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PROCEEDINGS
OF THE
SOCIETY FOR
EXPERIMENTAL BIOLOGY AND MEDICINE
VOLUME XIII
1915-1916
EDITED BY THE SECRETARY
NEW YORK
1916
PRESS OF
THE NEW ERA PRINTING COMPANY
LANCASTER, PA.
CONTENTS.
SCIENTIFIC PROCEEDINGS (69th—76th meetings): PAGE
Communications of the sixty-ninth meeting, October 20, 1915. 5
Communications of the seventieth meeting, November 17, 1915 .
Communications of the seventy-first meeting, December 15, 1915
Communications of the seventy-second meeting, January 19, 1916
Communications of the seventy-third meeting, February 16, 1916
Communications of the seventy-fourth meeting, March 15, 1916 . : ;
Communications of the seventy-fifth meeting, April 19, 1916
Communications of the seventy-sixth meeting, May 24, 1916
RECAPITULATION OF THE NAMES OF THE AUTHORS AND OF THE TITLES OF THE
COMMUNICATIONS 3 é : : ; 3 ; . .
EXECUTIVE PROCEEDINGS (69th—76th meetings)
REGISTER OF NAMES AND ADDRESSES OF THE MEMBERS
List OF OFFICERS . : ; : -
CLASSIFIED LIST OF MEMBERS. j : ; : gi é F :
INDEX OF THE SCIENTIFIC PROCEEDINGS
POR te
oat 10 ALT
} oo
o +e :
Liveath:
SCIENTIFIC PROCEEDINGS.
ABSTRACTS OF COMMUNICATIONS.
Sixty-ninth meeting.
Cornell University Medical College, October 20, 1915. President
Lusk in the chair.
I (1065)
As to the cause of the dilatation of the subclavian artery in certain
cases of cervical rib.—Experimental Study.
By W. S. HaLstED, M.D. and Mont RErp, M.D.
[From the Department of Surgery, Johns Hopkins University.]
In twenty-four or more instances a circumscribed dilatation of
the subclavian artery has been observed in cases of cervical rib.
The dilatation in these cases is distal to the site of pressure made
by the rib.
As to the cause of these aneurisms there has been considerable
conjecture, usually prefaced by the comment that their occurrence
would be comprehensible if they presented on the proximal instead
of on the distal side of the compression.
Weakening of the wall of the artery from erosion or trauma,
variable or intermittent pulse pressure, and vasomotor dis-
turbances in nutrition are the suggestions which have been offered
to explain the phenomenon.
For several years my experiments in arterial compression have
had more or less in view the determination of the cause of this
dilatation. For the past year they have been continued by Dr.
Mont Reid and myself almost exclusively with the object of
shedding light on this problem. In 1906 we (Dr. Richardson,
I
2 SCIENTIFIC PROCEEDINGS (69).
Dr. Dawson and myself) made the observation! that after partial
occlusion of the thoracic aorta the maximum pressure may be
permanently lowered as much as 46 mm. Hg, and the minimum
pressure actually increased distal to the constricting band of metal.
The dilatation of the artery observed in arterio-venous fistula,
might, it seemed to me, have a bearing on the interpretation of the
aneurisms in cases of cervical rib. Might not both phenomena,
I asked myself, be due to degenerative changes in the arterial
wall consequent upon lowered pressure—in the case of the cervical-
rib-aneurisms, upon lowering of the pulse pressure.
Now, inasmuch as dilatation of the subclavian artery has rel-
atively so seldom been observed with cervical rib (perhaps 24 times
in about 400 cases) it seemed to me that if it were due merely to
the lowered pulse pressure then only a very definite absolute or
relative amount of reduction of the systolic pressure would suffice
to produce it.
In June, 1914, I observed, in a dog, for the first time an un-
questionable dilatation of the three arteries below the constricting
band which had been placed just above the aortic trifurcation.
The constriction exercised by the band was sufficient to greatly
lessen, if not, indeed, to obliterate the palpable thrill produced by
the constriction, but not enough to shut off the palpable pulse.
With this observation as fresh incentive, Dr. Reid and I have con-
tinued the experiments for the past year and a half with encourag-
ing results: in only one additional instance, however, was there a
very striking dilatation. In this, as in the one of the preceding
year, the occlusion of the aorta by the band was almost total.
If the occlusion must be so nearly complete in order to effect
a pronounced dilatation it will assist to explain not only the dif-
ficulty we have had in producing it in dogs, but also the fact that
it has been observed relatively so seldom in the human subject
from compression of the subclavian artery by a cervical rib. For
when in dogs the aortic pulse is occluded beyond the stage of
1DoG 96. PARTIAL OCCLUSION OF THORACIC AORTA.
Operation, 22/5/1906. Sacrificed 7 months later.
Maximum Mean Minimum Pulse
Pressure. Pressure. Pressure. Pressure.
Femoraliocc cscs ce 116 93 88 28
(OhigyaGlangeaonaooedo 160 113 83 77
DuRATION OF LIFE IN RENAL INSUFFICIENCY. 3
palpable thrill the lumen is in danger of becoming obliterated—
as by the formation of a cylindrical fibrous cord beneath the band
—and thus cancel the experiment; and in the cervical rib cases
we may assume, argumentatively, that the subclavian artery,
compressed to the stage sufficient to produce an aneurism, is likely
to become totally occluded in the presumably considerable time
required for the manifestation of the dilatation. Thus, in dogs a
number of months must apparently elapse after the application
of the band before a dilatation in striking degree can occur. In
the two cases, observed just one year apart, 5 months and 20
days, and 6 months and 19 days, respectively, had elapsed. In
the second of these, however, a dilatation of less than one mm. was
found at the expiration of 2 months.
2 (1066)
A comparison of the effects of glucose and of meat administration
upon the non-protein blood nitrogen and the duration
of life in experimental renal insufficiency.
By J. H. AustTIN and S. S. LEOPOLD.
[From the John Herr Musser Department of Research Medicine and
the William Pepper Clinical Laboratory.]
The following study was undertaken to determine (1) whether
those dietary factors that tend to increase the non-protein blood
nitrogen in acute insufficiency also tend to shorten the duration
of life, and (2) the value of glucose in prolonging life in acute renal
insufficiency. The method adopted was to observe the daily
curve of non-protein blood nitrogen and the duration of life after
complete renal insufficiency had been induced by bilateral ureteral
ligation in a series of dogs; half the animals being given glucose,
the other half, meat.
Six dogs were divided into two groups of three each. One
group was fed upon beef heart, the other upon glucose dissolved
in water. On the third day, 5 c.c. of blood was taken from the
jugular vein for estimation of the total non-protein nitrogen by
the Folin method, and immediately thereafter each dog was
etherized and both ureters ligated. On the following day, blood
4 SCIENTIFIC PROCEEDINGS (69).
was again taken in the same manner and the diet resumed as
before operation. Every day thereafter blood was taken before
feeding. When the stomach became unretentive, the glucose
dogs were given ten per cent. solution of dextrose in distilled
water intraperitoneally. The meat dogs when they refused the
meat were allowed water as desired.
At autopsy, all the ureters were found satisfactorily ligated,
the pelvis of the kidney was filled but not materially distended and
there was no evidence of infection. In the animals receiving
glucose solution intraperitoneally, there was no evidence grossly
of free fluid in the peritoneal cavity.
The following facts were noted. After forty-eight hours on
the diets, but before ureteral ligation, the blood nitrogen of the
two groups was the same. The day after operation, neither meat
nor glucose having been given the day of the operation, the blood
nitrogen in both groups was almost the same, possibly a little
lower in the glucose group; the loss of weight at this time was
the same in the two groups. Forty-five hours after operation,
the meat animals having retained their feeding the day previous,
while the glucose animals had vomited, the loss of weight was
distinctly greater in the glucose group; the blood nitrogen was
distinctly lower in the glucose group. Sixty-nine hours after
operation only one of the meat group was surviving. All of the
glucose group were surviving, and exhibited blood nitrogens com-
parable with those of the meat group of twenty-four hours earlier.
Duration after operation was practically the same in animals 39
(meat) and 34 (glucose); the two other meat animals survived a
distinctly shorter time than the two other glucose animals.
In these experiments, the condition that makes for a lower
blood nitrogen also appears to make for a longer duration of life
after acute renal insufficiency, but for confirmation of these findings
and for further investigation of the factors involved, additional
studies are in progress.
NEPHRITIS. 5
3 (1067)
The significance of the uric acid, urea and creatinine of the blood
in early and late nephritis.
By V. C. MyYERs, M. S. FINE and W. G. LoucH.
[From the Laboratory of Pathological Chemistry and the Department
of Medicine, New York Post-Graduate Medical School and
Hospital.]
Typical cases of gout show, as a rule, blood uric acid values
from 2 to 5 times the normal. The amounts of urea and creatinine
are normal or in the case of urea, only slightly above normal.
Many early cases of nephritis, especially of the interstitial type,
give blood pictures which differ little from those of gout. The
uric acid findings are quite as high and the urea content varies
from only slightly above to more than double the normal amount.
The creatinine is only slightly increased. As the condition of
cases of this type becomes more severe, the retention of urea
increases, until we have high values for urea as well as for uric acid.
If improvement takes place the concentration of urea gradually
falls until the picture is that of the preceding group. If, on the
other hand, the case goes on to a fatal termination, the retention
of uric acid and urea is followed by that of creatinine, the concen-
tration of which may reach twenty times the normal. Here the
phthalein output is practically zero.
From the foregoing it would appear that as the permeability
of the kidney is lowered it becomes evident in the blood, first,
by an increase in the uric acid, second, by that of urea and lastly,
by that of creatinine. That this should be the case seems quite
plausible when we consider the ease of excretion of these con-
stituents, as determined from a comparative nitrogen partition
of normal urine and blood. Uric acid nitrogen forms 2 per cent.
of the non-protein nitrogen of both urine and blood, urea nitrogen
about 85 per cent. in urine but 50 per cent. in blood and creatinine
nitrogen 5 per cent. in urine but only 2 per cent. in blood.
6 SCIENTIFIC PROCEEDINGS (69).
4 (1068)
The movements of the mitral valve flaps studied by a new method.
By A. L. DEAN, JR. (by invitation).
[From the Physiological Laboratory, Cornell University Medical
College.|
The desirability of a more accurate knowledge of the move-
ments of the auriculo-ventricular valves led to the adoption of
the following method of study in the perfused cat’s heart: The
margins of an opening in the left auricle are stitched to the
bottom tube of a small cylindrical reservoir in which the height
of pressure is regulated and varied by a set of lateral overflow-
tubes. To one of the mitral valve flaps is attached a human hair
communicating with a short and delicate lever of straw, held up-
ward by slight spring tension. Whenever the valves move up
the lever is elevated and a small mirror attached to the axis of
the lever system is depressed. Upon this mirror is projected a
beam of light and through its reflection the movements of the
mirror are recorded upon a moving bromide surface. In this
way the oscillations of the valve flaps may be optically recorded
and compared with simultaneous optical tracings of auricular and
ventricular activity. To prevent the transfer of ventricular
movements to the threads connecting with the valves, the heart
is fastened by stitches, carefully placed around the auriculo-
ventricular ring to a neatly fitted ring of metal.
A study of the records thus far obtained indicates that the
following movements of the auriculo-ventricular valves occur in
every cardiac cycle: A very short interval after the onset of
auricular systole, the cusps are slightly depressed toward the
ventricle, but before the end of auricular systole they quickly
ascend toward the auricle. With the onset of auricular diastole
(which in these experiments began a distinct interval before sub-
sequent ventricular systole) the valves move ventricle-ward to
their former position. With the onset of ventricular systole the
cusps immediately move upward and close completely. So they
remain until ventricular relaxation begins. During ventricular
diastole the valves move ventricle-ward to a position that is lower
BLoop FaT IN RELATION TO HEAT PRODUCTION. 7
than that occupied previous to either auricular or ventricular
systole. In the latter portion of diastole (diastasis) they gradually
float upward but to a slight extent only. Superimposed upon the
main curves of closure are found oscillations of much smaller
amplitude and of shorter period. Further investigation is neces-
sary before it may be justly concluded that these correspond to
the vibrations responsible for the heart sounds.
The conclusion is reached that the mitral valves undergo two
movements toward closure in each cardiac cycle, the first near
the end of auricular systole, which is transient and incomplete,
and the second lasting throughout ventricular systole which is
complete and insures the effective closure of the valves.
5 (1069)
Blood fat in relation to heat production and depth of narcosis.
By J. R. MurLIN and J. A. RICHE.
[From the Physiological Laboratory of the Cornell University Medical
College, New York City.]
Experiments on dogs have been designed to answer the ques-
tions: (1) whether fat injected directly into the circulation can
be oxidized at once, and (2) what is the relation between the
concentration of fat in the blood and the heat production. Inci-
dentally it has been necessary to determine whether the depth
of narcosis had any effect on the amount of fat in circulation, and
on the heat production.
The following experiments may be cited as typical of the
effect on percentage of blood fat and on heat production, of a
single intravenous injection of 100 c.c. 3 per cent. emulsion of
lard oil. It will be seen that the heat production rises and the
R. Q. falls as the fat becomes more concentrated in the blood,
indicating, therefore, that the injected fat burns.
In order to insure complete muscular rest chloretone was
administered to a number of the animals and it was while con-
trolling the effect of this narcotic that the following observations
were made: (1) The percentage of blood fat runs parallel with the
depth of narcosis, 7. e., the deeper the narcosis the lower the blood
8 SCIENTIFIC PROCEEDINGS (69).
Dog Liters CO; | Liters O, Per Cent. wots
No, | Wt.Kgm.| Hour, eres lie R.Q. Blood Fat | Cal. per
(Carotid). Hour.
102] 7.0 I 2.171 3-013 a2 0.45 14.17
II 2.478 3.108 “719 0.47 14.92
100 c.c. jemulsion (3 per cent.) by jugular vein
III 2.327 3.100 “75 0.51 14.69
IV 2.627 3-653 -72 0.58 17.18
IIr| 7.0 I 2.581 3.016 85 14.66
II 2.578 3.023 | 85 14.70
100 c.c. jemulsion (3 per cent.) by jugular vein
III 2.435 3.008 -81 14.48
IV 2.442 3-251 75 15.41
Vv 2.494 3-412 -73 16.09
fat; (2) following a single injection (intraperitoneal, in mineral oil)
the heat production rises as the narcosis wears off, independently
of muscular motions, but parallel to the percentage of fat in the
blood. The following experiment illustrates the point.
Liters i Per Cent. Heat Produc-
Dog} wt. Kgm. Hour. CO: Liters O2 | R.Q. Blood Fat tion Calories
No per Hr. per Hr. (Carotid). per Hr.
0.41
117 9.0 I 1.880 2.223 85 10.81
II 1.812 2.038 -89 10.01
0.44
III an 2.08 ‘geen 8 eee
IV 2.08 2.401 ae 11.74
0.46
Vv 2.093 2.458 85 II.95
VI 2.081 2.589 .80 12.43
0.51
This relationship as regards depth of narcosis and percentage
of fat in the blood has been confirmed with morphine. With
ether a second narcosis had the effect of raising the blood fat to a
higher point than the first.
Blood fat has been determined by the nephelometric method
and heat production by indirect calorimetry.
1A single two-hour period.
PosITIVE NITROGEN BALANCE IN NEPHRITIS. 9
6 (1070)
The interpretation of a positive nitrogen balance in nephritis.
By HERMAN O. MOSENTHAL, M.D.
[From the Medical Clinic of the Johns Hopkins Hospital, Baltimore.]
In studying nitrogen metabolism in certain cases of nephritis,
a retention of this substance was observed. The conception of
the retention of nitrogen in nephritis, as understood by the
clinician, generally implies two facts: Firstly, that a positive
nitrogen balance is usually due to kidney insufficiency; secondly,
that the retained nitrogen is present in the body as waste-nitrogen
and circulates in the blood, in part, at least, as non-protein nitro-
gen. It is known from the work of Marshall and Davis! that urea
is evenly distributed throughout the body, except in certain
tissues, as the fat, bone, cartilage, etc., which do not take up urea.
In calculating the theoretical amounts of non-protein nitrogen to
be expected in the blood, it has been assumed that all the nitro-
gen which the body has metabolized and is about to excrete, in
TABLE 1.
THEORETICAL AND ACTUAL VALUES OF NON-PROTEIN NITROGEN
OF THE BLOOD RESULTING FROM NITROGEN RETENTION
IN CERTAIN CASES OF NEPHRITIS.
N. of Blood—Mg. per, 100 c.c.
N. Grams Retained
At Beginning of At End of Theoretical Value at | During Observation.
Observation. Observation, End of Observation.2
Case I 30 37 152 92.0
poe 30 38 116 65.0
ee a5 34 93 51.0
a 30 37 165 101.4
nes 29 27 119 69.1
oe 6
71 74 117 35-3
contradistinction to the nitrogen which the tissues are storing is
evenly distributed throughout the body as is the case with urea.
1 Marshall and Davis: Jour. Biol. Chem., 1914, XVIII, 53.
3 These figures represent the values obtained for non-protein nitrogen of the
blood at the beginning of the observation plus the theoretical value due to retained
nitrogen.
10 SCIENTIFIC PROCEEDINGS (69).
Applying these principles to the total non-protein nitrogenous
products, it is found that in a subject of average weight, for every
gram of nitrogen retained, the non-protein nitrogen of the blood
should be increased 1.33 mg. per 100 c.c. According to these
calculations, in the cases presented here, if none of the retained
nitrogen were assimilated or stored, and all of it circulated as
waste-nitrogen because the kidneys did not excrete it, the figures
shown in the table would be obtained.
This table shows that a positive balance of nitrogen in cases
of nephritis on a mixed diet is not necessarily followed by a
corresponding increase in the non-protein nitrogen of the blood.
It is evident that discretion must be exercised in interpreting a
normal figure for non-protein nitrogen of the blood as indicating
that no nitrogen retention has taken place, and in considering a
positive nitrogen balance as an absolute indication of the inability
of the kidney to excrete this substance.
7 (1071)
On the occurrence and distribution of potassium in normal and
nephropathic kidney cells.!
By Wo. DEB. MAc NIDER.
[From the Laboratory of Pharmacology, The University of North
Carolina.]
The observations which are contained in this summary are
based on the microchemical demonstration of potassium in the
kidney cells of thirty-four dogs. The animals have varied in
age from four months to something over ten years. Four of the
animals may be grouped as “normal animals.” They did not
receive any nephrotoxic substance and neither were they subjected
to the action of an anesthetic. After a period of three days of
observation these animals were killed by shooting.
The remaining thirty animals were rendered nephropathic
by uranium nitrate in the dose of 4 mg. or 6.7 mg. per kilogram.
They were anesthetized by either Gréhant’s anesthetic in 60 per
cent. strength, or by morphine-ether.
1 Aided by a grant from the fund for scientific research of the American Medical
Association.
PoTASSIUM IN KIDNEY CELLS. II
At the termination of the experiment small pieces of kidney
tissue were removed, and frozen sections not over 20 micra in
thickness were made. The sections were treated at once with
Erdmann’s! reagent as modified by Macallum? and used by him
in his studies ‘‘On the Distribution of Potassium in Animal and
Vegetable Cells.”
The reagent which consists in a solution of the hexanitrite of
cobalt and sodium serves as a complete precipitant of potassium
from its solutions, in the form of an orange-yellow precipitate of
the triple salt. If the salt is present in minute quantities the
crystalline form is absent. To render the detection of small
quantities of the salt possible, Macallum’ used ammonium sulphide
to react with the cobalt of the salt and form the black sulphide of
cobalt which is easily detected. This suggestion of Macallum’s
has been employed in the demonstration of potassium in all of
the sections.
The results which have been obtained are as follows.
1. The epithelial cells of the normal dog kidney show only
traces of potassium. The potassium is most marked in the loops
of Henle and is fairly evenly distributed throughout the cytoplasm
of the cells. It has never been demonstrated within the nucleus
of the normal cell.
2. The epithelium of the nephropathic kidney shows an increase
in potassium over that of the normal. The potassium differs in
distribution within the cytoplasm of the cell and has been demon-
strated within the nucleus of the cell.
3. The potassium in the nephropathic organs has been espe-
cially marked in the cells of the convoluted tubules. In the cyto-
plasm of the cells forming these tubules the potassium is not
uniformly distributed but is found to collect along the free margin
of the cells bordering the lumen of the tubule. A similar observa-
tion on the distribution of potassium salts was first made by
Macallum* in his studies of the frog kidney in which a decinormal
solution of potassium chloride was injected into the dorsal lymph
sacs of frogs.
1“ Anorganische Chemie,’’ 1898, p. 630. Reference given by Macallum.
2 Jour. Phys., Vol. XXXII, No. 2, p. 98.
8 Loc. cit.
4 Science, Vol. XXXII, No. 824, p. 497.
12 SCIENTIFIC PROCEEDINGS (69).
4. Such accumulations of potassium salts are as marked in the
kidney epithelium of nephropathic animals which are polyuric,
as they are in the nephropathic animals which have been rendered
anuric.
5. The above observation would tend to minimize the impor-
tance of potassium in being responsible for a lack of function on
the part of the kidney.
6. The age of the animal has apparently no constant influence
on the amount of potassium microchemically demonstrable.
However, the oldest animal of this series showed the most marked
potassium precipitate. In this animal, and one other of the
series, which were anuric from uranium, and in which the epi-
thelium of the convoluted tubules had undergone a severe swelling
and partial necrosis, not only did the cytoplasm of these cells
give the potassium reaction but potassium was also demonstrated
in the nucleus of the cell.
8 (1072)
The action of animal extracts upon the flow of bile.
By Isaac OTT, M.D., and Joun C. Scott, M.D.
[From the Laboratory of Experimental Research, Medico-Chirurgical
College of Philadelphia.]
Our experiments were made upon etherized cats. We injected
secretin at intervals and determined that equal doses of secretin
were followed by equal increments in the bile secretion. The
cystic duct was previously ligated close to the common duct into
which a glass cannula was inserted. After determining the effect
of a dose of secretin, we waited some time and then injected an
equal dose of secretin plus the infusion of the animal extract.
The drops of bile were counted for five-minute periods. We
found that adrenalin and the hypophysin of Fiihner (pituitrin)
greatly slowed the secretion. Pancreas slightly diminished the
secretion. Thyroid extract had hardly any effect. Tonsil extract
caused a marked increase. Thymic extract decreased it. Para-
thyroid, mammary and corpus luteum had no action.
THE AMMONIA OF THE GASTRIC JUICE. 13
9 (1073)
Gastro-intestinal studies XI. Studies on the relative digesti-
bility and utilization by the human body of lard and
hydrogenated vegetable oil.
By C. A. SMITH, RAYMOND J. MILLER and PHILIP B. HAWK.
[From the Department of Physiological Chemistry, Jefferson Medical
College, Phila.]
Two normal men were the subjects of the experiment, which
was conducted in two periods of eight days each, separated by
an interval of three days. The diets were so arranged that the
fat, ingested during the first period, was mostly lard, while that
of the second period was mostly hydrogenated vegetable oil.
The daily feces were analyzed for total fat, fatty acid, and neutral
fat by the Saxon method. The average percentagenof digestion
of lard was 96.75, and of the hydrogenated vegetable oil, 96.3,
while the average utilization percentages were 94.7 and 93.35
respectively. It is thus apparent that the hydrogenated vegetable
oil used in this experiment was as satisfactorily digested and
utilized by normal men as was lard.
10 (1074)
The ammonia of the gastric juice. (Preliminary communication.)
By Harry L. Huser. (by invitation).
[From the Hull Biological Laboratories of the University of Chicago.]
Recently Carlson! reported some observations on the occur-
rence of NH; in the gastric juice of man and of dogs. Further
observations have been made and a few of the results are given
below.
Three series of experiments were conducted: (a) On dogs with
Pawlow stomachs; (0) on normal human individuals, and (c) on
human individuals with gastric disturbances.
The NH;-content of the gastric juice of normal dogs varied
1Carlson, Am. Journal of Physiology, 1915, XXXVIII, p. 248.
14 SCIENTIFIC PROCEEDINGS (69).
in different dogs between 0.5-3.5 mg. NHs per 100 c.c. of juice,
there being some variation from day to day, and at different
times during the same day in the same dog. The juice was
collected for a period before feeding and one-hour periods after
feeding. The addition of NH,Cl to the dogs’ food caused in each
instance an increase in the NH;-content of the gastric juice col-
lected from the Pawlow pouch. Three dogs, in which gastric
ulcers had been produced experimentally showed an increase in
the NH;-content at irregular intervals. After a time two of
these dogs refused to eat and the juice collected during this time
showed a progressively increasing content of NH;. At autopsy
these dogs showed either active or healed ulcers, usually located
in the Pawlow pouch.
b. The experiments on normal healthy individuals were con-
ducted on men who were connected with the laboratory. The
juice was collected by means of the Rhefus stomach tube before
and after feeding on Ewald meal. With three exceptions the
NH;-content of the juices from these men ranged between 0.5-3.5
mg. of NHs per 100 c.c. of juice. In these three men the NH;-
content ranged between 10-15 mg. NH; per 100 c.c. of juice.
A series of experiments was then conducted on one of these
men with high NH;-content. The diet was so arranged that there
were periods of low protein and of high protein ingestion, and the
NH; content of the gastric juice and the total NH3; of each day’s
urine were noted. During low-protein ingestion the NH; content
of the gastric juice fell in five days from 12-3.5 mg. and the NH3
of the urine also showed a marked decrease. During the high-
protein ingestion the NH;-content of the gastric juice rose in 3
days from 12-28 mg. NH; and the NH; of the urine also showed
a marked increase. In both cases the NH; in the gastric juice
and in the urine came back to the original level within two days
after resuming the usual diet.
In the second experiment the diet was kept uniform through-
out the period and during certain periods an excess of alkalis or
of acids was added. During the period of excess alkali ingestion
the NH; of the gastric juice remained the same while the total
NH; of the urine fell to 1/10 its former level. During the ingestion
of acid the NH; of the gastric juice again remained the same while
ACTION OF HEAvy METALS ON ISOLATED INTESTINE. 15
the total NH; of the urine was increased. When the alkali and
the acid were left out of the diet the normal level for the NH; of
the urine was reached in a short time.
c. Estimation of the NH3-content of Ewald meal juice from
26 individuals with gastric disturbances, supposedly ulcers, was
made and in only five of these was the NH-content markedly
increased. In two of these cases the diagnosis of carcinoma of
the stomach was made with certainty; in another case a diagnosis
of ulcer with obstruction was made and in the remaining 2 cases
a diagnosis of gastric ulcer was made. Further work is being
conducted along this line with the view of determining the source
and the significance of the gastric juice ammonia.
II (1075)
The action of heavy metals on the isolated intestine.
By WILLIAM SALANT and C. W. MITCHELL.
[From the Pharmacological Laboratory of the Bureau of Chemistry,
Washington, D. C.]
In experiments with zinc which was used in the form of the
malate and carried out on isolated segments of the intestines of
cats and rabbits by the method of Magnus it was found that even
low concentrations may produce depression of muscular activity.
Asolution N/20,000 zinc malate, proved to be quite active in some
experiments. NV/10,000 and N/5,000 zinc malate produced, after
a brief preliminary stimulation, considerable decrease and some-
times irregularity of the force of rhythmic contractions. Occa-
sionally decrease of frequency and tonus were also observed.
When the segments of the intestine were suspended in pure
Locke solution again, some improvement occurred, although
it had been acted upon by zinc 45-70 minutes. In experi-
ments with concentrations of N/1,000 and N/500 and sometimes
even with NV/2,000 permanent injury to the tissues may be caused,
by the metal as no recovery could be observed when pure Locke
solution was substituted for one containing zinc.
The action of nickel employed in the form of the acetate was
also tested. Dilute solutions, N/10,000 and N/5,000 produced
16 SCIENTIFIC PROCEEDINGS (69).
temporary depression followed by recovery and sometimes stimu-
lation while the intestine was still in the solution of the salt.
Complete abolition of rhythmic contractions and decrease of
tonus were observed when much higher concentrations were used,
but the effect was not permanent in these experiments as recovery
in pure Locke solution took place after the intestine had been
suspended in N/500 nickel acetate for twelve minutes. The
reaction to pilocarpine and of barium was studied in experiments
with both of the metals. The evidence obtained points to injury
to nerve endings as well as of the muscle fiber, but the latter was
in some experiments much more resistant.
12 (1076)
A note on the failure of pituitrin to sensitize the sympathetic
system.
By R. G. Hoskins (by invitation).
[From the Laboratory of Physiology of the Northwestern University
Medical School.]
In 1912 Kepinow published the conclusion that the injection
of small quantities of pituitary extract “‘sensitizes’”’ the point of
attack of epinephrin.!_ His observations were that a given dose
of epinephrin produced a greater mydriatic effect in rabbits and
cats, a greater vasoconstriction in the Loewi-Trendelenberg frog
preparation and a greater vasomotor effect in rabbits and dogs if
immediately before hand a minimal dose of pituitary preparation
had been injected. Kepinow’s work has been quoted as the
basis for a rather far reaching conclusion that the pituitary gland
has normally the function of promoting the activity of the sympa-
thetic nervous system.
In various researches on the vasomotor system we have had
occasion to use pituitary extract in connection with epinephrin
and nicotin and it became important to know to what extent
sympathetic sensitization occurs. As our previous work has been
done exclusively on dogs we have investigated the matter in this
1 Kepinow, Archiv fiir experimentelle Pathologie und Pharmakologie, 1912, LXVII,
247.
ATRIOVENTRICULAR RHYTHM IN MAN. 17
species only. Kepinow used two kinds of pituitary material,
an extract prepared by himself, and the commercial preparation
“Pituitrin’? made by Parke Davis and Co. Kepinow states that
both preparations gave similar effects. Our experiments were
made with ‘‘Pituitrin” only.
Since the results are negative they may be reported very
briefly. The procedure was to determine the effects of injections
of given quantities of ‘‘adrenalin”’ and of nicotin, selected to give
a moderate rise of blood pressure. About 1 c.c. of adrenalin,
I : 100,000, and I c.c. of nicotin I : 4,000 are suitable for medium-
sized dogs. Having determined the reactions to these drugs,
pituitrin was injected by vein in quantity to give a slight rise of
pressure, €. g., .05 c.c. At various intervals from one half to
several minutes after this injection, the adrenalin and nicotin
injections were repeated. In no case was a significant change of
_ reaction noted. In most of the animals the vagi were cut, but
this procedure made no apparent difference in results.
In view of the restricted value of negative results some hesi-
tancy is felt in offering them for publication. In further con-
sideration, however, of the vast number of unjustified generaliza-
tions in the literature of internal secretion they are offered for
what they may be worth.
13 (1077)
The production of atrioventricular rhythm in man after the
administration of atropin. (Preliminary Communication.)
By FRANK N. WItson, M.D. (by invitation).
[From the Department of Internal Medicine, University of Michigan.]
During the past year two patients with cardiac complaints
were given repeated injections of I mg. atropin sulphate and on
each occasion atrioventricular rhythm was observed. This usually
appeared in from eight to ten minutes after the drug was given,
it persisted for only a few minutes, and disappeared before the
maximum effect of the drug was reached.
In order to determine whether this tendency to A-V rhythm
was peculiar to these patients or whether it exists normally, a
18 SCIENTIFIC PROCEEDINGS (69).
series of experiments on normal individuals was carried out.
Eighteen subjects, all under twenty-eight years of age and all
with apparently normal hearts, were given hypodermic injections
of 1 mg. of atropin. The effect of deep breathing and of ocular
pressure upon the cardiac mechanism was studied both before and
during the atropin action. Before the injection, A-V rhythm was
not produced in any of the subjects tested. Between eight and
fifteen minutes after the injection, however, A-V rhythm could
be produced by ocular pressure or deep respiration in the majority
of the subjects. After the atropin effect had reached its height
A-V rhythm could no longer be produced.
Three types of A-V rhythm were observed. In the first, which
occurred most frequently, the P—R interval was reduced and P
was inverted; in the second, the P—R interval was zero; while in
the third there was an R-P interval. The last was observed in
only two subjects. These differences evidently depended upon
the level of the pacemaker in the junctional tissues.
These observations may be explained on the assumption that
atropin releases the A-V tissues from vagus control somewhat
before it releases the sinus node. At this time stimulation of
the vagus slows the sinus rhythm without a correspondingly
great effect upon the inherent rhythms of the lower centers. The
latter therefore tend to usurp the pacemaking functions of the
heart. After full atropin action on the other hand, both the sinus
and the lower centers are released and the more rapid sinus rate
controls the cardiac rhythm. Before atropin vagus stimulation
probably slows the inherent rhythms of the lower centers as well
as that of the sinus so that the former do not ordinarily escape.
STUDIES ON SO-CALLED PROTECTIVE FERMENTS. 19
14 (1078)
Studies on so-called protective ferments VIII. On the mechanism
of anaphylaxis and antianaphylaxis.
By J. BRONFENBRENNER.
[From the Research Laboratories of the Western Pennsylvania
Hospital, Pittsburgh, Pa.]
As we have reported a little over a year ago,! the interaction be-
tween an immune serum and its corresponding substratum is fol-
lowed by a formation of toxic split products. We found that the
toxic material originated not from the substratum or antigen, but
from the serum itself.2 These findings threw light on some of the
unsettled questions in the theory of anaphylaxis and antianaphy-
laxis. Experiments conducted with the view of correlating our
findings with the accepted views on this subject, suggested a
following hypothesis about the nature and mechanism of ana-
phylaxis.
Blood serum contains normal proteolytic ferments which
require special conditions of the medium in order to exhibit their
activity. Normally the degree of concentration of colloids
in the serum offers an obstacle to the activity of these ferments.
In the experiments in vitro it is possible to change the degree of
concentration of colloids in the serum, thus diminishing its anti-
tryptic inhibiting power and setting free the ferments.*
This activation of normal serum can be accomplished by
mechanical adsorption, as in experiments of Plant, Peiper and
others, or by the dissolution of some of the serum colloids, as in
the experiments of Jobling.* In either case the degree of disper-
sion of remaining colloidal particles is increased and thus ferments
are allowed to act.
1 Bronfenbrenner, Pennsylvania State Journal, 1914, October, p. 20.
2? Bronfenbrenner, Proc. Soc. Exp. BIOL. AND MED., 1914, XII, p. 7-8; also
Journ. Exp. Med., 1915, Vol. XXI, No. 5, p. 480.
3 Bronfenbrenner, Journ. Exp. Med., 1915, XXI, No. 3, p. 221.
4 Jobling and Peterson, Journ. Exp. Med., 1914, Vol. XIX, p. 239. Though the
authors find it necessary to remove the lipoid in order to activate the enzyme, our
own experiments show that the removal of lipoid is not necessary. Mere bubbling
of ether vapor through the serum accomplishes the activation.
20 SCIENTIFIC PROCEEDINGS (69).
Our experiments have shown that also the physico-chemical
changes following the specific interaction between the antigen and
antibody influence the colloidal conditions of the medium in the
same manner.! Our records show that both stalagmometer and
refractometer register the increase of dispersion in the immune
serum following the addition of the specific antigen and parallel
with it the actual measurements of the antitryptic titer of the
serum show a steady diminution of the power of this serum to
check the activity of its own proteolytic ferments.?
In anaphylaxis the latter process takes place, namely, if a
suitable amount of antigen is injected into a sensitized animal,
the interaction between the specific antibodies and the antigen
produce a physico-chemical change in the serum, followed by a
diminution of its antitryptic activity. Once the balance between
the tryptic and antitryptic powers of the serum is destroyed, the
proteolytic ferments may attack the protein of the serum with the
production of toxic split products, and anaphylactic shock follows.’
That the mechanism of anaphylaxis rests on the disruption
of balance between the tryptic and antitryptic properties of the
serum is especially evident from our experiments in which we
succeeded in preventing anaphylactic shock in experimental
animals by increasing the antitryptic power of their serum at
will before subjecting them to shock.* In doing so we found that
practically any substance which caused the rise in antitryptic
titer of the serum of experimental animals, protected them also
from the subsequent anaphylactic shock. We found also that
all such substances are toxic by themselves if injected in sufficient
quantity. The mechanism of this protection seems to be as
follows.
The introduction of poisons in quantities not sufficiently large
to kill the animal outright is followed by the death of the tissues
immediately affected by the poison. With the death of the tissues
1 Bronfenbrenner, Proc. Soc. Exp. BIOL. AND MED., 1914, XII, p. 4.
2 Bronfenbrenner, Mitchell and Titus, Biochemical Bulletin.
* Bronfenbrenner, Penna. State Med. Journ., October, 1914; also Journ. Exp.
Med., 1915, Vol. XXI, p. 480. Inacurrent number of the Journ. of Exp. Med., this
view of anaphylaxis is corroborated by Jobling, Peterson and Eggstein.
‘ Bronfenbrenner and Schlesinger.
STUDIES ON SO-CALLED PROTECTIVE FERMENTS. a1
the intra cellular ferments are set free.' These ferments possibly
with the collaboration of the ferments thrown out from the sur-
rounding fixed cells as well as from blood serum and leucocytes
proceed to dispose of the dead material. Some of these split prod-
ucts of protein constituents of digested tissue cells, together with
some non-protein constituents (lipoids?) of these cells, exert antag-
onistic antitryptic action, and retard or stop the activity of
proteolytic ferments.
Since, as it was suggested before by us,? the specific ana-
phylactic shock is due to the intoxication of the animal following
the liberation of proteolytic enzyme in its blood, it is possible that
the preliminary injection of a suitable amount of poison causes
the increase of the amount of protein split products in the
circulation of the animal and the resulting change in the degree
of colloidal dispersion paralyzes the activity of proteolytic fer-
ments which are liberated upon the subsequent introduction
of a lethal dose of antigen into a sensitized animal.
The effect of the vaccinating injection of a sublethal dose of
antigen into sensitized animals, or of the vaccinating injection of
a sublethal dose of anaphylatoxin into normal animals, is evi-
dently due to the same mechanism of partial proteolysis followed
by the output of split products acting as antitrypsin and not to
the exhaustion of antibody.
Usually the anaphylactic state is taken to be the opposite to
the state of immunity. The above theory makes both the active
immunity and anaphylaxis a part of the same process. The
difference between the two reactions being only in the rapidity
and extent of proteolysis induced by the specific combination of
antigen with its antibody in vivo.
1 In the experiments which are to follow we will show the actual changes in the
blood and urine following the liberation of ferments during the specific anaphylactic
shock, as well as during nonspecific proteolysis due to poisoning.
§ Bronfenbrenner, Bioch. Bull., March, 1915, p. 87.
22 SCIENTIFIC PROCEEDINGS (69).
15 (1079)
Icterus. A rapid change of hemoglobin to bile pigment in the
pleural and peritoneal cavities.
By C. W. Hooprr, M.D., and G. H. WuIppie, M.D.
[From the George Williams Hooper Foundation for Medical Research,
University of California, San Francisco.]
In an earlier communication we have been able to show that
bile pigment could be formed from hemoglobin without the
agency of the liver. Solutions of hemoglobin were introduced
into the blood vessels of dogs whose livers had been excluded
from any part in this reaction. There was a prompt formation
of bile pigment from hemoglobin with no possible direct liver
action. This transformation can take place within a space of
two hours when active circulation is maintained in the head
and thorax alone. It seemed probable that the endothelium might
be the tissue whose activity was responsible for this change of
hemoglobin to bile pigments. This work has received confirma-
tion from experiments of McNee.
All our experiments were performed on normal dogs. Hemo-
globin in crystalline form dissolved in salt solution or obtained
from freshly laked red blood corpuscles was introduced into the
pleural or peritoneal cavities. The fluid was withdrawn after
different intervals varying from eight hours to three days. Care-
ful tests showed at times some bile pigment formation in eight
hours but always in twenty-four hours—often in sufficient amounts
to be estimated quantitatively. The amount of bile pigment
formation is considerable after an interval of two, three, or four
days—even more than five milligrams in some cases. It is to
be recalled that a dog of thirty pounds in weight may excrete
normally about 25 milligrams of bile pigment in six hours.
There is very good evidence that bile pigments may be formed
from hemoglobin by the agency of endothelial cells. There is
conclusive evidence that bile pigments can be formed by the
mesothelium of the serous cavities. It is possible that this
capacity of transforming hemoglobin into bile pigments may be
a general property of living protoplasm.
23
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24 SCIENTIFIC PROCEEDINGS (69).
16 (1080)
On a colorimetric method of adjusting bacteriological culture
media to any optimum hydrogen ion concentration.
By S. H. Hurwitz, K. F. MEYER and Z. OSTENBERG.
[From the George Williams Hooper Foundation for Medical Research,
Universtiy of California.]
In most bacteriological laboratories of this country adjustment
in the reaction of culture media by titration has largely replaced
all other methods. The indicator most commonly employed is
phenolphthalein, and the results are expressed in terms of the
amount of normal alkali necessary to bring one liter of the medium
to the desired reaction (Fuller Scale).
Recent studies have shown that the titrimetric method in its
present form is inaccurate. The results of titrations done in
this laboratory support the observations of Clark! that media
titrated to the end point of phenolphthalein and corrected to
definite degrees on the Fuller scale have different hydrogen ion
concentrations.
An exact knowledge of the reaction of a medium can be gained
only from a determination of its hydrogen ion concentration. It
is Our purpose to present a simple colorimetric method which
makes possible the accurate and rapid determination of the hydro-
gen ion concentration of culture media and their adjustment to
any optimum concentration of ionized hydrogen.
For our work we have made use of a set of standard solutions
recommended by Levy, Rowntree, and Marriott? for determining
the hydrogen ion concentration of the blood. These consist of
standard phosphate mixtures containing phenolsulphonephthalein.
The advantages of this indicator have been set forth by these
workers.
The medium is tested first to ascertain what its ionization is
before adjustment. This preliminary test can be carried out
quickly: to 3 c.c. of fluid is added 0.3 c.c. of a 0.01 per cent.
1 Clark, W. M., Jour. Infect. Dis., 1915, XVII, 109.
2 Levy, Rowntree, and Marriott, Arch. Int. Med., 1915, XVI, 389.
ADJUSTING BACTERIOLOGICAL CULTURE MEDIA. 25
solution of phenolsulphonephthalein, the fluid being read directly
in the comparator.!_ In most instances the culture fluid has been
roughly adjusted by the usual methods so that its reaction falls
ithin the lmnits of the tacale @il’= 6.4 46lpet = 84):
If the medium has not received a preliminary adjustment of
reaction, it may be too acid or too alkaline to be read directly.
In that event a specimen of the medium is titrated as follows:
to a 3 c.c. sample is added /20 acid or alkali solution? depending
upon the initial reaction of the medium, until a color is obtained
which corresponds to the hydrogen ion concentration desired.
The conversion of the amounts of /20 solutions read on the
pipette into m/1 solutions is made by referring to a curve plotted
for an average medium in which the amounts of the 7/20 solutions
required are plotted as abscisse and the corresponding amounts
of n/t solutions as ordinates. Final adjustment in reaction must
be made with sterile acid or alkali in order to avoid the change in
ionization caused by sterilization.
To illustrate the accuracy of the method the results of the
titration of five media are given in the table.
TABLE.
5 [x
Titra- a Stan- | 2/20 | Alkali |Value of
No. of | Date. Medium. | tionby| Preliminary | dard | Alkali | Added | pH Ob-
Exp. Fuller Test. Desired.| inc.c. | per | tained.
Scale. 25 C.C.
I Aug. 6 |Veal infusion 1.0 Below 6.4 ACEO On Or225|Nl774 5
2 Aug. 7. |Plain broth 0.8 6.9 720m WOsliy 205073) 1 7255,
3 Aug. 12 |Liebig’sbroth| 0.8 6.9 7.6 | 0.195 | 0.087 | 7.55
Tn 0:20), |\O0:005 | 7-7
4 Aug. 16 |Plain broth 0.8 7.15 | 76) ||| 0.13) 10.06 7.6
7.9 | 0.195 | 0.088 | 7.9
8.15
5 Extract I.0 6.9 7.5 | 0.08 | 0.04 7.55
7.9 0.21 0.096 | 7.9
1 In order to make direct comparisons possible even in the presence of the natural
color of the fluid tested, we have constructed a simple device whereby the medium
tested serves as a background for the standard test color to which it imparts its own
characteristic quality of color.
2 In order to keep the concentration of indicator during titration the same as its
concentration in the standard comparison tubes (0.3 c.c. to 3 c.c. or I to 11) the
solutions of 2/20 acid and alkali are so made up that one eleventh of their volume is
indicator solution. The solutions are kept in vessels protected against light, air,
and moisture, and the apparatus so arranged that the solution can be delivered
directly into a graduated one cubic centimeter pipette provided with a ground glass
stopcock on the principle of a burette.
26 SCIENTIFIC PROCEEDINGS (69).
The colorimetric method will be found of great value in the
adjustment of the hydrogen ion concentration of media for organ-
isms which are sensitive to the reaction of their culture fluids.
The method is comparable in a way to the fine adjustment of a
microscope. The method of titratable acidity serving only to
adjust media coarsely for the growth of the average organism.
17 (1081)
Sarcoma occurring in a guinea-pig.
By Ernest C. Dickson, M.B.
[From the Division of Medicine of Stanford University Medical
School.
On May 10, 1915, a large male guinea-pig which had seemed to
be in good health in the morning became suddenly ill in the
afternoon and died within a short time. The animal had been
injected some months previously with a culture of what was
supposed to be diphtheria bacilli, but it had survived the injection,
At autopsy a large freely movable mass was found in the mid-line
of the neck on the ventral surface, which was adherent to the
underlying tissues about mid-way between the lower jaw and the
shoulder girdle. The tumor was apparently encapsulated and
showed no attachment to the skin. It measured 34x2144x2%
cm. in the various diameters, was yellowish in color and quite firm.
The capsule was fibrous and cut with some difficulty, but the
central portion was quite friable. The cut surface was yellow
with many mottled patches which were dark red in color.
Surrounding the tumor and in the right axilla were a number
of metastatic nodules, the largest of which measured 144x1x%
cm., and the smallest being about the size of a grain of wheat.
Section of the larger nodules showed a cut surface which was
identical with that of the large tumor.
The thoracic and abdominal organs showed nothing unusual.
On microscopic examination the body of the tumor is seen to
consist of round and ovoid cells which vary considerably in size.
In places the cells are closely packed together but in others they
are separated by a reticulum of connective tissue. There are
SARCOMA OCCURRING IN A GUINEA-PIG. 27,
many large areas of necrosis in which practically all cellular
structure is lost. There is evidence of an unsuccessful attempt at
encapsulation, but the tumor has invaded the connective tissue
of the capsule as well as the adjacent areolar tissue.
The majority of the cells of the tumor resemble lymphocytes
in size and in appearance. The nuclei are relatively large and
are placed excentrically in the cell body. There are a few cells
of about the same size in which two nuclei are present, but there
are no large, multinuclear giant cells. In addition to the round
cells there are other larger cells which vary in size and shape and
which contain large, clear vesicular nuclei, and some very large,
round cells with small nuclei which are apparently phagocytes.
The stroma of the tumor consists of a reticulum of connective
tissue which contains typical spindle-shaped fibroblasts. In some
places there is little fibrous tissue, but in others the fibrosis is
quite marked. In the portions which have not undergone necrosis
there is a rich blood-capillary network in the stroma.
In the necrotic portions of the tumor the cells show varying
degrees of disintegration, and the stroma is studded with nuclear
debris. In many places the large phagocytic cells are filled with
nuclear fragments.
The appearance of the largest of the metastatic nodules is
identical with that of the more cellular portions of the tumor, and
there is marked necrosis and extensive invasion of the connective
tissue of the capsule. The smaller nodules are of much more
frequent development and show little evidence of necrosis.
Smears from the tumor and from the metastases showed no
eosinophiles and no bacteria. Tissue cultures were not made.
An attempt was made to transplant the tumor into a small
number of young guinea-pigs but it was unsuccessful. However
as I knew nothing of the tumor until about two hours after the
animal had died it was not possible to make the inoculations soon
enough to justify any hope that they would be successful.
SCIENTIFIC. PROCEEDINGS:
ABSTRACTS OF COMMUNICATIONS.
Seventieth meeting.
New York Post-Graduate Medical School, November 17, 1915.
President Lusk in the chair.
18 (1082)
Production of pneumonic lesions by intrabronchial insufflation of
unorganized substances.
By B. S. KLINE and S. J. MELTZER.
[From the Department of Physiology and Pharmacology of the
Rockefeller Institute for Medical Research.]
Earlier communications from this department have shown
that intrabronchial insufflations of virulent pneumococci in dogs
produce lobar pneumonia similar in every respect to the lesions
of lobar pneumonia as observed in human beings. In later experi-
ments, Wollstein and Meltzer demonstrated that, at least macro-
scopically, typical pneumonic lesions can be produced also by
avirulent pneumococci and by the saprophytic bacillus megather-
ium. Microscopically it was established that the lesions produced
by the virulent pneumococci contained a great amount of fibrin,
while the lesions produced by the last-named organisms (avirulent
pneumococci and bacillus megatherium) contained only very
little fibrin.
In the present series of experiments various unorganized sub-
stances were insufflated into the bronchi of dogs which were
killed after twenty-four or forty-eight hours. The substances
were: aleuronat suspension in starch solution (autoclaved), starch
solution, egg yolk, lecithin, egg white and cholesterin. The
results were striking and are as follows: aleuronat, starch, egg
29
30 SCIENTIFIC PROCEEDINGS (70).
yolk and lecithin produced lesions which macroscopically could
not be distinguished from those produced by insufflation of
pneumococci. Egg white never produced pneumonic lesions of
any extent and even in the occasional patches many of the bron-
chioles and alveoli containing this protein showed microscopically
no nucleated cells whatever in their lumina. The few patches
produced by cholesterin were associated with the bronchioles and
adjoining alveoli. We shall not describe here the nature of the
histological pictures of the lesions under discussion; we shall
merely mention the fact that those lesions produced by aleuronat
and starch were similar microscopically to those produced by
virulent pneumococci, that is, the lesion contained in addition
to an exudate of similar cells, etc., a good deal of fibrin; while
lesions produced by egg yolk and lecithin resembled more those
produced by avirulent pneumococci and bacillus megatherium.
In brief, we may say that our experiments have demonstrated
definitely that the hepatization of the lungs, similar to the one
observed macroscopically in lobar pneumonia, can be produced
by such unorganized substances as aleuronat, starch, egg yolk
and lecithin, while egg white does not produce such an effect.
The following chart illustrates our results:
Lesion,
: 1;
pee Patches of | Lobar Type Culiee.
Consolida- | of Consoli-
tion. dation.
Substance Injected Intrabronchially.
+ 8 sterile.
Aleuronat in starch solution........... 8
iD Cea Ra Aso Ono MOMmdrr COU Oda Gn OS 2 2 + Diao in
Saige Collbiatyls 5 ouobaudoouu 990995" 7 + 6 sterile.
IDF42 540) | Sp eo ood boo UtIO Tb ODO RO NTO mS II + Ge XG)P
1D Yolen Sisto Hic An GnOU OO AOU .O Goo 0 0.0 0-5 I + re
Teecithines 2h cece coos Mee ees Rieter 6 + AN
ES opuuhiteceremicieieierste tree reieestalenet rs 5 - - Stunt
1B oa Oca Aon Utd oad TOA ae 4 + Aiae 2
CASTRATION IN A HEN-FEATHERED COCKEREL. 31
19 (1083)
Demonstration of the appearance after castration of cock-feather-
ing in a hen-feathered cockerel.
By T. H. MorGan.
[From the Department of Zoology of Columbia University.]
In the Seabright race of fowls the male is hen-feathered, 7. e.,
the feathers on the back of the neck (the hackles) and those on
the posterior portion of the back (the saddle) are short and less
elongated, like those of the female. When the Seabright male or
female is crossed to fowls of another race in which the male has
the characteristic male-feathering, the F; males are hen-feathered,
or at least the dominance of hen-feathering is more or less complete.
In the second generation there are three hen-feathered to one
cock-feathered male.
It has been shown by Goodale that removal of the ovary of
the hen or of the duck leads to the development of the male-
feathering. I tried to discover whether the removal of the testes
in the hen-feathered males would cause them to develop the
hackles and saddle feathers of ordinary cocks. My first operations
were unsuccessful, owing to failure to completely remove the testis.
This autumn Dr. H. D. Goodale performed the operation for me
on F, hen-feathered birds that I had reared. At the time of
operation some of the saddle feathers were removed. The new
feathers that appeared were like those on the ordinary cock bird;
not only did they have the characteristic shape but were bright
red also. The result leads to the apparently paradoxical conclu-
sion that the removal of the testes of the hen-feathered cock
caused him to develop certain characteristic feathers peculiar to
the ordinary male.
The most probable interpretation of the effects of removal of
the ovary of the hen (an operation that leads her to develop the
male plumage) is that the ovary secretes some substance that
holds in check the development of the male plumage. Likewise
in the hen-feathered male it would seem probable that the testis
produces some substance that inhibits the development of the
complete male plumage. Possibly this substance is the same as
32 SCIENTIFIC PROCEEDINGS (70).
that produced in the hen that brings about in her the same result,
although there is no direct evidence to show that this is the real
explanation.
20 (1084)
Agglutination of bacteria in vivo; its relation to the destruction of
bacteria within the infected host and to septicemia.
By C. G. BULL.
[From the Laboratories of the Rockefeller Institute for Medical
Research.]}
An intravenous injection of immune serum causes an abrupt
disappearance of the bacteria from the circulating blood of animals
having a bacteremia or a septicemia. This is due to an immediate
agglutination of the bacteria and to an accumulation of the
bacterial clumps in the lungs, liver, spleen, etc. The clumps of
bacteria are phagocyted and destroyed mainly by the polymorpho-
nuclear leucocytes which accumulate in the internal organs
following an intravenous injection of foreign protein substances.
The septicemia or bacteremia does not recur as long as the immune
serum is kept in the blood in a sufficient concentration.
These phenomena occur very typically following intravenous
administration of specific immune sera in rabbits infected with
pneumococci or Shiga dysentery bacilli. If the rabbits are actively
immune to these bacteria, the same phenomena follow an intra-
venous injection of the bacteria. If the immune animals are
given sufficient quantities of the bacte ia, death may be caused
by intoxication in the absence of a septicemia.
In natural immunity the above described phenomena follow
immediately upon an intravenous injection of the bacteria.
Rabbits have a comparatively high natural immunity to many
bacteria, of which the following have been studied in this respect:
typhoid bacilli, colon bacilli, dysentery bacilli of the Flexner group,
Staphylococcus aureus and albus, non-virulent bacilli of the mu-
cosus capsulatus group, and non-virulent influenza bacilli. All of
these are agglutinated, phagocyted, and destroyed in normal
rabbits as pneumococci are in immune rabbits and none of them
causes a true septicemia in these animals.
NITROGEN IN TISSUES IN RENAL DISEASE. 33
Rabbits exhibit little or no resistance towards a virulent
strain of Bacillus avisepticus while dogs are not affected by a
subtoxic dose. These bacteria are not agglutinated in the circula-
tion of rabbits and soon begin to multiply and produce a fatal
septicemia. In dogs, on the other hand, they are agglutinated
and rapidly disappear from the circulation and no true septicemia
follows, and as stated, a subtoxic dose causes no symptoms in
these animals.
Hence, in the several instances studied, agglutination of
bacteria within the circulation of the infected animal is followed
by an abrupt disappearance of the bacteria from the blood stream,
by accumulation of the agglutinated bacteria within the internal
organs, and by phagocytosis of the bacterial clumps by the
polymorphonuclear leucocytes, and a true septicemia does not
arise.
21 (1085)
Accumulation of nitrogen in the tissues in renal disease.
By HELEN Davis and NELLIs B. FosTEr, M.D.
[From the Department of Medicine, Cornell Medical College, and the
New York Hospital.)
The retention of nitrogen as a manifestation of certain types
of renal disease is a well-recognized phenomenon. When the
conditions of study are carefully controlled with accurate analyses
of the food and excreta the amounts of nitrogen retained in the
body is, with severe cases, very large—two grams per day for
periods of two weeks is not an exceptional amount. Since these
patients are usually quite sick and commonly manifest no sign of
improvement so long as the retention persists it is inconceivable
that this nitrogen is retained for tissue growth in a physiological
sense. On the other hand while the blood of these patients
often shows on analysis an increase in non-protein nitrogen the
figure may not rse to any significant degree and never becomes
sufficiently high so as to account for more than a small fraction
of the nitrogen retained. We have also noted with several
patients during metabolism studies a flushing out of nitrogen so
that there resulted large minus balances during periods of improve-
34 SCIENTIFIC PROCEEDINGS (70).
ment in the patient’s condition. It seemed that the explanation
of these phenomena is that the tissues withdraw from the blood a
large part of the katabolic products which compose the retained
nitrogen and that this would be disclosed by analyses of organs
and tissues obtained at autopsy.
TISSUE ANALYSES.
Extract
Water Total Extract Nitrogen,
No, Diagnosis. Content Nitrogen Nitrogen | per 100 g.
Per Cent. | Per Cent. Per Cent. | Dry Sub-
stance.
Psoas Muscle.
A;))|VPancreatitis s.4crocecter isle Sat 72.9 2.92 0.24 0.88
TOs VW Peritonitis! taser. 6 etterrs eer eee 72.8 3.06 0.23 0.84
tr. 5|/:Cerebral thrombus... ee eet 73.8 2.99 0.24 0.90
12: 4| Paeumoniany.ccew eco oe eeiste fore 77.2 3.08 0.32 0.97
Liver.
A; T| IPANCheAtitis:, Giclee tec bie cicieceraine 74.4 2.52 0.18 0.70
TO: || Peritonitis::. <7 | ac ste savaversrerolerere = 62.1 2.34 0.17 0.44
rr. | Cerebralithrombusa x C, in which A is the surface
area in square cm., W is the weight in kilograms and H the height
in cm. and Caconstant. Various formulas were tried, care being
taken that 3/a + 1/6 should always equal 2 in order that the for-
mula might remain bi-dimensional. The formula A = W'/
< H' x C gavean average error of + 3.3 per cent. The formula
A = W'”? x H' x C gave an average error of 2.2 percent. It
was evident that values for 1/a somewhere between 1/3 and 1/2
would give better results. By a rather lengthy process of calcula-
tion it was found that the average error could be reduced to 1.7
per cent. if the formula were made A = W125 x H1/138 x C,
The calculation is not difficult if logarithms be used but a chart
has been devised by means of which it is possible to estimate
the surface area at a glance. The ordinates represent the height
Hospital Medical College, to whom we here express our sincere thanks. He found
the macroscopic and microscopic evidences of emphysema, with the exception of
increase in connective tissue. There were also areas resembling infarctions probably
due to rhexis as a result of the increased blood pressure in the pulmonary circuit.
1D. Du Bois and E. F. Du Bois, Proc. Soc. Exp. Biot. AND MED., 1914, XII,
16; Arch. Int. Med., 1915, XV, 868.
78 SCIENTIFIC PROCEEDINGS (72).
in cm., the abscisse the weight in kilograms and curved lines
drawn diagonally across the chart give the readings for the surface
area in square meters. The details of the work with the chart
and values of the constants will appear shortly in the Archives of
Internal Medicine.
46 (I1I0)
Diabetic dietetics. Glucose formation from protein foods.
By N. W. Janney and F. A. CSonKA (by invitation.).
[From the Chemical Laboratory of the Montefiore Home and Hospital,
New York City.]
The carbohydrate content of foods has been usually accepted
as a gauge of their adaptability to the dietary of diabetics. It
has, however, been demonstrated that proteins yield in the glyco-
suric organism large amounts of glucose (50 to 80 per cent. of their
weight, Janney). An exact knowledge of how much sugar arises
in diabetic metabolism from protein food is therefore of some im-
portance.
By observing certain precautions it was found possible to
determine rather accurately the amounts of glucose yielded by
various meats fed to dogs made completely diabetic by phlorizin. °
The sugar formed from beef, chicken, chicken eggs, rabbit and fish
could thus be ascertained to represent from 9 to 12 per cent. of
the uncooked moist food. The solid substances of these materials
produced from 36 to 48 per cent. glucose. Broiling and frying
lead to considerable loss of water with corresponding increase of
the percentage formation of glucose. Broiled beefsteak would
yield 17.5 per cent. glucose.
Flour fed in preliminary experiments of the same nature gave
rise to 92.5 per cent. of sugar on an anhydrous basis of calculation.
From this data it was computed that in regard to sugar production
or liberation in the organism 100 gm. bread is equivalent to about
350 gm. broiled beefsteak. In formulating diets for diabetics,
glucose formation from the protein as well as the carbohydrate
content of the food should therefore be considered. The experi-
ments here alluded to will be later reported in detail.
GASTRO-INTESTINAL MOTILITY. 79
47 (1111)
Effect of fatigue upon gastro-intestinal motility.
By Lupwic Kast.
[From the New York Post Graduate Medical School.|
On fifty-four patients and healthy individuals tests were made
regarding the motor power of the stomach and intestines by means
of X-ray examinations, and the stomach tube. In addition car-
mine was used for the determination of time during which same
was entirely eliminated from the body. The tested subjects were
kept on a standard diet for a number of days and the motor effi-
ciency of the stomach and intestines was determined repeatedly
by these methods. By taking the average, it was determined how
long it took the stomach, and separately, how long it took the
intestines, to propel the given amount of food. During the period
of tests, the amount of physical activity was regulated and the
mental activity approximately limited. Without any change of
diet and mental activity, the physical activity was markedly in-
creased or decreased and the effect of same upon the evacuating
power of the stomach or intestines observed. At other times,
without change in the diet or the amount of physical exercise, the
amount of mental efforts was markedly increased or decreased and
the effect observed in the same manner. Summarizing the results
of such experiments, it appears that the healthy individual showed
very little variation as regards the evacuating power of the stomach
or intestines as long as the physical exercise was not excessive.
Mental efforts had no perceptible effect.
In patients with moderate and marked degrees of atony and
splanchnoptosis, the evacuating power of the stomach and in-
testines decreased in direct proportion to the amount of mental
and physical exertion. Physical exertion had the same effect in
such individuals if the patient was kept in a horizontal position
during these exercises and during the period of observation in
order to eliminate the effect of splanchnoptosis in the erect position.
In patients who clinically appeared susceptible to the effects of
fatigue, mental exertion was more marked in its delaying effect
than physical exertion.
80 SCIENTIFIC PROCEEDINGS (72).
Recognizing the difficulty of even approximate measurements
of physical and mental efforts during several days, it is evident
that only a large number of similar experiments are apt to reduce
the effect of accidental factors or of physiological fluctuations in
the motor efficiency of the gastro-intestinal tract.
48 (1112)
On the behavior of the mammalian ovary and especially of the
atretic follicle towards vital stains of the acid azo group.
By HERBERT M. EVANS.
[From the University of California, Berkeley, California.]
I have given elsewhere! a description of those cells of the mam-
malian body which react so predominantly even if not in a wholly
specific way with vital dyes of the acid azo series as to justify their
recognition as a great functional unit or cell class. For the cells
in question it is suggested that we retain the old term macrophage,
which although proposed by Metchnikoff without the kind or the
complete extent of evidence now available for delimiting the class,
nevertheless puts in the foreground their salient structural and
functional peculiarity and has the further advantage of enabling
us to codrdinate these studies with those long made by the com-
parative pathologist.
It is worthy of note that in some of those cases of local tissue
degeneration and death which one must regard as physiological
or normal, the macrophages must, in analogy with the experience
of pathologists, be actively concerned. This above all is exem-
plified by the cyclic changes undergone by the mammalian ovary.
The strange cells which since the time of Pfliiger have been known
to be of assistance in atresia of the follicle and whose derivation
from granulosa or theca or from leucocytes, 7. e., from practically
every available source, has in turn been championed—are picked
out by the azo dyes so brilliantly and so electively as to preclude
the denial of their alignment as typical macrophage cells
There will be demonstrated a series of drawings of these
colonies of macrophages in the atretic follicle of the mouse, rat,
1 Evans, H. M., ‘‘The Macrophages of Mammals,’’ Am. Jour. Physiol., Vol. 37,
No. 2, May, 1913.
VITAL STAINS OF THE ACID AZO GROUP. 8I
guinea pig, rabbit, dog and monkey, in the latter of which the
conditions are so similar as to stand for the case in man.
Striking as they are, these studies are not sufficiently indicative
of the altogether unusual affinity of the atretic ovum macrophages
for these dyes, a fact which forces itself on our attention when
small doses of the dyes are given. The preparations showing the
scanty macrophage content of the peculiar atresia of the dog
demonstrate also that although but little general staining resulted,
these cells have all accumulated dense deposits of the vital dye.
The macrophages are the cells which penetrate the zona pellu-
cida of the degenerate ovum and in late stages of atresia may be
present solely within the zona.
A different and more unique reaction in the ovarian follicle
must now be mentioned. Impending atresia in the good-sized fol-
licle has as its ear-marks a reaction never seen in the healthy state,
for before the nuclear disorganization seen by Fleming and
Schottander takes place the granulosa cells destined to perish
have suddenly become permeable to the vital stain which they
house in cytoplasmic granules frequent enough to mark out the
whole layer as deeply stained. On the downhill, as it were, these
cells never increase in size or function so as to often be confused
with the macrophages. This reaction of the granulosa is signifi-
cant one must feel, chiefly as proof not only of preliminary cyto-
plasmic as against nuclear change but of physical change in the
protoplasmic state. It will be well now to know whether the dif-
fering behavior of the granulosa cells is dependent on a changed
protoplasm into which now diffusion can take place (increased
permeability) or whether it be due essentially to electrical surface
changes which let adsorptive forces operate. The conditions
which bring about this reaction are typically seen in the atresia
which always overtakes the next succeeding crop of Graafian
follicles after fertilization of the preceding crop, but this behavior
is not repeated further in the pregnancy, where now other forms
of atresia may come in. Enough warrant consequently exists
for the recognition of types of atresia the occurrence of which is
related with certainty to what one may broadly term the cycles
undergone by the ovary in general. An examination of this
point in animals where with many individuals we have followed
the sexual cycles, is in progress.
SCIENTIFIC PROCEEDINGS
ABSTRACTS OF COMMUNICATIONS.
Seventy-third meeting.
College of the City of New York, February 16, 1916.
President Lusk in the chatr.
49 (1113)
Concerning the protein content of meat.
By N. W. JANNEY.
[From the Chemical Laboratory of the Montefiore Home and Hos-
pital for Chronic Invalids, New York.|
The commonly accepted modes of determining the protein
content of animal muscle are open to criticism. Thus in such
standard works as that of KGnig,! also in Atwater and Bryant’s?
extensively quoted tables, the protein material has been usually
calculated by multiplying the total nitrogen content of the fresh
meat in per cent. by the factor 6.25. This, as is known, intro-
duces a considerable source of error, for of the total nitrogen
about 13 per cent. is combined in non-protein substances. More-
over the factor 6.25 is of itself incorrect. It is obtained on the
basis of accepting 16 per cent. as the average nitrogen content
of meat proteins, whereas it has been recently established in this
laboratory that the correct value lies between 16.2 and 16.7 per
cent. of the pure muscle proteins of various species.
A second indirect method of calculating the “protein sub-
stances’’ of meat has also been recognized by Atwater and Bryant.
According to this procedure the combined weights of the ether
1 Konig, J., ‘Chemie der Menschlichen Nahrungs und Genussmittel,’’ Berlin,
1903.
2 Atwater, W. O. and Bryant, A. P., U. S. Dept. of Agriculture, Bull. 28 (rev.),
1906.
83
84 SCIENTIFIC PROCEEDINGS (73).
soluble substances plus the ash is deducted from the total solids
of the meat and the result considered as representing protein
substance. Likewise this scheme falls short of an accurate
determination of the muscle proteins. Indeed even the total
nitrogenous substances cannot be thus obtained with precision,
for ether removes from meat various bodies containing nitrogen.
The cause of these difficulties and uncertainty has been the
lack of an accurate analytical procedure for the direct deter-
mination of the proteins in muscle. It was however found
practicable to develop such a method,! an improved form of which
will appear elsewhere. The principle involved in the modified
procedure is coagulation of the muscle in alcohol and the removal
of the non-protein material by extraction. When this is carried
out with certain precautions it is possible to completely separate
the fatty and other non-protein material from the proteins,
which can be obtained in a high state of purity.
The results thus arrived at by direct analysis are compared
in the following table to those obtained by calculation as above
indicated. The computed values exceed that of the protein
actually present, by fifteen to twenty per cent. In dietetic and
metabolic studies this discrepancy must therefore be considered.
A revision of various tables of food analyses in this respect is
indicated.
PROTEIN CONTENT OF MUSCLE.
Protein Calcu-
: Total Nitrogen Protein Actuall Nitrogen? in
Species. in Muscle. Be Boar Present. fl Protein.
BOS: tieidievalvseruereera tes 8°33 20.8 17.9 16.3
RaADbIt ei. ua cies istevee ts Hee 3-39 20.8 16.7 16.3
Chiékereys i315 Ske raid es ae 3-43 21.4 17.5 16.4
Busho(halibup)eieene seer 3.10 19.4 16.7 16.4
1 Janney, N. W., and Csonka, F. A., Jour. Biol. Chem., 1915, XXII, 195.
2. Not calculated ash free.
EPINEPHRIC CONTENT OF THE BLOOD. 85
50 (1114)
The epinephric content of the blood in conditions of low blood
pressure and “shock.”
By E. A. BEDFORD and H. C. JAcKson.
[From the Department of Physiology, University and Bellevue
Hospital Medical College.|
The following is a brief record of experiments undertaken to
determine whether in low blood pressure there is an increased
activity of the adrenals.
Dr. Alfred R. Allen, who presented before this society the
results of an experimental study of the condition of the Purkinje
cells in low blood pressure and shock, stated in conversation with
the authors that he found marked histological changes in the
adrenals under similar conditions. In the present series of experi-
ments an attempt has been made to answer the question both
qualitatively and within limits, quantitatively by an examination
of the blood of animals (dogs) taken from the adrenal vein, before
and after a more or less prolonged condition of low blood pressure.
In order that blood from the adrenal might be obtained un-
diluted by the general venous blood, all veins, except the inferior
mesenteric and the left adrenal, entering the vena cava for some
distance on either side of the adrenal vein were ligatured. Through
the inferior mesenteric a canula was inserted into the vena cava,
in such a way that the flow of blood in the vena cava was unob-
structed. At the time of drawing blood, the vena cava above
and below the entrance of the adrenal vein was closed by clamps,
having rubber protected jaws. The first blood, that in the
enclosed segment of the vena cava, was discarded.
Care was taken to measure the rate of flow in order to elimi-
nate the possibility that results obtained might be due to a
greater concentration of epinephrin, because of a less rapid flow
of blood through the organ, although its activity might not be
increased. At the beginning of the experiment, blood to be
used as control was taken from the jugular vein.
Carotid blood pressure was taken.
For the determination of epinephrin in the blood, an adapta-
86 SCIENTIFIC PROCEEDINGS (73).
tion of Hoskins’s method was used. The essential part of the
method is that the tone of rabbit intestine, immersed in oxy-
genated blood at 37 degrees C., is lowered and the rhythmic
contractions more or less inhibited by the presence of epinephrin
in the blood.
To determine the quantitative relation of epinephrin in the
samples of blood tested, two methods were used. Jn one method,
the tracings obtained were compared with tracings obtained on
the addition of known amounts of adrenalin to control blood.
In the second method, the blood giving the reaction for epinephrin
was diluted with control blood until the reaction of this blood
was similar to that of the compared blood.
Low blood pressures were brought about by one of there
methods;
1. Handling of intestines.
2. Hemorrhage.
3. Occlusion of the thoracic inferior vena cava.
In most of the experiments, pressures of 30 to 40 mm. of mercury
were obtained.
In all three types of experiments, the epinephric content of
the adrenal blood was increased, provided that the pressure was
sufficiently low and the condition of low pressure was maintained
for a sufficient length of time. Since the blood was diluted with
control blood to compensate for the difference in the rate of flow
through the adrenal organ, an increased activity of these organs
was indicated.
In some cases, it was necessary to dilute the experimental
adrenal blood with thirty-two times its volume of jugular blood,
before a tracing could be obtained similar to that of adrenal
blood, drawn before low pressure was induced. In other cases
the reaction was similar to the reaction given by control blood
to which had been added adrenalin sufficient to make a I to
10,000,000 dilution.
In experiments in which samples were taken at intervals, it
was shown that the marked increase of epinephric content of
blood occurred only after a considerable duration of a condition
of low blood pressure, varying from one to two hours.
In these experiments, the later samples indicated an increasing
amount of epinephrin in the blood.
GASTROINTESTINAL MOVEMENTS. 87
Most of the experiments in which pressure was not permitted
to go below 50 or 60 mm. Hg gave negative results. A few of the
handling experiments were exceptions.
These negative experiments served as controls, indicating that
the anesthetizing and general operative procedure did not bring
about the results obtained.
To be certain that the results were due to the presence in the
blood of the secretion of the adrenal gland and not to the secretion
of some other organ, for example the pituitary body, the adrenals
were ligatured in such a way that while the blood from the lumbar
branch of the adrenal vein was permitted to enter the vena cava,
no material could pass from the adrenal organ into the circulation.
Only negative results were obtained under these circumstances.
These experiments, therefore, seem to indicate that an in-
creased activity of the adrenals accompanies a somewhat :pro-
longed low blood pressure condition.
51 (1115)
On the augmenting action of ergotoxine (Dale and Barger) on the
gastrointestinal movements.
By S. J. MELTZER and T. S. GITHENS
[From the Department of Physiology and Pharmacology of the
Rockefeller Institute for Medical Research.]
About ten years ago Meltzer and Auer! reported animal experi-
ments in which intravenous injection of ergot augmented strongly
the spontaneous movements of the gastrointestinal canal and
increased the motor responsiveness of the canal to vagus stimu-
lation. In these experiments a fluid extract of ergot (U.S. P.)
was used. At about the same time Dale and Barger succeeded
in isolating from ergot an alkaloid which they named ergotoxine.
In their interesting publication on that preparation a year later
they ascribed the characteristic physiological effects of ergot to
the presence of this alkaloid. With reference to the action upon
the gastrointestinal movements they emphatically state that the
effect is comparatively slight and inconstant, and believe that
1 Amer. Jour. of Physiol., XVII, 143, 1906.
88 SCIENTIFIC PROCEEDINGS (73).
the augmentation of the movements of the intestines observed
by Meltzer and Auer must not have been due to a principle
peculiar to ergot. ‘‘The effect on the intestinal movements,”
they state, “‘of a complex fluid such as the liquid extract, con-
taining, apart from principles the action of which is peculiar to
ergot, choline and various other vascular depressants (ergotoxinic
acid, etc.), seems to us to need a more critical analysis before any
great importance is attached to it as a specific action.’”!
On account of that statement the behavior of peristalsis was
studied by us under the influence of Dale and Barger’s specific
alkaloid of ergot, ergotoxine.2 We shall confine our present
communication to the results which we have obtained in the
experiments on rabbits. The gastrointestinal gut was observed
in a trough made by suspension of the incised abdominal wall
and kept filled with a warm Ringer solution. The animals
received artificial respiration during the entire experimental
observation. The results were unmistakable and easily demon-
strable. Against Dale and Barger we must insist that augmenting
action of their ergotoxine upon peristalsis is very pronounced and
constant. It is only indispensable that the animal should not be
too deep under the influence of ether, the only narcotic which
we have used in the present experiments. A trick which favors
further the augmenting action of ergotoxine upon peristalsis is the
injection of a warm isotonic solution (0.9) of NaCl into the fundus
of the stomach. After intravenous injections of Io mgr. of
ergotoxine, not only the pendular movements and the circular
constrictions become greatly intensified, but the contents of the
intestines are seen carried down by ‘“‘peristaltic rush’ (Meltzer
and Auer)* through large parts of the small intestines and even
through their entire length from the stomach to the cecum.
The movements are followed by a strong constriction of the
intestine extending over an inch and longer. Even the empty
parts of the intestines of a ribbon-like relaxed appearance show
unmistakable contractions after an injection of ergotoxine. The
1 Biochemical Journal, II, 287, 1907.
2It was obtained from Burroughs, Wellcome & Company. The alkaloid is
prepared in the Wellcome Physiological Research Laboratories, London, of which
H. H. Dale is the director.
3 Amer. Jour. of Physiol., XX, 259, 1907.
AN ALLERGIC SKIN REACTION TO DIPHTHERIA BACILLI. 89
peristaltic augmentation became manifest also in parts of the
colon and not infrequently even in the otherwise inert cecum.
The augmented waves of the stomach are not very pronounced
but the pyloric part of the stomach often contracts strongly as a
whole. The vagus nerves were stimulated within the thorax in
their course upon the lower part of the esophagus. Ergotoxine
unmistakably increases the motor responsiveness of all parts of
the gut to stimulation of the nerves even when their cardiac action
is in no way involved.
52 (1116)
An allergic skin reaction to diphtheria bacilli.
By J. A. Ko_MeEr, M.D.
[From the McManes Laboratory of Experimental Pathology of the
University of Pennsylvania and the Laboratory of the Phila-
delphia Hospital for Contagious Diseases.]|
While immunity in diphtheria may be regarded as being
principally antitoxic in nature, it is highly probable that antie
bodies of a lytic nature may be concerned. With this in view,
we have applied an allergic skin reaction in addition to the toxin
test of Schick, in studying immunity in diphtheria to the following
persons:
1. To 123 persons of various ages, most of whom were healthy
and well and had never had diphtheria or received an injection of
diphtheria antitoxin.
2. To 61 persons receiving curative or prophylactic doses of
diphtheria antitoxin.
The antigen for the allergic tests was prepared of 45 recently
isolated cultures of diphtheria bacilli of various types; each culture
was grown in glucose broth for four days and all mixed ina single
flask and shaken mechanically with glass beads to break up clumps.
To each 100 c.c. of the emulsion was added 5 c.c. of sterile horse
serum antitoxin (2,500 units) and the whole shaken at room
temperature for four hours. After this time the emulsion was
placed in sterile centrifuge tubes and the bacilli separated and
90 SCIENTIFIC PROCEEDINGS (73).
washed twice with large volumes of sterile salt solution. After
the final washing the bacilli were re-suspended in sufficient sterile
salt solution to make, after thorough shaking, about two billion
bacilli per cubic centimeter. This emulsion was heated at 60° C.
for an hour; cultured for sterility and preserved with 0.2 per cent.
tricresol. Subcutaneous injection of 1 and 2 cubic centimeters
into 250 gram guinea-pigs showed absolutely no evidences of local
reaction or general toxemia. In conducting the test, 0.1 c.c. of
the emulsion which we have called diphtherin, was injected intra-
cutaneously in the arm.
Reactions with the diphtherin were usually well marked and
of two types, papular and pustular reactions. The latter were
more severe than the former and both occurred with well-defined
zones of erythema. These reactions usually reached their height
within seventy-two hours and then began to recede.
The toxin tests were conducted with one-fortieth the M. L. D.
of toxin diluted with sufficient normal salt solution containing
0.2 per cent. tricresol to render the dose 0.1 c.c. which amount
was injected intracutaneously.
The throats and noses of a large number of persons were cul-
tured to study the relation between the occurrence of positive
reactions and the presence or absence of diphtheria bacilli in the
upper air passages.
The bacteriolytic power of the sera of persons reacting posi-
tively and negatively to the diphtherin test, for living diphtheria
bacilli were conducted toward throwing more light upon the
nature of the allergic antibody. Complement fixation and
agglutination tests were likewise conducted.
The following is a summary of the results of this investigation:
1. An allergic skin reaction was observed in about 70 per cent.
of children and 35 per cent. of adults following the intracutaneous
injection of a polyvalent antigen of washed, neutralized and heat-
killed diphtheria bacilli.
2. These reactions were regarded as allergic in character and
therefore entirely distinct from the toxin reaction of Schick.
3. About 53 per cent. of persons of various ages yielded positive
diphtherin and negative toxin (Schick) reactions. About Io per
cent. yielded negative diphtherin and positive toxin reactions,
AN ALLERGIC SKIN REACTION TO DIPHTHERIA BACILLI. 91
both tests agreeing therefore in about 63 per cent. of persons;
12.5 per cent. reacted positively and 24.1 per cent. negatively to
both tests.
If a positive diphtherin reaction may be regarded as an index
of lytic immunity, only Io per cent. of persons were found who
did not show the presence of either an antitoxic or lytic immunity,
while 53.3 per cent. showed both types of antibodies; 24.1 per cent.
showed antitoxic immunity only and 12.5 per cent. allergic, but
no antitoxic antibody.
4. The percentage of positive diphtherin reactions was slightly
greater among those who were convalescent from diphtheria.
5. There is no relation between the occurrence of positive and
negative diphtherin and toxin reactions and the presence or
absence of diphtheria bacilli; a negative toxin reaction in a person
presenting clinical evidences of infection indicates that the indi-
vidual does not require antitoxin but nothing more; he may be
infected with virulent diphtheria bacilli capable of disseminating
the disease.
6. The sera of persons yielding positive diphtherin reactions
were not found to possess demonstrable bacteriolytic properties
for diphtheria bacilli.
7. The sera of persons yielding positive diphtherin reactions
yielded weakly positive or negative complement fixation and
agglutination reactions with diphtherin as antigen.
8. Whether or not the diphtherin reaction will prove of practical
value in handling outbreaks of diphtheria from the standpoint
of passive immunization and diagnosis will depend upon future
experiences under such conditions and also upon the results of
experimental work bearing upon the broad question of allergic
reactions as an index of immunity; it would appear at least that
more attention should be paid the question of bacteriolytic im-
munity in diphtheria.
92 SCIENTIFIC PROCEEDINGS (73).
53 (1117)
Nitrogen retention in nephritis in children.
By Irvine S. CuTTER and Max Morse.
[From the Biochemical Laboratory, University of Nebraska, College
of Medicine, Omaha.]|
The writers have studied the urinary excretion of nitrogenous
products in nephritic children and conclude that retention, such
as is described for all of these components in adults by Mosenthal,
Folin, Foster and others for cases of nephritis does not obtain in
children. It is to be expected that as far as some of the com-
ponents are concerned, such as creatin, the adult and child con-
dition would be different for the metabolism of creatin in children
differs from that of the adult. We have found that retention
of creatin and creatinin is a matter of twenty-four hours or less
and that no retention occurs beyond that time in the cases ex-
amined. The excretion of these constituents was followed by
studying 24-hour specimens, supplemented by shorter time
specimens and temperature, food, etc., were checked in all cases.
In none of the cases studied were the creatin results vitiated by
the acetone nor by the aceto-acetic acid factor, attention to which
was duly paid. The figures obtained from the nephritic children
were lower than those obtained from a study of a number of
normal children from the Child Saving Institute, but this is not
interpreted as meaning retention, for immobility in children
involves lower nitrogen output in general. The marked divergence
in data of creatinin excretion in the nephritic children from day
to day is typically different from the figures obtained by others in
the adult, for the typical condition in the adult is constancy in
amount of excretion, whereas in the children the variation from
day to day was marked. The writers are not aware of a similar
study having been made previous to the present one.
EFFECT OF TEMPERATURES ON FORMATION OF HEMOLYSINS. 93
54 (1118)
The effect of moderately high atmospheric temperatures upon
the formation of hemolysins.
By C.-E. A. WINSLOW, JAMES ALEXANDER MILLER and
W. C. NoBLe.
[From the New York State Commission on Ventilation.]
The experiments which have been reported in regard to the
effect of high atmospheric temperatures upon susceptibility to
bacterial infections, or upon the immunity reactions in response
thereto, seem at first sight to be conflicting and unsatisfactory.
Some authors report increased resistance as a result of external
heat and others precisely the reverse. A more careful analysis
shows however that if the several factors at work in such experi-
ments and the various conditions employed by different investi-
gators be considered, the results are reasonably harmonious. A
moderate amount of heat may naturally be expected to produce
a different result from temperatures so severe as to lead to a
condition of fever in the experimental animals; and exposure to a
hot atmosphere may produce one effect on the susceptibility of
an animal to subsequent infection and quite another on the course
of an infection already established.
The majority of investigators have been chiefly interested in the
effect of the condition of fever upon recovery from infection, and
have therefore exposed their animals to atmospheric conditions
sufficiently extreme materially to increase the body temperature.
Experiments of this kind have quite uniformly indicated that the
progress of an infection already established is in greater or less
degree checked by an artificial fever due to a very high atmospheric
temperature, or produced by the Sachs-Aronson operation. Such
experiments have been made and such a conclusion reached by
Rovighi, Walther, Filehne, Hildebrandt, Loewy and Richter,
Kast, Engelhardt, and Rolly and Meltzer. In all these experi-
ments the high atmospheric temperatures used were 35°-41° C.
and the body temperatures of the animals 40°-42°. Vincent and
94 SCIENTIFIC PROCEEDINGS (73).
Sacquépée and Loiseleur on the other hand found resistance
lowered by high heating, but for the most part their experiments
were concerned with the lighting up of latent infection or the
invasion of bacteria from the digestive tract, a very different
phenomenon from the progress of the struggle for immunity against
an infection already established.
Finally there is another type of experiments in which the
effects upon vital resistance of a moderately high temperature
(30°-35°) have been studied; and these experiments yield results
quite different from those which have just been reviewed. While
a temperature approaching 40° by producing a state of fever
appears to favor recovery from an infectious disease, a somewhat
lower temperature seems to exert a lowering effect on general vital
resistance without the compensating stimulation of vital processes
which may accompany the development of fever. Five different
investigations, the only ones with which we are familiar bearing
on this point, all warrant the same conclusion. Fermi and
Salsano (1892) found that a strain of avian tubercle bacilli which
was incapable of producing a general infection in normal guinea
pigs could be found in abundance in the glands of animals kept
at 33°-35°. Similarly mice when heated showed many more
tubercle bacilli, of both avian and human types, in their glands
than did control animals; and the infection was still further
increased by combining high humidity with the high temperature.
Graziani (1906) studied the agglutinating power of the blood of
rabbits kept at various temperatures. At 2° to 4° the blood would
agglutinate at a dilution of I in 1541; at 18°, I in 854; at 32°,
I in 727. In another series the blood of rabbits kept at 32°
agglutinated at a dilution of I in 1250 while if the animals were
occasionally relieved by cold baths the agglutinating power rose
to I in 2425. Ritzmann (1907) kept guinea pigs, white rats and
mice at 35° and found that heated animals died from half a day
to three and a half days after injections of streptococci, control
animals after one and a half to eight days. Injections of toxin-
free tetanus spores and of tetanus spores plus streptococci yielded
similar results. Ritzmann also cites experiments of Wysso-
kowitsch leading to the same conclusion. Finally Ruata (1909)
kept guinea pigs at a temperature of 30° with a relative humidity
EFFECT OF TEMPERATURES ON FORMATION OF HEMOLYSINS. 95
of 85-95 per cent. and injected them with doses of typhoid,
paratyphoid, dysentery and colon bacilli and cholera spirilla
which were not fatal for normal animals.
All the guinea pigs thus
treated died in 4-26 hours, while, of control animals exposed to
the heat alone, without injections, 30 per cent. succumbed.
3.670
\ ee
2,000 EMOLYSIN FORMATION
b AVERAGES
\ FOR ALL ANIMALS
\
\
: HEATED
: CONTROLS
\
\
He
& (.006 \
re) \
7) \
q \
: \
Z \
Y
96 SCIENTIFIC PROCEEDINGS (73).
Our own experiments, which were undertaken as a part of the
extensive studies of the New York State Commission on Ventila-
tion, have dealt with this same problem of the effect of moderately
high temperatures and were carried out in the bacteriological
laboratores of the University and Bellevue Hospital Medical
College.
Normal healthy rabbits were kept (2-4 at a time) in an incu-
bator, 25’’x 48’ x 12’, at a temperature ranging from 29°-32° C.
Control animals were kept at room temperature (18°-21°). At
the beginning of the experiment each rabbit was bled (1 to 2 c.c.)
and then inoculated intravenously with 1% c.c. of a 50 per cent.
suspension of washed sheep erythrocytes. During the experi-
mental period each rabbit was bled once a week for trial titrations
of the hemolytic activity of the serum, and inoculations with the
sheep cells in increasing doses were made twice a week during
this period.
The hemolytic activity of the serum was determined as follows:
The rabbit serum was inactivated at 55° C. for one half hour.
A 5 per cent. suspension of sheep corpuscles was used, and for
complement, normal guinea pig serum diluted 1-10.
The rabbit serum was prepared in varying dilutions, as indicated
by the results of the previous titrations, and each dilution was then
titrated in the same way.
A series of ten test tubes was set up with 0.1 c.c. of sheep
corpuscle suspensionand 0.1 c.c. of diluted guinea pig complement,
and varying amounts of rabbit serum. The test tubes were then
placed in a water bath at 37° for 1 hour. At the end of that time
readings were made, and the smallest amount of rabbit serum of
the dilution which gave complete hemolysis was taken as the
hemolytic unit. Thus, if .06 c.c. of a dilution of I-500 was the
smallest amount of serum giving complete hemolysis, then .06 of
this solution was taken as the hemolytic unit (.06 x 1/500 =
1/8333 ¢.c. = .120 cubic millimeters) and this decimal, representing
the actual dilution of serum in cubic millimeters found effective
under the conditions of the experiment, was taken as the measure
of the hemolytic power of the serum. The figures in the table
(used as ordinates in the chart) are derived in this way. 1I0 in
the table means that 1/100 of a c.c. of serum (10 cubic mm.)
showed no hemolytic activity. The error introduced into the
EFFECT OF TEMPERATURES ON FORMATION OF HEMOLYSINS. 97
calculation of averages by calling this figure 10 when we only
know that it was greater than 10 will not materially affect the
results. Titrations were not performed during the second week
of Series II. Series I and III were stopped after six weeks,
Series V after five weeks and Series IV after four weeks. Other
blanks in the table are due to the death of the animals.
Hemolytic Power of Serum. (Cubic Millimeters of Serum
a Necessary to Hemolyze.)
Series. | Rabbit. Air
Temperature. |
I 2 3 4 5 6 7
Week. | Weeks. | Weeks. | Weeks. | Weeks. | Weeks. | Weeks.
I I 30° EO.000)| 2k18)|\) .O70)|)) a1 ws ni
2 30° .500] .083| .070] .059 | .044 | .022
51 20° T0/000)| LOE |) -073)| 0 =. dis a6
52 20° 10.000 .069 .050} .060 +105 -089
II 186 30° -145 bs -069| .020 | .040 | .047 | .067
187 30° aLOO) Nene .036| .034 | .032 | .054 | .084
183 20° .178 ae .075| .014 | .040 | .055 | .067
184 20° -189 ae -024] .040 :020 .033 .O61
iil 70 30° -588| .060| .028] .025 | .029 | .033
45 30° 1.000} .067| .042]| .022 | .033 | .o40
86 20° .213| .050] .024] .020 | .025 | .029
I10 20° -500| .067] .029] .o17 | .022 | .028
IV 88 30° -588| .075| .020] .026
171 30° -588| .069| .025] .032
4 20° -500| .027| .020|] .026
IQI 20° -200| .044| .015| .020
Vv 72 30° 10.000 |10.000| .172
17 30° 10.000] .100] .164| .. oe
100 30° 10.000 |10.000|10.000} .200 | .044
B73 30° -588| .400/10.000] .063 .044
83 20° 23) 002) ie a6 ae
136 20° -238| .270|10.000| .238 | .083
147 20° 2833)| -097'| 0763) .rr0 |) .083
148 20° -588| .588| .161| .047 | .o40
General average . 30° 3-670| 2.100] 1.720] .053 | .038 | .039 | .076
20° I.970| .144| .958| .059 | .052 | .047 | .064
The results as presented in the table appear to indicate a
distinct decrease in the rate of hemolysin formation on the part
of the heated rabbits. The hemolytic power of the blood of
individual animals of course varies within wide limits, yet the
averages show that in order to produce hemolysis it was uniformly
necessary to use larger quantities of serum from the heated rabbits
during the first three weeks. The influence of heat appears to
show itself in a delayed formation of hemolysins rather than in a
98 SCIENTIFIC PROCEEDINGS (73).
permanent inhibition, as later on the average curves for heated
and control animals are essentially the same. In Series III there
were only two occasions in which the lowest serum strength for a
heated rabbit fell below the highest for a control rabbit; and in
Series IV, not one.
The wide variations exhibited in individual animals preclude
the possibility of drawing definite and final conclusions from these
results, but their general tendency, as evidenced by averages,
agrees with the results of the other observers cited and they
strongly suggest that moderately high air temperatures (30°) do
not favor the development of immune bodies in the blood as
higher temperatures producing a condition of fever have been
reported to do, but on the other hand may be distinctly inimical
to such development.
SCIENTIFIC PROCEEDINGS
ABSTRACTS OF COMMUNICATIONS.
Seventy-fourth meeting.
Presbyterian Hospital, March 15, 19106.
President Loeb in the chair.
55 (1119)
The cytology of the exudate in the early stages of experimental
pneumonia.
By FRANK A. EVANS (by invitation.).
[From the Department of Pathology of the Presbyterian Hospital.]
The cells have been studied in the early exudate of pneumonia
produced in rabbits by intrabronchial injection of pneumococcus
group I, group IV, and by an attenuated strain of pneumococcus
furnished by Dr. Carrol G. Bull’s laboratory at Rockefeller
Institute; by streptococcus hemolyticus and by a streptococcus
isolated from the mouth of a normal individual by Miss Olmstead
of Presbyterian Hospital; and in the exudate in reaction to intra-
bronchial injection of 33 per cent. egg yolk in neutral broth. The
early exudate in three cases of human pneumococcus pneumonia
has also been available for study.
In all of these lesions, although many polymorphonuclears
were often present, in many of the alveoli the cytology of the exu-
date was predominantly mononuclear in character. These mono-
nuclear cells may be classified as follows: a few typical small
lymphocytes of the blood; a few epithelial cells from the alveolar
walls; relatively many oxydase-containing large mononuclears
greatly resembling the so-called transitional cells of the blood;
and almost as many non-oxydase containing large mononuclears of
99
100 SCIENTIFIC PROCEEDINGS (74).
the blood or closely related forms. In pneumonia induced in
animals heavily stained with lithium carmine, no cells stained with
carmine took part in the formation of the exudate. No plasma
cells were seen.
56 (1120)
Technique of cultivating human tissues in vitro.
By R. A. LAMBERT, M.D.
[From the Pathological Laboratory of the Presbyterian Hospital.]
Several difficulties have been encountered in the cultivation of
human tissue in vitro.
In the first place human fibrin is readily liquefied by fresh
tissue, so that when human plasma is used as a culture medium
the cells find no framework on which to grow. Losee and Ebeling
overcame this difficulty by transferring the tissue fragments at
frequent intervals before liquefaction took place. We have solved
the problem in another way which does not necessitate frequent
transfers. The method consists in using as a culture medium
chick plasma, the fibrin of which resists digestion, with the ad-
dition of an equal quantity or more of human serum. In this
medium the cells grow much more actively than in pure chick
plasma. Since there is no liquefaction it is not necessary to make
subcultures oftener than every 5 to 7 days.
That fresh human tissue cannot always be obtained when
wanted has appeared to be another difficulty in the study of human
tissues in cultures. We have found, however, that human tissues,
just as those of lower animals, may be preserved for 5 to 10 days
before using, if cut into small pieces, covered with salt solution
and put aside ina cool place. Serum and Ringer’s solution possess
no advantage over ordinary salt solution and a temperature of
15° C. appears to be as satisfactory as a lower temperature.
Tissues obtained at autopsy may be used though often infected.
We have obtained good growth of connective tissue from pieces
of liver and testis taken from a body six hours after death.
The sterilization of infected tissues constitutes a problem
which we haye not yet solved satisfactorily. Skin, which is
DEVELOPMENT OF IMMUNE REACTIONS. IOI
practically always infected superficially, may be partially sterilized
with little injury to the tissue by rinsing the surface quickly with
weak alcohol (60 per cent.). In a large number of preparations
from a piece of skin treated in this way, a fair percentage will
show no bacterial contamination, and some of the remainder will
show only occasional colonies. We have obtained a good growth
of epithelium from pieces of circumcision tissue thus treated.
A large number of antiseptics and disinfectants—toluol, chlo-
retone, tricresol, phenol, silver nitrate, hypochlorites (Dakin’s
solution), argyrol, iodine, potassium cyanide, and_bichloride
of mercury, have been tested on tissues more diffusely infected.
For nearly all of these the strength of solution necessary to kill
bacteria (staphylococcus aureus) also injures the cells.
Experiments carried out so far, however, indicate that potas-
sium cyanide and probably also bichloride of mercury are ex-
ceptions to this rule. For example, potassium cyanide in I-2,000
dilution is a very good disinfectant but injures cells very slightly.
More complete reports of these experiments will be presented in
a subsequent communication.
57 (1121)
Development of immune reactions in serum disease.
By W. T. LONGCOPE and F. M. RACKEMANN.
[From the Medical Clinic of the Presbyterian Hospital, Columbia
University, New York.]
The occurrence of immune reactions to horse serum and their
relationship to the development of serum disease in man, we have
studied by two methods: first, the sensitiveness of the skin to
intravenous injections of 0.02 c.c. of horse serum, undiluted or
diluted ten times or one hundred times with 0.85 per cent. NaCl;
and secondly, by determining the presence of anaphylactic anti-
body in the blood serum of the patient by transference to guinea-
pigs through passive sensitization.
Eleven patients have been studied, who have received for
therapeutic purposes from 4 c.c. to 350 c.c. of horse serum, in the
102 SCIENTIFIC PROCEEDINGS (74).
form of diphtheria antitoxin, antimeningococcus serum or anti-
pneumococcus serum, intravenously, intraspinally or intramuscu-
larly. Nine of the eleven cases developed serum sickness.
All of the cases, whether or not they developed serum disease,
showed sooner or later a positive specific reaction to the intracu-
taneous injection of horse serum. This was never obtained before
the seventh day following the first therapeutic injection of horse
serum and was first observed between this day and the eighteenth.
It was never demonstrable until after the appearance of serum
disease.
Anaphylactic antibodies could not be demonstrated in the two
cases that did not develop serum disease. In all of the other nine
cases these antibodies were found at some time in the serum of the
patient. In but one case did they appear before the onset of
serum disease and then on the fifth day after the therapeutic
injection of horse serum. Neither in this instance nor in any
other was the anaphylactic antibody demonstrable in the patient’s
serum during the early part of serum sickness. In all nine cases
the anaphylactic antibody was present in maximum concentration
at the close of the serum sickness and in one instance persisted for
sixty-eight days after the disease. In two cases in which the
original attack of serum sickness was followed by a relapse, the
antibodies could not be definitely demonstrated until the end of
the relapse, that is twenty-one and twenty-four days after the
therapeutic injection of horse serum. In several instances it was
possible to sensitize guinea-pigs both passively and actively to
horse serum with portions of the same specimen of blood serum
drawn from the patients towards the close of the serum sickness,
thus demonstrating that some of the proteins of horse serum and
antibodies for the proteins of horse serum may exist at the same
time in the circulation in man.
These experiments show that anaphylactic antibodies for horse
serum appear in maximum concentration in the blood serum
towards the close of serum sickness and suggest that their presence
in the circulation in large amounts determines the recovery from
this disease.
IMMUNIZATION WITH SENSITIZED BACTERIA. 103
58 (1122)
Immunization with sensitized bacteria.
By Homer F. Swirt and RALPH A. KINSELLA.
[From the Medical Clinic of the Presbyterian Hospital.]
The object of the present study was to determine the relative
immunizing property of various preparations of green strepto-
cocci. Two different strains of green-forming streptococci were
used, both isolated from cases of acute rheumatic fever. The lethal
dose of these organisms for mice, was from 0.1 to 0.5 c.c. of a
twenty-four hours broth culture. The sensitized vaccine was
prepared from a twenty-four hours broth culture, centrifugalized,
washed, killed at 56°, strong anti-serum added, incubated one hour,
washed and suspended in saline. Sensitized vaccines were always
freshly prepared. Three different antibodies have been studied,
agglutinins, complement fixing bodies and protective antibodies.
Rabbits were immunized by first injecting dead organisms, later
by living organisms and the comparative curve of antibody for-
mation studied. With unsensitized vaccine there was strong
formation of antibodies in from twelve to sixteen days, the curve
for agglutinins, complement-fixing antibodies and protective anti-
bodies running parallel. The animals immunized or rather in-
jected with sensitized vaccines showed at times a late formation
of weak agglutinins or complement-fixing antibodies. In no case
have animals injected with sensitized vaccines shown the presence
of protective antibodies. The protective antibodies we tested
by injecting diminishing quantities of the rabbit serum with
lethal doses of bacteria into mice.
Our conclusions from this experiment are that it is impossible
to demonstrate the presence of antibodies in rabbits immunized
with sensitized vaccines either living or dead. These results
cannot be applied to immunization with all varieties of bacteria,
because at present work in progress shows that agglutinins may
be induced by the injection of sensitized pneumococci.
104 SCIENTIFIC PROCEEDINGS (74).
59 (1123)
The effect of sodium citrate on blood coagulation in hemophilia.
By REUBEN OTTENBERG.
[From the Pathological Department of Mt. Sinai Hospital]
The question of the effect on blood coagulation of the injection
of sodium citrate into the circulation was raised immediately
after the introduction of the citrate method of obtaining blood
for transfusion.
Weil found that in cases with normal coagulation, the coagu-
lation time immediately after citrate transfusions was slightly
shortened instead of lengthened. As the question is one of
particular importance in the hemorrhagic diseases and as there
have been no observations recorded on the ultimate effect of
citrate administration on the coagulation of blood, I wish to
present some experiments in a case of hemophilia, whose prolonged
coagulation time made it particularly suitable for this study.
The patient, an adult male, had nearly bled to death at least
six times and presented all the typical features of the disease
excepting the family history. The blood count showed nothing
abnormal and the blood platelets were within the normal range or
slightly above it (490,000 per cu. mm. counted in metaphosphate
solution in a counting chamber). The coagulation time of his
blood obtained at various intervals within the preceding three
years had always been between one and two and a half hours.
The method of determining the coagulation time consisted in
obtaining approximately three cubic centimeters of blood with a
hollow needle direct from an arm vein. The blood was received
into a clean five cubic centimeter test tube and observed
at regular intervals, being kept at approximately body tempera-
ture. Complete coagulation was recorded when it was possible
to turn the tube up-side down without the blood flowing. Begin-
ging or partial coagulation was noted by the retarded flow of the
blood when the tube was slanted. This method is far preferable
to all the methods which involve the taking of drops of blood from
the finger or ear as these methods, due to the admixture of fluids
EFFECT OF SODIUM CITRATE ON BLOOD COAGULATION. 105
from subcutaneous tissues, give notoriously inaccurate results.
With the present patient on several occasions, blood so obtained
and examined in capillary tubes coagulated in from ten to forty
minutes at times when the venous blood was known to coagulate
in one to two hours.
The injection of 150 cubic centimeters of normal blood from
another person mixed with 0.3 gram of sodium citrate shortened
the coagulation time of the patient’s blood taken ten minutes
after the transfusion from one hour fifteen minutes (beginning
coagulation at fifty minutes) to seventeen minutes (beginning
coagulation twelve minutes). Twenty-four hours later, however,
the coagulation time was found to be practically the same as
before the transfusion, namely one hour fifteen minutes for com-
plete coagulation (beginning coagulation forty-five minutes).
The coagulation time of blood obtained nine days later was one
hour, thirty-five minutes (beginning coagulation one hour and
twenty minutes).
The intravenous injection of 0.6 gram of sodium citrate (20 cubic
centimeters of 3 per cent. citrate solution) shortened the coagulation
time of the blood obtained ten minutes after the injection from
one hour thirty-five minutes (one hour twenty minutes beginning
coagulation) to twenty-five minutes. Forty-eight hours later,
however, the coagulation time was found to have been lengthened
out to two hours and fifty minutes (beginning coagulation one
hour and twenty-five minutes).
At this time when the coagulation of the blood was at its
longest, an experiment was made to see whether there was any
immediate effect of citrate on coagulation after the citrate was
injected into a muscle. The result was negative; the coagulation
time taken thirty-five minutes after the intragluteal injection of
0.72 grams of sodium citrate was two hours and fifty-three minutes
(beginning coagulation two hours). The coagulation time of the
patient was not determined again for two weeks when it was
found to have returned to approximately the same level as had
been usual before the citrate injections, namely one hour (be-
ginning coagulation forty-five minutes).
The citrate injections and the blood transfusion produced no
ill effects whatever. The patient continued to have occasional
106 SCIENTIFIC PROCEEDINGS (74).
slight ecchymoses as before. Two months after the citrate injec-
tions he had another one of his attacks of severe hemorrhage.
CONCLUSIONS
I. In hemophilia the intravenous injection of sodium citrate
produces an immediate and great shortening of coagulation time
which is followed, twenty-four to forty-eight hours later, by a
return of coagulation time to its former prolonged period, or by a
much greater prolongation of coagulation time than before.
II. The intramuscular injection of sodium citrate seems to
have practically no immediate effect on coagulation time.
60 (1124)
The influence of intravenous injections of magnesium sulphate
upon the activities of the center of deglutition.
By J. AUER and S. J. MELTZER.
[From the Department of Physiology and Pharmacology of the
Rockefeller Institute for Medical Research.]
In order to understand our experimental results the following
physiological facts have to be recalled. Three different phe-
nomena which are under the reflex control of the center of deglu-
tition must be distinguished: (1) The transmission of food from
the mouth through the pharynx into the esophagus. This is a
complex process which comprises the execution, in a codrdinate
and stable manner, of three separate activities: the closure of the
entrances into the post-nasal cavity and into the larynx, and the
rapid transportation of the contents of the mouth into the proper
direction. We shall designate the entire action as the initial act
of deglutition. The reflex mechanism which controls it, is more
resistant to anesthesia than the two reflex mechanisms of the
phenomena to be mentioned next. (2) The peristaltic move-
ments of the esophagus. This is dependent only upon the occur-
rence of the first mentioned mechanism, the initial act of the
deglutition, and is independent of the actual passing of some con-
tents through the esophagus or of the anatomical continuity of the
latter. Transection of the esophagus or complete removal of a
ACTIVITIES OF CENTER OF DEGLUTITION. 107
great part of it does not prevent the contraction in the lower part
of the esophagus or of the cardia in due time after the initial de-
glutition, a time which varies with different species of animals
(Mosso, Kronecker and Meltzer). The initial sensory impulse,
after reaching the center of deglutition, passes consecutively
through a number of sections of that center, sending, while thus
passing, motor impulses to the corresponding sections of the esoph-
agus. This reflex mechanism of primary peristalsis (Meltzer) is
very resistant to fatigue, but is less resistant to anesthesia than
any other of the reflexes with which we are here concerned.
(3) Local reflexes of secondary peristalsis (Meltzer). A mechanical
stimulus applied to any part of the mucosa of the esophagus (dis-
tension) will cause a contraction of the corresponding part of that
canal, and, if this stimulus is brought about by some movable
mass within the lumen of the esophagus, this mass will be driven
down into the stomach by a wave which is difficult to distinguish
from a wave of primary peristalsis. If the mass is mechanically
prevented from being moved downward, no contraction takes
place at any other part of the esophagus below the stimulating
mass. That the secondary peristalsis is due to a central reflex and
not to a peripheral mechanism, is proven by the fact that it disap-
pears as soon as the vagi are cut. This central reflex is readily
fatigued, but is, on the other hand, more resistant to central
anesthesia than the transmission of the impulse from section to
section within the center.
For several years we were engaged, at various times, in bringing
forward evidences for the central nature of the inhibitory action
of magnesium salts. With this object in mind, we studied in the
present series of experiments the action of these salts upon the
primary and the secondary peristalsis of the esophagus. The
animals, dogs, received for anesthesia, three milligrams of morphin
per kilo body weight. This permitted the operative procedures
needed for our experiments, which consisted in exposing the trachea
and making a window in it below the larynx; the transection of
the esophagus and tying a glass tube in the upper end of it, the
exposing of one superior laryngeal nerve and of tying a cannula
in the external jugular vein. A short time after the operation the
initial act of deglutition could be brought on by either of the three
108 SCIENTIFIC PROCEEDINGS (74).
following methods: by tickling the pharynx with a probe intro-
duced through the window in the trachea, by injecting water or
saline solution into the pharynx by the same route, or by electrical
stimulation of the superior laryngeal nerve. The occurrence of
peristaltic or local contractions of the various parts of the thoracic
esophagus were observed by means of a catheter introduced into
the stomach end of the esophagus. The catheter had around its
esophageal end a small balloon of thin rubber; its outer end was
connected with a water manometer. Magnesium sulphate was
used in M/4 solution and was infused into the jugular vein from a
Mariotte burette.
We shall report now our results very briefly. Before mag-
nesium was given each initial act of deglutition was followed, as a
rule, by a primary peristaltic contraction of every part of the
thoracic esophagus. Further, stimulation of the esophagus, by
moving of the catheter within the esophagus to a new place, or
by a temporary distension of the balloon by air, brought on, as a
rule, several consecutive contractions of the part in which the
balloon was located (secondary peristalsis).
Some time after magnesium was permitted to run into the
jugular vein we met first a phase in which the primary peristalsis
disappeared, that is, no contraction of any part of the thoracic
esophagus was observed to follow the initial act of deglutition.
During this early stage the secondary peristalsis was in nearly all
cases still present and quite normal; nor was the primary act of
deglutition noticeably affected. When, however, more of the
solution was infused, a stage was encountered in which also the
secondary peristalsis was practically gone, while the initial act of
deglutition was still only moderately weakened, and stimulation
of the vagus still caused a fairly good contraction of the esophagus.
A still further inflow of the magnesium solution finally greatly
weakened, or even completely abolished, the initial act of deglu-
tition also.
From these observations it is evident in the first place, that the
first effect of the magnesium consists in a weakening or complete
abolition of the primary peristalsis, which means that the in-
hibitory action of magnesium was exerted during this first phase
exclusively or essentially upon the transmission of the sensory
ACTIVITIES OF CENTER OF DEGLUTITION. 109
impulse from section to section within the center of deglutition.
The occurrence of efficient initial acts of deglutition and the
presence of the secondary peristalsis testify that during this
early stage the local reflexes within the center controlling the
primary act of deglutition and the secondary peristalsis are little
affected. This is in harmony with the fact that the mechanism in
control of the two mentioned local reflexes are more resistant to
anesthesia than the mechanism which controls the primary peris-
talsis. In the second phase also the local reflexes, controlling the
secondary peristalsis, are abolished, while the initial act of deglu-
tition is still fairly active. In this phase stimulation of the vagus
causes a fairly good contraction of the esophagus. The facts ob-
served during this phase permit the following two conclusions: (I)
That the inhibitory action of magnesium in this phase is exerted
essentially on the center and but little, if any, upon the motor nerve
endings, and (2) that the local reflex of secondary peristalsis, which
comes only occasionally into play, is more readily affected than the
local reflex of the mechanism of the initial act of deglutition which is
frequently in action and which has to be of a stable and resistant
character. In the third phase, when the initial act of deglutition
is also abolished, the inhibitory action of magnesium is probably
exerted upon the center as well as upon the motor nerve endings.
For our present purpose, however, it is of no interest to us to
analyze the conditions prevailing during this phase.
The chief results of our experiments, so far as the action of
magnesium is concerned, consists in the following conclusions:
that a graded intravenous injection is capable of causing a com-
plete central depression of the mechanism of deglutition before a
peripheral effect can be ascertained; that the transmission of
impulses from section to section within the center is more readily
affected than reflex actions, and that reflexes of an important
function in frequent action are more resistant than local reflexes
of an incidental character.
110 SCIENTIFIC PROCEEDINGS (74).
61 (1125)
Diabetes of maximum severity with marked improvement.
By H. RAWLE GEYELIN (by invitation).
[From the Medical Clinic of the Presbyterian Hospital, Columbia
University, New York.]
Case on whom the following interesting observations were
made is a man, nineteen years of age, with history of diabetes
of six weeks’ duration accompanied by extreme loss of weight
(fifty pounds) and other classical symptoms of diabetes. Ad-
mitted to the hospital in condition bordering on coma.
Sugar output stationary for five fasting days. Symptoms
slightly worse. Alternate fast and protein feedings accompanied
disappearance of sugar in three weeks. During this period ex-
hibited excessive nitrogen loss (from 25 to 38 gm. daily). Extreme
acidosis and a dextrose: nitrogen ratio for three consecutive days
of over 3.65.
Subsequently a tolerance of 250 gm. carbohydrate was ob-
tained and four months after onset patient was tolerating 100 gm.
carbohydrate on a mixed diet of protein, fat and carbohydrate,
aggregating 2,500 to 3,000 calories daily. There was no acetone
in the urine and the blood sugar remained normal (below 0.1
per cent.), the percentage when patient was first sugar-free having
been .195 per cent., and on admission 0.312 per cent.
Just before discharge from hospital developed a peritonsillar
abscess. Tolerance for carbohydrate markedly diminished in this
period but rapidly returned after infection had subsided.
Points of unusual interest:
1. Most excessive continued nitrogen waste.
2. Highest D:N ever seen with recovery.
3. Acute onset of diabetes of great intensity, subsequent rapid
development of high food tolerance with normal blood sugar.
DIABETES. III
62 (1126)
The control of acidosis and its relation to impaired sugar meta-
bolism in human diabetes.
By FRANK P. UNDERHILL.
[From the Sheffield Laboratory of Physiological Chemistry, Yale
University, New Haven.]
Acidosis exerts a distinct influence upon carbohydrate meta-
bolism. This assertion is supported by the observation of
Elias,! who demonstrated that the introduction of acid into dogs
and rabbits leads to hyperglycemia and glycosuria. Moreover,
the same author? has concluded that the so-called “hunger dia-
betes’’ of young dogs? is in part, at least, a condition due to acidosis,
as determined by the carbon dioxide content of the blood and an-
alysis of the alveolar air. Observations upon human diabetes
teach that acidosis obtains in this condition also.
A state of alkalosis is likewise potent in exerting an action upon
carbohydrate metabolism but this influence is contrary to that of
acidosis. Pavy and Godden‘ showed that the glycosuria provoked
by ether and chloroform disappears after the intravenous injection
of sodium carbonate. Given by mouth or intravenously sodium
carbonate will abolish the hyperglycemia of ‘‘hunger diabetes”
and glycosuria will either entirely disappear or be greatly dimin-
ished, according to Elias.’ After removal of the pancreas sodium
carbonate introduced into the blood stream causes diminution in
the excretion of sugar.6 Later work by Murlin (reported at the
December meeting of the Society for Biological Chemists) has
shown that under the influence of sodium carbonate the respira-
tory quotient is increased in depancreatized dogs. At the Decem-
ber meeting of the Society of Biological Chemists Underhill re-
ported that in the hyperglycemia produced by epinephrine the
1 Biochem. Zeit., 1913, 48, p. 120.
2 Elias, Biochem. Zeit., 1913, 52, p. 331.
3 Hofmeister, Arch. f. Exper. Pathol. u. Pharm., 1890, 26, p. 355.
4 J. Physiol., 1911-12, 43, Proc., p. vii.
6 Biochem. Zeit., 1913, 52, Pp. 331.
6 Murlin and Kramer, J. Biol. Chem., 1913, 15, p. 365.
112 SCIENTIFIC PROCEEDINGS (74).
intravenous administration of sodium carbonate will significantly
lower the excretion of sugar in the urine, the hyperglycemia being
correspondingly decreased in height and duration. It was also
stated that the intravenous injection of sodium carbonate into
normal animals will sometimes although not invariably cause a
distinct fall in the blood sugar content.
From these illustrative observations it may be concluded that
a condition of acidosis tends toward the elimination of carbohy-
drate from the body whereas alkalosis shows a tendency to conserve
the carbohydrate. Otherwise expressed it seems tenable that
carbohydrate metabolism of the organism is maintained in equili-
brium by a balance between the acids and bases of the body.
Applying these ideas to human diabetes one gains the following
conception of its chemical pathology: without reference to what
may initiate the abnormal condition, a state of acidosis unques-
tionably develops and must tend to become aggravated, if any-
thing, by the characteristic acid-producing foods that characterize
the conventional diabetic dietary. From what has already been
pointed out, however, it seens reasonable to conclude that anything
which will counteract or neutralize the continuous stream of acid
entering the body should benefit the individual. One is led to
ask, what influence would this have upon the excretion of sugar if
the organism were once saturated, so to speak, with alkali and
enough alkali continually supplied to neutralize the exogeneous
and the endogenous acid? These considerations have been put
to the test in a young diabetic, 26 years of age, with a very severe
type of diabetes. When first seen by me fifteen months ago there
was a sugar excretion of 151 grams per day. Ona restricted diet
the output of sugar was reduced to 25-50 grams, acetone and
diacetic acid always being present in relatively large quantities.
After a year’s interval in spite of very stringent dietary restrictions
the sugar excretion suddenly increased to 70-80 grams daily.
Gradually increasing doses of sodium bicarbonate to a maximum
of 120 grams per day resulted in a gradual diminution of sugar
output until the urine became sugar-free. The dosage of sodium
bicarbonate was thereupon decreased at the rate of 7 grams per
day until the intake amounted to 42 grams which has been main-
tained to the present time. Under the alkali treatment the urine
AcIDOSIS AND CREATINE ELIMINATION. 113
has remained free from sugar for a period of seventeen days during
which the food ingested has been augmented little by little to the
point where about 10 grams of carbohydrate in addition to that
present in the previous strict diet are being ingested daily.
Throughout the entire course of his treatment the patient has
continued at his duties as an instructor in the university.
63 (1127)
Possible inter-relations between acidosis and creatine elimination.
By FRANK P. UNDERHILL.
[From the Sheffield Laboratory of Physiological Chemistry, Yale
University, New Haven.]
Current views associate the elimination of creatine with some
perversion of carbohydrate metabolism. The probability of a
close relationship of this sort is indicated by the well known fact
that a deficiency of carbohydrate in the body leads to creatine
elimination which may be checked promptly by ingestion of car-
bohydrate. There are experimental facts which the familiar
hypothesis fails to explain. McCollum and Steenbock! found
that in the pig a diet of corn products led to the appearance of
relatively large quantities of creatine in the urine. Similar
experiments of Folin (reported at the December meeting of the
American Society of Biological Chemists) with oat feeding yielded
comparable results. The dietaries employed can scarcely be
regarded as lacking in carbohydrate.
Deficiency of carbohydrate usually means an accompanying
acidosis, not necessarily caused by ketogenic substances, which
presumably involve the tissues associated with creatine-creatinine
metabolism. At any rate nearly every instance in which there is
creatine in the urine is accompanied by an acidosis—generally a
ketonuria also. These facts suggest the hypothesis that a con-
dition of acidosis in the tissues is responsible for the appearance of
creatine in theurine. To test it the following questions demand an
answer.
1J, Biol. Chem., 1912-13, 13, p. 209.
114 SCIENTIFIC PROCEEDINGS (74).
1. Will creatine appear in the urine, even in the presence of an
abundant carbohydrate supply, if acidosis is induced?
2. Will the elimination of creatine disappear if the acidosis is
abolished, quite independently of the factor of carbohydrate
supply?
Upon a diet of oats and corn, containing an adequate supply
of carbohydrate, creatine promptly appears in the urine of the
rabbit. A marked condition of acidosis, as measured by the hy-
drogen ion concentration of the urine, is always associated with
this phenomenon. Oats and corn are pronounced acid-producing
foods. On the other hand, if a base-producing food, such as
carrots, is fed to rabbits with creatinuria this symptom rapidly
disappears as the urine becomes strongly alkaline.
The protein per se is without special significance in the phe-
nomenon under discussion; for upon a diet consisting of oats, corn
and carrots creatine fails to appear in the urine, and the reaction
of the latter remains alkaline. Equally significant is the further
fact that the ingestion of HCl in addition to the mixed diet causes
the appearance in the urine of significant quantities of creatine.
Simultaneously the hydrogen ion concentration of the urine is
markedly increased.
The conclusion seems inevitable that there is an inter-relation-
ship between acidosis and creatine elimination. Creatine ex-
cretion may prove to be an index of a condition of acidosis in the
organism.
64 (1128)
On the production of soap jellies, and the physical conditions under
which jelly formation takes place.
(Preliminary communication.)
By G. H. A. CLOWEs.
[From the Biological-Chemical Laboratory of the State Institute for
the Study of Malignant Disease, Buffalo, N. Y.]
In the course of experiments regarding the influence exerted by
various electrolytes on the equilibrium of emulsions, published in
the year 1913, the writer noted that NaCl, when used at a concen-
tration in excess of .4M, caused a precipitation of some constituent
PRODUCTION OF SOAP JELLIES. 115
of the aqueous phase of an emulsion of oil dispersed in water, and
that the emulsion subsequently broke down, the oil and water
layers separating. This effect was believed to be attributable to
the precipitation of the surface film of soap on which the stability
of the emulsion depended. To test this question Na oleate was
treated with salt at different concentrations, and it was found that
at .4 to .45M NaCl complete precipitation of the soap took place.
It was noted, however, that prior to precipitation a tendency to
jelly formation was exhibited in the zone from .2M NaCl to .4 or
.45M NaCl.
An attempt to repeat this experiment with a soap, which had
been slightly acidified either by the addition of a minute quantity
of oleic acid or of mineral acid, gave an entirely different result, an
opalescence with increasing cloudiness and tendency to precipita-
tion was noted between .2M and .4M NaCl, followed by com-
plete precipitation at .45M NaCl. Further tests using varying
proportions of soap, varying proportions of NaOH, and of NaCl
and other salts of Na, brought out the remarkable fact that, as
long as the soap employed was not too greatly diluted and was
slightly alkaline, a jelly would be formed at all points between
.2M Na and .45M Na regardless of whether the Na was derived
from NaOH, from NaCl or other salts of Na.
In very concentrated soap solutions or in very strong alkali
the jelly formation commences at a somewhat higher concentration
and continues also somewhat above .45M. But it may be stated
as a general principle that a zone of jelly formation obtains within
these ranges, provided the original concentration of OH ions is in
excess of the amount required to produce a strong pink coloration
of the soap solution with phenolphthalein. If insufficient alkali
is present as a result of the addition of small amounts of organic or
mineral acids to the system, precipitation instead of jelly formation
is observed. Since jelly formation commences and ends at a
given strength of the Na salt almost regardless of the nature of the
anions present provided there is a sufficient initial concentration
of OH ions, it seems probable that the explanation is as follows:
A dispersion of Na oleate in water represents a dispersion of
particles of oleic acid by means of NaOH. Further additions of
NaOH lead to a more perfect dispersion of the soap particles,
116 SCIENTIFIC PROCEEDINGS (74).
owing to the fact that the OH ion is more readily adsorbed than
the Na ion. NaCl exerts a similar effect to NaOH, the Cl ions
exerting a dispersing effect analogous to that of the OH ions, but
since they are far less readily adsorbed than the OH ions their
effect is considerably smaller. This point may be demonstrated
by adding NaCl in increasing amounts to a soap solution contain-
ing enough alkali to give a strong pink color with phenolphthalein.
A discharge of the color takes place, and the amount of alkali re-
quired to compensate for the effect of the NaCl introduced follows
a logarithmic curve indicating clearly that the added NaCl either
promotes the adsorption of OH ions already present, or that the
Cl ions are more readily adsorbed than the Na ions, thus leading
to a reduction in the OH ion concentration in the water phase.
The soap particles possess a negative charge attributable pre-
sumably to adsorbed anions. This charge prevents their coales-
cence until the concentration of the Na ions reaches such a point
that they also come into play and by adsorption on the particles
tend to counteract or diminish the negative charge conveyed by
the previously adsorbed OH or Cl ions.
When a certain concentration of the cation is reached, a criti-
cal zone commences, in which jelly formation or precipitation
appears to depend entirely upon the relative proportions of ad-
sorbed cations and anions. If at the commencement of this critical
zone the residual negative charge carried by the particles resulting
from an adsorption of anions in excess of cations is sufficient to
maintain a perfect dispersion of the particles throughout the
system, as indicated by an examination of the suspensions for
Brownian movement by means of the ultramicroscope, jelly
formation will ensue at higher concentrations. If this residual
negative charge on the particles is insufficient, if they no longer
exhibit perfect dispersion when examined by means of the ultra-
microscope, if agglutination, aggregation and sedimentation under
the influence of gravity has already commenced, precipitation
necessarily ensues at higher concentrations. It is obvious, there-
fore, that the formation or non-formation of the jelly in this
critical zone is dependent simply upon the relative concentration
in the system at the lower critical point of anions like OH, which
are more readily adsorbed and anions like Cl which are less readily
PRODUCTION OF SOAP JELLIES. T17.
adsorbed, and more or less readily adsorbed cations. If at this
critical point, the sum total of adsorbed anions is not sufficiently
in excess of that of adsorbed cations to insure perfect dispersion,
precipitation instead of jelly formation ensues. This explains the
necessity for a certain minimum concentration of NaOH, with its
readily adsorbed OH ions, to insure jelly formation in the case
cited above.
It must be presumed that at the moment at which the particles
suffer a sufficient loss of charge no longer to repell one another, they
tend to coalesce with one another, and also become distorted and
elongated into films and rods under the influence of changing sur-
face tension conditions. It is obvious if they are sufficiently
finely dispersed at this point that each particle will coalesce with its
neighbor to form a jelly-like structure (analogous in a sense to a
honey-comb) enclosing globules of water between the coalescing
particles of the original dispersed phase, the structure retaining
the form of the original containing vessel. If on the other hand
the particles were not sufficiently dispersed at the time at which
coalescences commenced, if they were already partly aggregated
and no longer exhibiting perfect Brownian movement, they would
no longer be perfectly distributed throughout the entire mixture,
they would be further apart and, as a result of the diminution of
their charge, would tend to aggregate and precipitate to the bottom
of the vessel.
There are obviously a large variety of possible intermediary
structures between the most perfect jelly formation, resembling
a honeycomb, which would be impermeable to water, and the
precipitated structure which would be absolutely permeable.
Various degrees of permeability would result from the production
of systems analogous to a sponge in which two continuous phases
exist side by side, and the permeability of such systems would de-
pend upon the extent to which intercommunication between ad-
jacent partially enclosed aqueous phases has been maintained.
Further experiments with CaCl, and soap suspensions confirmed
this theory and afford a satisfactory explanation for the phe-
nomena of blood coagulation, the production of the casein clot,
and other cases of jelly formation actuated by salts of Ca. The
conversion of a system consisting of particles of fibrinogen dis-
118 SCIENTIFIC PROCEEDINGS (74).
persed in water, into a system consisting of a more or less perfect
dispersion of water in an external or continuous fibrin phase may
be further explained in a manner analogous to the explanation
offered for the transformation of emulsions of oil in water into
emulsions of water in oil, by considering the surface tension rela-
tions on both sides of a concentration film formed at the interface
between the dispersed fibrinogen particles and the surrounding
water. This phase of the question will be discussed in a subsequent
paper on the process of blood coagulation.
This theory, that jelly formation depends on the extent of dis-
persion of colloidal aggregates when exposed to the effect of a
precipitating agent, offers an explanation for the variations in
permeability of a hypothetical protoplasmic membrane, or for that
matter of tissues as a whole, under the influence of suboxidation
products. A reduction in the concentration of OH ions available
for adsorption resulting from the presence of acids would render
the dispersion of certain colloidal aggregates less perfect than is
normally the case. These aggregates would then tend to pre-
cipitate rather than to undergo jelly formation when subjected to
the influence of coagulating agents. The structure formed would
necessarily be more permeable and would possess less strength
and elasticity than that formed under normal conditions of jelly
formation. The destruction of emulsions and jellies, with result-
ing precipitation of the soap present when the concentration of
NaCl exceeds .4M, probably bears some relation to the observation
of Jacques Loeb that marine organisms are rapidly destroyed when
exposed to that strength of NaCl, unless CaCl, or some other
antagonistic salt is added.
The principle involved in the case of soap jellies considered
above applies equally well to the reverse type of jelly formation
where cations promote dispersion and anions exert an aggregating
or precipitating effect.
The writer wishes to express his indebtedness to Miss Ruth
Theis for her assistance in carrying out certain of the experiments
referred to in this paper.
THE HuNGER MECHANISM IN BIrRDs. 119
65 (1129)
The hunger mechanism in birds.
(Preliminary report.)
By F. T. RoGeErs (by invitation).
[From the Hull Physiological Laboratory of the University of Chicago.]
Both normal and decerebrate pigeons have been used in this
study. Hunger is marked by the appearance of restlessness.
This restlessness appears before the crop is completely empty.
Lack of water even though the crop be distended with dry food is
marked by restlessness of the bird. These things are true of
birds with cerebrum intact, partially, or wholly removed.
During hunger, changes occur in the behavior of the crop. In
the normal bird with “‘appetite’’(?) satisfied or at least in the
bird which does not of its own accord eat of an abundant food
supply, the crop is very much distended. In this condition only
occasional contractions of the organ can be detected by means of a
rubber balloon; none are visible to the eye (after removal of the
feathers over the crop). An hour or two after feeding there begins
to appear in the crop contractions in groups of three or four at
intervals of 15-20 minutes. The activity of the crop is gradually
augmented and 8-12 hours later there occur groups of 8-20 con-
tractions at intervals of 10-30 minutes. Still later in some birds
(probably young) the crop is in a state of almost continuous activ-
ity. When the content of the crop has been lessened to about one
third of its capacity these contractions are directly visible. At this
time they may be seen to involve principally the lower part of the
crop. When it is completely empty these contractions are periodic
in groups of 8-16 occurring at intervals of 10-60 minutes. Each
contraction may be seen to begin at the upper part of the crop
and sweep as a deep constriction, preceded by a marked bulging
or relaxation, over the entire crop (and probably down to the
gizzard). Each wave requires a time interval of 12 to 15 seconds
to complete its cycle.
This visual evidence justifies the balloon method of recording
the contractions. Unless the pressure used is excessive the
balloon does not initiate the contractions.
120 SCIENTIFIC PROCEEDINGS (74).
In the crop which contains plenty of food and water a sudden
distension of the balloon has little effect. Sudden distension of a
balloon in an empty crop initiates a group of contractions. Using
too big a balloon or using too much pressure so as to cause excessive
dilatation of the crop causes sideways shifting of the neck and
crop (shrugging of the shoulders so to speak) evidently an effort
on the part of the animal to remove the obstruction. Similar
movements may be seen in normal birds which have stuffed them-
selves with corn. By mechanical manipulation of the crop with
the fingers isolated peristaltic contractions of the crop may be
caused. Mere stretching movements of the neck are not sufficient
to account for these contractions for they occur when the bird is
held quietly in the hand.
In the normal bird these contractions may be inhibited by
external influences such as light and noise. Light and sound do
not inhibit them in decerebrate birds but rough handling may do
so. Such disturbances of body coérdination as those following
extirpation of the semicircular canals or lesions of the cerebellum
inhibit the contractions of the crop. Incidentally, during the
period of marked incodrdination following lesions of the semi-
circular canals or cerebellum the crop is emptied much more
slowly than in normal birds.
Tonus changes undoubtedly occur but tracings are likely to
be deceptive on this point because of the close relation of the crop
to the cervical muscles. Any shifting in the position of the head
will be registered by the recording balloon in the crop. Hence
tracings may be meaningless. But in the hungry bird the crop
can be seen to be constricted into a much smaller area. It can
hardly be believed that the crop is simply folded and fallen
together. (Histological study of the crop distended and empty
is being made.)
A small fistula in the crop does not cause any visible difference
in the contractions. The contractions may be inhibited by putting
water into the crop through the fistula or by feeding the bird.
Water given by mouth does not immediately inhibit the contrac-
tions for the peristaltic waves from the throat spread downward
over the entire crop. No visible difference can be made out be-
tween contractions of the empty crop initiated by swallowing
OxYGEN CONSUMPTION IN REGENERATING TISSUE. I2I
water or those occurring perioically without swallowing, except as
to their point of origin.
Restlessness of the starved decerebrate bird may be clearly
periodic or more or less continuous. If it tends to be continuous
picking up the bird and holding it in the hand for a moment and
then freeing it will end the restlessness, unless contractions of the
crop are occurring at the same time. If the crop is actively con-
tracting the bird will continue his fruitless wanderings.
66 (1130)
Oxygen consumption in regenerating tissue.
By G. G. Scott.
[From the United States Fisheries Biological Station, Woods Hole,
Mass.*}
Little knowledge has been obtained as to the rate of metabolism
of regenerating tissue as compared with that of normal tissue.
Child, ’15,? has found that susceptibility or physiological resistance
of organisms varies directly with the rate of metabolism. He
found, in practice, that a measure of the resistance to cyanide
poison was an efficient method for determining the rate of meta-
bolism. In experiments of regenerating tissue of Planaria (flat-
worm) he concluded that immediately after operation, the rate of
metabolism fell below normal, remained there for a few days,
then arose above normal where it remained for some time after
regeneration was complete, when it gradually approached normal.
I obtained the same result with Sagartia, a small anemone (Coelen-
terate). In my method the rate of metabolism was measured by
determining the amount of oxygen consumed by the regenerating
animals as compared with the normal animals. Oxygen deter-
minations were made by means of the Winkler method. The
experiment continued for twelve days. Determinations were
made every twelve hours. Table I shows percentage consumption
1 Published by permission of Commissioner of Fisheries.
2 Child, C. M., “Senescence and Rejuvenescence,”’ University of Chicago Press,
I9I5.
122
SCIENTIFIC PROCEEDINGS (74).
of oxygen by regenerating animals as compared with normal
animals for each twelve hour period.
BOS GN Ce
% &% YS NO 4H HH AH HH HH HH HB HH H
WNH HOO MNT AMN HRW NH OS
Reg.
Reg.
Reg.
Reg.
Reg.
Reg.
Reg.
Reg.
Reg.
Reg.
- Reg.
. Reg.
. Reg.
. Reg.
. Reg.
Reg.
. Reg.
. Reg.
. Reg.
. Reg.
. Reg.
. Reg.
. Reg. Sagartia consumed 122%
Sagartia consumed 109%
Sagartia consumed 96%
Sagartia consumed 82%
Sagartia consumed 95%
Sagartia consumed 97%
Sagartia consumed 93%
Sagartia consumed 99%
Sagartia consumed 111%
Sagartia consumed 165%
Sagartia consumed 142%
Sagartia consumed 140%
Sagartia consumed 135%
Sagartia consumed 117%
Sagartia consumed 130%
Sagartia consumed 187%
Sagartia consumed 135%
Sagartia consumed 125%
Sagartia consumed 124%
Sagartia consumed 144%
Sagartia consumed 129%
Sagartia consumed 125%
Sagartia consumed 119%
TABLE I.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
of amount of oxygen consumed by nor.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
Sagartia.
The result is parallel to that found by Child with Planaria.
While extensive morphological
studies on regeneration have
been made, it is necessary that a more complete study of the physi-
ological processes involved should also be made.
SCIENTIFIC PROCEEDINGS
ABSTRACTS OF COMMUNICATIONS.
Seventy-fifth meeting.
College of Physicians and Surgeons, April 19, 1910.
President Jacques Loeb in the chair.
67 (1131).
An active expiratory muscle in the chicken which is inhibited by
stimulation of the central end of the vagus. A demonstration.
By A. L. MEYER and S. J. MELTZER.
[From the Department of Physiology and Pharmacology of the
Rockefeller Institute for Medical Research.]|
In mammals expiration is passive under ordinary conditions.
It is only under abnormal conditions that certain muscles become
active during the expiratory phase of respiration. At the last
meeting of the Federation of American Societies for Experimental
Biology! we made the statement that in the fowl normal expiration
js active. We wish to demonstrate the truth of this statement by
a graphic method. We have found that the innermost of the ab-
dominal muscles in the chicken when carefully isolated contracts
regularly with each expiration. When the contractions of this
muscle are recorded simultaneously with the movements of the
thorax it will be observed that the muscle contracts during ex-
piration and suddenly relaxes during the onset of inspiration.
The literature concerning the effect upon the respiration of
stimulation of the central end of the vagus in mammals is very
extensive and full of conflicting opinion as to the nature of this
effect. In fowls stimulation of the central end of the vagus causes
an unmistakable inhibition of the contractions of this muscle.
When the movements of the thorax and the contractions of the
123
124 SCIENTIFIC PROCEEDINGS (75).
expiratory muscle are registered simultaneously, stimulation of
the central end of the vagus brings out an instructive picture.
Throughout the period of stimulation the thorax remains quiescent
in an inspiratory position, while the expiratory muscle remains
completely relaxed.
This phenomenon is another instance of the general law of
“contrary innervation’? (Meltzer), or ‘‘reciprocal innervation”’
(Sherrington). Inhibition of the expiratory group of muscles
during inspiration was suggested by one of us over thirty years
ago.!
68 (1132)
A demonstration of the effects of some lesions of the nervous
system.
By J. GoRDON WILSON and F. H. PIKE.
[From the Department of Otology, Northwestern University, and the
Department of Physiology, Columbia University.]
The effects of the lesions were shown in cinematograph films
of three different animals. A rabbit which was brought into the
laboratory some months ago presented constant marked torsion of
the head to the left. There was no nystagmus, but merely a
constant deviation of the eyes. The animal could move about on
rough surfaces if it went slowly and carefully, or if its left side was
supported by the side of the cage. If put on a smooth surface
with the left side unsupported, any attempt on the part of the
animal to move was followed by rolling movements to the left,
about the long axis of the body. If no obstacle was placed in its
way, the animal might roll for several yards before regaining its
upright position. The animal was said to be about eight months
old at the time it was brought into the laboratory, and to have
been in the same condition from birth. The only gross changes
visible at autopsy were in the left otic labyrinth. The nature of
these changes was not determined by inspection. The histological
report will be presented later. One interesting point in the de-
portment of the rabbit was its lack of compensation for the loss
1 Arch. fiiy Physiol. (DuBois-Reymond’s) 1883, 216.
SEPARATION OF SERUM. 125
of the labyrinth, as compared with the deportment of cats or dogs
after loss of one labyrinth.
Two cats were subjected to experimental ablation of the vermis
and left lateral lobe of the cerebellum. The eye movements were
different from those following labyrinthine lesions. One marked
motor defect was the trembling and uncertainty of movement of
the head when attempting to take food. Two different stages in
recovery from the effects of the cerebellar lesion were shown, in
one of the cats, with the gradual amelioration of the symptoms in
the second stage taken at an interval of about one month after the
first.
The film of the rabbit was made through the courtesy of Pathe
Freres. The films of the cats were paid for out of the Patton
Fund of Northwestern University Medical School.
69 (1133).
A separation of serum into coagulative and non-coagulative
fractions.
By ALFRED F. HEss.
[From the Research Laboratory, Depariment of Health, New
York City.]
As is well known, it is possible, by means of salting out with
appropriate percentages of ammonium sulphate or sodium chloride,
to almost entirely separate the albumin from the globulin and the
pseudo-globulin of serum. This has been done in the preparation
of diphtheria antitoxin, where it has been found that the antitoxin
is closely associated with the pseudo-globulin fraction.
A similar procedure was carried out to determine the associ-
ation of the coagulative principles of the serum. It was found that
in human plasma as well as in horse plasma, these substances are
linked with the euglobulin fraction. If these three proteid frac-
tions of the serum are separated and dissolved in normal salt
solution and added to plasma (with the addition of a small amount
of calcium) the euglobulin will markedly hasten coagulation,
whereas the two other fractions will have either no effect or a
slightly inhibitory action. It is possible in this way to prepare a
126 SCIENTIFIC PROCEEDINGS (75).
refined serum containing only 14 to 1 per cent. proteid, that is to
say, one tenth to one fifth the quantity present in normal serum,
but possessing an equal potency as regards coagulation. This
preparation may be passed through a Berkefeld filter so as to be
rendered sterile.
This euglobulin would seem to be of value for subcutaneous or
intravenous use in hemorrhage, particularly on account of its
small quantity of proteid.
70 (1134)
Comparative distribution of urea, creatinine, creatine, uric acid,
and sugar in blood and spinal fluid.
By M. S. FINE and V. C. Myers.
[From the Laboratory of Pathological Chemistry, New York Post-
Graduate Medical School and Hospital.|
Comparative analyses of blood and spinal fluid were carried
out in 15 cases. These patients were suffering from nephritis of
various stages of severity, and gave chemical blood pictures varying
from practically normal to the excessive retention of uremia.
The concentration of urea in the spinal fluid averaged 88 per cent.
of that in the blood; the concentration of creatinine, 46 per cent.;
of creatine, 22 per cent.; and of uric acid, 5 per cent. of the re-
spective concentrations in the blood. If these differences in con-
centrations may be regarded as representing the relative perme-
ability of the cells separating the blood and spinal fluid, one notes
that the extent of passage into the spinal fluid is greatest for urea,
less for creatinine, still less for creatine and least for uric acid. It
is of interest to note that this represents also the order of their
solubility in water, and, in part, the relative ease with which
these substances appear to be eliminated by the kidney.
It may be further observed that the sugar concentrations of
the spinal fluid in these fifteen cases averaged 57 per cent. as much
as that of the blood.
ANTAGONISM BETWEEN ATROPIN AND CENTRAL EMETICS. 127
71 (1135)
Antagonism between atropin and certain central emetics.
By Cary EGGLESTON.
[Laboratory of Pharmacology, Cornell Medical College, New York
City.]
The minimal certainly emetic vein dose of pilocarpin alkaloid
(hydrochloride used) was determined for dogs as 0.7 mg. per kilo.
It having previously been observed that atropin was capable of
antagonizing the emetic action of pilocarpin, experiments were
made to determine the smallest dose of this alkaloid (sulphate
used) which was just sufficient to prevent emesis from the minimal
emetic dose of pilocarpin. The antagonistic dose of atropin
alkaloid was then determined for twice, four, eight and sixteen
times the minimal dose of pilocarpin.
The results showed that it requires about 1/75th as much
atropin base as of pilocarpin base to antagonize the emetic action
of the smallest effective dose of the latter. About the same ratio
was found for twice the dose of pilocarpin. For four times the
minimal dose of pilocarpin 1/95th as much atropin was required;
for eight times about 1/125th; and for sixteen times about 1/200th.
Similar experiments were made with nicotin and atropin, but
the toxicity of the former drug prevented the use of amounts
larger than the minimal emetic dose. Atropin was found to an-
tagonize nicotin in the proportion of about I : 70 (both in terms
of base).
Other emetics previously shown to cause vomiting through
central action were tested with atropin in doses up to 5.0 mg. of
the base per kilo, or 500 times the effective dose against pilocarpin
and 1,000 times that against nicotin. In no case was there any
antagonism demonstrable. The drugs used were apomorphin,
morphin, ouabain and emetin.
It has been shown! that pilocarpin produces emesis through a
direct central action and since section of the vagi does not increase
the minimal emetic dose, a local action of the drug in producing
emesis seems very improbable. The antagonism of atropin,
1 Eggleston, C. and Hatcher, R. A., Jour. Pharm. and Exp. Ther., 1915, VII, 225.
128 SCIENTIFIC PROCEEDINGS (75).
therefore, would seem to be a central one, probably in the nature
of a depression of certain central structures concerned with the
vomiting act, or of certain paths to or from the central mechanism.
It should be stated that the dose of atropin required to antagonize
the minimal emetic dose of pilocarpin is insufficient to dilate the
pupil and does not appreciably diminish the salivation or diarrhea
produced by the pilocarpin. The mechanism of antagonism
between atropin and nicotin is apparently the same as between
atropin and pilocarpin, and it is interesting to recall the fact that
nicotin and pilocarpin—the only central emetics which we have
found so far to be antagonized by atropin—are very closely related
in their pharmacologic actions.
Atropin is stated to be capable of preventing the emesis often
seen following the therapeutic use of morphin in man and that
induced in dogs. The mechanism of this action is usually given
as involving a local action of both drugs on the stomach, morphin
emesis being ascribed largely to a marked stimulation of the motor
endings of the vagus in the stomach, which are depressed by
atropin. In dogs, at least, morphin has been shown! to produce
emesis through a central action and we have not been able to
prevent this action by atropin in any dose. This failure con-
firms the observations of Guinard,” who, however, conceded some
antagonistic action between atropin and morphin in man, which
he thought due to a synergistic central depressant action of the
two drugs.
The failure of atropin to antagonize the central emetics studied,
other than pilocarpin and nicotin, raises several interesting points
regarding the physiology of vomiting. We are all aware of the
number and diversity of ways by which vomiting may be induced
and of the existence, therefore, of many afferent paths for the
stimulation of the central vomiting mechanism. It is suggested,
on the basis of the present observations, that atropin antagonizes
nicotin and pilocarpin on the one hand by depressing some limited
portion of the vomiting center, and on the other hand fails to
antagonize the other centrally acting emetics used since these
may possibly influence the central mechanism through other and
different portions.
1 Loc. cit.
2 Lyon Medical, 1895, LXXX, pp. 37 and 40.
DISTRIBUTION OF FAT-SOLUBLE A. 129
Experiments were also conducted using hyoscyamin in place of
atropin, and others are now under way covering some of the other
drugs with central emetic actions. The results of all of these will
be detailed in the complete paper to be published later.
72 (1136)
The distribution of the fat soluble A, the growth-promoting sub-
stance of butter fat, in the naturally occurring foodstuffs.!
By E. V. McCo.titum, Nina SIMMONDS, and WALTER PITz
(by invitation).
[From the Laboratory of Agricultural Chemistry of the Wisconsin
Experiment Station.]
That butter fat and egg yolk fats contain a substance whose
chemical nature is unknown, which is indispensable for growth or
prolonged maintenance of health was first pointed out by McCol-
lum and Davis. Later they showed the presence of this substance
in the maize kernel and in wheat embryo, and presented some
evidence that if it is found in the oat kernel it is in very small
amount.? Our further studies have confirmed these observations.
McCollum and Kennedy? have discussed the desirability of
employing the term “fat-soluble A”’ for this, to distinguish it from
the ‘water-soluble B,”’ a substance which is widely distributed in
the natural foodstuffs of both animal and vegetable origin and is
likewise indispensable for growth or prolonged maintenance. The
water-soluble B only is concerned with the production and cure of
polyneuritis in pigeons.
Our experimental work with the grains has shown that the
content of the fat-soluble A is greater in the maize kernel than in
wheat, and greater in wheat than in the oat kernel. In all three
the content is too low to induce growth at the maximum rate
even though all other factors in the diet be near the optimum.
1 Published with the permission of the Director of the Wisconsin Experiment
Station.
2 McCollum and Davis, Jour. Biol. Chem., Vol. 15, p. 167 (1913); Vol. 21, p. 179
(1915); Vol. 23, pp. 181 and 231 (1915).
3 McCollum and Kennedy, ibid., vol. 24, p. 491 (1916).
130 SCIENTIFIC PROCEEDINGS (75).
We have much experimental evidence indicating that the unknown
A is principally confined to the germ of the seed. Sunflower seed
appears to be fairly rich in this substance.
We have also found that the leaves of certain plants, especially
alfalfa and cabbage are very rich in the fat-soluble A as compared
with the grains. It is probable therefore that it is universally
associated with metabolizing plant cells. We have rats in our
colony which have grown to very near the normal adult size at
slightly below the normal rate on a simple mixture of polished rice
sixty and powdered alfalfa leaves forty per cent. They are in an
excellent condition after eight months on this diet and one female
has produced young.
We wish to call attention to the importance of having found a
good source of the fat-soluble A in foodstuffs containing but little
fats and other substances soluble in lipoid solvents. We shall
report later on methods of isolating this substance from such
sources.
73 (1137)
The effect of exercise on the blood sugar of depancreatized dogs.
By GEORGE M. MACKENZIE (by invitation.)
[From the Department of Pathology of the College of Physicians and
Surgeons, Columbia University, New York.]
Blood sugar curves of dogs made to run on the treadmill one to
four days after extirpation of the pancreas showed:
1. That after 20 to 30 minutes of such exercise, in animals
which were being fed 200 grams of meat and bread daily, there
occurred a fall in the amount of reducing substance in the blood,
sometimes amounting to as much as 100 mgm. per 100 c.c.
2. That in starved animals such exercise caused a rise in the
amount of reducing substance in the blood, amounting in one case
to 85 mgm. per 100 c.c. during 30 minutes of exercise.
The conclusions suggested by these results are that, even after
complete extirpation of the pancreas the power of sugar consump-
tion is not entirely lost, and that there may be a difference in the
power of such animals to utilize sugar according as it is derived
from tissue proteins or by absorption from the intestinal tract.
STUDIES ON THE BLOOD OF THE ALBINO Rat. 131
74 (1138)
Studies on the blood of the albino rat.
Its normal cellular constituents. Their reaction to sarcoma
growth and to benzol treatment.
By KENNETH TaytLor, M.A., M.D. (by invitation.)
[From the Laboratories of the Department of Medicine, University of
Minnesota.]|
While working with a transmissible sarcoma of the white rat
it was found desirable to note the changes in the blood picture
during the growth of the tumor. With the idea of establishing the
normal, a careful search of the available literature on the blood of
the albino rat (and also the common wild rat) was made. No
reports on the blood plates and only a few on the leucocytes of
this animal could be found. For this reason, and because of the
growing importance of the white rat as a laboratory animal and its
availability for tumor work, it has seemed advisable to contribute
this small series of studies on the blood of albino rats, with special
reference to the blood plates in the normal animal. The changes
in the blood picture due to sarcoma growth and benzol treatment
have been observed.
Hans Hirschfeld! in a paper on the differential morphology of
the white cells of the blood reported the usual types of cells to be
present in the blood of rats of mixed and albino breeds. He noted
especially a cell with annular nucleus and fine eosinophilic granules.
He did not report the usual number of white cells or their differ-
ential count.
Pappenheim? was the first to report in brief the changes in the
blood of the albino rat coincident with the progress of fatal trans-
missible sarcoma. His conclusions were that, except for a very
slight secondary anemia and a great degree of polychromatophilia
and granular degeneration of the red cells, there was little reaction
on the part of the blood until the tumor ulcerated, when a marked
leucocytosis appeared.
1 Virchow Archiv, 1897, CXLIX, p. 22.
2 Folia Hematol., Vol. X, 2, p. 393.
132 SCIENTIFIC PROCEEDINGS (75).
Hirschfeld! reached much the same conclusions, but reported
finding normoblasts in the blood. He recorded a leucocyte count
of 180,000 in one rat with an ulcerating sarcoma. No normal
standard of number or of differential count of leucocytes was re-
ported in either of these papers. The blood plates were not
enumerated or described.
In the work presented here the leucocytes and blood plates
were counted after the method of Wright and Kinnicutt? designed
for plate counting. It was modified by the procedure of first
drawing the fixing fluid to the mark .5 in the stem of the white
cell pipette in order to prevent the blood from coming in contact
with a dry surface. A 1-20 dilution was invariably used. Two
pipettes were used for each count and five drops counted from each
to determine the number of leucocytes. One hundred small
squares (Turk’s stage) from the dilutions in each pipette were
counted to determine the number of blood plates. The following
table shows the results of eighteen studies on the blood of eight
apparently healthy albino rats. For the blood plates the figures
given represent the nearest 50,000; for the leucocytes the nearest
500.
Average mumberiblood platesiperccmien. eerie icicle ieisiels 1,000,000
(Variation 850,000 to 1,200,000)
Average number of leucocytes per ccm............s+e-se+e-ee 19,000
(Variation 12,000 to 30,000)
Average number of red blood cells per ccm..................- 10,000,000
(Variation 9,000,000 to 10,500,000)
Differential leucocytes (300 cells counted) :
Average polymorphontclearcellsapeemanicneinieiiorincineeeae 50 per cent.
(Variation 30 per cent.—60 per cent.)
Average mononuclear cells (small 30 per cent., large 14 per cent.) . 44 per cent.
(Variation in proportion great)
Averageitransitionallcells) Vira «herelereheleterslencteve leletoisietetarcrersieiaiete 5 per cent.
(Variation 2 per cent.-14 per cent.)
Averageleosinophil icellaiyanecwie ic rtaicheleteneneeheeeisienc ie terol stere taliarevieNs I per cent.
(Variation 0-3 per cent.)
It will be seen from the table for normal rats that the blood
plate count shows an unusually small degree of variations. The
1 Fol. Hemat., Vol. X, 2, p. 393.
27. A.M.A., Tort, LVI, p. t457-
STUDIES ON THE BLOOD OF THE ALBINO Rat. 133
plates themselves, however, differ greatly in size, measuring from
two to five micra in long diameter. In the diluting fluid they
usually appear slightly oval and cup-shaped. They are granular.
Varied forms, however, may be seen in the dry and stained pre-
parations. The red blood cells, even in healthy rats, always
show considerable granular degeneration and polychromatophilia.
The total number of leucocytes is high as compared with the human
count and, as shown in the table, has a wide variation in apparently
normal animals. The reaction to an inflammatory process com-
monly results in a high leucocytosis. Several counts of over
45,000 per c.c. were made. Two were in cases of large subcu-
taneous abscesses, two in cases of ulcerating sarcoma.
The differential count of leucocytes in normal animals shows a
fairly constant percentage of polymorphonuclear cells. The total
number of mononuclear cells is also fairly definite, but there is
great variation in the proportion of large and small cells in different
animals. While the small mononuclear cells usually greatly
outnumber the large, two apparently normal rats showed more
large than small cells.
In five albino rats inoculated with sarcoma the blood was
observed. The results agree in general with those of Pappenheim
and of Hirschfeld. No marked change in the blood picture is
noticeable during the growth of the tumor except an increase in
the amount of polychromatophilia of the red cells and a slight
increase of small mononuclear leucocytes at the expense of the
large mononuclear and transitional cells. When the tumor was
of an ulcerating character, however, leucocytosis was marked:
45,000 in one, 50,000 in another rat. Blood plates remained
stationary.
Benzol leucopemia was readily produced in two rats, the
leucocyte count falling from 15,000 to 1,200 in one, and from
25,000 to 2,200 in the other; while the plates showed a correspond-
ing drop from 850,000 to 500,000, and from 1,000,000 to 500,000
respectively inside of ten days. The fall in leucocytes was pro-
portionately greater in the large mononuclear and transitional
cells than in the polymorphonuclear and small mononuclear
cells.
134 SCIENTIFIC PROCEEDINGS (75).
SUMMARY.
1. In the blood of the normal albino rat the blood plate count
approximates 1,000,000 per c.c. and is fairly constant.
2. The total leucocyte count is variable and reacts violently
to ulcerative processes.
3. The reaction of the cellular elements of the blood to the
growth of transmissible sarcoma is slight and probably in no way
characteristic.
4. Benzol injections produce a more rapid and proportionately
greater reduction in the leucocytes of the blood than in the plates.
75 (1139)
Gravimetric determination of beta-oxybutyric acid.
By Donatp D. VAN SLYKE.
[From the Department of Chemistry of the Rockefeller Institute.|
If beta-oxybutyric acid is oxidized with dichromate in the
presence of sulfuric acid and mercuric sulfate, a precipitate of the
acetone compound of mercury sulfate can be obtained in an
amount proportional to the beta-oxybutyric present. Thus, if
175 c.c. of a beta-oxybutyric solution containing 9 per cent. of
sulfuric acid, 2 per cent. of mercuric sulfate, and 0.25 gram of
potassium dichromate are boiled under a reflex for an hour, 7.7
milligrams of mercury-acetone compound are precipitated for each
milligram of beta-oxybutyric acid present. The beta-oxybutyric
acid may vary from I to 9 mg. without affecting the ratio, if the
concentrations of the other reagents are kept constant.
76 (1140)
Complement fixation in tuberculosis.
By H. R. MILLER, M.D. and HANs ZINSSER, M.D.
[From the Department of Bacteriology, College of P. and S., Columbia
University, New York.|
In a recent communication to the New York Pathological
Society, the material of which is to appear in the American Journal
COMPLEMENT FIXATION IN TUBERCULOSIS. 135
of Medical Sciences, the writers described work on complement-
fixation in tuberculosis, carried out with a very simple antigen
which had yielded and is still yielding results more satisfactory
than those hitherto reported by other workers who had used other
antigens. The work followed a study of culture-filtrate antigens,
such as those devised by Besredka and by Petroff, and the special
modifications of the Besredka medium employed by Bronfen-
Brenner and by Craig. These antigens did not in our hands react
with the regularity which we thought should attend a reaction of
specific diagnostic value. Owing to irregularities perhaps due to
constituents of the media, it was thought wise to return to the
bacillary substances themselves, work along this line having been
attended by considerable success within recent years—notably in
the hands of Radcliffe, Dudgeon, Weir, and Stimson. It should not
be forgotten that the same direction of investigation was followed
in the earlier work of Wassermann and Citron and in that of Cal-
mette.
The method employed is in general identical with that which
we have been using in this laboratory for the extraction of Tre-
ponema pallidum, typhoid bacilli and streptococci, and differs in
no essential particular from the so-called ‘‘endotoxin”’ extraction
method employed by Besredka in 1906 with organisms of the
typhoid-colon group. Since we feel that the procedure at present
in use in the Columbia laboratory should be thoroughly rein-
vestigated by other workers, we believe that it is proper to give
in great detail the method by which the antigen is made.
The bacilli which, so far, have been used for the production
of the antigen have been of the human type, some of them isolated
by Miller, some of them obtained from Professor Theobald Smith,
some from the laboratory of Professor William H. Park, and some
from the laboratory of Parke Davis & Co. They have been grown
mainly on the gentian-violet medium of Petroff and on Miller’s
modification of this medium; also on Petroff’s potato broth. It is
at present the impression of the writers that the medium on which
the bacilli are grown plays no great part in determining the use-
fulness of the antigen. It seems, however, to be important that a
number of different strains should be used—that is, that the
antigen should be polyvalent—and the use of relatively young
136 SCIENTIFIC PROCEEDINGS (75).
cultures is advisable. So far, in most of the reactions, unheated
bacteria have been used. Inasmuch as the method of production,
under these circumstances, is fraught with a not inconsiderable
element of danger, we have recently begun to use bacteria heated
to 60° for a half hour, and, in the one series so carried out, no
deteriorating effect of the heating was apparent. These problems
of detail, as well as many others, are being more thoroughly in-
vestigated.
20 mgm. of the moist tubercle-bacillus mass are weighed out,
placed in a conical 15 c.c. centrifuge tube, and to it are added 90
mgm. of table salt. With a glass rod, filed to roughness at the end,
this paste is ground by hand for about one hour. Distilled water
is then added to isotonicity; that is, 10 c.c. to the quantities above
described. This is the antigen. Just before using, it is shaken up
and the heavier particles are allowed to settle in the course of a
few minutes. Except for the removal of these larger elements,
the suspension is used as a whole without centrifugation and
without filtration.
The antigen so prepared has hardly ever been found anticomple-
mentary in quantities as large as I c.c. and has given fixation with
positive sera (the inactivated sera used in quantities of 0.1 c.c.) in
amounts as low as0.02c.c. The titrations, as well as the reactions,
have been done with one half the original Wassermann quantities,
using a sensitization of two units of amboceptor and two units of
complement. So far, we have used the anti-sheep rabbit hemolytic
system. As a routine, the 37° one-hour water-bath incubation
has been employed. A number of parallel series have been done
by the four-hour ice-box method, but, since this seems to make
little difference in the results, the time-saving 37° method was
decided upon as a routine procedure. The antigen appears to be
quite stable. We have used with satisfaction antigens as old as
six or seven weeks, kept on ice.
We are ready to report the results of 602 cases tested. Of
these 103 were negative for tuberculosis; that is to say, they
represent patients in whom, clinically, tuberculosis was excluded.
226 cases were clinically diagnosed as actively tubercular. Their
sera tested gave the following results:
“Stage one”’ cases: 32 in all with active clinical symptoms gave
COMPLEMENT FIXATION IN TUBERCULOSIS. 1a7,
positive fixations. Tubercle bacilli were not found in the sputum
from 16 of these cases. 7 of these 16 were suffering from the early
incipient type of the disease.
“Stage two’’: We tested the sera of 110 such cases. All but
12 in this series had positive bacteriological proof of infection.
The fixation test was positive in all but 2 cases.
“Stage three’’: 84 cases tested. One very advanced case gave
no fixation. The sputum was positive in all but one case. The
fixation test was positive in 83 of these 84 third-stage cases.
We have, then, 226 patients suffering from clinically active
tuberculosis, in whom the test was positive in 223 cases. —
The reaction was done with 88 sera from so-called healed
(arrested or inactive) cases. These were cases which, at one time
or another, had suffered from active tuberculosis but which were,
at present, apparently free from symptoms pointing to absorption
from any active focus. 54 cases were negative for the test; 13
cases were positive, however in 8 of these tubercle bacilli were found
in the sputum shortly before the test was performed; 21 cases
showed weak fixation. Here we have a group of 88 healed, or
better, arrested cases in which the reaction was negative in 54,
weak in 21, and positive in 13, 8 of these 13 being cases with positive
sputa.
Our next group consists of 140 doubtful cases where no diag-
nosis was established and where, obviously, no bacteriological
proof was present. 32 in this group gave positive fixation; 108
negative. In some of the 32 positive cases, a definite clinical
diagnosis of tuberculosis was subsequently established. In none
of the 108 negative cases has there been found, thus far, any
evidence of tuberculosis. This group of doubtful cases includes
84 pulmonary cases, 5 glandular cases (3 of which were diagnosed
as Hodgkin’s disease. In one of the Hodgkin cases, the serum
report was +-+-+-+, and an excised lymph node from this patient
turned out to be tubercular upon later pathological examination) ;
also there were 21 eye cases, I case of sepsis of the throat, and 12
miscellaneous cases.
45 positive Wassermann sera were tested. 2 gave positive
fixation with the tubercle bacillus antigen. One of these two
cases was a dispensary patient who at present can not be followed
138 SCIENTIFIC PROCEEDINGS (75).
up; the other was a patient with tuberculous peritonitis who had
had lues.
Almost all the healed cases had positive skin tests, yet 54 of
the total series of 88 showed negative fixation; 24 of the 103 cases
in which tuberculosis had been excluded gave marked intradermic
reaction to tuberculin. The fixation test was negative in these
24 also.
In the foregoing communication, we have reported an antigen
for complement-fixation in tuberculosis which has seemed to give
us results more regular and satisfactory than those reported by
other workers and which has the advantages of great simplicity.
The nature of the reaction and its results incline us to believe
that we are dealing with a specific reaction which depends
upon the presence or absence of antibodies to the tubercle bacillus
in the circulation of the patient. It is well to bear this in
mind in judging the results of the reaction, since it must not be
forgotten that specific complement fixation may not be a direct
measure of infection, but rather indirectly it may point to the
invasion of the body by a specific microérganism, by determining
the presence of antibodies. Thus, it may be too much to expect,
especially in a disease so chronic as tuberculosis, to find antibodies
circulating in all forms and in all stages of the disease. Perhaps
this will make it more easy to understand why the reaction has
given positive results in active cases only, nearly always negative
ones in inactive tuberculosis, and was occasionally negative when
tubercle bacilli were in the sputum but the clinical condition was
one of arrested disease. It is this aspect of the reaction particularly
which leads us to hope that it will be of clinical value in indicating,
not so much the existence of infection as of determining the activity
of the focus, and, incidentally, giving us a method of studying the
fluctuations of antibodies during the disease. The work will, of
course, have to be continued by multiplying the number of cases
already observed. We are also proceeding in our own laboratory
to study the specificity of antigens made with bovine cultures and
to study the relationship of this reaction to the diagnostic tuber-
culin tests.
ANTIGENIC PROPERTIES OF BACILLUS TyYPHOSUS. 139
77 (1141)
Preliminary studies on the antigenic properties of different
strains of bacillus typhosus.
By SANFORD B. HooKER. (by invitation.)
[From the Hearst Laboratory of Pathology and Bacteriology, Unt-
versity of California.]
A search through the literature reveals no report of special
work upon antigenic differences among typhoid strains, although
serologic methods have frequently been used in differentiating
typhoid from closely allied organisms. The demonstration of the
severally specific antigenic individualities, notably of strains of
pneumococci, streptococci, gonococci, meningococci, and influenza
bacilli; the fluctuant epidemiologic severity of typhoid fever from
time to time; the observation that antityphoid inoculation confers
no protection against paratyphoid infection; the growing list of
instances in which antityphoid inoculation has been unsuccessful,
the previously known and personally confirmed fact that a poly-
valent antigen is essential for good alexin fixation reactions in
typhoid fever are the main facts which have led to this investiga-
tion.
Attention has been focused chiefly upon the delicately specific
method of alexin fixation as a means of detecting antigenic dif-
ferences. A considerable number of confirmatory agglutinin
absorption experiments have also been performed.
Materials and Technic.—Of the 48 strains which have been used
21 are laboratory strains two to fifteen years old, and the rest have
been isolated and authenticated during the past year by Gay and
Chickering in the course of studies of local cases of typhoid fever.
The antigens used were washed, formalized suspensions of
typhoid bacilli. These suspensions have been used also for im-
munizing rabbits, being eminently satisfactory for this purpose as
agglutinogens for agglutination and absorption tests. The total
volume of the fixation test has been one cubic centimeter, one fifth
that of the classical Wassermann. Sheep cells, rabbit anti-sheep
hemolysin, and guinea-pig serum make up the hemolytic system
140 SCIENTIFIC PROCEEDINGS (75).
which has daily been balanced by simultaneous alexin and he-
molysin titrations. Pooled serum from six or more guinea pigs
has been preserved by salting for alexin, a method which has
many advantages.
Serum cross-titrations with standardized antigens have de-
veloped the fact that different strains of typhoid bacilli fall into
different groups, somewhat analogous to the groupings of pneu-
mococci. This is evidence which must be seriously considered in
the explanation of the causes of failure in typhoid vaccination,
since it is not unlikely that subjects may have been infected with
strains dissimilar to those used for prophylactic inoculation. The
strains tentatively placed in Group I cross-fix with all antigens;
those in Group II cross-fix with each other but not with Group I
strains. Group I-A strains give irregular results. There is no
apparent relation between the virulence of the organisms as in-
dicated by the severity of the disease which they caused, or between
their toxicity as indicated by the reactions produced in immunizing
rabbits, and this grouping. All of the Group II strains, however,
have been under artificial cultivation for a number of years.
Number 13 is the Rawlins strain, so extensively used in prophy-
lactic immunization. It seems to possess a lesser antigenic com-
plexity than does any organism in the other groups. On this
evidence it would seem to be theoretically of less value as an
immunizing strain.
The results of absorption tests, while somewhat less consistent,
are confirmatory of the findings obtained with the alexin fixation
reaction. Group I and Group II serum absorbed respectively
with Group I and Group II strains give usually negative results
with all organisms. Absorption of Group I sera with Group II
strains results in the removal of all agglutinins for Group II while
agglutinins for Group I still remain.
It is considered that the evidence of antigenic differences thus
far discovered among typhoid strains is sufficiently valid to warrant
the presentation of these data, and sufficiently encouraging to
justify similar more extensive work, especially with regard to the
comparative protection which is aroused by strains of different
character. Although the principle of polyvalency has been used
empirically in the past, it would seem advisable now on more
ANTIGENIC PROPERTIES OF BACILLUS TYPHOSUS. 141
certain data to employ a polyvalent typhoid vaccine for im-
munizing and therapeutic purposes compounded in accordance
with these groups as tentatively suggested and which may be
confirmed or extended in the future.
78 (1142)
Note on ‘ Salt fever.”
By THEO. C. BURNETT and GEO. H. MarrTIN, Jr.
[From the Rudolph Spreckels Physiological Laboratory of the Uni-
versity of California.]
About six years ago one of us! published a short account of the
rise of temperature which follows the injection of sodium chloride
into rabbits, either intravenously or subcutaneously. This fact
seems to have been completely overlooked by subsequent inves-
tigators, of whom there are many (Bingel, Freund, Samelson, Hort
and Penfold, McIntosh, Fildes and Dearborn, and others).
Samelson? claims that the rise of temperature is due to bacterial
toxins contained in the distilled water, and not to the sodium
chloride. The observations were made on nursing children.
Freund,? on the other hand, maintains that the sodium chloride
is the cause of the rise of temperature, at the same time admitting
the fact that contaminated water may also cause fever.
As we wished to make use of this fact in another connection,
it became necessary for us to be sure that the rise of temperature
was due to the injection of the salt, and to that end we have re-
peated the earlier work. Sterile sodium chloride was put in a
flask that had been throughly sterilized, and the water, redistilled
in glass, was received directly into the flask from the condenser.
The mouth of the flask was closed with sterile cotton, and the
solution (m/6 concentration) was used as soon as it had cooled down
to the proper temperature. There can be no doubt, therefore, of
the purity of the solution as far as bacterial contamination is
concerned. Antiseptic precautions were observed in making the
injections.
1 Burnett, Univ. Calif. Publ., Vol. 4, 1910, p. 5.
2Samelson, Monatsch. f. Kinderheilk, Vol. 11, p. 3.
3 Freund, Arch. f. exp. Path. u. Pharm., Vol. 74, 1913, p. 311.
142 SCIENTIFIC PROCEEDINGS (75).
The results confirmed our earlier work. The temperature rose
steadily after injection, the height being roughly proportional to
the dose given. The maximum was attained in from three to
five hours, after which the temperature gradually returned to
normal. As an example, a rabbit weighing 2,500 grams was
injected subcutaneously with 25 c.c. sterile sodium chloride
solution, m/6. In five hours a maximum temperature of 40.4° C.
was recorded. The next day 35 c.c. were injected, with a maxi-
mum of 41° C. in five hours. The following day 20 c.c. gave a
maximum of 40° C.in three hours. 25 c.c. Ringer’s solution caused
a slight rise, but not so marked as the pure sodium chloride (39.8°
as against 40.4°). This is not in accord with our original findings,
and it is possible that the distilled water used at that time was not
perfectly pure. Our present results are more in harmony with
Loeb’s theory of balanced solutions, and with the results of other
workers.
As intravenous injections become more and more general, it
would seem wise, when sodium chloride is used as a menstruum
for other substances, that the amount injected should be so gradu-
ated as to fall below that which will cause a febrile reaction; or
better still, Ringer’s solution should be used.
79 (1143)
The influence of morphin upon the elimination of intravenously
injected dextrose.
By I. S. KLEINER and S. J. MELTZER.
[From the Department of Physiology and Pharmacology of the Rocke-
feller Institute for Medical Research).
In a series of eight experiments dextrose was injected into
dogs which had received 10 mg. of morphin per kilo of body
weight. Ten other dogs received similar dextrose injections but
no morphin, the slight operation having been performed under
local anesthesia produced by cocain or ethylchloride. The dosage
of dextrose was 4 gm. per kilo of body weight, injected in a 20
per cent. solution in about one hour. The difference in the urinary
INFLUENCE OF MORPHINE UPON INJECTED DEXTROSE. 143
and blood findings in these two series of experiments was quite
striking. In the eight morphinized animals the average quantity
of sugar in the urine secreted in two hours and a half (that is,
from the beginning until one and a half hours after the end of the
injection) amounted to 63 per cent. of the injected sugar, 80
per cent. being the largest and 50 per cent. the smallest quan-
tity. The average quantity of sugar in the urine of six non-
morphinized dogs in two hours and a half, amounted only to about
17 per cent. of the injected sugar, 30 per cent. being the highest
and 4 per cent. the lowest quantity. There was, also, however,
a difference between the two series of dogs in the volume of urine
secreted. In the morphinized dogs the average amount of the
injected sugar solution was 137 c.c. and of the urine 197 c.c.; in
the non-morphinized dogs the average of the injected sugar solu-
tion was 187 c.c. and of the urine only 83 c.c. On this account
experiments were made on four non-morphinized dogs in which
the dextrose was dissolved in 144 M solution of sodium sulphate,
and there resulted a reversal in the relation of the volumes of the
injected sugar and the urine: 212 c.c. of dextrose solution injected
and 281 c.c. of urine secreted. Nevertheless, the elimination of
sugar in the urine was not increased. In fact, in these four
experiments the elimination of sugar in the urine was even less;
it amounted on the average only to about 9 per cent. of the in-
jected sugar, 13 per cent. being the highest and 7 per cent. the
lowest quantity.
As to the sugar content of the blood, we may state briefly that
in the non-morphinized dogs the original level was reached in half
an hour after the end of the injection, while in the morphinized
dogs that level was reached only one hour and a half after the
end of the injection.
Summarizing briefly our results with regard to the effect of
morphin we may say that, on the one hand, it increases consider-
ably the elimination through the kidneys of intravenously injected
dextrose, while, on the other hand, it perceptibly retards the re-
turn of the sugar content of the blood to its previous level.
“ i" ,
rd ah es
ai cy ae
SCIENTIFIC PROCEEDINGS
ABSTRACTS OF COMMUNICATIONS.
Seventy-sixth meeting.
Yale University, New Haven, May 24, 1916. Vice-President Gies
in the chatr.
80 (1144)
The therapeutic effect of wheat germ and of yeast in infantile
scurvy.
By ALFRED F. HEss.
[From the Bureau of Laboratories, Department of Health,
New York.]
As is well known, yeast is a specific therapeutic agent in the
cure of beri beri or its prototype, polyneuritis gallinarum. Studies
upon infants showed, however, that when autolyzed yeast was
given in daily quantity of 15 to 30 cc. a day, it was unable to
cure moderate cases of infantile scurvy, even when taken for a
period of two to three weeks. Yeast was however able to bring
about growth in infants.
Wheat germ was found to possess antiscorbutic power, which
however cannot be compared to that of orange juice. In some in-
stances it was able to prevent the occurrence of the subacute
scurvy which follows the use of pasteurized milk; in one instance
this disorder developed notwithstanding the fact that the infant
had received wheat germ and the watery extract of the germ for
many weeks.
Scurvy can develop while an infant is making steady gain in
weight for weeks or months, and, on the other hand, the symptoms
can disappear under antiscorbutic treatment, although no gain
is manifested. It is therefore evident that growth is not an es-
145
146 SCIENTIFIC PROCEEDINGS (76).
sential factor connected with the scorbutic condition. This
should be borne in mind, and the results of experiments on growth
should not be considered as directly transferable to infantile
scurvy or similar dietary diseases.
81 (1145)
Oxygen utilization by fishes and other aquatic animals.'
By Gero. G. Scott.
[From the United Siates Fisheries Biological Station, Woods
Hole, Mass.]
A. Lowering of the temperature causes a reduction in the rate
of oxygen consumption. In one case, while one lot of fishes
consumed 78 per cent. of the available oxygen supplied at 12° C;
a similar lot of fishes in water 4° colder consumed but 60 per cent.
In a second case, a fish in water at 20° C. consumed 94 per cent.
of the oxygen present while a similar fish at 3° C. consumed but
57 percent. Breathing had ceased in this case but was resumed
on return to warmer water.
B. It was noted that oxygen was consumed more rapidly in
tall, narrow vessels of water than in broad shallow ones. Fishes
moving about in shallow vessels of water tend to reaerate the
same. In one experiment to test this, one lot of fishes in tall
narrow vessels of water consumed 80 per cent. of the oxygen
supply, while in the broad shallow water at the end of the same
period, the analysis indicated a reduction of but 20 per cent. of
the original oxygen supply.
C. Fishes kept in dark vessels apparently consume oxygen at
a less rapid rate than those exposed to light. Thus in the light
one fish consumed oxygen at the rate of 0.12 c.c. per gm. per hr.
while in the dark the rate was D. II c.c. per gm. perhr. But
there was no evidence as to rate of oxygen consumption being
less at night than in the daytime the rate being approximately the
same.
D. Some forms show more resistance to low oxygen supply
1 Published by permission of the Commissioner of Fisheries.
NUTRITIVE VALUE OF SOME COTTON-SEED PRODUCTS. 147
than others. This is particularly true of invertebrates. Res-
piration ceases altogether, and returns if the specimen is re-
turned within certain time limits, to aerated water. The toad
fish and killifish live in water with low oxygen content while
butterfish and menhaden quickly succumb to reduction in oxygen
supply.
E. The average rate of oxygen consumption for two species
of marine worms is about 0.0205 c.c. per gm. per hr.; while that
of two mollusks is about the same, 7. €., 0.0215 c.c. O2 per gm.
per hr. That of the fish, tautog, was 0.088 c.c. per gm. per
hr. Most marine invertebrates consume oxygen at a very low
rate; fishes at a much higher rate; with amphibia the rate is be-
tween that of invertebrates and fishes; the rate with mammals
and birds is relatively high, that of birds being extremely high
as compared with anatomically lower forms.
82 (1146)
The nutritive value of some cotton-seed products in growth.
By THoMAS B. OSBORNE and LAFAYETTE B. MENDEL.
[From the Laboratory of the Connecticut Agricultural Experiment
Station and the Sheffield Laboratory of Physiological Chem-
istry in Yale University, New Haven, Connecticut.]
When certain animals are fed on a ration containing an abun-
dance of cotton-seed meal they frequently give evidence of so-
called cotton-seed injury. This has been attributed to irritation
from the indigestible husks, the oil, harmful microdrganisms, and
specifically toxic chemical compounds. The possibility suggests
itself that the rations are frequently far from ideal or adequate
in respect to the various essential nutrients, inorganic salts and
““accessories.”’ Richardson and Green! have found that when the
ration of rats is otherwise suitable, toxic symptoms do not follow
the use of cotton-seed meal. With their approval we refer to our
own experiments, which are still in progress. To ascertain whether
1 This has since been published: Richardson and Green, Jour. Biol. Chem., June,
1916, XXV, 307.
148 SCIENTIFIC PROCEEDINGS (76).
the cotton-seed proteins are notably deficient for the purposes of
nutrition, we have conducted feeding experiments on rats in which
these proteins furnished practically all of the food nitrogen and in
which the other essential dietary components were supplied by
adding to the products to be tested a suitable mixture of ‘‘pro-
tein-free milk,’ butter fat and starch which, with the addition of
adequate protein, has been shown in hundreds of experiments to
be sufficient for perfect growth. In this way we have found that
satisfactory growth can be made by rats when either cotton-seed
globulin or the total cotton-seed protein precipitated from alkali
extracts of cotton-seed meal is employed without other significant
protein sources in the mixture. No toxic symptoms have ap-
peared, even when the supposedly harmful meal also was used,
during a period in which the animals reached a large size. In ex-
periments in which the inorganic components were furnished by
our ‘‘artificial protein-free milk’’ there was no failure of growth
when the cotton-seed meal was used, thus suggesting that the
latter contains the equivalent of the “determinant,” ‘‘food ac-
cessory,”’ or ‘‘vitamin’’ deemed essential for nutrition and furnish-
ed in fat-free milk. These results corroborate the conclusions
of Richardson and Green! soon to be published.
83 (1147)
The early responses of frog embryos to tactile stimulation.
By DAVENPORT HOOKER.
[From the Anatomical Laboratory of the Yale University School of
Medicine, New Haven, Conn.|
In the course of some experiments on the regeneration of the
spinal cord of frog embryos, it became necessary to establish
certain facts in regard to their early tactile responses, as has been
done for Diemyctylus and Amblystoma by Coghill. The results
of this study are briefly summarized here.
The frog embryo exhibits a reaction toward the side stimulated
as its first response to tactile stimulation with a fine human hair.
This occurs so constantly that it must be regarded as normal for
the frog, though only an occasional and aberrant reaction in the
1 Loc. cit.
RESPONSES OF FROG EMBRYOS TO TACTILE STIMULATION. 149
salamanders. This first response is followed by an avoiding-,
a double-coil-, an S-reaction and the swimming movement, in
order.
In a number of embryos, the cord was cut at different levels to
determine the location in the cord where stimuli are transferred
from one side of the body to the other. The results are, briefly,
as follows: (1) when the cut passes through the brain, the por-
tion anterior to the cut never responds to stimuli, while that
posterior to it exhibits the usual series of reactions; (2) when the
cut passes through the medulla, the same results are obtained;
(3) when the cut passes through the middle of the body at a point
just behind the medulla, both parts usually go through the normal
series of reactions; (4) when the cut passes just anterior to the
tail or (5) through the tail itself, the part of the body anterior to
the cut goes through the normal series of responses, while that
posterior to it remains negative. From these results it is evident
that in the middle of the embryo there is a region about a milli-
meter in length which includes the upper part of the spinal cord
and the lower part of the medulla, in which the decussations of the
primary spinal nerve-paths take place, enabling the transfer of
stimuli from one side of the body to the other.
A large series of experiments on this particular region show that
the crossing does not take place at any one easily localizable point,
but rather throughout the entire region. When the cord has been
cut here, the two portions of the body thus isolated from each
other go through the normal series of responses to stimuli inde-
pendently of one another, the part in front of the cut usually being
somewhat in advance of that behind it. Further, reversal of this
region in no way affects the appearance of the responses, nor is it
possible to differentiate between the time of appearance of reac-
tions in the two extremities of the reversed piece.
Coghill suggests that the reaction toward the side stimulated,
which appears as an aberrant form cf response in Amblystoma, may
be due to the transmission of the stimuli along the collaterals to
the muscles of the same side before the main path to those of the
opposite side is fully awakened. That this is actually the case in
the frog is apparently demonstrated by the nature of the re-
sponses obtained as the embryo enters the second or avoiding-
150 SCIENTIFIC PROCEEDINGS (76).
reaction stage. At this time the embryos when first stimulated
almost always give a reaction toward the side stimulated and
only exhibit the avoiding reaction after several responses of the
more primitive type. This would seem to indicate that connec-
tions across the body occur only as a summation of stimuli.
In conclusion, it may be stated that the early tactile responses
in the frog embryo are very similar to those of Amblystoma ex-
cept that they are preceded by a constant response toward the
side stimulated. The localization of the decussation in the cord
seems to cover a wider region than that described by Coghill for
the salamander and in this region to appear simultaneously over
a length of one half to one millimeter.
84 (1148)
Permeability vs. tolerance of the kidneys for sugar in diabetes
mellitus.
By ALBERT A. EPSTEIN.
[From the Department of Physiological Chemistry, Mt. Sinai
Hospital.]
In the study of the relation of hyperglycemia to glycosuria in
diabetic and non-diabetic conditions, the following facts have
been elicited:
1. In diabetic individuals possessing healthy kidneys the
glycosuria bears a definite relationship to the hyperglycemia.!
2. Cases of diabetes with definite renal disease, frequently
show no relationship between the hyperglycemia and the gly-
cosuria.2,. The hyperglycemia in such individuals is usually
greater in proportion to the glycosuria than it is in those with
normal kidneys. Means which promote renal secretion, increase
the urinary output of sugar, with a consequent reduction of the
hyperglycemia.
3. Acute impairment of renal function in clinical and experi-
1 Epstein, Albert A., ‘Studies on Hyperglycemia in Relation to Glycosuria,”
Monograph, 1916, New York. Proc. Soc. Exper. Biot. AND MeEp., Vol. XIII,
p. 67, 1916.
21d.
SUGAR IN DIABETES MELLITUS. I5I
mental diabetes leads to a diminution or cessation of the glyco-
suria with a progressive rise in the sugar content of the blood.
The removal of both kidneys in animals, previously made dia-
betic by pancreatectomy, causes a progressive increase in the
hyperglycemia.!
4. Operative procedures in non-diabetic individuals, involving
the use of anesthetics (nitrous oxid and ether) lead to the develop-
ment of a hyperglycemia, and rarely a glycosuria.
5. Cases of diabetes are frequently encountered showing no
evidence of renal disease, in which the glycosuria disappears
spontaneously or as the result of treatment, but in which a hyper-
glycemia persists. The hyperglycemia may be of high degree,
and show slight or no variation.
When tests to ascertain the functional activity of the kidneys
are instituted on the different types of cases represented above,
the following phenomena are observed:
1. In diabetic individuals in whom the glycosuria is propor-
tionate to the hyperglycemia the response of the kidneys to the
phenolsulfonephthalein test is normal.
2. That when the hyperglycemia and the glycosuria in dia-
betic individuals do not show any relationship (the hyperglycemia
being greater than one would expect to find with a limited gly-
cosuria) there is a delayed excretion of phenolsulfonephthalein.
This group of cases, as stated above, is usually demonstrably
nephritic.
3. Non-diabetic cases, subjected to surgical procedures (under
anesthesia) which develop a hyperglycemia but no glycosuria,
show a delayed elimination of phenolsulfonephthalein2
4. Diabetic individuals, who lose their glycosuria spontan-
eously or as a result of treatment, but retain a hyperglycemia,
show a normal excretion of the dye.
5. From the observations thus accumulated, the following
deductions are made:
1. When a diabetic process is active (as a result of disease or
experimental procedures) actual disease or defective function of
1 Epstein, Albert A., and Baehr, George, J. Biol. Chem., Vol. XXIV, p. 1, 1916.
* Epstein, A. A., Reiss, J., and Branower, J., Soon to be published. Jour. Biol.
Chem.
152 SCIENTIFIC PROCEEDINGS (76).
the kidneys leads to diminution or cessation of the glycosuria
with a ‘“‘progressive’’ accumulation of sugar in the blood. The
hyperglycemia in such instances does not remain stationary, but
rises steadily—and often very rapidly.
2. Surgical procedures (under anesthesia) cause a disturbance
in the carbohydrate metabolism, with the consequent accumu-
lation of sugar in the blood (hyperglycemia). A glycosuria in
such cases is usually absent, evidently because the function of the
kidneys is impaired.
3. Cases of diabetes which become “‘a-glycosuric’’ spontaneously
or following treatment, retaining a hyperglycemia, reveal the
fact that disturbance of renal function has no part in the process.
Diminution or cessation of glycosuria through impairment of
renal function leads, as a rule, to a progressive increase in the
sugar content of the blood; but the glycosuria in these cases is
not “‘progressive.”’” The hyperglycemia in such cases may be of
high degree, and remains uninfluenced by starvation. Further-
more, such of the cases as are relieved of their glycosuria by
treatment, may upon liberal administration of carbohydrate,
develop a glycosuria, with further increase in the hyperglycemia,
and show a definite relationship between the two.
These facts are interpreted as signifying that a shifting in the
plane of carbohydrate metabolism may take place, in diabetes so
that the utilization of sugar by the tissues proceeds at a higher
level. Whereas there is no a priori reason to believe that the
utilization of sugar in the kidney differs in any way from that of
any other organ or tissue, it is concluded, that renal permeability
for sugar, is constituted of two phases: (1) a negative phase,
2. €., diminished permeability due to impairment of renal function;
and (2) a positive phase, diminished permeability due to increased
tolerance of the kidney for sugar.!
1 Tests of renal function by means of lactose, according to Schlayer and Hedinger,
are in progress, and the results will be reported later.
ALIMENTARY HYPERGLYCEMIA AND GLYCOSURIA. 153
85 (1149)
Studies in alimentary hyperglycemia and glycosuria.
By C. V. BAILEY.
[From the Department of Medicine, New York Post-Graduate
Medical School and Hospital, Dr. Edward Quintard,
Director.|
Using a modification of the Lewis and Benedict method! for
the estimation of sugar in the blood the normal value seems to be
between 0.09 and 0.12 per cent. Blood was examined in the
morning before the patients had anything to eat or drink, the urine
from a simultaneous half-hour period being tested for sugar.
Applying the above procedure it was found that in uncom-
plicated nephritis the blood sugar ranged from 0.12 per cent. in
mild cases to 0.26 per cent. in severe cases with marked nitrogen
retention. Cases of glycosuria upon admission were excreting
anywhere from a mere trace to 6 or 7 per cent. sugar in the 24-hour
specimen of urine. These cases seemed to fall into two distinct
classes; (1)those having a normal or nearly normal morning blood
sugar with urine sugar free by ordinary tests; (2) those having a
high morning blood sugar (0.3 per cent. or over) and a compara-
tively small amount of sugar in the urine. In the former class
were found the cases of ‘‘mild diabetes” and cases of hyperthy-
roidism; the latter class included cases with marked constitutional
symptoms and definite signs of nephritis—‘‘severe diabetes.”
Tests of alimentary hyperglycemia and glycosuria were begun
in the morning on an empty stomach. A specimen of blood and a
half-hour specimen of urine were collected preceding the adminis-
tration of a small quantity of glucose (30 to 90 grams in 400 c.c.
weak tea). Following this the blood was tested at 15-minute
intervals for the first 114 hours and at 14 hour intervals for the
succeeding 414 hours. Half-hour specimens of urine were col-
lected. The percentage of sugar was determined in the whole
blood, plasma, unwashed corpuscles and urine. The units hemo-
globin, percentage of corpuscles to whole blood and urinary se-
1 Myers, V. C., and Bailey, C. V., J. Biol. Chem., 1916, XXIV, 147.
154 SCIENTIFIC PROCEEDINGS (76).
cretion in cubic centimeters per minute were also determined in
each specimen.
In an apparently normal subject the whole blood contained
0.12 per cent. glucose, the percentage in the plasma being slightly
lower, and that in the unwashed corpuscles, slightly above that
in the whole blood. The sugar in the urine was apparently about
the same as in the plasma.!_ Following the ingestion of 75 grams
glucose in 400 c.c. fluid the sugar in the blood rose evenly and
rapidly, reaching its highest point in about 1 hour, returning to
normal by the end of 214 hours, falling below normal at the third
hour, and from the fourth to the sixth hour retaining its normal
level. The increase and decrease in the plasma seemed to be a
little more rapid than in the whole blood, although the difference
was very slight. The hemoglobin dropped 3 to 5 per cent. in
from 15 to 70 minutes, then increased rapidly, later more slowly,
reaching its normal in from 14% to 3 hours. Urinary secretion
decreased during the development of the hyperglycemia, increas-
ing as the blood sugar decreased. The sugar in the urine apparent-
ly increased at the same rate as in the blood up to a concentration
of 0.17 per cent. From this point the increase was much more
rapid in the urine, so that when the blood sugar had reached its
highest point, 0.23 per cent. at the end of one hour, the urine
contained 0.9 per cent. sugar. The decrease in the urine sugar
was rapid for the succeeding hour, then much slower, so that the
normal concentration was not reached until about 6 hours after
the ingestion of the sugar.
In a case of renal diabetes there was an initial hypoglycemia
with a marked glycosuria (3 per cent.). The blood sugar curve
was of the normal type, but the urine sugar curve abnormally high.
In a case of early diabetes the initial blood sugar and urine
sugar values were normal. Alimentary hyperglycemia was rapid,
the highest point being reached in about one-half hour, return to
normal taking place in less than 2 hours. The urine sugar curve
was abnormally high with a sluggish return to normal.
In diabetes of long standing without signs of nephritis, the
initial blood sugar value was high (0.2 per cent.), the urine value
normal. Blood sugar and urine sugar curves were of the previous
1See Myers, V. C., Proc. Soc. Exper. BIOL. AND MED., 1916, XIII, 180.
DIGESTIBILITY AND UTILIZATION OF EGG-PROTEINS. 155
type, but the blood sugar curve was higher and of longer duration.
Cases of diabetes with signs of nephritis showed an initial high
blood sugar with comparatively low urine sugar. The blood sugar
curve increased at about the normal rate but return to normal did
not take place before 414 to 6 hours. The urine sugar curve was
low, the highest concentration being 1.5 per cent., although the
blood at that time contained 0.31 per cent. sugar.
Cases of chronic nephritis showed an initial high blood sugar,
0.16 per cent., with urine normal. Alimentary hyperglycemia
was delayed and prolonged, the highest point being reached in
2 hours and return to normal not taking place before 4 to 6 hours.
The highest point in the urine sugar curve was 0.5 per cent., the
blood at that time containing 0.37 per cent.
A case of chronic parenchymatous nephritis showed a constant
glycosuria of 0.5 per cent. This was independent of the blood
sugar up to the latter’s concentration of 0.21 per cent. In a
second test where the blood sugar reached 0.4 per cent. the urine
sugar increased to 1.0 per cent., later decreasing and continuing
at 0.5 per cent., the blood containing 0.2 per cent.
Cases of myxedema and hypopituitarism were also studied.
In these cases the initial blood sugar and urine sugar values were
normal. Alimentary hyperglycemia was delayed and prolonged
as in nephritis and kidney permeability was greatly decreased.
86 (1150)
The digestibility and utilization of egg-proteins.
By W. G. BATEMAN. (By invitation.)
[From the Sheffield Laboratory of Physiological Chemistry, Yale
University, New Haven, Connecticut.]
Raw egg-white is found to be a decidedly indigestible sub-
stance. It may cause diarrhea in dogs, rats, rabbits and men
when ingested in any large quantity. Its utilization by the body
is poor since it is used only to the extent of from 50 to 70 per cent.
Subjects can acquire a certain tolerance for the native protein
after ingesting it for several days so that it no longer causes
diarrhea and is somewhat better utilized.
156 SCIENTIFIC PROCEEDINGS (76).
Raw egg-white can be made digestible through coagulation
by heat; by precipitation with alcohol, chloroform, or ether; by
incubation with dilute acids or alkalies; by partial digestion by
pepsin; by conversion into alkali meta-protein.
The indigestibility of native egg-white probably lies either in
its antitryptic content or in its chemical constitution. Its physical
texture appears to play a minor part in its behavior.
Of the individual proteins constituting egg-white, the albumin
fraction appears to be the indigestible component.
The whites of the hen’s egg and duck’s egg act alike in causing
diarrhea and in being poorly utilized.
Egg-yolk either raw or cooked is excellently utilized. It
sometimes causes digestive disturbances in dogs, apparently
because of its high fat content.
A review of the literature shows that dietitians have relied,
in general, upon the early observations of Beaumont as support
for the use of raw eggs. These observations were in the main
exact; but, so far as the digestibility of raw egg-white is concerned,
were misinterpreted.
In current dieto-therapy raw whole eggs, raw egg-white and
albumen-water are extensively prescribed. There appears to be
little in their conduct as foodstuffs, however, to warrant such
faith in their nutritive value or ease of assimilation.
87 (1I51)
The position of the head after experimental removal of the otic
labyrinth.
By A. L. PRiIncE. (By invitation.)
[From the Physiological Laboratories of Columbia University, and
the Yale Medical School.|
In the vertebrates usually employed in the physiological
laboratory, unilateral destruction of the otic labyrinth is imme-
diately followed by a permanent torsion of the head to the in-
jured side.!_ In a series of experiments on cats, I have found that
this posture is associated with diminished tonus in the cervical
1 Wilson and Pike, Philosophical Transactions of the Royal Society, London,
1912, series B, Vol. 203, pp. 127-160.
EXPERIMENTAL REMOVAL OF OTIC LABYRINTH. 157
musculature on the side of the lesion. Although this investigation
is as yet incomplete, it is considered desirable to report the fol-
lowing facts.
Asa means of determining the effect of impaired tonus on torsion
of the head, apart from that arising from destruction of the laby-
rinth, the following procedure was adopted:
I. UNILATERAL SECTION OF THE DORSAL ROOTS OF THE CER-
VICAL NERVES.
Accompanying the impairment of muscular tonus occasioned
by this procedure, there is torsion of the head to the side of the
injury. This torsion can only be attributed to the unbalanced
activity of the neck muscles on the intact side. It is to be noted
that the character of torsion following section of the dorsal roots
of the cervical nerves does not differ greatly from that seen after
unilateral removal of the labyrinth. In a dog, which has not
yet come to autopsy, section of the dorsal roots was followed by
torsion of the head to the side away from the lesion.
The remaining series illustrate the torsional effect of various
combined lesions.
2. UNILATERAL REMOVAL OF THE LABYRINTH AND SECTION OF
THE DORSAL ROOTS OF THE CERVICAL NERVES ON
THE OPPOSITE SIDE.
As stated before, unilateral removal of the labyrinth is fol-
lowed by torsion of the head to the side of the injury. Upon
subsequent section of the dorsal roots of the cervical nerves on the
opposite side this torsion is greatly reduced and in some cases
entirely disappears. Reversal in the order of the experiment
does not affect the end result; the torsion resulting from the first
procedure is always decreased or abolished by the second.
3. UNILATERAL REMOVAL OF THE LABYRINTH AND SECTION OF
THE DORSAL ROOTS OF THE CERVICAL NERVES ON THE OPPOSITE
SIDE FOLLOWED BY EITHER (@) REMOVAL OF THE RE-
MAINING LABYRINTH, OR (b) SECTION OF THE RE-
MAINING DORSAL CERVICAL ROOTS.
In these experiments the final procedure is followed by a
reappearance of the head torsion. The direction of the torsion,
however, is always to the side on which two lesions are combined.
158 SCIENTIFIC PROCEEDINGS (76).
4. UNILATERAL REMOVAL OF THE LABYRINTH AND SECTION OF
THE DORSAL ROOTS OF THE CERVICAL NERVES
ON THE SAME SIDE.
When these procedures are successively applied to the same
side, the degree of head torsion brought on by the first procedure
is always accentuated upon application of the second. As in the
experiments of the series 2, the order of the experiment can be
reversed without influencing the results.
5. UNILATERAL REMOVAL OF THE LABYRINTH AND SECTION OF
THE DORSAL ROOTS OF THE CERVICAL NERVES ON THE SAME
SIDE, FOLLOWED BY EITHER (@) REMOVAL OF THE RE-
MAINING LABYRINTH, OR (b) SECTION OF THE RE-
MAINING DORSAL CERVICAL ROOTS.
In this series the accentuated torsion appearing after the second
procedure is decreased on applying the third.
These results indicate that the torsion of the head resulting
from destruction of the labyrinth is caused by an impairment of
tonus in the neck muscles on the side of the lesion. The cervical
musculature involved in torsion of the head is influenced by two
distinct tonus mechanisms. The afferent impulses of the first
mechanism arise in the otic labyrinth; those of the second arise
in the peripheral endings of the fibers of the dorsal roots of the
cervical nerves. Injury to either of these two mechanisms does
not result in absolute loss of tonus in the cervical musculature, for
when destruction of the labyrinth and section of the dorsal roots
of the cervical nerves are combined on the same side, the degree of
head torsion brought by the first lesion is somewhat accentuated
by the second.
The relation of possible cerebellar paths to the labyrinthine
head torsion is now under investigation. The data available at
present are outlined below.
6. UNILATERAL REMOVAL OF THE LABYRINTH AND SECTION OF
THE INFERIOR CEREBELLAR PEDUNCLE ON EITHER SIDE.
The torsion resulting from destruction of the labyrinth is not
modified to any considerable extent by section of the posterior
cerebellar peduncles.
Is UTERINE ACTIVITY SUBJECT TO CEREBRAL CONTROL? 159
CONCLUSIONS.
The torsion of the head after unilateral removal of the laby-
rinth is due to the preponderating activity of the muscles of the
intact side. The afferent impulses concerned come largely from
the labyrinth, the muscles, the tendons of the neck, and the artic-
ulations of the cervical vertebre.
88 (1152)
Is uterine activity subject to cerebral control?
By H. G. BARBouR and N. H. COPENHAVER. (By invitation.)
[From the Depariment of Pharmacology, School of Medicine, Yale
University.]
Although morphin is known to delay the progress of labor we
have hitherto been unable to detect any inhibitory influence of
this drug upon the tone or activity of the uterus in animals. It
causes rather an increase in tone in the isolated uterus of cat and
guinea pig,! and often in the intact uterus of the decerebrate cat
or anesthetized rabbit.2, The only inhibition of the uterus by
morphin which we have observed previous to the present work has
been accounted for by circulatory collapse.
Conditions of anesthesia or decerebration under which the
morphin was given in our previous work have, by exclusion, led
us to the belief that morphin, in clinical doses, inhibits uterine
activity by a purely cerebral action. Desiring more direct evi-
dence on this point we were led to inquire into the nature of cere-
bral control of the uterus, if any exists.
To this end we have begun by the employment of a method
subjecting a part of the cortex and basal ganglia to the influence
of cold and heat. This is done by means of a double metal tube
fixed in the skull of a rabbit, on one side, anterior to the coronal
suture and passing through the anterior portion of the corpus
striatum to the base of the skull. The lateral ventricle is usually
entered. This procedure, which was first employed by one of us
1 Barbour, H. G., and Copenhaver, N. H., Journ. Pharm. and Exp. Ther., 1915,
VII, 529.
2 Barbour, H. G., Journ. Pharm. and Exp. Ther., 1915, VII, 547.
160 SCIENTIFIC PROCEEDINGS (76).
in the study of cerebral control of body temperature,! is performed
aseptically under light ether anesthesia. As soon as the animal
is free from the narcotic certain cerebral functions may be in-
fluenced by the passage through the tube of hot or cold water.
The advantages of such a method are (1) the confinement of
the effective agent entirely to the cerebrum (or a portion of it),
(2) the absence of an anesthetic, and (3) the ease with which the
functional activity of the brain can be quickly altered in either
of two opposite directions.
The uterine activity was recorded by air conduction from a
finger cot inflated within the rabbit’s uterus. Most of the animals
used were in early pregnancy. The results obtained by this
method show that under a few minutes of cerebral cooling (10° C.)
the cavity of the uterus becomes much diminished in volume
and there is a tendency to an increase in the amplitude and fre-
quency of the individual contractions; on the other hand a change
to heating (45° C.) soon causes a reversion to original conditions.
Although voluntary limb movements are sometimes a disturbing
factor we have been able to exclude these entirely as the cause of
the changes described.
There is however no doubt that the changes in volume of the
uterine cavity are largely dependent upon changes in tone of the
abdominal musculature. One can readily follow with the hand
the contraction and relaxation of the recti, for example, which
are associated respectively with cooling and heating of the cere-
brum. Furthermore the uterine changes were not observed in
two curarized animals, nor were they obtainable in an animal
with cord completely transected between the sixth and seventh
dorsal vertebrae. However, under both of the latter conditions the
normal activity of the uterus was very feeble.
The method of excluding the voluntary abdominal muscles
by suspending the intact uterus, surrounded by warm oil, in a
cylinder has failed to give very positive evidence of a direct cere-
bral control of the uterus. This method has always been pursued
under light anesthesia however. In one of six animals there was
under cerebral cooling a marked increase in tone which was not
diminished by cessation of the cooling process. In another the
1 Barbour, H. G., Arch. exp. Path. u. Pharm., 1912, 70, I.
ENDOMIXIS IN DIVERSE RACES OF PARAMECIUM AURELIA. I6I
amplitude of the individual contractions increased markedly under
cooling and diminished under heating. The other four experi-
ments were negative.
Thus far then we have established a definite cerebral in-
fluence over the volume of the uterine cavity. The fact that this
appears to be largely if not entirely a control of the voluntary
musculature of the abdomen does not detract from its importance
in connection with the birth process.
Returning to the morphin question, we have now given small
subcutaneous doses of this substance in two animals which had
responded well to cerebral cooling and heating in the manner
above described. Here the morphin, given to unanesthetized
animals, resulted in a depression of the uterine activity, although
the dose was so small in one case (.0o1 gram per kilo) that
the animal remained sitting upright and occasional normal limb
movements continued to occur. Cerebral cooling now had no
effect upon the volume of the uterine cavity of these morphinized
animals, showing clearly how morphin can influence labor by a
central action.
89 (1153)
Endomixis in diverse races of Paramaecium aurelia.
By LORANDE Loss WoopRvUFF.
[From the Osborn Zoélogical Laboratory, Yale University.]
Woodruff and Erdmann in 1914! described a normal periodic
reorganization process without cell fusion, which they termed
endomixis, in Paramaecium aurelia. This study was based chiefly
on pedigreed cells from Woodruff’s 5,000-generation race of Para-
maecium aurelia, though specimens of a race of this organism
isolated by Erdmann in Germany showed the same phenomenon.
The present communication is to prove the general occurrence
of endomixis in races of Paramaecium aurelia, since this has been
questioned, on a priori grounds, by certain authors.
The following races of Paramaecium aurelia, in addition to
those mentioned above, have now been studied:
Oberlin Race. Isolated at Oberlin, Ohio. Carried in pedi-
1 Loc. cit.
162 SCIENTIFIC PROCEEDINGS (76).
greed culture from October 8, 1914, to date, during which time it
has attained 951 generations.
Bryn Mawr Race. Isolated at Bryn Mawr, Pa. In pedigreed
culture from January 7, 1915, to February 8, 1916, when it was dis-
continued at the 650th generation.
Oxford Race. Isolated at Oxford, Ohio. Pedigreed culture
started on July 16, 1915, and has to-day (May 24, 1916) attained
the 779th generation.
Woods Hole Race. Isolated at Woods Hole, Mass. Pedi-
greed culture begun on August I1, 1915, and discontinued on
January 14, 1916, at the 305th generation.
Each of the above races has shown endomixis at the regular
rhythmic periods throughout its culture and therefore this ad-
ditional data from races from diverse sources fully corroborates
the statement of Woodruff and Erdmann! that ‘‘this reorganiza-
tion process is a normal phenomenon and probably occurs in all
races of the species Paramaecium aurelia.”’
go (1154)
Further investigations on the cyclic changes in the mammalian
ovary.”
By LEO LOEB.
[From the Department of Comparative Pathology, Washington
University, St. Louis, Mo.]
In former investigations I have described cyclic changes in the
ovaries of the guinea pig which depend largely upon injurious
influences exerted upon the ovaries in the period directly pre-
1 Woodruff and Erdmann, ‘‘Complete Periodic Nuclear Reorganization without
Cell Fusion ina Pedigreed Race of Paramaecium,"’ PRoc. Soc. FOR EXPER. BIOLOGY AND
MeEp., Vol. 11, 1914 (preliminary paper). Erdmann and Woodruff, ‘‘ Vollstandige
periodische Erneuerung des Kernapparates ohne Zellverschmelzung bei reinlinigen
Paramaecien,”’ Biol. Centr., Bd. 34, 1914 (preliminary paper). Woodruff and
Erdmann, ‘‘A Normal Periodic Reorganization Process without Cell Fusion in
Paramaecium,” Journal of Exper. Zoology, Vol. 17, No. 4, 1914 (complete paper).
2 During the summer 1915 serial sections of a number of ovaries were made for
me in the department of anatomy of Washington University. I wish to express my
appreciation to Dr. R. T. Terry for placing the facilities of his laboratory at my
disposal.
CycLic CHANGES IN MAMMALIAN OVARY. 163
ceding ovulation. Inasmuch as ovulation depends upon degenera-
tive changes having previously taken place in the corpora lutea,
the cyclic changes in the life of the follicles are correlated with
the cyclic changes in the corpus luteum. It was our aim to
determine whether the same ovarian cycle existed in all mammalian
ovaries; we examined for this purpose ovaries of the rabbit and
of the ferret at various periods of sexual activity.
Summarizing our observations we may state that neither in
the rabbit nor in the ferret do cyclic changes in the follicular ap-
paratus of the ovaries, comparable to those of the guinea pig,
occur. In the period immediately preceding or following ovula-
tion no marked degeneration of the follicles takes place. If any
follicles degenerate at all (in consequence of the circulatory
changes in the ovaries during this period?), such a degeneration
can only affect a few large follicles, while in the guinea pig a sudden
degeneration of all the follicles, with the exception of the smallest
ones, takes place during this period. The other changes sub-
sequent to this sudden disintegration of follicles in the guinea pig
are likewise absent in the rabbit and ferret.
The ovaries of the guinea pig also differ in other respects from
those of the rabbit and ferret:
1. In the guinea pig a so-called “interstitial gland”’ is absent,
while it is present in the ovaries of the rabbit and ferret.
2. In the guinea pig during heat a spontaneous ovulation
usually takes place. This ovulation in no way depends upon a
preceding copulation; while in the rabbit and, as far as we could
determine, also in the ferret, heat as such is not sufficient, but a
copulation needs to take place in order to insure ovulation.
It would be of interest to determine whether there exists a
general correspondence of these various factors in such a way that
animals which, like the guinea pig, do not have an interstitial
gland and ovulate ‘‘spontaneously,’’ show a very marked cycle
in the ovarian follicular apparatus, while animals that possess an
interstitial gland and do not ovulate ‘‘spontaneously”’ do not
possess such a cycle.
It shall be determined in subsequent investigations whether
cyclic or other changes occur in the interstitial gland of rabbit
and ferret. We may, however, state here that during the winter
164 SCIENTIFIC PROCEEDINGS (76).
period (December and first half of February) the ovaries of the
ferret are small and differ from the ovaries in the period of sexual
activity especially through the diminution in the number of good
follicles. While the ovaries as a whole are smaller, the interstitial
gland is well preserved during the winter months.
QI (1155)
The cyclic changes in the mammary gland of the guinea pig.
By Cora HESSELBERG and LEO LOEB.
[From the Department of Comparative Pathology, Washington
University, St. Louis, Mo.]
Our interest in the character of the cyclic changes of the
mammary gland and in their mechanism was twofold. (1) In
former investigations Loeb has shown that an early extirpation of
the ovaries reduces to a very marked extent the incidence of
cancer of the breast in mice. It was therefore of interest to in-
quire more closely into the relations between ovaries and mammary
gland, and (2) we wished to determine whether there exists a paral-
lelism between the cyclic changes in the mammary gland on the
one hand and in the ovaries and uterus on the other hand. One of
us had formerly shown that in the cyclic changes of ovaries and
uterus we could distinguish two phases: the first, comprising ovu-
lation and the heat changes in the uterus, depends upon the ab-
sence of the corpus luteum. These are prevented by a substance
secreted by the lutein cells. This phase is, however, dependent
upon another constituent of the ovaries. The second phase,
comprising the further cyclic changes in the uterus as well as the
production of decidua and deciduomata, requires a substance
secreted by the corpus luteum. Do corresponding phases exist
in the case of the mammary gland?
Relatively little is known concerning the cyclic changes in the
mammary gland. Bouin and Ancel, as well as Frank and Unger,
have shown that in the rabbit, even in the absence of pregnancy,
but in the presence of corpora lutea, proliferation takes place in
the mammary gland. Proliferation also occurs regularly during
pregnancy. Frank and Unger have furthermore demonstrated
CycLic CHANGES IN MAMMARY GLAND OF GUINEA PIG. 165
that the experiments of Starling and others concerning the source
of the growth substance which acts on the mammary gland are not
conclusive.
Our investigations concern the cyclic changes in the mammary
gland of the guinea pig. We studied the mammary glands as well
as uterus and ovaries in 262 animals, in almost all of which the
time of ovulation had been ascertained prior to the experiment.
In many of these animals the effect of ovaries and uterus on the
cyclic changes was analyzed by various experimental procedures.
Without going into a detailed discussion of our results, we
may state our principal conclusion as follows: The normal sexual
cycle of the guinea pig (the period between two ovulations), has
a duration of approximately 16-18 days. Wecan also, in the case
of the mammary gland, distinguish two phases in this cycle—one
comprising the time of heat and ovulation and two or three days
following ovulation; in the large majority of cases the mammary
gland proliferates mitotically during this phase. In the second
phase, comprising the remainder of the sexual cycle, proliferation
is as a rule absent. Only toward the end of this phase, from the
fifteenth to the twentieth day, we find again in some cases pro-
liferation. The first proliferating phase depends upon the ab-
sence of the corpus luteum. We can accelerate it by an early
extirpation of the corpora lutea, in a way similar to the acceleration
of ovulation and uterine heat changes by the same procedure.
The corpus luteum of the ordinary sexual period in the guinea pig
does not usually produce proliferation of the mammary gland.
Also during pregnancy, which lasts in the guinea pig about twice
as long as in the rabbit, proliferation of the mammary gland
occurs regularly only after the twenty-fourth day of pregnancy.
If through certain experimental procedures we prolong the
sexual cycle, we find usually a proliferating gland in cases in which
well-developed living deciduomata and good corpora lutea, or
in which strongly developed, not degenerated corpora lutea with-
out deciduomata, are present. In those cases in which during the
period of prolongation deciduomata and corpora lutea are degen-
erating, proliferation of the mammary gland as a rule is absent.
In case of castration and of the presence of hypotypical
ovaries, proliferation of the mammary gland is not found. Con-
166 SCIENTIFIC PROCEEDINGS (76).
sidering all the facts we may conclude that while proliferation
in the first phase depends upon the absence of the corpus luteum
and upon the activity of another constituent of the ovaries, the
proliferation which is found following the first period is in all
probability due to substances secreted by the corpus luteum.!
In the guinea pig, however, the effect of this substance becomes
apparent only at a much later period than in the rabbit. The
adaptive character of this phenomenon is clear if we remember
that in the rabbit the functioning of the mammary gland is re-
quired at a much earlier period than in the guinea pig. Repeated
intraperitoneal injections of corpus luteum of the cow does not
produce a proliferation of the mammary gland in the guinea pig.
92 (1156)
The chlorides of the plasma in uremia.
By FRANKLIN C. McLean. (By invitation.)
[From the Hospital of the Rockefeller Institute for Medical Research,
New York.]
Previous investigations relating to the chlorid content of the
blood or plasma in uremia have yielded conflicting results, some
figures much lower than the lowest normal limit having been
reported.2. It is known that the chlorid content of nephritic
plasma is usually somewhat higher than that of the average nor-
mal plasma, but the findings in uremia have apparently so far not
been explained.
We have been able, in several cases, to make frequent obser-
vations of the chlorid content of the plasma of nephritic individuals
during life, and up to the time of death in uremic coma. We have
found a diminution of chlorids in the plasma to be the usual ac-
companiment of uremia, and we have found this decrease of
chlorids in the plasma to accompany the increased H + ion con-
centration frequently observed in the blood of uremic patients
shortly before death.
1 The latter part of this conclusion depends in part at least upon the correctness
ot the observations of Bouin and Ancel and Frank and Unger.
2 Strauss, H., ‘‘ Die chronischen thieren entziindungen,”’ Berlin, 1902, 51.
a
THE CHLOR DES OF THE PLASMA IN UREMIA. 167
That increased acidity of the blood causes a diminution of the
plasma chlorids, and an increase in the chlorid content of the cells
had been shown experimentally both in vitro and in vivo by
Hamburger.!. That a similar change occurs with the increased
acidity of the blood in uremia is illustrated by the following case
of nephritis, terminating in uremia.
Case 1. 'P. W. M., male, age 44, chronic interstitial nephritis,
uremia. Patient admitted June 17, suffering with chronic in-
terstitial nephritis, hypertension and secondary cardiac failure
with edema. With rest in bed the heart condition rapidly im-
proved and the edema disappeared. Following this the patient
felt well and the condition remained stationary, until October 12,
when an impending uremia first became manifest by an increase in
the blood urea and a diminished urea excretion. From June 17
to October 6 there were made twenty blood analyses, with simul-
taneous urine analyses, and the results showed very slight varia-
tion. The average of the figures for this period is shown below.
The maximum concentration of NaCl in the plasma during this
period was 6.53 grams per liter and the minimum was 6.06. The
phthalein elimination, shortly after admission, was twice found
to be 8 per cent. in two hours?
It will be seen from the table that the diminished urea function,
beginning about two weeks before symptoms of uremia appeared,
was accompanied by a diminution in the reserve alkalinity of the
plasma, as shown by Van Slyke’s method. On October 29 an
actual increase in acidity, as shown by the P, of the blood, was
present and there had been a sudden fall in the chlorid concentra-
tion of the plasma. This concentration remained low thereafter
until death, and the change was far greater than could possibly be
accounted for by the diminished diet. The change in NaCl con-
centration in the plasma is exactly that produced by intravenous
injection of any acid, in quantities sufficient to change the Py, of
the blood, and is accounted for by the increased acidity of the
blood occurring during the disease.
1 Hamburger, H., ‘‘Osmotischer Druck und Ionenlehre,’’ Wiesbaden, 1902, J, 317.
2 The observations on urea and chlorid elimination were made by the methods
previously described by the author (Jour. Exper. Med., 1915, XXII, 212 and 366).
Plasma COz was determined by the method of Van Slyke (Proc. Soc. Exp. BIoL.
AND MEb., 1915, XII, 165). For the determination of hydrogen ion concentration
in the whole blood by the gas chain method, I am indebted to Mr. G. E. Cullen.
168 SCIENTIFIC PROCEEDINGS (76).
TABLE I.
re ; a
= Plasma Sodium ° 2 &
: q § Chlorid per Liter. | > Ea
= : Ss ips By | 83
2 4 ee los So) as
g S 85 Ms - : 5 O8 58 Remarks
A = |ga|seleal ge la” | ae
ro} a S01 o0 o a Fa
June 17
to
Oct. 6 1.176) 5.5 | 5.86] 6.27/+0.41 | 49.1
Octisi2 1.542| 2.9 | 5.86| 6.25| +0.39 Patient feels quite
well
Oct. 21 2.125| 1.5 | 5.92| 6.34) +0.42 Appetite slightly di-
minished
Oct. 26 3-005| 0.7 | 5.84) 6.28| +0.42 | 34.8 Complains of head-
ache and muscular
cramps
Oct. 27 3.285) 0.4 | 5.84| 5.89 +0.05 | 19.0 Nausea and vomiting
Oct. 28 3-430) 0.27| 5.78| 5.83) +0.05 | 20.9 Severe headache;
typical air hunger
Oct. 29 3-935) 0.17, 5.73) 5.02) —0.71 | 17.6| 6.86) Headache; stertorous
breathing; mental
condition good
Oct. 30] 9
In Experiments 19, 20, 25, and 31 the picric acid method was
employed, and in 27 and 30 that of Bertrand.
Control experiments in which the dextrose solution was made
faintly acid, or contained an excess of sodium chloride (16 per cent.)
did not reveal any such difference in the reducing power of the
two bloods. Neither did injections of acid or alkali alone cause
any difference. Many other details remain to be further investi-
gated. For the present, however, the results clearly demonstrate
that, when dextrose is injected in moderate amounts into the
ENDOTHELIAL OPSONINS. 171
blood of the portal system, a large proportion of it becomes re-
tained in the liver provided alkali is simultaneously injected in
sufficient amount to produce a distinct lowering of the H-ion
concentration of the portal blood. A similar retention can not
be demonstrated by the above method when the dextrose solution
is neutral or, acid, or when it is made markedly hypertonic with
sodium chloride.
94 (1158)
Endothelial opsonins.
By W. H. MANwWaARING and Harry C. CoE.
[From the Department of Bacteriology and Immunity, Leland Stan-
ford, Jr., University.|
If the blood-free liver of a normal rabbit is repeatedly perfused
with a sample of Ringer’s solution containing a known number of
pneumococci, no diminution in the pneumococcic count of the
perfusion fluid is observed, even after a dozen passages through
the liver.
If the liver of an actively immunized rabbit is similarly per-
fused, the pneumococcic count is rapidly decreased. After three
or four passages, the perfusion fluid usually becomes sterile.
Histological study of the perfused liver now shows numerous
pneumococci adherent to the capillary endothelium. Few if any
agglutinated masses are seen.
Normal rabbit serum added to the perfusion fluid in amounts
not exceeding Io per cent. causes no appreciable retention of the
pneumococci by normal livers. Immune serum similarly added
causes a quantitive retention of the pneumoccoci.
Immune serum will cause this retention when tested in less
than a hundredth of the concentration necessary to cause ag-
glutination.
The serum component causing the pneumococcic retention is
thermo-stable (60° C., 30 min.).
Unagglutinated pneumococci sensitized by exposure to im-
mune serum and then washed free from serum, are retained
quantitatively by normal livers.
The serum component responsible for the retention is therefore
172 SCIENTIFIC PROCEEDINGS (76).
evidently an opsonin or bacterio-tropin so altering the pneumo-
cocci as to cause their adhesion to the capillary walls.
This opsonin is relatively inactive for the extrahepatic capil-
laries. The hind-quarters, lungs, kidney and intestines of normal
rabbits can be repeatedly perfused with Ringer’s solution con-
taining as much as I per cent. immune serum, with only a slight
retention of the pneumococci by these organs, while 0.001 per
cent. immune serum will cause their quantitative retention by the
liver. (Spleen and bone-marrow not yet tested.)
Defibrinated normal rabbit blood used as the perfusion fluid
will cause a slight deposit of the pneumococci in all organs.
95 (1159)
Specific receptors of fixed tissues.
By W. H. MANWARING and YOSHIO KUSAMA.
[From the Department of Bacteriology and Immunity, Leland Stan-
ford, Jr., University.]
If Ringer’s solution containing I per cent. goat serum is re-
peatedly perfused through the blood-free liver of a normal, ana-
phylactic or immune rabbit, no diminution in the amount of goat
serum in the perfusion fluid is produced, that can be detected by
titration with a specific precipitating serum.
If defibrinated normal, anaphylactic or immune rabbit blood
is added to the perfusion fluid, diminutions in the amount of goat
serum are observed after repeated liver passage; but in all cases
these diminutions are identical with diminutions observed in
control samples of the fluid kept at incubator temperature and
not passed through the liver.
The perfusion experiments therefore furnish no evidence of the
existence of a specific receptor apparatus (Ehrlich) for goat pro-
teins, in normal, anaphylactic, or immune rabbit livers.
PROTEIN ABSORPTION BY BLOOD CORPUSCLES. 173
96 (1160)
Protein absorption by blood corpuscles.
By W. H. MANWARING and YOSHIO KUSAMA.
[From the Department of Bacteriology and Immunity, Leland Stan-
ford, Jr., University.]
If I per cent. goat serum is added to freshly drawn defibrinated
normal rabbit blood, the mixture incubated for one hour, and then
separated by centrifugation into serum and corpuscle fractions,
a titration of the serum fraction by specific precipitin methods
will usually show but 25 per cent. of the goat protein originally
added to the blood.
If the serum and corpuscle fractions so obtained are allowed
to undergo independent autolysis (10 hrs., 37° C.), a distinct
restoration of the goat protein is observed in each fraction. The
restoration of the protein in the corpuscle fraction, however, is
usually much more pronounced than that in the serum fraction,
and may amount to as much as 50 per cent. of the total protein
originally added to the blood.
If goat serum is slowly injected intravenously into normal
rabbits in amounts not exceeding I per cent. of the total blood
volume, and blood is withdrawn from 1 to 4 hours later, a distinct
restoration of the goat protein can be brought about by allowing
the centrifuged but unwashed corpuscles so obtained to undergo
autolysis.
Parenterally introduced proteins, therefore, are apparently
absorbed in large measure by the circulating blood corpuscles.
97 (1161)
Toxicity of foreign sera for the isolated mammalian heart.
By W. H. Manwarinc, ARTHUR R. MEINHARD and HELEN
L. DENHART.
[From the Department of Bacteriology and Immunity, Leland Stan-
ford, Jr., University.]
Seven per cent. to 10 per cent. goat serum in Locke’s solution
perfused under constant pressure and temperature through the
174 SCIENTIFIC PROCEEDINGS (76).
coronary arteries of an isolated normal rabbit heart, usually pro-
duces the following series of phenomena:
1. An initial tachycardia, lasting about three minutes, suc-
ceeded by
2. A period of apparently normal heart action, lasting about
five minutes, succeeded by
3. A secondary tachycardia, lasting about two minutes,
ushering in
4. A period of decreasing rate and strength of heart action,
increasing irregularities, etc., usually ending in inactivation of the
heart in about ten minutes.
If goat serum is separated into diffusible and non-diffusible
fractions by dialysis through a celloidin membrane, and the two
fractions are tested independently, the following results are usually
obtained:
1. The diffusible substances tested in 7 per cent. to Io per cent.
dilution usually produce an initial tachycardia indistinguishable
from the tachycardia from the whole serum. This is succeeded
by a period of regular rate and rhythm usually lasting for over
an hour.
2. The non-diffusible substances (serum colloids) similarly
tested usually give no initial tachycardia, the rate and rhythm
continuing unchanged for about ten minutes. There is then usual-
ly a slight secondary tachycardia, ushering in a period of decreasing
heart action, usually ending in inactivation in about fifteen min-
utes.
The secondary tachycardia is always accompanied by a pro-
gressively decreasing rate of perfusion through the coronary
arteries, and beginning myocardial edema. We are therefore
inclined to attribute the secondary tachycardia and subsequent
heart-death to a breaking down of the capillary defenses (increased
capillary permeability), allowing the foreign colloids to pass out
of the capillaries into the tissue spaces, thus coming into direct
contact with the essential myocardial cells.
ANAPHYLACTIC AND IMMUNE REACTIONS. 175
98 (1162)
Analysis of the anaphylactic and immune reactions by means of
the isolated mammalian heart.
By W. H. Manwarinc, ARTHUR R. MEINHARD and HELEN
L. DENHART.
[From the Department of Bacteriology and Immunity, Leland Stan-
ford, Jr., University.]
The heart of a rabbit sensitized to goat serum, tested in a
blood-free condition by perfusion with 7 per cent. to 10 per cent.
goat serum, is more resistant than a normal rabbit heart similarly
tested. The increased resistance is shown by the absence of the
initial tachycardia, the absence or delayed development of the
secondary tachycardia, and a prolongation of the life of the isolated
organ.
Hearts of rabbits sensitized or immunized by repeated in-
jections with goat serum, are more resistant than those sensitized
with a single injection.
Normal rabbit serum, corpuscles or defibrinated blood, added
to the perfusion fluid, decreases its toxicity. The antitoxic action
of defibrinated blood is apparently equal to the sum of the anti-
toxic actions of its serum and corpuscles.
Anaphylactic rabbit serum similarly added usually markedly
increases the toxicity of the perfusion fluid. Such an anaphylactic
serum mixture may completely inactivate anormal heart within
from two to four minutes. Hearts of anaphylactic and immune
rabbits are more resistant than normal hearts to such mixtures.
The active principle of the anaphylactic serum responsible for
this increased toxicity is thermo-labile, the toxin-increasing or
toxin-producing power being completely lost, if the serum is
heated to 60° C. for 30 minutes.
The active principle is not complement, since such inactivated
anaphylactic sera cannot be reactivated by the addition of un-
heated normal serum.
The active principle is presumably not precipitin, since the
specific precipitins of rabbit serum are not destroyed, or at least
not completely destroyed, by heating the serum to 60° C. for 30
minutes.
176 SCIENTIFIC PROCEEDINGS (76).
Such inactivated anaphylactic sera are strongly antitoxic.
The presence of a thermo-stable antitoxin in the unheated ana-
phylactic serum is apparently masked by the relatively strong
thermo-labile toxin-increasing or toxin-producing substance.
This thermo-stable antitoxin is present in larger amounts in
the sera of rabbits sensitized or immunized by multiple injections,
than in rabbits sensitized by a single injection.
Sera of partially immunized rabbits (3-5 injections) added to
the perfusion fluid, usually give a non-fatal shock with normal
hearts. The heart may come to a complete standstill by the end
of four minutes, may remain inactive! for from two to four min-
utes, and then recover completely within two or three minutes.
A heart that has passed through such a non-fatal shock will
usually continue to beat strongly and regularly for an hour or more.
Sera of highly immunized rabbits (8-12 injections) added to
the perfusion fluid, usually give no shock, and show only a marked
antitoxic action.
99 (1163)
Autolysis of anaphylactic and immune tissues.
By W. H. MANWARING and RUTH OPPENHEIMER.
[From the Department of Bacteriology and Immunity, Leland Stan-
ford, Jr., University.]
The post-mortem autolysis of normal, anaphylactic and
immune guinea pig livers was followed by determining the changes
in the relative amounts of coagulable and non-coagulable ni-
trogen (Kjeldahl method). The anaphylactic guinea pigs had
been sensitized by a single injection of egg-white or goat serum.
The sensitizing dose varied from 0.1 c.c. to 2 c.c. Analyses were
made from 11 to 17 days after the injection. The immunized
guinea pigs had been injected at 4-7 day intervals with from
3 to 7 doses of the same antigens. They were analyzed from
8 to 12 days after the final injection. A summary of the data so
obtained is shown in the following table:
1 The coronary perfusion is made under constant pressure, and is only partially
dependent upon heart action. The perfusion of the myocardium, therefore, con-
tinues during the inactive period,
AUTOLYSIS OF ANAPHYLACTIC AND IMMUNE TISSUES. 177
Percentage of Non-coagulable N.
__| Post Mor-
Total N per Gram. Tonto | tem Autoly-
dintes 6 Hrs. 24 Hrs. 3 Days. Ser
INOTMaleeeeieies 0.034 gr. 10.5 13.5 20 23 12.5%
Anaphylactic... 0.032 gr. T1355 16 21.5 26 12.5%
Immunesn eee 0.029 gr. I4.5 16.5 22 30.5 16 %
Selected cases ..| .022-.026 gr. 16-18 18-20 25-30 40-45 26 %
The table shows a slight decrease in the average total N per
gram of liver tissue in the anaphylactic animals, and a distinct
decrease in the immune animals, the decrease being particularly
marked in certain selected cases.
The table also shows a distinct increase in the average per-
centage of non-coagulable N in both anaphylactic and immune
animals, confirming data recently published by Pick and Hashi-
moto.!
Contrary to their findings, however, the anaphylactic livers
showed no increase in the amount of post-mortem autolysis.
A distinct increase in post-mortem autolysis, however, was
observed in the immune livers, the phenomenon being particularly
marked in certain selected cases. The selected animals were for
the most part guinea pigs in which a marked Arthus phenomenon
had been produced.
100 (1164)
Hepatic bacteriolysins. (Preliminary report.)
By W. H. Manwarinc and Harry C. Coe.
[From the Department of Bacteriology and Immunity, Leland Stan-
ford, Jr., University.]
If pneumococci are deposited by perfusion methods in the
liver of a normal rabbit, in the presence of normal rabbit blood,
and the infected organ is now incubated at 37° C., a slight multi-
plication of the deposited pneumococci takes place. After 5 or 6
hours, the tissues begin to be distinctly overgrown by the mic-
roorganisms.
1 Pick and Hashimoto, Arch. f. exper. Path. u. Pharm., 76, 1914, p. 89; Zeit. f.
Immunitilsf., 21, 1914, p. 237. Compare also Barger and Dale, Biochem. Jour., 8,
I9QI4, p. 670.
178 SCIENTIFIC PROCEEDINGS (76).
If pneumococci are similarly deposited in the liver of an actively
immunized rabbit, in the presence of immune rabbit blood, a
gradual decrease in the deposited pneumococci is observed. By
the end of 5 or 6 hours’ incubation, the tissues have usually become
relatively sterile. The few remaining microérganisms usually
multiply later to form distinct colonies. The microérganisms
in the larger hepatic blood vessels, not in contact with the specific
parenchyma cells, are not so destroyed.
This hepatic destruction of the pheumococci is not associated
with leucocytic accumulations, nor is it necessarily accompanied
by phagocytosis by the endothelial cells. There is apparently
an hepatic mechanism in the immune animals for the extra-
cellular destruction or digestion of the microérganisms. Pneu-
mococci taken up by the endothelial cells are apparently protected
to a certain extent from this destruction.
IOI (1165)
A method for the determination of small amounts of sugar in urine.
By V. C. MYERs.
[From the Laboratory of Pathological Chemistry, New York Post-
graduate Medical School and Hospital.)
All human urines probably contain small amounts of sugar,
as has quite recently been pointed out by both Cole! and Folin,”
who have described tests for the detection of this small amount of
sugar. It has been found possible to determine this reducing
substance by precipitating the creatinine and uric acid, and
probably other interfering substances with picric acid as suggested
by Folin for his qualitative test, and then employing a technique
similar to that introduced by Benedict and Lewis® for the esti-
mation of the sugar of the blood.4 It is presumed that the re-
1 Cole, S. W., Lancet, 1913, II, 861.
2 Folin, O., J. Biol. Chem., 1915, XXII, 327.
3 Lewis, R. C., and Benedict, S. R., J. Biol. Chem., 1915, XX, 61. See also
Myers, V. C., and Bailey, C. V., J. Biol. Chem., 1916, XXIV, 147.
4In a recent conversation with Professor S. R. Benedict, he informed me that
Mr. Oesterberg, of the Cornell Chemical Laboratory, had likewise utilized this.
method for urine.
SMALL AMOUNTS OF SUGAR IN URINE. 179
ducing substance in question is glucose, although this has been
found difficult of positive proof. This question is being further
investigated.
The method is carried out as follows: About 2 grams of dry
picric acid are added to 10 c.c. of urine in a test tube and the
tube vigorously shaken. The tube is now stoppered and placed
in an ice box at o° C. After the tube has stood for an hour, it is
again shaken and then allowed to stand over night in the ice box,
after which the mixture is filtered through a small filter paper into
a dry test tube. The filtrate now contains less than 0.1 mg. of
creatinine per c.c., a quantity too small to invalidate the estimation
of the sugar. If the urine has reacted negatively to Benedict’s
qualitative reagent, the filtrate is diluted 1-5 or I-10 with sat-
urated picric acid solution. If the qualitative test has shown a
small amount of sugar, a greater dilution is made. The following
rule may be followed: for 0.1 per cent. of sugar dilute 1-5, for
0.2 per cent. dilute 1-10, for 0.3 per cent. dilute 1-15, etc. To
develop the color, pipette 3 c.c. of the diluted fluid into a tall,
narrow tube graduated to 10, 15 and 20 c.c., add 1 c.c. of saturated
sodium carbonate solution and heat in a beaker of boiling water
for 15 minutes. The tube is now thoroughly cooled and diluted
with water to the mark most satisfactory for colorimetric compari-
son. Either pure glucose in saturated picric acid or a standardized
picramic acid solution may be used as standard.1
INFLUENCE OF THE INGESTION OF GLUCOSE ON THE SUGAR OF THE URINE
Time 2 Sugar of Blood, Sugar of Urine, Benedict’s Qualitative Reaction
A. M.-P. M. Per Cent. Per Cent. for Sugar in the Urine.
0.12 0.09
9-10 0.15 0.10 } Negative
0.17 0.17
IO-II 0.23
0.22 0.90 Strongly positive
II-12 0.16 0.41 Positive
0.15 0.32 Slight cloudiness
I2- I 0.14
0.14 0.27 Slight cloudiness
I- 2 0.13
0.13 0.25 Very slight cloudiness
2-3 0.09 0.17 Negative
1See Myers and Bailey, J, Biol. Chem., 1916, XXIV, 150.
275 grams of glucose by mouth at 9.15.
180 SCIENTIFIC PROCEEDINGS (76).
Normal urine appears to contain between 0.08 and 0.2 per cent.
sugar. Urines which give only a slight reaction with Benedict’s
qualitative reagent give higher figures with this method, generally
between 0.25 and 0.35 per cent. The data on the previous page
from a human adult, kindly loaned by Dr. Bailey,! nicely illustrate
several of the points in question. -
The above results scarcely appear to support the recent con-
clusions of Taylor and Hulton? regarding the assimilation limit
of glucose. If, however, only the twenty-four hour specimen of
urine had been examined as in their experiments, the result would,
no doubt, have been negative.
102 (1166)
Regeneration in the mesencephalon of Amblystoma.
By H. SAXTON Burr. (By invitation.)
[From the Department of Anatomy of the Yale University School
of Medicine, New Haven, Conn.]
In April of the present year the writer published a report of an
experimental study of regeneration in the forebrain of Amblys-
toma. ‘The results showed that the removal of the cerebral hemi-
sphere together with the end-organ normally connected with it
(the nasal placode), was not followed by a regeneration of nervous
tissue. On the other hand, when the cerebral hemisphere was
was removed, leaving the nasal placode in place as a functionally
active organ, complete regeneration of the hemisphere occurred.
It was concluded that the functional activity of the nasal placode
provided the requisite stimulus, at first through some hormone
reaction and later through the active ingrowth of the olfactory
fibers, for the regeneration of the hemisphere.
This spring the same type of experiment has been performed
with the ocular complex. Amblystoma larve were subjected to
two series of operations. In the first the right eye and the under-
lying mesencephalon was removed. In the second the right eye
was turned back with a flap of skin and the underlying brain re-
moved, the eye being then returned to its normal position.
1See Bailey, C. V., Proc. Soc. ExPER. BIOL. AND MED., 1916, XIII, 154.
2 Taylor, A. E., and Hulton, F., J. Biol. Chem., 1916, X XV, 173.
REGENERATION IN THE MESENCEPHALON. 181
The results are briefly these. The removal of the eye and the
brain results in the formation across the gap of the wound of a
curtain of tissue in all probability derived from the ependymal
lining of the neural tube. The ingrowing fibers of the optic nerve
from the left eye apparently stimulate the tissue thus formed to
regenerate to a considerable extent. At the same time forward
growing fibers from lower centers also afford some stimulus for
regeneration, as was shown in the case of the primitive pallium of
the telencephalon. The tissue thus regenerated is very similar in
its organization to that normally found, except that important
optic areas are lacking. An analysis of the fiber tracts involved
must be deferred until later.
The removal of the mesencephalon leaving the eye in its nor-
mal position results in an almost complete regeneration of the
optic lobes. In one larva only a very slight defect in the right
mesencephalon distinguishes it from a normal unoperated individ-
ual. The process is apparently a much faster one than it is in the
case of the olfactory system for the complete regeneration has
occurred at the end of some three weeks, while in the case of the
cerebral hemispheres complete regeneration did not occur until
the end of as many months. This is really not so strange as
would seem on the face of it, because, as the writer has shown
elsewhere, the optic sense becomes functionally active some time
before the olfactory. The early activity of the eye would then
result in an early stimulus to regeneration.
These results show, as in the former experiments, that func-
tional activity of the end-organ normally connected with the
brain affords the necessary stimulus to regeneration of the part
of the brain removed.
103, (1167)
Conduction, excitability and rhythm-forming power of the atrio-
ventricular connection in the turtle.
By HEnry LAURENS.
[From the Osborn Zoélogical Laboratory, Yale University.]
As in the heart of the turtle Clemmys lutaria and of the lizards
Lacerta viridis and agilis (Laurens!) the right and left parts of the
1 Laurens, Pfluger’s Archiv, 1913, 150, p. 139.
182 SCIENTIFIC PROCEEDINGS (76).
atrio-ventricular funnel of Malacoclemmys geographica are the
portions which are most efficient in conducting the contracting
impulse from the auricles to the ventricle. When the auricles
are partially separated from the ventricle by a series of cuts
leaving only a narrow connection, and in consequence of which
atrio-ventricular block (complete or incomplete) has been brought
about, it is these parts which are later most capable of conducting
the impulse from the auricles to the ventricle so that the contrac-
tions of the latter follow those of the auricle codrdinatedly, or so
that the incomplete block is decreased.
Stimulating the funnel of beating (in situ and excised) and still
hearts (first Stannius ligature) with single shocks (quick make
and break) and with interrupted currents of short duration
have shown (1) that the funnel is more easily excited than the
base of the ventricle, (2) that the right and left parts of the funnel
are more easily excited than other parts (dorsal and ventral) and
(3) that the excitability of the funnel increases as one approaches
the auricle.
The stimulation of the funnel just below the level of the A-V
boundary of beating hearts with interrupted currents, even when
these are strong and of long duration, can only occasionally pro-
duce a ‘‘fibrillation’”’ of the ventricle or a V-A rhythm (funnel
rhythm) which lasts over after the stimulation is discontinued.
This is possible, however, and curves have been obtained from
excised hearts showing a duration for several seconds of a funnel
rhythm following a ventricular ‘‘fibrillation.”’ In the still heart
the setting up of a funnel rhythm is more easily and frequently
accomplished, and several cases have been registered showing
a funnel rhythm lasting for several minutes.
104 (1168)
The influence of the vagi and of the sympathetic nerves on the
rhythm-forming power of the atrioventricular con-
nection in the turtle.
By Henry LAURENS and C. C. GAULT.
[From the Osborn Zoélogical Laboratory, Yale University.]
The investigations here reported were undertaken to deter-
mine the action of the vagus and sympathetic nerves upon the
ATRIO-VENTRICULAR CONNECTION IN THE TURTLE. 183
V-A rhythm produced by electrical stimulation of the atrio-
ventricular funnel. In Malacoclemmys geographica the two
nerves are not fused into a single trunk, but run separately in the
neck just median to the carotid artery. The turtles were decere-
brated, and the plastron removed, the circulation being kept
intact to a large degree. The vagus was stimulated just above
the thoracico-abdominal ganglion, and the sympathetic, between
the median cervical and the first thoracic ganglion.
Stimulation of the vagus nerves alone gave the usual results.
The effects of sympathetic stimulation were, however, not so
clearly marked. The general effect was a slight augmentation
of the auricular contractions. Acceleration of the heart beat
was less frequently obtained, the average being from 2 to 3 beats
per minute, although an acceleration of as many as 6 beats per
minute was registered.
Conjoint stimulation of the vagus and the atrio-ventricular
funnel just below the A-V boundary with relatively strong in-
terrupted currents produces a V-A rhythm which lasts over,
in different experiments for varying lengths of time, after the
stimulation has been discontinued. In these cases stimulation
of the vagus nerves with a current of sufficient strength to still
the normal heart causes only a decrease in the height of the auri-
cular contraction with no effect on the rate of beat. Stimulation
of the sympathetic with strong currents stops the funnel rhythm,
after which a normal atrio-ventricular beat begins.
105 (1169)
Changes in form and position of the retinal elements of normal
and transplanted eyes of Amblystoma larve
occasioned by light and darkness.
By Henry LAURENS and J. W. WILLIAMS.
[From the Osborn Zoélogical Laboratory, Yale University.]
In order to investigate the changes occasioned by light and
darkness in the retinal elements of a Urodele a series of experi-
ments on large (37 to 45 mm.) larval and on recently metamor-
phosed individuals of Amblystoma was carried out. It was found
184 SCIENTIFIC PROCEEDINGS (76).
that the pigment of these eyes undergoes a decided forward move-
ment when the animals are transferred from darkness to light.
In darkness most of the pigment is massed near the base of the
epithelial cells, and only comparatively few needles extend into
the protoplasmic processes between the visual cells. In light a
decidedly greater amount of pigment moves toward the external
limiting membrane so that the basal layer is thinner. Measure-
ments of the distance from the external limiting membrane to the
nearest pigment needle (or from the choroid edge of the epithelial
cells to the farthest pigment needle) are practically the same in
light and dark eyes, so that this kind of measurement gives no
indication of the extent of movement of the pigment.
The cones in the light eye are 4.2 uw shorter than those in the
dark eye, the total expanded length of the cones being 25 yw. The
rods seem to be longer in the light eyes than in the dark, but the
increase is too slight to permit of satisfactory measurement.
Optic cups were transplanted at the tail bud stage to various
parts of the body, where they developed to form more or less
perfect eyes. The region of the auditory vesicle seemed to offer
a particularly advantageous spot for the transplant. In the
transplanted eyes the movement of the pigment is fully as great
as in the normal eyes. The cones also contract in the light but
only to the extent of about 2.5 p.
Pigment migration and cone contraction therefore do take
place in a Urodele retina and can do so independently of the cen-
tral nervous system.
106 (1170)
The alleged exhaustion of the epinephrin store in the adrenal by
emotional disturbance.
By G. N. STEWART and J. M. RoGorr.
[From the H. K. Cushing Laboratory of Experimental Medicine of
Western Reserve Unwersity.]
1. It has been stated that a marked diminution in the store of
epinephrin in the adrenal gland is associated with various kinds of
emotional excitation. Thus Elliott! speaks of morphin-“‘ fright”
in cats causing exhaustion of a gland whose splanchnic nerve
1 Journal of Physiology, 1912, 44, p. 374.
EXHAUSTION OF THE EPINEPHRIN STORE. 185
supply is intact, as compared with the other adrenal whose splanch-
nic supply has been previously severed. We can confirm his
statement as to the difference in the content produced under the
influence of morphin but we do not think that fright has anything
to do with the result since it is also obtained in dogs where there
are no signs of fright.
2. The signs of morphin-“‘fright”’ can all be elicited by ad-
ministering morphin to a cat in which one adrenal has been re-
moved and the splanchnic supply of the other cut and in which
accordingly no demonstrable liberation of epinephrin through the
splanchnics takes place. A cat in this condition behaves identi-
cally in the same way as a cat whose adrenal splanchnic supply
has been cut on one side but left intact on the other. The pupils
are widely dilated and there is the same characteristic restlessness
and incessant movement. The content of epinephrin in the re-
maining adrenal of the first cat is found to be practically the same
as that of the adrenal removed before the administration of mor-
phin while the content of the adrenal with intact splanchnic
supply in the second cat is definitely diminished.
3. When a cat with the splanchnic supply of one adrenal cut is
frightened for many hours by a dog in which also the splanchnic
supply of the adrenal has been divided on one side both animals
undoubtedly experience emotions of great intensity. Neverthe-
less the content of epinephrin in the gland whose nerve supply is
intact is not sensibly diminished as compared with the other.
4. We can confirm the statement that §-tetrahydronaphthy-
lamine causes in cats extreme exhaustion of the epinephrin store
of an adrenal whose nerve supply is intact as compared with its
fellow whose nerve supply has been previously severed.! Elliott
associates this with the emotional ‘“‘alarm.’’ We have attempted
to test this interpretation by making observations on rabbits.?
1 Elliott, loc. cit.
2 Division of the nerves to one adrenal is complicated in the rabbit by the fact
that the right adrenal seems to derive a portion of the nerve supply concerned in
changes in the epinephrin store from the left splanchnic (Kahn, Archiv fiir die
gesammle Physiologie, 1911, CXL, 209; Nishi, Archiv fiir Exper. Path. u. Pharmakol.,
1909, LXI, 401). We therefore tried to eliminate the nervous connections of the
left adrenal by dividing all branches going to it from the celiac ganglion and in
addition cutting any strands from the lumbar sympathetic and the sympathetic
itself below the diaphragm.
186 SCIENTIFIC PROCEEDINGS (76).
We have not seen nearly as great a degree of exhaustion in this
animal as in the cat. This might be interpreted as in favor of
Elliott’s view, since signs of ‘‘emotional”’ disturbance are also less
marked in the rabbit, although great dilatation of the pupil, in-
creased respiration and other symptoms are present, which, ac-
cording to Mutch and Pembrey! ‘‘give the impression that the
drug produces a state of increased psychic activity accompanied
by muscular action appropriate to the emotions.’’ It seems to
us, however, more natural, considering our results with morphin
and ‘‘frightening’’ without drugs to interpret the greater effect on
the epinephrin content in the cat as due to some other action of
the drug than the hypothetical emotional disturbance.
We determined the epinephrin content by the colorimetric
method of Folin, Cannon and Denis, which we found to agree
sufficiently well with blood pressure observations on the pithed cat.
107 (1171)
The liberation of epinephrin from the adrenals.
By G. N. STEWART and J. M. RoGorr.
[From the H. K. Cushing Laboratory of Experimental Medicine,
Western Reserve University.]
The solution of the question of the liberation of epinephrin
into the adrenal veins and the estimation of the amount so liber-
ated in the absence of artificial stimulation of the splanchnics are
complicated by the fact that after withdrawal of blood pressor
substances are quickly developed in it, which give the same effect
as epinephrin on such objects as the vessels of a frog’s legs.?_ It is
therefore desirable to demonstrate the fact of its liberation and to
assay its amount without the necessity of withdrawing blood.
We have done this (in the cat) by means of the denervated eye
reactions (of Meltzer),? and by the effect on the blood pressure
curve.
1J. of Physiology, 1911, 43, p. 109.
2 Cf. Trendelenburg, Archiv f. Exper. Path. u. Pharmakol., 1915, 79, p. 154.
3 Experiments on the liberation of epinephrin by stimulation of the splanchnics,
in which the eye reactions were used, have been described by us elsewhere, Journal
of Pharmacology and Experimental Therapeutics, 1916, 8, p. 205.
LIBERATION OF EPINEPHRIN. 187
1. For the eye reactions all that is necessary is to clamp off
temporarily a pocket of the inferior vena cava so that only adrenal
vein blood enters it. A clamp is applied just above the iliac veins.
The renal veins are then clamped and the segment of cava emptied
of blood by gently stripping it upwards. Finally a clamp is put on
the cava above the adrenal veins. Only a few seconds are oc-
cupied in the adjustment of these clamps. Small branches of
the segment of cava have been previously tied. The pocket is
allowed to fill with blood from the adrenals. When the clamps
are removed, the eye reactions are elicited at practically the same
time interval as when the splanchnics are stimulated with the
vessels free.
2. After section of both splanchnics (above the diaphragm)
the reactions can no longer be obtained. Section of the splanch-
nics has therefore greatly diminished, if not abolished, the libera-
tion of epinephrin. This is not due to the low blood pressure
caused by division of the nerves. For if only the right splanchnic
is cut there is little, if any, fall of blood pressure. Nevertheless
when the cava pocket is closed off as described, and in addition a
clamp is put on the left adrenal vein, the right being free, no eye
reaction is elicited on allowing the pocket to empty itself. When
the experiment is repeated with the left adrenal vein free the
reaction is obtained, although of course less strongly than with
both splanchnics intact and both adrenal veins open, since only
half the amount of epinephrin is discharged.
3. To demonstrate the effect of epinephrin liberated into a
cava pocket upon the blood pressure of the same animal, a some-
what different procedure must be adopted, in order to avoid undue
disturbance of the blood pressure curve on forming and on re-
leasing the pocket. The lower end of the cava segment is tied
permanently after previous ligation of the abdominal aorta and
squeezing of blood from the legs. The renal arteries and veins
are also tied. When the eye reactions are available to compare
with the blood-pressure curve and manipulation of the intestines
is avoided during the application of the upper clamp to the cava
segment, it is not always necessary that the circulation through
the intestines and liver should be interfered with. Even when
the blood pressure curve is somewhat irregular the rise of pres-
188 SCIENTIFIC PROCEEDINGS (76).
sure caused by the liberated epinephrin, occurring at a definite
interval after release of the pocket, can be identified by the fact
that the eye reaction also commences at or about this moment.
However, to further strengthen the evidence we have made experi-
ments in which the celiac and superior mesenteric arteries are first
tied off, then the renal arteries, and then the abdominal aorta just
below the kidneys. As much blood as possible is got into the
anterior end of the animal, and then the inferior cava is tied above
the iliacs. The renal veins are then ligated, and the cava pocket
now represents only a blind pouch upon the circulation, the filling
of which from the adrenal veins, or the emptying of which after
removal of the upper clamp produces relatively little mechanical
effect upon the blood pressure. The lower end of the animal is
kept raised throughout the experiment. This facilitates emptying
of the pocket without manipulation.
4. In different experiments we have assayed, by the injection
of known quantities of adrenalin, the amount of epinephrin
liberated without artificial stimulation of the splanchnics, under
our experimental conditions (narcosis with urethane alone, and
with urethane supplemented with ether). For example, in one
experiment we found 0.0005 mg. and in another 0.0009 mg. per kg.
of animal, per minute. When the pocket is allowed to fill during
stimulation of the splanchnics, with intervals of rest, the effect
on release is distinctly greater than when it is allowed to fill for
the same time without artificial stimulation of the nerves.
5. We have endeavored to measure the amount of blood
collected in the pocket, without bringing it into contact with any
foreign substance, in the following way: One of the iliac veins is
tied near its distal end and the other near the cava. Both iliacs
are then divided distal to the ligatures. By means of the ligature
on the first iliac it is suspended vertically, while the greater part
of the cava segment lies undisturbed. The iliac vein thus serves
as the neck of a measuring flask, so to say, the body of which is
composed of the cava segment. It is not difficult to determine
the moment when the blood, entering the pocket practically
without resistance, the walls of the vein being scarcely at all
distended so long as the vertical portion of the pocket is empty,
just reaches the proximal end of the iliac. If undue exposure of
ATTENUATION OF LivING AGENTS OF CYANOLOPHIA. 189
the vein is prevented, a comparison of the flow from the adrenals
in successive observations is made possible by noting the intervals
of time necessary for the pocket to fill up to this point. The
quantity of blood required to fill the pocket can be determined
once for all in each animal. The vertical position of a portion of
the pocket helps to empty it without manipulation when the
clamp is removed.
6. The sensitiveness of the eye-reactions to epinephrin dis-
charged from the adrenals, for example in response to stimulation
of the splanchnics, can be increased notably by temporarily clamp-
ing off alternative arterial paths. This must be done at such an
interval of time after the beginning of stimulation as is not more
than sufficient to allow the epinephrin to reach the beginning
of the aorta. A larger proportion of the blood containing the
epinephrin is thus forced to take the path to the eye whose re-
actions are being studied. If, for instance, the left iris is the
denervated one, clamping at the proper moment of the thoracic
aorta and the innominate markedly increases the reaction. It can
be further increased by tying off all accessible branches of the left
carotid except those through which the eye must obtain its blood
supply.
108 (1172)
Attenuation of the living agents of cyanolophia.
By RHODA ERDMANN. (By invitation.)
[Osborn Zoélogical Laboratory, Vale University, New Haven, Conn.]
Paschen! (1911) says that leucocytes must play an important
part in the process of immunization. This remark seems partly
justified in the attenuation process of cyanolophia. The living
agents of cyanolophia are differently affected in tissue cultures
‘of red bone marrow from white bone marrow.
1 Paschen, O., ‘‘Handbuch der Technik und Methodik der Immunititsfor-
schung,” 1911.
190 SCIENTIFIC PROCEEDINGS (76).
EXPERIMENT I, I9Q15.
SERUM B TAKEN 36 Hours AFTER INOCULATION OF VIRULENT BRAIN A IN A
CHICKEN, SHORTLY BEFORE ITS DEATH, WAS INOCULATED IN A TISSUE
CULTURE OF RED BONE MARROW AND CHICKEN PLASMA.
Length of Time in which Virulent Serum B was | Record No. Length of
Cultivated in Tissue Culture at 30° C. of Chicken, | Date of Death.) Incubation.
NOViiT2=NOV. DS cicain crn ao oe Cee ena No. 1 Nov. 18 48 hours
NovertZeNovs iS iinc cevere acter No. 3 Nov. 17 38 hours
Novi T2=NOViiScci cere eerie No. 2 Nov. 21 72 hours
This proves that the virus can be kept alive six days at a
temperature of 38° C. in a tissue culture of ved bone marrow.
Chickens No. 1 and No. 3 died in 48 and 38 hours. Chicken
No. 2, which had been inoculated with serum B that had been
kept 6 days in the tissue culture, died twenty-four hours later
than the chicken which had been inoculated with serum B that
had been only three days in a tissue culture of red bone marrow.
This proves a certain attenuation by the cultivation of virulent
serum in red bone marrow.
The living agents, which probably cause cyanolophia, can be
cultivated in red bone marrow tissue cultures even longer than
six days without losing their virulence.
EXPERIMENT XII, 1916.
SERUM V TAKEN 36 HourS AFTER INOCULATION OF VIRULENT BRAIN U IN A
CHICKEN, SHORTLY BEFORE ITS DEATH, WAS INOCULATED IN A TISSUE
CULTURE OF RED BONE MARROW AND CHICKEN PLASMA.
Length of Time in Which Virulent
Serum V was Cultivated in Record No. of Date of Death. Length of
Tissue Culture at 38° C. Chicken, Incubation.
hebs23—Marchisie acacitecie cece No. 17 March 4 48 hours
Febs2a—MarchiO-re ee eee eee No. 18 Remained alive
Beb.23=Mareh Gili. cpersstspecctatece wee No. 19 Remained alive
So we can keep virulent the living agents of cyanolophia outside
of the chicken 6-8 days at 38° C., yet in the tissue culture of red
bone marrow the virus dies after 12 days. This is a perfect
analogue to the experiment of Marchoux,! who cultivated the virus
of cyanolophia in a culture medium which contained red blood
corpuscles. He even believed that the living agents of cyanolo-
phia had multiplied and produced a much stronger virus than that
1 Marchoux, ‘‘Cultures in vitro du virus de la peste aviaire,’’ Compt. rend.
Acad. Sc., T. 147, p. 357, 1908.
ATTENUATION OF LivING AGENTS OF CYANOLOPHIA. I9QI
he inoculated in his cultures. In tissue cultures of red bone
marrow no multiplication of the virus was observed, but a certain at-
tenuation, as proved by the prolongation of the incubation period.
Different results were obtained by using white bone marrow.
EXPERIMENT II, 1915.
SERUM C TAKEN 36 Hours AFTER INOCULATION OF VIRULENT BRAIN B IN A
CHICKEN, SHORTLY BEFORE ITS DEATH, WAS INOCULATED IN A TISSUE
CULTURE OF WHITE BONE MARROW AND CHICKEN PLASMA.
===
Length of Time in Which Virulent Serum B was| Record No. of Length of
Cultivated in Tissue Culture at 38° C. Chicken. Date of Death. | Jncubation.
IN OWS OD) CCL Aa cute. oransie Siorsncve che «10 6.6 Sievers No. 3a Living
INOW SOD CCHOR) craneretslereisters sorta. s, sleescels No. 4 Living
SERUM C, THE SAME AS USED IN EXPERIMENT BEFORE, IN RED BONE MARROW
AND PLASMA.
Length of Time in which Virulent Serum C Record No. Length of
was Cultivated in Tissue Culture at 30°C. of Chicken, | Date of Death.) Incubation.
INOVESORD ECHO sooo seein No. 5 Dec. 9 | 72 hours
This experiment proves that after 4 or 6 days in white bone
marrow the virus is attenuated. After inoculation of virus in
red bone marrow the animal died. A controlling experiment,
in which serum C was used, which was kept 7 days on ice, showed
that this serum killed the chicken after 38 hours, the usual time
in which this strain of cyanolophia killed the animal. It was
perfectly attenuated by the cultivation in white bone marrow,
partially by cu'tivation in red bone marrow, and not at all by keeping
the serum on ice.
The inoculation of attenuated serum and white bone marrow
protected, to a certain degree, the chicken against a new inocula-
tion, as the following experiments prove.
EXPERIMENT III, 1915-1916.
SERUM Y TAKEN 36 Hours AFTER INOCULATION OF VIRULENT BRAIN H IN A
CHICKEN, SHORTLY BEFORE ITS DEATH, WAS INOCULATED IN A TISSUE
CULTURE OF WHITE BONE MARROW AND PLASMA.
Length of Time in which Virulent Serum y Record No, Length of
was Cultivated in Tissue Culture at 38° C. of Chicken, Date of Death. Incubation,
DECHZ6—DeC R20 reeicten cerning articles No. 3a Living
WGC: /26—D EG 2915, telateie a. etausieie epicres No. 4 Living
WeCh26—DEGr 20s. erence cele nein a ys No. 6 Died Jan. 1 72 hours
192 SCIENTIFIC PROCEEDINGS (76).
Chickens Nos. 3a and 4 had been inoculated with serum B and
white bone marrow before, but not chicken No. 6. The same re-
sults were attained in experiments IV, V, VI, VIII, and X. All
chickens which were treated with attenuated serum did not die
when inoculated with a 2d or 3d dose of attenuated serum. All
chickens which were not treated succumbed to the first doses of
attenuated virus when it was kept only 2 or 3 days’ time in tissue
culture of white bone marrow. Always controlling experiments
with the same untreated serum kept on ice were started, which
killed the animals in due time.
It is possible to keep virulent the living agents of cyanolophia
in plasma alone at a temperature of 38° for six days and
longer, but the same virus dies in plasma, in which living
white bone marrow is kept, in six days (experiment IX).
The controlling experiment with serum, which was kept on ice,
was positive, so it is true that the virulence of living agents of
cyanolophia will be attenuated, and later die through the activity
of the leucocytes. It was possible to shorten the length of time
in which virulent serum was kept in tissue cultures of white bone
marrow, and still inoculate it without success when the treated
animals were used again. Animals 3a, 4, 7 and 8 survived after
inoculation with serum M, which had been only 2 days in tissue
culture (Experiment VI). Serum M, killed chicken My, after
it was kept 5 days on ice, in due time. A shortening of the at-
tenuation period to twenty-four hours was not sufficient to weaken
the serum Mz. Chicken 7, which again was used, died in forty-
eight hours, after having veen inoculated with serum Mz, that
had only been one day in plasma and white bone marrow.
Steinhardt and Lambert! cultivated the living agents of vac-
cinia in tissue cultures of rabbit cornea. They report a definite
increase of the virus, as measured by the effects of successful
reinoculations. Growth of the virus could be observed in tissue
cultures of the rabbit’s cornea only, while heart, kidney and
liver gave no results. My experiments, previously reported,
prove a rapid attenuation of the virus in white bone marrow tissue
1 Steinhardt, E., and Lambert, R. A., ‘‘Studies on the Cultivation of the Virus
of Vaccinia, II,’ Journ. of Inf. Diseases, 1914, Vol. 14, pp. 87-92.
DIGESTION IN BLATTID. 193
culture. This is quite remarkable, because the living agent of
cyanolophia is not surpassed in virulence by any other virus.
The next series of experiments will deal with the attenuation
of the living agents of cyanolophia in brain and liver tissue cul-
tures and with the importance of these and the white bone marrow
tissue cultures for active immunization.
109 (1173)
Experiments on the physiology of digestion in Blattide.
By ELpon W. SANFORD. (By invitation.)
[From Osborn Zoélogical Laboratory, Yale University.|
The question as to whether fat is digested and absorbed in the
crop of the cockroach was answered in the affirmative by Professor
Petrunkevitch in 1898, but in the negative by more recent authors.
My investigations, which were done under the direction of Pro-
fessor Petrunkevitch, show that fat is split to soluble products
and absorbed in large amount in the crop, the process being ob-
servable as gradually more and more in the crop’s epithelial cells
at successive intervals up to forty-eight hours, and gradually less
afterward. Some cells absorb so much that they appear solid
black when stained with osmic acid. Ligation of the crop from
the stomach does not hinder or modify the process. Fatty acids
are absorbed like fats.
At certain intervals after fat feeding much fat is found in the
tracheal tubes, sometimes filling them, sometimes in a thin layer
on their walls, sometimes only on the supporting spirals, and
sometimes mingled with chyme. This chyme resembles that
normally present in the crop lumen; it is regularly present in some
of the trachee, and in it leucocytes are often found. The chyme
is evidently a normal content. The fat enters the tubes through
the tracheal end cells, after being absorbed by them from the
lumen of the crop.
194 SCIENTIFIC PROCEEDINGS (76).
110 (1174)
On the transformation of the plasma clot.
By GEORGE A. BAITSELL. (By invitation.)
[From the Osborn Zoélogical Laboratory, Yale University, New
Haven, Connecticut.]
It has previously! been shown by the author that in tissue
cultures and in wound healing in the frog a fibrous tissue which is
apparently identical with normal connective tissue may be formed
by a direct transformation of a plasma clot. In an endeavor to
analyze this reaction, plasma clots made from centrifuged blood
plasma have been subjected to various conditions of tension and
pressure. The results obtained show that with the aid of these
mechanical factors it is possible to directly transform a typical
fibrin net into a fibrous tissue. Judged from its histological
structure when stained with Mallory’s connective tissue stain,
this new fibrous tissue is apparently identical with normal con-
nective tissue of the frog. By varying the conditions it is possible
to obtain preparations which will show various stages in the trans-
formation ranging from a typical fibrin net to a fibrous tissue made
up of bundles of wavy fibers such as is characteristic of normal
connective tissue.
III (1175)
The effect of moderately high atmospheric temperatures upon
the formation of agglutinins.
By C.-E. A. WINSLOW, JAMES ALEXANDER MILLER, and W. C.
NOBLE.
[From the New York State Commission on Ventilation.]
In an earlier communication? we have pointed out that previous
experiments on the effect of atmospheric temperature upon the
development of various immunity reactions suggest two general
conclusions: (1) That very high atmospheric temperatures,
over 35° C., tend to produce a condition of fever and to hasten
1(a) Jour. Exp. Med., Vol. 21, 1915, pp. 455-479; (b) Jour. Exp. Med., Vol. 23,
1916, pp. 439-456.
2 Proc. Soc. Exp. BIOL. AND MED., 1916, Vol. XIII, p. 93.
FORMATION OF AGGLUTININS. 195
the production of antibodies of various sorts, while (2) moderately
high atmospheric temperatures (30°-35° C.), apparently tend to
decrease the power of producing antibodies, presumably by a
lowering of general vital resistance without the stimulus which
accompanies the production of fever. We reported certain
experiments of our own which were in harmony with the last
conclusion, inasmuch as they showed an apparent diminution
in hemolysin production in rabbits kept at an atmospheric tem-
perature of 29°-32° C. The present report deals with similar
experiments upon the effect of moderately high temperature
upon the formation of agglutinins.
This particular immunity reaction has been studied in relation
to temperature by several observers. Rolly and Meltzer! kept
rabbits in an incubator at 34°-38° under which condition their
body temperature rose to 40°, they lost weight and showed a de-
crease in hemoglobin; yet when injected with typhoid bacilli they
showed a marked increase both in bactericidal and agglutinating
power. On the other hand Graziani? studied the agglutinating
power of the blood of rabbits kept at lower temperatures and
found, as workers on other immunity reactions have done, that
moderately high heat was harmful and not helpful. The blood of
rabbits kept at 2°-4° C. would agglutinate at a dilution of 1 in
1,541; at 18°, 1 in 854; at 32°, I in 727. In another series the
blood of rabbits kept at 32° agglutinated at a dilution of I in 1,250,
while if the animals were occasionally relieved by cold baths the
agglutinating power rose to I in 2,425.
Studies on the agglutinating power of the blood of human
beings after hot baths are conflicting. Leube* reports that typhoid
convalescents showed a material increase in the agglutinin content
of the blood after hot baths (40° for 30 minutes); while Moon*
could not find any such increase after Turkish baths (30 minutes
in a dry room at 82° C. and 20 minutes in a steam room at 54°).
In our own experiments, which were carried out in the bacterio-
logical laboratories of the University and Bellevue Hospital Medi-
cal College, five series of rabbits, including 14 animals in all, were
1 Deut. Arch. f. klin. Med., XCIV, 1908, p. 335.
2 Centr. f. Bakt. Orig., XLII, 1906, p. 633.
3 Verhandl, d. Deutschen Kongresses f. innere Med., XXVII, 1910, p. 218.
4 Jour. Infect. Dis., XIV, 1914, p. 56.
196 SCIENTIFIC PROCEEDINGS (76).
kept (2-4 at a time) in a large incubator (12’ x 2’ x 4’) at a tem-
perature ranging between 29° and 32° C. A similar series of 13
control animals was kept at room temperature (18°-21°). The
animals were immunized by giving them intraperitoneally suc-
cessively increasing doses of a suspension of killed typhoid bacilli.
The injections were given twice a week. Bleedings were taken
at weekly intervals and serum was drawn off from the clot and
diluted with sterile salt solution.
For microscopic agglutination tests, one loopful of diluted
serum and one loopful of a twenty-four hour broth culture of the
typhoid bacillus were mixed on a cover glass, and a hanging-drop
mount made. This was incubated at 37° Centigrade and readings
made after one hour. In every case the figures in the table
represent the actual dilution of serum found effective; the larger
the fraction, the weaker the agglutinating power.
The results of the experiments are given in full in Table I and
the averages by series in Table II. No general average can be
fairly calculated because the agglutinating power of all the rabbits
in Series V was so low that in a general average these Series V
figures swamp all the rest and the net result depends simply on
the number of Series V results included in a given week.
TABLE I.
EFFECTIVE DILUTION OF SERUM EXPRESSED IN DECIMALS.
|
Heated Animals, Control Animals.
mit} ae st =
2/3 Week. a Week,
a | 3 Gy |
wn | %
I 2 3 4 5 I 2 3 4 5
Te 3].00500].00066] ——— | ——— | —— |I01] .02000) .00025; ——— | ——— | ——
4|.02000|.00100] ——— | —— | —— |102] .00100; .00067, ——- | —— | ——
1, 5|.00200) died I5] .00050| .OOOI2) .00003| .00005| ——
9}.00050 00050 16| .00020) .00012| .ooor0| died | ——
6|.00200|.00100|.00010 .00010| died | 17] .00050) .00010! .oOOTO} .oooLo| ——
7}.00200|.00012|.00010|.00010|.00012| 18] .00050] .00012| .oooTO) .ooor0] ——
8|}.00500).00012|.00010|.00010}.00012
III ./172/.01000!.00025|.00020 .00020].00050|170| .01000 .00010) .00013! .o0017| .00025
173 -02000|.00025|.00029 .00033|.00050/171| .01000| .00017| .00020| .00025] .00045
IV .| 76|).00018).00017|.00017|.00010| ——— | 93] .00500) .00029| .00200} .00008)} .oooIO0
176 .00050|.00017|.00017 .0001I| —— |141| .00500| .o1000) .o5000| died
V ..| 92).10000].01000).01000}.02000 44| .I10000| .02000) .01000} .o1000
143 02000 O1000].02000) 54 01000} .01000| .02000] .02000
145, —— 02000).02000).02000, 72| ——— | .01000) .o1000| .00400
QUANTITATIVE DETERMINATION OF PLASMA PROTEIN. 197
TABLE II.
RESULTS AVERAGED BY SERIES. EFFECTIVE DILUTION OF SERUM EXPRESSED IN
DECIMALS.
Week.
Series.
I 2 3 4 5
ee AIOE eatedtsveycss ie itceste -01250 .00083
Control sn sere -01050 .00046
if) ent Nemes oe Fleatedsi< is. clepesis .00520 .00078 .00020 .00010 00012
Controle: .00380 .00023 .00008 .00008
TIThes Svese ts Freatednrars svereracts .01500 .00025 -00024 .00027 -00050
Controle erie -01000 .000I3 .OOOI7 .0002T -00035
EWireesslere Heated. 2 c.0 etc ert. .00034 .OOO17 .OO0O17 .000IO0
Controlaare cect -00500 -00514 .02600 .00008 .0O00IO
Widijciavesete Eleatiediays sie cierers « .10000 .01500 -01500 .01660 .02000
Controls ecic< .10000 .01400 -01000 .OII00 .02000
In general our results confirm those of Graziani and suggest
that a moderately high atmospheric temperature (29°-32° C.)
tends slightly to decrease the power of agglutinin formation in the
rabbit. In Series IV alone this was not indicated. Here both
control rabbits gave abnormal results. No. 93 showed a marked
drop in agglutinating power during the third week; while No. 141
never formed any powerful agglutinins and died after the third
week. With the exception of this series there are sixteen weekly
averages of heated and control rabbits compared in Table II. In
these sixteen cases the effective dilution for heated and control
animals was on two occasions the same while in the other fourteen
instances a consistently larger amount of serum was needed to
produce agglutination in the case of the heated animals.
112 (1176)
Improved methods for the quantitative determination of plasma
proteins.
By GLENN E. CULLEN and Donatp D. Van SLYKE.
[From the Hospital of the Rockefeller Institute for Medical Research.]
The blood is drawn into a tube containing an amount of po-
tassium oxalate sufficient to make 0.2 or 0.3 per cent. oxalate
solution, and is centrifuged twenty minutes.
198 SCIENTIFIC PROCEEDINGS (76).
Fibrin.—5 c.c. of plasma are run into a beaker containing
100-150 c.c. 0.8 per cent. NaCl and 2-5 c.c. of a 2.5 per cent.
CaCk solution. The CaCl, may be in amounts from 2-25
equivalents of the oxalate, but about five equivalents are best.
When coagulation is complete, the fibrin is filtered, the clot washed
with 0.8 per cent. NaCl, and the nitrogen determined by Kjeldahl.
The above is an adaptation of Howell’s method for determining
the activity of thrombin.1. The filtrate from the clot may be
tested for complete precipitation by addition of a solution con-
taining thromboplastic substances.
Albumin and Globulin are calculated from the following three
determinations:
Total nitrogen is determined on a 1-2 c.c. sample.
Non-protein nitrogen is determined in the filtrate obtained
after precipitating the plasma with nine volumes of trichloracetic
acid.
Nitrogen of Globulin Filtrate:—Globulin and fibrin are pre-
cipitated by adding to 5 c.c. of plasma 20 c.c. of H:O and 25 c.c.
of saturated ammonium sulfate solution. 20 c.c. of the filtrate
are mixed in a Kjeldahl flask with 3 gm. MgO “‘ Merck’s Reagent”’
and 350 c.c. of 50 per cent. alcohol. The solution is distilled until
the distillate gives a negative test to red litmus. This takes about
one hour and reduces the volume to about 20 c.c. The nitrogen,
representing albumin plus non-protein nitrogen, is then deter-
mined by Kjeldahl, using 25 c.c. HxSO,. When the digestion
mass becomes light brown, the sides of the flask are washed down
with a few c.c. of water and ten more c.c. H:SO, added.
Calculation:
Filtrate N—Non-protein N = Albumin N.
Total N—(Filtrate N + Filbrin N) = Globulin N.
1 Am. Jour. Physiol., 1910, XXVI, 453.
RESPONSE OF SINGLE CELLS TO STIMULATION. 199
113 (1177)
The response of single cells to electrical stimulation.
By R. A. SPAETH. (By invitation.)
[From the Osborn Zoélogical Laboratory, Yale University, New
Haven.|
There is an accumulation of embryological, morphological and
physiological evidence at hand showing that the melanophores of
vertebrates are to be considered highly modified smooth muscles
cells. By means of a simple recording device the responses of
single melanophores of Fundulus heteroclitus to faradic and gal-
vanic stimulation have been studied in some detail.
In faradic stimulation it appears that as regards the duration
of the latent period, the quantity of current necessary to bring
about a response in the cell, the increased height of the contraction
curve with an increase in the strength of stimulus and the develop-
ment of tetanus by properly spaced single break shocks, the con-
traction curves for a single melanophore show a striking resem-
blance to smooth muscle graphs obtained from the bladder of the
cat (Stewart) and the stomach of the frog (Howell).
A constant current, which has previously been supposed to
produce an expansion of the melanophores, causes a contraction
when applied through non-polarizable electrodes of the Zn-ZnSO,
type. An expansion of the melanophores may be produced by
galvanic stimulation if platinum electrodes are used but this has
been shown to be due to hydroxyl ions liberated at the cathode.
Both the make and the flow of the constant current are effective
contracting stimuli. With currents of moderate strength there
is, at first, a rapid rise in the contraction curve due to the combined
effects of make and flow but subsequently a partial falling off of
the contraction giving a typical plateau. Stewart has found
precisely the same conditions in the bladder of the cat. No
response to the breaking of the constant current has thus far been
observed in the melanophore.
The evidence obtained from these experiments with the re-
sponses of single melanophores to electrical stimulation, appears
to strengthen and corroborate the writer’s contention that in the
200 SCIENTIFIC PROCEEDINGS (76).
melanophore we are dealing with a modified and disguised type
of smooth muscle cell.
114 (1178)
Characteristics of the precipitation reaction.
By RICHARD WEIL.
[From the Department of Experimental Medicine, Cornell Medical
College, New York City.]
In a previous communication I showed that when a chemically
pure protein, such as crystallized egg albumin, is used as antigen,
it combines with the precipitin of immune serum to the complete
exhaustion of either factor from the mixture. From these ob-
servations the conclusion was drawn that an equilibrium subject
to the laws of mass action, such as had been previously described
in precipitation reactions, does not exist in these reactions, those
results being attributable to the use of impure antigens, such as
complex native sera. Further study has shown that chemically
pure antigen unites with the precipitin in proportions that are
definite and constant. The same amount of precipitinogen
always ‘‘binds’’ an equivalent amount of precipitin, regardless of
the relative excess of the latter substance in the mixture. The
reverse of this statement likewise holds true. Hence it follows
that it has not been possible to demonstrate the Danyz-Dungern
phenomenon in the precipitation reaction when carried on with
pure reagents. It appears likely, therefore, that the reaction
conforms to the type of quantitative chemical reactions, and is
not comparable to the adsorption phenomena exhibited by mutu-
ally precipitating colloids.
RECAPITULATION OF THE NAMES OF THE
AUDHORS AND) OF Shy JITLES OF
THE COMMUNICATIONS.
VOLUME XIII.
Auer, J.
1095. The action of the depressor nerve on the pupil.
1124. [with S. J. Meltzer.] The influence of intra-
venous injections of magnesium sulphate upon the activities
of deglutition.
Austin, J. H.
1066. [with S.S. Leopold.] Effects of glucose and of
meat on the blood nitrogen and the duration of life in ex-
perimental renal insufficiency.
Bailey, C. H.
1099. Observations on cholesterol-fed guinea pigs.
1100. Lesions produced in rabbits by repeated in-
travenous injections of living colon bacilli.
Bailey, C.V.
1149. Studies in alimentary hyperglycemia and
glycosuria.
Baitsell, George A.
1174. On the transformation of the plasma clot.
Barbour, H. G.
1152. [with N. H. Copenhaver.] Is uterine activity
subject to cerebral control?
Barringer, Theodore B.
1093. The circulatory reaction to graduated work as
a test of the heart’s functional capacity.
Bateman, W. G.
1150. The digestibility and utilization of egg proteins.
Bedford, E. A.
1114. [with H. C. Jackson.] The epinephric content
of the blood in conditions of low blood pressure and ‘‘shock.’’
Bergeim, Olaf.
1097. See Fowler, C. C.
201
202 SCIENTIFIC PROCEEDINGS (76).
Bronfenbrenner, J.
1078. On the mechanism of anaphylaxis and antiana-
phylaxis.
tog2. Antitryptic index in its relation to the clinical
manifestations of anaphylaxis.
1096. Some suggestions as to the etiology and treat-
ment of eclampsia.
Bull, C. G.
1084. Agglutination of bacteria in vivo; its relation
to the destruction of bacteria within the infected host and
to septicemia.
Burnett, Theodore C.
1142. [with George H. Martin, Jr.] Note on “Salt
fever.”’
Burr, H. Saxton.
1166. Regeneration in the mesencephalon of Am-
blystoma.
Clowes, G. H. A.
1r28. On the production of soap jellies, and the
physical conditions under which jelly formation takes place.
(Preliminary communication.)
Coe, Harry C.
1158. See Manwaring, W. H.
1164. See Manwaring ,W. H.
Copenhaver, N. H.
1152. See Barbour, H. G.
Csonka, F. A.
1110. See Janney, N. W.
Cullen, Glenn E.
1176. [with Donald D. Van Slyke.] Improved meth-
ods for the quantitative determination of plasma proteins.
Cutter, Irving S.
1117. [with Max Morse.] Nitrogen retention in
nephritis in children.
Davis, Helen.
1085. [with Nellis B. Foster.] Accumulation of
nitrogen in the tissues in renal disease.
NAMES OF AUTHORS. 203
Dean, A. L.
1068. The movements of the mitral valve flaps,
studied by a new method.
Denhart, Helen L.
1161. See Manwaring, W. H.
1162. See Manwaring, W. H.
Dickson, Ernest C.
1081. Sarcoma occurring in a guinea pig.
Dresbach, M.
1089. See Williams, J. R.
DuBois, Delafield.
110g. [with E. F. DuBois.] A height-weight for-
mula to estimate the surface area of man.
DuBois, E. F.
1109. See DuBois, Delafield.
Eggleston, Cary.
1135. Antagonism between atropin and certain cen-
tral emetics.
Epstein, A. A.
1104. On the relation of blood sugar to glycosuria
in diabetes mellitus.
1148. Permeability vs. tolerance of the kidneys for
sugar in diabetes mellitus.
Erdmann, Rhoda.
1172. Attenuation of the living agents of cyanolophia.
Evans, Frank A.
1119. The cytology of the exudate in the early stages
of experimental pneumonia.
Evans, H. M.
1112. On the behavior of the mammalian ovary and
especially of the atretic follicle towards vital stains of the
acid azo group.
Ewing, E. M.
1105. The relation of the sugar content and concen-
tration of the blood to urine formation. Preliminary report.
Fine, M. S.
1067. See Myers, V. C.
204 SCIENTIFIC PROCEEDINGS (76).
1134. [with V. C. Myers.] Comparative distribution
of urea, creatinine, creatine, uric acid, and sugar in blood
and spinal fluid.
Fitzpatrick, C. B.
1086. The utilization of ‘‘reactor’”’ milk in tuberculo-
medicine.
Foster, Nellis B.
1085. See Davis, Helen.
1088. The isolation of a toxic substance from the
blood of uremic patients.
Fowler, C. C.
1097. [with Olaf Bergeim and P. B. Hawk.] The
availability of certain indicators in the determination of
gastric acidity.
Friedman, E. D.
1108. [with H. C. Jackson.] The carbon dioxide
content of blood and alveolar air in obstructed expiration.
Gault, C. C.
1168. See Laurens, Henry.
Geyelin, H. Rawle.
1125. Diabetes of maximum severity with marked
improvement.
Githens, T. S.
1115. See Meltzer, S. J.
Halsted, W. S.
1065. As to the cause of the dilatation of the subcla-
vian artery observed in certain cases of cervical rib.
Hawk, P. B.
1073. See Smith, C. A.
1097. See Fowler, C. C.
Hess, Alfred F.
1094. The influence of infantile scurvy on growth
(length and weight.)
1098. An interrelationship between calcium and
antithrombin in blood coagulation.
1133. A separation of serum into coagulative and
non-coagulative fractions.
1144. The therapeutic effect of wheat germ and of
yeast in infantile scurvy.
NAMES OF AUTHORS. 205
Hesselberg, Cora.
1155. [with Leo Loeb.] The cyclic changes in the
mammary gland of the guinea pig.
Hooker, Davenport.
1147. The early responses of frog embryos to tactile
stimulation.
Hooker, Sanford B.
1141. Preliminary studies on antigenic properties
of different strains of bacillus typhosus.
Hooper, C. W.
1079. [with G. H. Whipple.] Icterus. A rapid
change of hemoglobin to bile pigment in the pleural and
peritoneal cavities.
Hoskins, R. C.
1076. A note on the failure of pituitrin to sensitize
the sympathetic system.
Huber, H. L.
1074. The ammonia of the gastric juice.
Hurwitz, S. H.
1080. [with K. F. Meyer and Z. Ostenberg.] Ona
colorimetric method of adjusting bacteriological culture
media to any optimum hydrogen ion concentration.
Jackson, H. C.
1108. See Friedman, E. D.
1114. See Bedford, E. A.
Janney, N. W.
1110. [with F. A. Csonka.] Diabetic dietetics. Glu-
cose formation from protein food.
1113. Concerning the protein content of meat.
Jones, F. S.
1107. See Rous, Peyton.
Kast, Ludwig.
1111. Effect of fatigue upon gastro-intestinal mo-
tility.
Kinsella, Ralph.
1122. See Swift, Homer F.
Kleiner, I. S.
1103. [with S. J. Meltzer.] On the production of
hyperglycemia and glycosuria by magnesium salts.
206 SCIENTIFIC PROCEEDINGS (76).
1143. [with S. J. Meltzer.] The influence of mor-
phin upon the elimination of intravenously injected dextrose.
Kline, B. S.
1082. [with S. J. Meltzer.] Production of pneu-
monic lesions produced by intrabronchial insufflation of un-
organized substances.
Kolmer, J. A.
1116. An allergic skin reaction to diphtheria bacilli.
Kusama, Yoshio.
1159. See Manwaring, W. H.
1160. See Manwaring, W. H.
Lambert, R. A.
1120. Technique of cultivating human tissues in
vitro.
Laurens, Henry.
1167. Conduction, excitability and rhythm-forming
power of the atrioventricular connection in the turtle.
1168. [with C. C. Gault.] The influence of the vagi
and of the sympathetic nerves on the rhythm-forming power
of the atrioventricular connection.
1169. [with J. W. Williams.] Changes in form and
position of the retinal elements of normal and transplanted
eyes of Amblystoma larve occasioned by light and darkness.
Leopold, S. S.
1066. See Austin, J. H.
Loeb, Leo.
1154. Further investigations on the cyclic changes
in the mammalian ovary.
1155. See Hesselberg, Cora.
Loewe, Leon.
1106. A method for the estimation of levulose in
presence of glucose.
Longcope, W. T.
1121. [with F. M. Rackemann.] Development of
immune reactions in serum disease.
Lough, W. G.
1067. See Myers, V. C.
NAMES OF AUTHORS. 207
MacKenzie, George M.
1137. The effect of exercise on the blood sugar of
depancreatized dogs.
Macleod, J. J. R.
1157. The stimulating influence of alkali on hepatic
glycogenesis.
MacNider, W. deB.
1071. On the occurrence and distribution of potas-
sium in normal and nephropathic kidney cells.
Manwaring, W. H.
1158. [with Harry C. Coe.] Endothelial opsonins.
1159. [with Yoshio Kusama.] Specific receptors of
fixed tissues.
1160. [with Yoshio Kusama.] Protein absorption
by blood corpuscles.
1161. [with Arthur R. Meinhard and Helen L. Den-
hart.] Toxicity of foreign sera for the isolated mammalian
heart.
1162. [with Arthur R. Meinhard and Helen L. Den-
hart.] Analysis of the anaphylactic and immune reactions
by means of the isolated mammalian heart.
1163. [with Ruth Oppenheimer.] Autolysis of ana-
phylactic and immune tissues.
1164. [with Harry C. Coe.] Hepatic bacteriolysins.
Martin, Jr., George H.
1142. See Burnett, Theodore C.
McCollum, E. V.
1136. [with Nina Simmonds and Walter Pitz.] The
distribution of the fat soluble A, the growth promoting
substance of butter fat, in the naturally occurring foodstuffs.
McLean, Franklin C.
1156. The chlorides of the plasma in uremia.
Meinhard, Arthur R.
1161. See Manwaring, W. H.
1162. See Manwaring, W. H.
Meltzer, S. J.
1082. See Kline, B. S.
1102. See Meyer, A. L.
208 SCIENTIFIC PROCEEDINGS (76).
1103. See Kleiner, I. S.
1115. [with T. S. Githens.] On the augmenting
action of ergotoxine (Dale and Barger) on the gastrointes-
tinal movements.
1124. See Auer, J.
1131. See Meyer, A. L.
1143. See Kleiner, I. S.
Mendel, Lafayette B.
1146. See Osborne, Thomas B.
Meyer, A. L.
1102. [with S. J. Meltzer.] On continuous insuffla-
tion in fowls. A demonstration.
1131. [with S. J. Meltzer.] An active expiratory
muscle in the chicken which is inhibited by stimulation of
the central end of the vagus.
Meyer, K. F.
1080. See Hurwitz, S. H.
Miller, H. R.
1140. [with Hans Zinsser.] Complement fixation
in tuberculosis.
Miller, James Alexander.
1073. See Smith, C. A.
1118. See Winslow, C.-E. A.
1175. See Winslow, C.-E. A.
Mitchell, C. W.
1075. See Salant, William.
Morgan, T. H.
1083. Demonstration of the appearance after cas-
tration of cock-feathering in a hen-feathered cockerel.
Morse, Max.
1117. See Cutter, Irving S.
Mosenthal, H.
1070. The interpretation of a positive nitrogen bal-
ance in nephritis.
Murlin, J. R.
1069. [with J. A. Riche.] The fat of the blood in
relation to muscular activity and heat production.
NAMES OF AUTHORS. 209
Myers, V. C.
1067. [with M. S. Fine and W. G. Lough.] The
significance of the uric acid, urea and creatinine of the blood
in early and late nephritis.
1134. See Fine, M. S.
1165. A method of the determination of small amounts
of sugar in urine.
Noble, W. C.
1118. See Winslow, C.-E. A.
1175. See Winslow, C.-E. A.
Oppenheimer, Ruth.
1163. See Manwaring, W. H.
Osborne, Thomas B.
1146. The nutritive value of some cotton-seed prod-
ucts in growth.
Ostenberg, Z.
1080. See Hurwitz, S. H.
Ott, Isaac.
1072. [with John C. Scott.] The action of animal
extracts upon the flow of bile.
Ottenberg, Reuben.
1123. The effect of sodium citrate on blood coagu-
lation in hemophilia.
Pike, F. H.
1132. See Wilson, J. Gordon.
Pitz, Walter.
1136. See McCollum, E. V.
Prince, A. L.
1151. The position of the head after experimental
removal of the otic labyrinth.
Rackemann, F. M.
1121. See Longcope, W. T.
Riche, J. A.
1069. See Murlin, J. R.
Rogers, F. T.
1129. The hunger mechanism in birds. Preliminary
report.
210 SCIENTIFIC PROCEEDINGS (76).
Rogoff, J. M.
1170. See Stewart, G. N.
1171. See Stewart, G. N.
Rous, Peyton,
1107. [with F. S. Jones.] A method for obtaining
suspensions of living cells from the fixed tissues, and for the
plating out of individual cells.
Salant, William.
1075. [with C. W. Mitchell.] The action of heavy
metals on the isolated intestine.
Sanford, Elden W.
1173. Experiments on the physiology of digestion in
Blattide.
Scott, G. G.
1130. Oxygen consumption in regenerating tissue.
1145. Oxygen utilization by fishes and other aquatic
animals.
Scott, John C.
1072. See Ott, Isaac.
Simmonds, Nina.
1136. See McCollum, E. V.
Smith, C. A.
1073. [with Raymond J. Miller and P. B. Hawk.]
Studies on the relative digestibility and utilization by the
human body of lard and hydrogenated vegetable oil.
Spaeth, R. A.
1177. The response of single cells to electrical stimu-
lation.
Stewart, G. N.
1170. [with J. M. Rogoff.] The alleged exhaustion
of the epinephrin store in the adrenal by emotional disturb-
ance.
1171. [with J. M. Rogoff.] The liberation of epineph-
rin from the adrenals.
Swift, Homer F.
1122. [with Ralph A. Kinsella.) Immunization with
sensitized bacteria.
NAMES OF AUTHORS. 211
Taylor, Kenneth.
1138. Studies on the blood of the albino rat. Its
normal cellular constituents. Their reaction to sarcoma
growth and to benzol treatment.
Underhill, Frank P.
1126. The control of acidosis and its relation to
impaired sugar metabolism in human diabetes.
1127. Possible interrelations between acidosis and
creatine elimination.
Van Slyke, Donald D.
1101. The determination of amino nitrogen in urines
containing glucose and albumin.
1139. Gravimetric determination of betaoxybutyric
acid.
1176. See Cullen, Glenn E.
Weil, Richard.
1087. Anaphylatoxin and the mechanism of ana-
phylaxis.
tog0. Equilibrium in the precipitation reaction.
togi. Equilibrium in the dissociation of precipitates.
1178. Characteristics of the precipitation reaction.
Whipple, G. H. ;
1079. See Hooper, C. W.
Williams, J. R.
1089. [with M. Dresbach.] The possible association
of diabetes mellitus and splenohepatomegaly, Goucher;
report of a case.
Williams, J. W.
1169. See Laurens, Henry.
Wilson, Frank N.
1077. The production of atrioventricular rhythm
in man after the administration of atropin.
Wilson, J. Gordon.
1132. [with F. H. Pike.] A demonstration of the
effects of some lesions of the central nervous system.
Winslow, C.-E. A.
1118. [with James Alexander Miller and W. C.
Noble.] The effect of moderately high atmospheric tem-
peratures upon the formation of hemolysins.
212 SCIENTIFIC PROCEEDINGS (76).
1175. [with James Alexander Miller and W. C.
Noble.] The effect of moderately high atmospheric tem-
perature upon the formation of agglutinins.
Woodruff, Lorande Loss.
1153. Endomixis in diverse races of paramecium
aurelia.
Zinsser, Hans.
1140. See Miller, H. R.
EXECUTIVE PROCEEDINGS.
MAIN SOCIETY.
Sixty-ninth Meeting.
Cornell University Medical College, October 20, 1915. Presi-
dent Lusk in the chair.
Members present: Auer, Austin, Barber, Benedict, Draper,
J. W. DuBois, Gettler, Gies, Githens, Greenwald, Halsted, Harris,
Howe, Kleiner, Kober, Lee, Lusk, Mandel, J. A., Meltzer, Mosen-
thal, Murlin, Myers, Pepper, Pike, Riddle, Ringer, Senior,
Swift, Wallace, Wiggers.
Members elected: B. S. Kline, H. Plotz.
Seventieth Meeting.
New York Post Graduate School, November 17, 1915. Presi-
dent Lusk in the chatr.
Members present: Auer, Austin, Bull, Coca, Eggleston,
Ewing, James, Fine, Fitzgerald, Foster, Gies, Githens, Goldfarb,
Harris, Hatcher, Howe, Jackson, Kline, Kober, Lambert, Lee,
Lusk, Meltzer, Morgan, Murlin, Myers, V. C., Pepper, Pike,
Ringer, Senior, Wasteneys, Weil, Wollstein.
Members elected: W. E. Dandy, R. G. Hoskins, E. B. Krumb-
haar, Shiro Tashiro.
Seventy-first Meeting.
Rockefeller Institute for Medical Research, December 15, I915.
President Lusk in the chair.
Members present: Auer, Benedict, Cole, R. I., Draper, J. W.,
Eggleston, Gies, Githens, Hartwell, Hatcher, Hess, Howe, Jackson,
Kleiner, Lambert, Lee, Loeb, Lusk, Meltzer, Pappenheimer,
Wallace, Weil, Wollstein.
Member elected: R. M. Taylor.
The Secretary read the following report from the Council:
““At a special meeting of the Council of the Society for Ex-
perimental Biology and Medicine, held Saturday, November 20,
213
214 SCIENTIFIC PROCEEDINGS (76).
1915, the following motion was offered, seconded and unanimously
passed, seven members of the Council having been present and
voting:
“‘On the basis of Article III, Section 2, of the Constitution,
dealing with forfeiture of membership, and which specifies that
‘Any member of this Society who may consent to the use of his
name in any way that would aid in increasing the sale of any patent
medicine, proprietary food preparation, or any similar product
for which, in the opinion of the Council, inaccurate or misleading
claims are made, shall forfeit his membership, it 1s moved that,
in the opinion of the Council of this Society, meeting in special
session, Dr. S. P. Beebe did consent to the use of his name in such
a way that it did aid in increasing the sale of a patent medicine or
similar product, for which inaccurate or misleading claims have
been made.”
It was moved by Dr. Meltzer and seconded by Dr. Auer that
this report of the Council be accepted as representing the attitude
of the Society in this matter. This motion was passed unani-
mously without discussion, twenty-two members present and
voting.
Seventy-second Meeting.
University and Bellevue Hospital Medical College, January 19,
1916. President Lusk in the chair.
Members present: Atkinson, Auer, Benedict, Cole, R. I.,
DuBois, Edwards, Ewing, E. M., Fine, Gies, Githens, Greenwald,
Harris, Hartwell, Jackson, Kleiner, Loeb, J., Lusk, Mandel, A. R.,
Mandel, J. A., Mayer, Meltzer, Murlin, Myers, Oppenheimer,
Pike, Ringer, Rous, Wasteneys, Wiggers.
Members elected: Harold Amoss, A. A. Epstein, N. W. Janney,
F. W. Peabody, Louise Pearce.
Seventy-third Meeting (Thirteenth Annual Meeting.)
College of the City of New York, February 16, 1916. President
Lusk in the chair.
Members present: Auer, Bull, Edwards, Githens, Goldfarb,
Jackson, Kleiner, Kober, Lusk, MacNeal, Meltzer, Myers, Noble,
Scott, E. L., Scott, G. G., Winslow.
Member elected: Casimir Funk.
EXECUTIVE PROCEEDINGS. 215
The meeting was held 5.00 P. M., and was followed by a dinner
at 7.00 P. M. Election of officers occurred for the ensuing year
after the dinner and resulted as follows:
President, Jacques Loeb; Vice-President, William J. Gies;
Secretary-Treasurer, Holmes C. Jackson; Council members of
the Society, J. Auer and E. F. DuBois.
Seventy-fourth Meeting.
Presbyterian Hospital, March 15, 1916. President Jacques
Loeb in the chatr.
Members present: Atkinson, Auer, Bull, Cohn, Cole, DuBois,
Epstein, Fine, Githens, Greenwald, Harris, Hess, Howe, Jackson,
Janney, Kirkbride, Klein, Lambert, Loeb, J., Longcope, Lusk,
Meltzer, Murlin, Myers, Ottenberg, Pappenheimer, Pepper,
Swift, Underhill, Wadsworth, Weil, White.
Members elected: Carl Ten Broeck, Edward Uhlenhuth.
Seventy-fifth Meeting.
College of Physicians and Surgeons, April 19,1916. Vice-Presi-
dent Gies in the chair.
Members present: Auer, Berg, Eggleston, Fine, Funk, Gies,
Githens, Hess, Howe, Jackson, Kleiner, Lee, Meltzer, Oppen-
heimer, Pike, Scott, E. L., Uhlenhuth, Zinsser.
Seventy-sixth Meeting.
Yale University, New Haven, Conn., May 24,1916. Vice-Presi-
dent Gies in the chair.
Members present: Atkinson, Benedict, Epstein, Fine, Gies,
Githens, Harrison, Harris, Hess, Hooker, Jackson, Kleiner,
Kober, Lee, Lusk, Meltzer, Mendel, Myers, Scott, G. G., Uhlen-
huth, Underhill, Wadsworth, Winslow, Woodruff.
Members elected: Walter Eddy, Rhoda Erdmann, Reynold
Albrecht Spaeth, Carl Vernon Weller.
The meeting was held at 4.30 P. M., in the Osborn Zodlogical
Laboratory. At the conclusion of the meeting an informal dinner
was held at the Hotel Taft. Twenty-two members attended
with New Haven guests.
Pacific Coast Branch.
Tenth Meeting.
San Francisco, California, October 6, 1915.
216 SCIENTIFIC PROCEEDINGS (76).
Members present: Addis, Burnett, Cooke, Dickson, Kocher,
Lucas, Maxwell, Meyer, Ophiils, Walker.
Eleventh Meeting.
San Francisco, California, December 1, 1915.
Members present: Burnett, Cooke, Dickson, Evans, Lucas,
Meyer, Ophiils, Swain, Walker, Whipple.
Twelfth Meeting.
San Francisco, California, April 18, 1916.
Members present: Addis, Burnett, Cooke, Crawford, Dickson,
Evans, Gay, Kellogg, Kocher, Meyer, Morgan, Robertson,
Whipple.
REGISTER OF NAMES AND ADDRESSES OF
THE MEMBERS OF THE SOCIETY FOR
EXPERIMENTAL BIOLOGY AND
MEDICINE.
IABBOLL AL ICKAND PRs GC sa: .a7o eve fol aisyensieveveistevsiers) sleveve: shies: University of Pennsylvania.
PAUSE ee OPINGE Jresers ate. 5 siettoret/erta hier chor sy-a1crapeven seo. nie bia av everenaval ears Johns Hopkins University.
(ADAMI ely GEORGE 5.40 o.2605 10 5 brecciale s Alsreisieishniua gion os McGill University, Montreal.
PADI DIS hs DHOMASS avers, cnc otessisusseveve'e) shaseiacejevs Leland Stanford University, San Francisco.
PADI R A ELE AN IV estore) ayehetarehe elekekelielevalteve) osteo: cve\ctetons) oi tecieterarevehe, eps Harvard University.
FADED LS AUVAG soles cich oh vor ayvere ora tera ovicva erotatate suchovs New York Polyclinic Medical School.
AULEENG Als IRE GINALD e070 susp eiciare sieleie wee ofbvceunisue: Serres University of Pennsylvania.
IAUSBERG; CART) :sreiesassieneiste arse U. S. Department of Agriculture, Washington, D. C.
AMOSS; VAR OLD oats ecie aleceisisle s ioiatets Rockefeller Institute for Medical Research.
ANDERSON): JOHN he -crs: oie em © seb ereebue eioeretd University of California.
COUNCIEMAN, *WIPLIAM: “Ty s.c.osc 2 uk tinrow ene eee toe kie aie sees Harvard University.
CRAMPTON, GC. WARD.0 20. ciecscce cass Department of Education, New York City.
CRAMPTON; HENRY, Eo. cies niece ates! neve aie eel bieidele rere mieeiee Columbia University.
CRAWFORD, ALBERT (C7. odien,vocciscieee ee nee eee Leland Stanford University.
GRite; GEORGEOW. .5. gee sie nen oS eRe Western Reserve University, Cleveland.
CUSHING, HUAR VEY 5 co: 503 Sis so aie ce eee Oe Ce eee eee Harvard University.
DAKAIN, (He -Ds 5 Scie he tae on ole bia eee ee 819 Madison Avenue, New York City.
DANDY; ' WAUTER:E. 2 0 voces one re ine ae Meee ie eraiens Johns Hopkins University.
DAVENPORT, CHARLES Bilicc gman sccee = Carnegie Institution, Station for Experi-
mental Evolution, Cold Spring Harbor, Long Island, N. Y.
DICKSON; JEG ued eee eee Stanford University Medical School.
DOCHEZ) Ap Roa cise ws os eae eee Rockefeller Institute for Medical Research.
DONALDSON, (Hi: SH oe Senin. ee elaieia eeeioie Wistar Institute of Anatomy, Philadelphia.
DRAPER; (GEORGE ieee oe eee eee Presbyterian Hospital, Columbia University.
DRAPER; Jo. Wok, o5:5 «acca Se alsin e Salo orale ath oeiviere aie eel New York University.
DRESBACH, M255 c:s2piescecpmanisine oes we sais cemieoee eee ee Cornell University.
DuBOIS;, EAR er coettisee iad ee eee Cornell University Medical College.
DUNHAM, ‘EDWARD -Koi7..2..Ke aia = Gisjore a re ae ise ieee aie oe New York University.
DUVALL, CHARLES (Wi, <0: cause norte nies or ane 2 oe cieleleierare erste Tulane University.
EDDY, WALTERSE =. a-tale elects arate High School of Commerce Annex, New York City.
EEDMUNDS, (@o9W too n0oa.0 0 ale tis crcre Sie eit ee te Dee aleie Mictereate University of Michigan.
Epsatr, DAVID ic. celes et vs stte vents cis eels else eis ers ieieie oe eine Harvard University.
EDWARDS Dat] ike cca elses eite sete College of the City of New York.
ISENBREY> GAD sajeieicie cleo oetes astern seule sieks St. Luke’s Hospital, New York City.
EGGLESTON; ‘GARY: F ecce.oe site eeaiccaetetcelce Cornell University Medical College.
EISBERG, (CHARLES"A.) <.-2 2 Eee eee corer ence ee Mount Sinai Hospital.
EELSER, “(WiILETAMIT 22.0 27008 oS tenet ee Cornell University Medical College.
EMERSON, EIAVEN Sd). . ccc & Se eee me eieiors Health Department, New York City.
JEPSTEEN, ALBERT IAL sulin sk ston ico semicon Mt. Sinai Hospital, N. Y.
ERDMANN;, "RHODA <5.