jacob Bronfenbrenner, E. E. Butterfield, Alexis Carrel, P. F. Clark, A. E.
Cohn, Rufus Cole, A. R. Dochez, Simon Flexner, T. H. Githens, Walter A.
Jacobs, I. S. Kleiner, Richard V. Lamar, P. A. Levene, Jacques Loeb, W.
H. Manwaring, S. ). Meltzer, Gustave M. Meyer, F. M. McCrudden,
James B. Murphy, Hideyo Noguchi, G. Canby Robinson, Peyton Rous, H.
F. Swift, Donald D. Van Slyke, C. J. West, Martha Wollstein.

Brooklyn Botanic Garden.—C, Stuart Gager.

819 Madison Avenue.—H. D. Dakin.

30 West gath Street.—Horst Oertel.

Non-Resident.

Agricultural Experiment Stations. Connecticut (New Haven)—Thomas
B. Osborne. Kentucky (Lexington).—J. H. Kastle. Maine (Orono)—Raymond
Fearl.

Carnegie Institution of Washington.—Francis G. Benedict (Vutrition
Laboratory, Boston), A. M. Banta, Charles B. Davenport, R. A. Gortner
(Station for Experimental Evolution, Cold Spring Harbor, N. Y.), D. T.
MacDougal (Washington), AlfredG. Mayer (Marine Laboratory, Tortugas,
Fla.). y

Colleges. Baltimore Medical.—Charles E. Simon. Aryn Mawr.—
Don R. Joseph. Medico-Chirurgical (Philadelphia).—Isaac Ott. Jefferson
Medical (Philadelphia).—P. B. Hwak. Philippine Medical (Manila).—R.
B. Gibson, A. O. Shaklee. Trinity (Hartford).—Max W. Morse.

Georgia State Board of Health (Atlanta). Katharine R. Collins.

Flospitals. Barnard Skin and Cancer (St. Louis),—Leo Loeb. Mercy
(Pittsfield, Mass.). Thomas Flournoy. Michael Reese (Chicago).—James
W. Jobling.

Institutes. Massachusetts, of Technology.—Percy G. Stiles. Phipps
(Philadelphia).—Paul A. Lewis. Wistar (Philadelphia),—H. H. Donaldson,
Shinkishi Hatai, E. B. Meigs.

U. S. Departments. Agriculture (Washington, D.C.).—Carl L. Als-
berg, William N. Berg, F. J. Birchard, William Salant; Treasury (Public
Health and Marine-Hospital Service, Washington, D. C.).—John F.
Anderson, W. H. Frost, W. H. Hale, Reid Hunt and W. H. Schultz.

Universities. Buffalo.—G.H. A. Clowes, Herbert U. Williams. Ca/z-
fornia. —J. V. Cooke, F. J. Fitzgerald, F. P. Gay, T. Brailsford Robertson,
H. Wastenays. Chicago. —A. J. Carlson, Ludvig Hektoen, Edwin O. Jordan,
Frank R. Lillie, H. Gideon Wells. Czuctnatt2.—Martin H. Fischer. Corned.
Melvin Dresbach, Andrew Hunter, Sutherland Simpson. Harvard.—Her-
man M. Adler, Walter B. Cannon, W. T. Councilman, Harvey Cushing,
David L. Edsall, Otto Folin, H. T. Karsner, G. H. Parker, Franz Pfaff, W. T.
Porter, Joseph H. Pratt, M. J. Rosenau, Theobald Smith, E. E. Southard,
Richard P. Strong, E. E. Tyzzer, S. Burt Wolbach, Robert M. Yerkes.
Japan.—Naohidé Yatsu. Johns Hopkins.—John J. Abel, W. W. Ford, W. S.
Halsted, A. D. Hirschfelder, William H. Howell, John Howland, H. S.
Jennings, Walter Jones, Adolph Meyer, E. A. Park, William H. Welch.
Leland Stanford.—A. C. Crawford, E. C. Dickson, W. Ophils, McG:#l/
(Montreal).—J. George Adami, John L. Todd. Michigan.—C. W.
Edmunds, Otto C. Glaser, A. W. Hewlett, Carl G. Huber, Warren P.
Lombard, Frederick G. Novy, Aldred S. Warthin. Minnesota.—F. W.
Schlutz. Vebraska.—A. E. Guenther. orth Carolina.—W. de B. Mac-
Nider. orthwestern.—John B. Murphy. Fennsylvania.—J. H. Austin,
Alexander C. Abbott, D. H. Bergey, Ralph S. Lillie, Richard M. Pearce,
Edward T. Reichert, Alfred N. Richards, A. I. Ringer, J. Edwin Sweet,
A.E. Taylor. ittsburgh.—C. C. Guthrie, Oskar Klotz, Jacob Rosenbloom.
Princeton.—Edwin G. Conklin. Southern California (Los Angeles).—Lyman
B. Stookey. S¢. Zouts.—E. P. Lyon. Toronto.—J. B. Leathes, A. B.
Macallum. Zulane.—Charles W. Duval. Virginia. —H. E. Jordan.
Washington (St. Louis).—Joseph Erlanger, Eugene L. Opie, Ernst Sachs,
Philip A. Shaffer. W7sconsin.—Charles R. Bardeen, C. H. Bunting, L. J.
Cole, Arthur S. Loevenhart, Mazyck P. Ravenel. Western Reserve (Cleve-
Jand).—George W. Crile, Paul J. Hanzlik, J. J. R. Macleod, Torald Soll-
mann, G. N. Stewart. Va/e.—R. H. Chittenden, Ross G. Harrison, Daven-
port Hooker, Lafayette B. Mendel, Frank P. Underhill, Lorande Loss
Woodruff.

Cambridge, England.—C. G. L. Wolf.
Switzerland (Lucern).— Fritz Schwyzer.
Yonkers, N. Y.—Isaac F. Harris.

Members present at the fifty-fourth meeting:

Atkinson, Auer, Austin, Bronfenbrenner, Butterfield, Calkins, Eisenbrey,
Ewing, J., Field, Fitzpatrick, Jackson, Kleiner, Meigs, Meltzer, Morgan,
Murlin, Myers, Norris, Oppenheimer, Rous, Salant, Scott, E. L., Stockard,
Terry, Wiggers, Wilson, Wood.

Members elected at the fifty-fourth meeting:

A. H. Caulfeild, J. W. Churchman, H. R. Gaylord, H. H. Janeway,
J. R. Mohler, A. R. Moore, R. Pemberton, G. B. White.

Dates of the next two regular meetings:
October 15, 1913.—December 17, 1913.

CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR
EXPERIMENTAL BIOLOGY AND MEDICINE.

Resident (Greater New York).

College of the City of New York.— A. J. Goldfarb, Thomas A. Storey, C.
E. A, Winslow.

Columbia University.—Russel Burton-Opitz, Gary N. Calkins, Henry
E. Crampton, Haven Emerson, Nellis B. Foster, William J. Gies, P. E.
Howe, Theodore C. Janeway, R. A. Lambert, Frederic,S. Lee, Isaac Levin,
C. C. Lieb, W. F. Longcope, W. G. MacCallum, Thomas H. Morgan, H.
O. Mosenthal, B. S. Oppenheimer, Alwin M. Pappenheimer, F. H. Pike,
E. L. Scott, Henry C. Sherman, M. Sittenfeld, H. B. Williams, Augustus PB.
Wadsworth, Edmund B. Wilson, Francis C. Wood.

Cornell University Medical College.—S. P. Beebe, Stanley R. Benedict,
M. J. Burrows, B. H. Buxton, A. F. Coca, E. F. DuBois, C. Eggleston,
William J. Elser, James Ewing, Robert A. Hatcher, Graham Lusk, J. F.
McClendon, John R. Murlin, Chas. R. Stockard, Oscar Teague, John C.
Torrey, Richard Weil, C. J. Wiggers.

Flospitals. Bellevue.—Cyrus W. Field, Charles Norris. <zug’s County.
—B. T. Terry. J%. Sinaz.—Charles A. Elsberg, R. Ottenberg. Roosevelt.
—G. K. Falk. Presbyterian.—R. A. Cecil, George Draper. S¢. Lukes.—
A, B. Eisenbrey.

New York City Departments. Education.—C. Ward Crampton.
Health.—James P. Atkinson, Edwin J. Banzhaf, L. W. Famulener, C. B.
Fitzpatrick, Alfred F. Hess, Edna Steinhardt, Anna W. Williams,

New York Polyclinic Medical School.—Isaac Adler.

New York Post-Graduate Medical School.—Ludwig Kast, W. J. Mac-
Neal, V. C. Myers. y

New York University. — Harlow Brooks, J. W. Draper, Edward K. Dun-
ham, E. M. Ewing, Holmes C. Jackson, Arthur R. Mandel, John A. Mandel,
William H. Park, H. D. Senior, Douglas Symmers, George B. Wallace.

Rockefeller Institute for Medical Research.—John Auer, F. W. Bancrott,
Jacob Bronfenbrenner, E. E. Butterfield, Alexis Carrel, P. F. Clark, A. E.
Cohn, Rufus Cole, A. R. Dochez, Simon Flexner, T. H. Githens, Walter A.
Jacobs, I. S. Kleiner, Richard V. Lamar, P. A. Levene, Jacques Loeb, W.
H. Manwaring, S. J. Meltzer, Gustave M. Meyer, F. M. McCrudden,
James B. Murphy, Hideyo Noguchi, G. Canby Robinson, Peyton Rous, H.
F. Swift, Donald D. Van Slyke, C. J. West.

Brooklyn Botanic Garden.—C, Stuart Gager.

$19 Madison Avenue.—H. D. Dakin.

30 West ggth Street.—Horst Oertel.

Non-Resident.

Agricultural Experiment Stations. Connecticut (New Haven)—Thomas
B, Osborne. Kentucky (Lexington).—J. H. Kastle. Mazne ( Orono)—Raymond
Fearl.

Carnegie Institution of Washington.—Francis G. Benedict (Vutrition
Laboratory, Boston), A. M. Banta, Charles B. Davenport, R. A. Gortner
(Station for Experimental Evolution, Cold Spring Harbor, N. Y.), D. T.
MacDougal (Washington), Alfred G. Mayer (Marine Laboratory, Tortugas,
Fia.).

Colleges. Bryn Mawr.—Don R. Joseph. Medico-Chirurgical (Phila-
delphia).—Issac Otto. Jefferson Medical (Philadelphia).—P. B. Hawk.
Philippine Medical (Manila).—R. B. Gibson, A. O. Shaklee. Trinity (Hart-
Jord).—Max W. Morse.

Georgia State Board of Health (Atlanta), Katharine R. Collins.

Flospitals. Barnard Skin and Cancer (St. Louis).—Leo Loeb. Mercy
(Pittsfield, Mass.). Thomas Flournoy. Michael Reese (Chicago).—James
W. Jobling.

Institutes. Massachusetis, of Technology.—Percy G. Stiles. Phipps
(Philadelphia).—Paul A. Lewis. Wistar (Philadelphia).—H. H. Donaldson,
Shinkishi Hatai, E. B. Meigs.

U. S. Departments. Agriculture (Washington, D.C.).—Carl L. Als-
berg, William N. Berg, F. J. Birchard, William Salant; Treasury (Public
flealth and Marine-Hospital Service).—John F. Anderson, W. H. Frost, W.
H. Hale, Reid Huntand W. H. Schultz, Washington, D. C.

Universitics. Buffalo.—G. H. A. Clowes, Herbert U. Williams. Ca/i-
fornia. —J. V. Cooke, F. J. Fitzgerald, F. P. Gay, T. Brailsford Robertson,
H. Wastenays. Chicago. —A. J. Carlson, Ludvig Hektoen, Edwin O. Jordan,
Frank R. Lillie, H. Gideon Wells. Cimcinmattz.—Martin H. Fischer. Cornell.
Melvin Dresbach, Andrew Hunter, Sutherland Simpson. Aarvard.—Her-
man M. Adler, Walter B. Cannon, W. T. Councilman, Harvey Cushing,
David L. Edsall, Otto Folin, H. T. Karsner, G. H. Parker, Franz Pfaff, W. Tf.
Porter, Joseph H. Pratt, M. J. Rosenau, Theobald Smith, E. E. Southard,
Richard P. Strong, E. E. Tyzzer, S. Burt Wolbach, Robert M. Yerkes.
Japan.—Naohidé Yatsu. Johns Hopkins.—John J. Abel, W. W. Ford, W.S.
Halsted, A. D. Hirschfelder, William H. Howell, John Howland, H. S.
Jennings, Walter Jones, Adolph Meyer, E. A. Park, William H. Welch.
Leland Stanford.—A. C. Crawford, E. C. Dickson, W. Ophiils, Hans
Zinsser. McGill (Montreal).—J. George Adami, John L. Todd. Michigan.
—C. W. Edmunds, Otto C. Glaser, A. W. Hewlett, Carl G. Huber, Warren P.
Lombard, Frederick G. Novy, Aldred S. Warthin. Minnesofa.—F. W.
Schlutz. Mebvaska.—A. E. Guenther. orth Carolina.—W. de B. Mac-
Nider. Northwestern.—John B. Murphy. Fennsylvania.—J. H. Austin,
Alexander C. Abbott, D. H. Bergey, Ralph S. Lillie, Richard M. Pearce,
Edward T. Reichert, Alfred N. Richards, A. I. Ringer, J. Edwin Sweet,
A.E. Taylor. ittsburgh.—C. C.'Guthrie, Oskar Klotz, Jacob Rosenbloom.
Princeton.—EdwinG. Conklin. Southern California (Los Angeles).—Lyman
B. Stookey. S7. Zouis.—E. P. Lyon. Toronto.—J. B. Leathes, A. B.
Macallum. 7Zu/ane.—Charles W. Duval. Virginza.—H. E. Jordan.
Washington (St. Louzs).—Joseph Erlanger, Eugene L. Opie, Ernst Sachs,
Philip A. Shaffer. Wisconsin.—Charles R. Bardeen, C. H. Bunting, L. J.
Cole, Arthur S. Loevenhart, Mazyck P. Ravenel. Western Reserve (Cleve-
land).—George W. Crile, Paul J. Hanzlik, J. J. R. Macleod, Torald Soll-
mann, G. N. Stewart. Va/e.—R. H. Chittenden, Ross G. Harrison, Daven-
port Hooker, Lafayette B. Mendel, Frank P. Underhill, Lorande Loss
Woodruff.

Cambridge, England.—C. G. L. Wolf.
Switzerland (Lucern).—Fritz Schwyzer.
Yonkers, IV. Y.—Isaac F. Harris.

Members present at the fifty-third meeting:

Bancroft, Beebe, Bronfenbrenner, Brooks, Butterfield, Eisenbrey, Ew-
ing, E. M., Ewing, J., Field, Fitzpatrick, Githens, Goldfarb, Jackson, Kleiner,
Lambert, McClendon, Meigs, Meltzer, Murphy, Norris, Pike, Stockard,
Wastenays, Wiggers.

Members elected at the fifty-third meeting:
A. O. Gettler, J. A. Hartwell, G. G. Scott, Martha Wollstein.

Dates of the next two regular meetings:
May 21, 1913.—October 15, 1913.

I sbit Be

CLASSIFIED LIST OF MEMBERS OF THE SOCIETY FOR
EXPERIMENTAL BIOLOGY AND MEDICINE.

Resident (Greater New York).

Bellevue Hospital.—Cyrus W. Field, Charles Norns.

Columbia University.—Russell Burton-Opitz, Gary N. Calkins, Henry
E. Crampton, Richard H. Cunningham, Haven Emerson, Nellis B. Foster,
William J. Gies, A. E. Guenther, Philip H. Hiss, P. E. Howe, Theodore
C. Janeway, R. A. Lambert, Frederic S. Lee, Isaac Levin, C. C. Lieb, W.
F. Longcope, W. G. MacCallum, Thomas H. Morgan, H. O. Mosenthal,
Alwin M. Pappenheimer, B. S. Oppenheimer, F. H. Pike, E. L. Scott,
Henry C. Sherman, M. Sittenfeld, Hugh A. Stewart, Augustus CD. Wadsworth,
Edmund B. Wilson, Francis C. Wood.

Cornell University Medical College.—S. P. Beebe, Stanley R. Benedict,
M. J. Burrows, B. H. Buxton, A. F. Coca, E. F. DuBois, C. Eggleston,
William J. Elser, James Ewing, Robert A. Hatcher, Graham Lusk, J. F.
McClendon, John R. Murlin, Chas. R. Stockard, John C. Torrey, Richard
Weil, C. J. Wiggers.

Mt. Sinat Hospital.—Charles A. Elsberg, R. Ottenberg.

New York City College. —A. J. Goldfarb, Thomas A. Storey, C. E. A.
Winslow.

New York City Departments. Education,—C. Ward Crampton.
Health.—James P. Atkinson, Edwin J. Banzhaf, L. W. Famulener, C. B.
Fitzpatrick, Alfred F. Hess, Edna Steinhardt, Anna W. Williams.

New York Polyclinic Medical School.—Isaac Adler. |

New York Post-Graduate Medical School.—Ludwig Kast, W. J. Mac-
Neal, V. C. Myers.

New York University. — Harlow Brooks, J. W. Draper, Edward K. Dun-
ham, E. M. Ewing, Holmes C. Jackson, Arthur R. Mandel, John A. Mandel,
William H. Park, H. D. Senior, Douglas Symmers, George B. Wallace.

Rockefeller Institute for Medical Research.—John Auer, F. W. Bancrott,
Jacob Bronfenbrenner, E. E. Butterfield, Alexis Carrel, P. F. Clark, A. E.
Cohn, Rufus Cole, A. R. Dochez, Simon Flexner, T. H. Githens, Walter A.
Jacobs, I. S. Kleiner, Richard V. Lamar, P. A. Levene, Jacques Loeb, W.
H. Manwaring, S. J. Meltzer, Gustave M. Meyer, F. M. McCrudden, J. B.
Murphy, Hideyo Noguchi, G. Canby Robinson, Peyton Rous, H. F. Swift,
Donald D. Van Slyke, C. J. West.

Sage Institute of Pathology.—Horst Oertel.

King’s County Hospital.—B. T. Terry.

Roosevelt fospital.—G. K. Falk.

Presbyterian Hospital. —R. A. Cecil, George Draper.

St. Francis Hospital.—Fritz Schwyzer.

St. Luke’s Hospital—A. B. Eisenbrey.

Brooklyn Botanic Garden,—C, Stuart Gager.

819 Madison Avenue.—H. D. Dakin.

Non-Resident.

Bryn Mawr College.—Don R. Joseph.

Carnegie Institution of Washington.—Francis G. Benedict (Nutrition
Laboratory, Boston), A. M. Banta, Charles B. Davenport, R. A. Gortner
(Station for Experimental Evolution, Cold Spring Harbor, N. Y.), D. T.
MacDougal (Washington), Alfred G. Mayer (Marine Laboratory, Tortugas,
Fia.).

Connecticut Agricultural Experiment Station (New Haven).—Thomas
B. Osborne.

Geergia State Board of Health ( Atlanta).—Katharine R. Collins.

Kentucky Agricultural Experiment Station (Lexington).—J. H. Kastle.

Maine Agricultural Experiment Station (Orono).— Raymond Fearl.

Massachusetts Institute of Technology.—Percy G. Stiles.

Medico-Chirurgical College (Philadelphia).—Isaac Ott.

Mercy Hospital (Pittsfield, Mass.).—Thomas Flournoy.

Michael Reese Hospital (Chicago).—James W. Jobling.

Jefferson Medical College.—P. B. Hawk.

Philippine Medical School (Manila).—R. B. Gibson, A. O. Shaklee.

Trinity College (Hartford ).—Max W. Morse.

Yonkers, NV. Y.—Isaac F. Harris.

U.S. Departments. Agriculture (Washington, D.C.).—Carl L. Alsberg,
William N. Berg, F. J. Birchard, William Salant; Jnterior (Philippine
Islands, Bureau of Science, Manila).—Richard P. Strong, Oscar Teague.
Treasury (Public Health and Marine-Hospital Service).—John F. Anderson,
W. H. Frost, W. H. Hale, Reid Huntand W. H: Schultz, Washington, D. C.

Universitics. Buffalo.—G. H. A. Clowes, Herbert U. Williams. Ca/z
fornia. —F¥. P. Gay, J. V. Cooke, F. J. Fitzgerald, T. Brailsford Robertson,
H, Wastenays. Chicago. —A. J. Carlson, Ludvig Hektoen, Edwin O. Jordan,
Frank R. Lillie, H. Gideon Wells. Cimcznatti.—Martin H. Fischer. Corned].
Melvin Dresbach, Andrew Hunter, Sutherland Simpson. AHarvard.—Her-
man M. Adler, Walter B. Cannon, W. T. Councilman, H. Cushing, David
L. Edsall, Otto Folin, H. T. Karsner, G. H. Parker, Franz Pfaff, W. T. Porter,
Joseph H. Pratt, M. J. Rosenau, Theobald Smith, E. E. Southard, E. E.
Tyzzer, S. Burt Wolbach, Robert M. Yerkes. /afan.—Naohidé Yatsu.
Johns Hopkins.—John J. Abel, W. W. Ford, W. S. Halsted, A. D. Hirsch-
felder, William H. Howell, John Howland, H. S. Jennings, Walter Jones,
Adolph Meyer, E. A. Park, William H. Welch. Leland Stanford.—
A. C. Crawford, E. C. Dickson, W. Ophiils, Hans Zinsser. McGii/
(Montreal ).—J. George Adami, John L. Todd. A%ichigan.—C. W. Edmunds,
Otto C. Glaser, A. W. Hewlett, Carl G. Huber, Warren P. Lombard,
Frederick G. Novy, Aldred S. Warthin. Minnesota.—F.W. Schlutz. North
Carolina.—W.de B. MacNider. orthwestern.—John B. Murphy. Penn-
sylvania.—J. H. Austin, Alexander C. Abbott, D. H. Bergey, Ralph S.
Lillie, Richard M. Pearce, Edward T. Reichert, Alfred N. Richards, A. I.
Ringer, J. Edwin Sweet, A. E. Taylor. Pittsburgh. —C. C. Guthrie, Oskar
Klotz, Jacob Rosenbloom. /rinceton.—Edwin G. Conklin. Southern Calz-
fornia (Los Angeles).—Lyman B. Stookey. S¢. Louis.—E. P. Lyon.
Toronto.—J. B. Leathes, A. B. Macallum. TZz/ane.—Charles W. Duval.
Virginia. —H. E. Jordan. Washington (St. Louis).—Joseph Erlanger,
Eugene L. Opie, Ernst Sachs, Philip A. Shaffer. Wzsconsin.—Charles R.
Bardeen, C. H. Bunting, Arthur S. Loevenhart, Mazyck P. Ravenel.
Western Reserve (Cleveland).—George W. Crile, Paul J. Hanzlik, J. J. R.
Macleod, Torald Sollmann, G. N. Stewart. Ya/e.—R. H. Chittenden, Ross
G. Harrison, Davenport Hooker, Lafayette B. Mendel, Frank P. Underhill,
Lorande Loss Woodruff.

Cambridge, England.—C. G. L. Wolf.

Phipps Institute (Philadelphia).—Paul A. Lewis.

Wistar Institute of Anatomy (Philadelphia).—H. H. Donaldson, Shin-
kishi Hatai, E. B. Meigs.

Barnard Skin and Cancer Hospital (St. Louzs).—Leo Loeb.

Members present at the fifty-second meeting:

Atkinson, Auer, Austin, Bancroft, Beebe, Bronfenbrenner, Butterfield,
Calkins, Clowes, Cohen, Cole, Draper, J. W., DuBois, Dunham, Eisenbrey,
Elsberg, Emerson, Ewing, J., Famulener, Fitzpatrick, Flexner, Gies, Githens,
Goldfarb, Halstead, Harris, Hess, Jackson, Jacobs, Joseph, Kast, Kleiner,
Lambert, Lee, Levin, I., Levene, P. A., Longcope, Lusk, MacCallum, W. J.,
MacNeal, McCrudden, Mandel, J. A., Manwaring, Meltzer, Murphy, Myers,
Norris, Oppenheimer, Pappenheimer, Park, Pearce, Pike, Ringer, Robinson,
Rous, Scott, Senior, Sittenfeld, Stewart, H. A., Steinhart, Terry, Wadsworth,
Wallace, Weil, Wiggins, Williams, Anna, Williams, H. B., Wood.

Members elected at the fifty-second meeting:

RA, Cecil ue: Eggleston, G. K. Falk, Davenport Hooker, P. E. Howe,
C. J. West.

Officers elected at the fifty-second meeting:
President, James Ewing ; Vice-President, Cyrus W. Field; Treasurer,
Charles Norris ; Secretary, Holmes C. Jackson.

Dates of the next two regular meetings:
April 16, 1913—May 21, 1913.

NT

0 750