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r*t
4*%
Studies from the Rockefeller
Institute for Medical Research
Rockefeller Institute for Medical Research, Rockefeller Institute
fe<.
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STUDIES
FROM
THE ROCKEFELLER INSTITUTE
FOR MEDICAL RESEARCH
REPRINTS
VOLUME XVI
NEW YORK
The Rockefeller Institute for Medical Research
1913
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i^MB^
^
r APR 1 1913
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Contents
v^. On the Combined Action of Muscle Plasma and Pan-
creas Extract on Some Mono- and Disaccharides.
By P. A. Levene and G. M. Meyer. (From the
Laboratories of The Rockefeller Institute for
Medical Research, New York.)
V^. On the Action of Various Tissues and Tissue Juices
on Glucose.
By P. A. Levene and G. M. ^yer. (From the
Laboratories of The Rockefeller Institute for
Medical Research, New York.)
V^. The Action of Leucocytes on Glucose.
By P. A. Levene and G. M. Meyer. (From the
Laboratories of The Rockefeller Institute for
Medical Research, New York.)
v<2|.. On the Action of Tissue Extracts Containing Nucleo-
sidase on a and fi Methylpentosides.
By P. A. Levene, W.' A. Jacobs, and F. Medigre-
ceanu. (From the Laboratories of The Rocke-
feller Institute for Medical Research, Nciv York,)
y^5. The Toxicity of Sugar Solutions upon Pundulus and
the Apparent Antagonism between Salts and
Sugar.
By Jacques Loeb. (From the Laboratories of
The Rockefeller Institiite for Medical Research,
New York.)
V^6. Pharyngeal Insufflation, a Simple Method of Artificial
Respiration. A Preliminary Note.
By S. J. Meltzer. (From the Laboratories of The
Rockefeller Institute for Medical Research, New
York.)
ill
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iv Contents
yfj. On the Adaptation of Fish (Fundulus) to Higher
Temperatures.
By Jacques Loeb and Hardolph Wasteneys.
{From fhe Laboratories of The Rockefeller In-
stitute for Medical Research, New York.)
y6. A Further Report on the Production of Experimental
Chronic Nephritis in Animals by the Administra-
tion of Uranium Nitrate.
By Ernest C. Dickson. {Prom the Pathological
Laboratory of Cooper Medical College and Le-
land Stanford Jr, University,)
Jg. Malarial Pigment (Hematin) as a Factor in the Pro-
duction of the Malarial Paroxysm.
By Wade H. Brown. {Prom the Pathological
Laboratory of the University of North Carolina,)
/.
lo. A Biochemical Study of the Phenomena Known as
Complement-Splitting. First Paper: Splitting of
the Complement Associated with Globulin Precipita-
tion.
By Jacob Bronfenbrenner and Hideyo Nog^chi.
{Prom the Laboratories of The Rockefeller In-
stitufe for Medical Research, and the Laboratory
of Biological Chemistry of Columbia University,
Neiv York.)
Vii. A Biochemical Study of the Phenomena Known as
Complement-Splitting. Second Paper: Splitting of
the Complement without a Visible Alteration of the
Proteid Constituents.
By Jacob Bronfenbrenner and Hideyo Noguchi.
{From the Laboratories of The Rockefeller In-
stitute for Medical Research, and the Laboratory
of Biological Chemistry of Columbia University,
New York.)
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Contents v
^2. A Quantitative Study of the Effects of Adrenalin on
the Pupils of Rabbits after Removal of a Superior
Cervical Ganglion.
By Don R. Joseph. {From the Laboratories of
The Rockefeller Institute for Medical Research,
New York.)
/li. Transplantable Tumors of the Fowl: A Neglected
Material for Cancer Research.
By Peyton Rous, James B. Murphy, and W. H.
Tytler. {From the Laboratories of The Rocke-
feller Institute for Medical Research, New York.)
vf4. On Sphingosine.
By P. A. Levene and W. A. Jacobs. {From the
Laboratories of The Rockefeller Institute for
Medical Research, New York.)
A
s/\e.
/>
On the Transmission of Immunity from Mother to
Offspring. A Study upon Serum Hemolysins in
Goats.
By L. W. Famulener. {From the Research Lab-
oratory of the Department of Health, New York.)
The Role of Injury in the Production of a Chicken
Sarcoma by a Filterable Agent.
By Peyton Rous, James B. Murphy, and W. H.
Tytler. {From the Laboratories of The Rocke-
feller Institute for Medical Research, New York.) '
The Relation between a Chicken Sarcoma's Behavior
and the Growth's Filterable Cause.
By Peyton Rous, James B. Murphy, and W. H.
Tytler. {From the Laboratories of The Rocke-
feller Institute for Medical Research, New York.)
V i8. The Nature of the Filterable Agent Causing a Sar-
coma of the Fowl.
By Peyton Rous and James B. Murphy. {From
the Laboratories of The Rockefeller Institute for
Medical Research, New York.)
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VI
Contents
^19. A Clinical Study of Acute Poliomyelitis.
By Francis W. Peabody, George Draper, and A.
R. Dochez. (From the Hospital of The Rocke-
feller Institute for Medical Research, New York.)
y^o. Picrolonates of the Monoamino-Acids.
By P. A. Levene and Donald D. Van Slyke.
(From the Laboratories of The Rockefeller In-
stitute for Medical Research, New York,)
y^i. The Preservation of Tissues and Its Application in
Surgery.
By Alexis Carrel. (From the Laboratories of
The Rockefeller Institute for Medical Research,
New York,)
(22, Some Aspects of Anaphylaxis.
By John Auer. (From the Laboratories of The
Rockefeller Institute for Medical Research, Nezv
York,)
y23. Intratracheal Insufflation im physiologischen Labo-
ratorium.
By S. J. Meltzer. (From the Laboratories of The
Rockefeller Institute for Medical Research, New
York,)
/'
24. t)ber die Potentialdifferenzen an der unversehrten
und verletzten Oberflache pflanzlicher und tieri-
scher Organe.
By Jacques Loeb and Reinhard Beutner. (From
the Laboratories of The Rockefeller Institute for
Medical Research, New York,)
y/25. Anaphylaxie als eine Ursache von Koordinations-
storungen des Herzschlags beim Kaninchen.
By John Auer. (From the Laboratories of The
Rockefeller Institute for Medical Research, New
York,)
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Contents
VII
1/26. -Zur Photometric des Blutfarbstoffes.
By E. E. Butterfield. (From the Laboratories of
The Rockefeller Institute for Medical Research,
New York.)
1/27. tJber die Hemmung der Giftwirkung von NaJ,
NaNOg, NaCNS und anderen Natriumsalzen.
By Jacques Loeb. (From the Laboratories of
The Rockefeller Institute for Medical Research,
New York,)
^8. Uber Kriterien der Anaphylaxie.
By John Auer. {From the Laboratories of The
Rockefeller Institute for Medical Research, New
York.)
(/$9. Chemische, physikalische und biologische Studien uber
die aus den drei Monoaminomonocarbonsauren :
GlykokoU, d-Alanin und 1-Leucin darstellbaren
strukturisomeren Tripcptide.
By Emil Abderhalden and Andor Fodor. {Aus
dem physiologischen Institut der Universitdt
Halle a. S.)
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Repilntcd from the jq|&{^l of BioLOOMALt^lmTRT, Wl. xi, no. 4, 1912
ON THE COMBINED ACTION OF MUSCLE PLASMA
. AND PANCREAS EXTRACT ON SOME MONO- AND
DISACCHARIDES.
By p. a. LEVENE and G. M. MEYER.
(From the Rockefeller Institute for Medical Reaearch^ New York.)
(Received for publication, March 14, 1912.)
Through the experiments reported in a previous communication*
the present writers have demonstrated that by the combined
action of muscle plasma and pancreatic extract d-glucose was con-
verted into a disaccharide. On the other hand, under the same
conditions of experiment maltose was cleaved into glucose. Nat-
urally it became important to make clear whether the action was
applicable also to other sugars. Of the previous writers only
HalP allowed the muscle plasma to act on other sugars than glu-
cose. This investigator extended his experiments to d-levulose,
/-arabinose and d-xylose,.and was led to the conclusion that the
glycolytic action of his enzjone mixture was limited to glucose only.
In the course of our previous work the optimal conditions for the
action of muscle plasma and pancreatic extract were determined
with greater certainty, and this made it urgent to repeat and to
extend the experiments of Hall; all the more since the action of the
enzyme mixture is viewed at present in a different light.
Of the hexoses levulose, mannose and galactose were employed.
The mannose was obtained through the courtesy of Dr. Hudson of
Washington, and we wish to express our appreciation for his kind-
ness. Of the pentoses I-arabinose, d-xylose and rf-ribose were used
for the experiments. Lactose was the disaccharide tested.
In regard to hexoses it was foimd that mannose remained un-
changed under the conditions of our experiments, but rf-levulose
showed under the influence of muscle plasma and of pancreatic
* Levene and Meyer: this Journal, ix, pp. 97-107, 1911.
» Hall: Amer. Journ. of Physiol., xviii, pp. 283-294, 1907.
347
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348 Action of Muscle Plasma and Pancreas Extract
extract a diminution of reducing power which could be restored to '
nearly the original power by means of hydrolysis with dilute min-
eral acid.
Ill this respect our conclusions differ from those of Hall. The
analysis of tables published by Hall, however, shows some disappear-
ance of levulose through the action of the enzyme mixture, but the
writer is inclined to explain these changes by faults in technique,
namely by bacterial contamination. On the other hand, there is
no record in his report of the concentration of the sugar employed
in his experiments, and the importance of this factor has been
emphasized in our previous publication. All our experiments
were controlled by bacteriological examination, and only those
experiments were taken into consideration which proved free from
any bacterial growth. Hence the disagreement between our re-
sults and those of Hall in regard to d-levulose is probably caused
by the difference in the sugar concentration employed in the experi-
ments of Hall and in ours.
Regarding the pentoses our experiments are in full accord with
those of Hall, and there was never observed a diminution in the
reducing power of the pentose even when the concentration of the
sugar solutions employed in the experiments was verj' considerable.
Also in regard to lactose the results of our experiment harmonize
with those of Hall.
On the basis of all this experience one is led to the conclusion
that the muscle plasma combined with pancreatic extract possesses
the power to cause condensation of only two closely related hexoses,
namely of rf-glucose and of d-levulose, and that it remains without
action on mannose, xylose, ribose, and lactose. The same enzyme
mixture also has the power to bring about the hydrolysis of mal-
tose, but not of lactose.
EXPERIMENTAL PART.
The enzyme mixtures were prepared in the manner described in
the previous communication.
All the details of sugar analysis were the same as there described.
The condensed levulose was hydrolyzed by heating on a water
hath for two hours with 5 per cent hydrochloric acid.
The results of the analysis are given in the following tables.
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p. A. Levene and G. M. Meyer
349
d'LevtUose
if Mdl
|3 !|S^|«28
a. At beginning of experiment | 0.5
After thirty-six hours 0.5
b. At beginning of experiment ; 0.5
After thirty-six hours I 0.5
After hydrolysis I 0.5
c. At beginning of experiment | 1.0
After thirty-six hours ' 1.0
d. At beginning of experiment | 2.0
After thirty-six hours i 2.0
28.7
25.8
24.1
22.4
23.2
27.9
26.3
31.2
29.4
Galactose.
a
s
57.4
51.6
48.2
44.8
46.4
27.9
26.3
15.6
14.6
8 go
Sign
Sa52;
20.09
18. Oa
17.2 !
16.0 I
16.6 I
10.0 I
9.4
5.46|
5.21
J. 03
L2
§1
a
10.1
7.0
I
0.6 6.0
0.25 I 4.5
I
K U
^•M2g
3 o« y
, At beginning of experiment 0.5
After thirty-six hours 0.5
At beginning of experiment 0.5
After thirty-six hours 0.5
At beginning of experiment 1.0
After thirty-six hours 1.0
. At beginning of experiment 2.0
After thirty-six hours 2.0
l-Arabinose.
I iS .
o ^
23.55
23.60
19.0
19.1
22.8
22.6
22.9
22.9
47.1
47.2
38.0
38.2
22.8
22.6
11.45
11.45
17.23
17.28
13.93
13.95
8.34
8.34
4.17
4.17
5 go
5aZ
I . « Ml
â– ' u U U â–º
a. At beginning of experiment » i 0.5
After thirty-six hours ' 0.5
6. At beginning of experiment » i 1.0
After thirty-six hours i 1.0
26.4
26.4
21.6
21.6
52.8 il4.8
52.8 14.8
21.6 6.37
21.6 6.37
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350 Action of Muscle Plasma and Pancreas Extract
d-Xylose.
a. At beginning of experiment
After thirty-six hours
6. At beginning of experiment
After thirty-six hours
n H
M a ^
Is 2S^ MpS *ea
gS gsJ5 g£8 ^ggg' I
0.5 I 24.1 I 48.2 j 12.35
0.5 I 24.1 ! 48.2 12.35
1.0 24.3 I 24.3 , 6.22
1.0 . 24.3 24.3 6.22
Lactose.
a. At beginning of experiment
After thirty-six hours
b. At beginning of experiment
After thirty-six hours
3.0
3.0
4.0
4.0
18.6
18.5
17.0
17.2
6.20
6.17
12.0
12.0
4.25 I 8.0
4.30 ; 8.0
d-Riboae,
H
e
I
^ ml S H I N O " I
At beginning of experiment.
After thirty-six hours
0.5 22.2 44.4 | 15.8
0.5 22.1 I 44.2 15.8
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p. A. Levene and G. M. Meyer
351
b.
Mannose.
' Q
Ji, 00
" a
B_
At beginning of experiment ! 0.5
After thirty-six hours 0.5
At beginning of experiment 1 .25
After thirty-six hours 1 .25
At beginning of experiment 1.0
After thirty-six hours 1.0
S CO
24.4
24.2
31.8
31.7
36.5
36.5
NH«CNS
PER CUBIC
CENTIMETER
â– hi
a
!
48.8
15.05
48.4
15.05
25.44
7.84
25.4
7.84
36.5
11.25
36.5
11.25
In calculating the grams sugar for 100 cc. solution the following
values for 1 cc. -ft^ NH4CNS equivalent to milligrams sugar were
used.
Mgs.
Glucose 3.58
Laevulose 3.58
Maltose 5.78
Galactose 5.76
Mg».
Arabinose 2.95
Xylose 2.56
Ribose 2.56
Lactose 1 . 89
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Reprinted from The Journal op Biolooical Chcmibtrt. Vol. XI. No. 4, 1912.
ON THE ACTION OF VARIOUS TISSUES AND TISSUE
jmCES ON GLUCOSE.
Br P. A. LEVENE and G. M. MEYER.
(From the Rockefeller Institute for Medical Research , New York.)
(Received for publication, March 14, 1912.)
The literature on the glycolytic action of various animal organs
contains most contradictory and confusing statements. While
some writers claimed the presence of sugar-destroying enzjines in
all organs and tissues, other observers detected such enzymes only
in few organs, and only under very definite conditions, namely,
in the presence of some auxiliary substance. Thus, in recent years,
Amheim and Rosenbaum* and Stoklasa^ and his co-workers claimed
a general distribution of sugar-destroying enzymes in all animal
tissues. Rapoport* observed gylcolytic action only in blood and
fibrin and obtained negative results from his experiments with
other organs. Finally, R. Hirsch^ and 0. Cohnheim*^ observed that
the glycolytic action is brought about by the combined action of
pancreas extract and of the liver, or by pancreas extract and mus-
cle plasma.
It is possible that the observations reported by every one of
the writers are correct, and the apparent contradictions were
brought about by the different conditions of the respective experi-
ments. It became evident from our work on the combined action
of muscle plasma and pancreatic extract that alone variations in
the sugar concentration may change the results of the experiments
to such an extent that a marked disappearance of glucose will
1 Zeilschr.f. physiol. Chem,, xl, p. ^20, 1C03-1904.
« Pfltiger's Archiv, ci, p. 311, 1904.
» Zeilschr.f. klin. Med., Ivii, p. 208, 1905.
* Hofmeister's Beilrdge, iv, p. 535, 1903.
*ZeiUchr.f. physiol. Chem., xxxix, p. 336, 1903; xlii, p. 401, 1904; xliil,
p. 547, 1904-1905; xlvii, p. 253. 1906.
353
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354 Action of Tissues on Glucose
take place in one instance, and no change in the sugar content in
the other. An analogous observation was made in the work on
leucocytes, which will be reported subsequently.
Another factor determining the result of the experiment lies in
the degree of alkalinity or acidity of the solutions used in the experi-
ment. The importance of this factor was pointed out first by the
work of Hall, and was corroborated by our own experiments. In
agreement with Hall we found that Henderson^s phosphate mixture
offers the best medium for the study of the so-called glycolytic
process. Still another cause for divergence in the characteristics
of the results one may find in the difference in the species of the
animal whose organs were employed in the experiment. Recent
work on the enzymes of animal tissues has brought to light the
great differences in the enzyme content of analogous organs of
animals belonging to different species. Of course some of the dis-
crepancies in the results of individual writers may have been caused
by the different degrees of antiseptic precaution exercised by them.
Only rarely were experiments controlled by bacteriological exami-
nations. Further, the analytical methods employed by different
workers varied greatly in their accuracy. And yet another source
for possible error may be found in the fact that rarely was there
made an attempt to search for the products of the disappearing
sugar.
The knowledge of the products formed in course of the experi-
ment is important not only for theoretical reasons but as a means of
detecting bacterial contamination. Thus in all our experiments of
the last two years carbon dioxide was only rarely detected in the
reaction mixture, and its presence always indicated bacterial con-
tamination; therefore, we are inclined to believe that in the experi-
ments of other writers, where there was reported the formation
of carbon dioxide from sugar, this resulted from bacterial activity,
and not through the actions of tissue enzymes.
All these considerations led us to subject to a revision all the
older observations on the presence in animal organs of either
"glycolytic'^ enzymes or of activators of the enzymes, all the more
since it became evident that the so-called glycolysis under the
combined action of muscle plasma and pancreas consisted in a con-
densation and not in a destruction of glucose. In course of this
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p. A, Levene and G. M. Meyer 355
work the organs of the rabbit and of the dog were employed. In
one series of experiments the enzymotic effect of the tissue pulp
was tested and in another of the tissue juices. A still different
series of experiments aimed to investigate the presence in vari-
ous organs of enzyme activators. For this purpose the action
of various organs aided by extracts of other organs was tested.
Every experiment was controlled by bacteriological examinations,
made by Dr. Bronfenbrenner; only experiments that proved free
from any bacterial growth were taken into consideration.
The results of the experiments were as follows:
A. In the experiments with the rabbit, without the aid of acti-
vators only the action of liver and of muscle tissues were tested.
The results in both experiments were negative. With the aid of
pancreas extract also only the same two tissues were tested. Posi-
tive results were obtained only with the muscle plasma.
B. In the experiments with the dog without the aid of activators
I'the following tissue juices were employed: muscle, lung, intestine,
kidney, pancreas and spleen. All experiments were negative.
As activators the extracts of the pancreas and of the spleen were
employed. With each activator were employed the same tissue
juices as in the first series. The results were the following. The
addition of pancreas extract did not alter the action of the tissue
juices; on the other hand, after the addition of spleen extract as
activator there was observed a fall of the reducing power of the
sugar solution in the experiments with muscle, lung, liver and pan-
creas. The action was of very moderate intensity.
In the experiments with tissue pulp the following organs were
used: muscle, spleen, liver, lung and pancreas. The results in all
experiments were negative, excepting the liver. The experiments
with liver tissue and additional glucose showed at the end of the
experiments no change in the original reducing power; on the other
* hand, in the control experiments with liver tissue alone there was
observed an increase in the reducing power. Hence it follows
that in the experiment with additional glucose there was a com-
pensation of phenomena so that the rise of glucose formation was
observed by a simultaneous disappearance of glucose.
With the addition of pancreas extract as activator the same
organs were tised. The results were the same as in previous series.
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356 Action of Tissues on Glucose
With the addition of spleen extract as activator the presence of
enzymotic action was observed in the experiments with muscle,
lung, liver and pancreas tissues.
Thus in the dog the spleen and not pancreas is the organ contain-
ing the activator for the enzyme causing the condensation of
glucose.
The general conclusion from the present experiments is that
under the conditions here reported, namely, in the presence of
antiseptics and under the conditions where access of oxygen is not
totally excluded, animal tissues or their juices, aided or unaided