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THE AMERICAN JOURNAL
OF
PHYSIOLOGY:
EDITED FOR
The American Pbhysiolomcal Society
BY
H. P. BOWDITCH, M.D., BOSTON FREDERIC S. LEE, Pu.D., NEW YORK
R. H. CHITTENDEN, PH.D., NEW HAVEN JACQUES LOEB, M.D., BERKELEY
W. H. HOWELL, M.D., BALTIMORE W. P. LOMBARD, M.D., ANN ARBOR
W. T. PORTER, M.D., BOSTON
TEE
AMERICAN JOURNAL
OF
hielo | Ore CG.
VOLUME X.
BOSTON, U.S.A.
GIN Ne UAND COMPANY
1904
QP
|
As
VviLO
cop. Z
Copyright, 1903
By GINN AND COMPANY
Gniversity ress
JOHN WILSON AND Son, CAMBRIDGE, U.S.A.
OB ise Dee S:
No. I, SEPTEMBER I, I903.
NOTES ON THE HEART ACTION OF MOLGULA MANHATTENSIS (VERRILL).
meeunee Vi Tica THUALEY, fhe 8 ee te ee ge wes
THE SKIN AND THE EYES AS RECEPTIVE ORGANS IN THE REACTIONS
SPOR GS TOP EMGHT:.~ 2090 Cr dteod PERO ea) gh Woe yes re) ta hoe ee hg
INFLUENCE OF RENNIN UPON THE DIGESTION OF THE PROTEID CON-
SUrEb Ets: ORDER, \ Arir iephe We ter es ne a we | eg
RESPIRATION EXPERIMENTS IN PHLORHIZIN DIABETES. By Arthur R.
BAGUIO LILI AGILE LGU oe tt Sod ey ie ee
THE EFFECT OF LECITHIN ON THE GROWTH OF THE WHITE Rat. By
ICN PLAINES, Meee ot Nees” ee ME eed ae ek lak
ON THE ACTION OF PHLORHIZIN. Sy Percy G. Stiles and Graham Lusk
No. II, OcroBer 1, 1903.
EXPERIMENTS ON THE DIGESTIBILITY OF VEGETABLES. Ly A. P. Bryant
Tare EN REE STINE EO TASC Usa De 5 5 3loy ease ell, Sony Thy Pi FALE es SE COG ERE [he
No. III, NoveMBer 1, 1903.
ON THE ACTION OF SALINE PURGATIVES IN RABBITS AND THE COUNTER-
ACTION OF THEIR EFFECT BY CALCIUM. By John Bruce MacCallum
THE CEREBRO-SPINAL FLUID IN HYDROCEPHALUS. By /sador H. Coriat
ON THE TIME RELATIONS OF PROTEID METABOLISM. By P. B. Hawk
ON THE DISTRIBUTION OF OssEOMUCOID. By Christian Seifert and
BA a) AGIOS. (sake iy 5 ese Ren war es OS PK) aL wae os
ON THE VARIATIONS OF BLOOD-PRESSURE DURING THE BREATHING OF
RABESIne Atm + By Tredericiid: BOGE. 6 as aha elte 6 Ge
PAGE
28
37
47
57
67
SI
149
v1 Contents.
No. IV, DECEMBER I, 1903.
PAGE
REACTIONS TO TEMPERATURE CHANGES IN SPIRILLUM, HYDRA, AND
FRESH-WATER PLANARIANS. By S. O. Mast . . . ». » + « « 165
THE HYDROLYSIS AND SYNTHESIS OF FATS BY PLATINUM BLACK. Sy
Hugh Netlson cee ee ey we es (srr
THE STATIC FUNCTION IN GontoNEMUS. By Louis Murbach . . . .° 201
No. V, JANUARY I, 1904.
THe EFFECTS OF VARIOUS SALTS ON THE TONICITY OF SKELETAL
Muscies. By W.D. Zoethout -. . + + +) = js oe) 5 ee
SomE EFFECTS OF THE RONTGEN RAYS ON THE DEVELOPMENT OF
Empryos. By P. K. Gilman and F. H. Baetjer . . «~~ % oes
Tue EFFECTS OF IONS ON THE DECOMPOSITION OF HYDROGEN PEROX-
IDE BY PLATINUM BLack. By C. Hugh Neilson and Orville H.
Brown 6 6 eR wee el a a Se) en
CONCERNING THE FORMATION OF SUGAR FROM LEuCIN. By /. T. Halsey 229
LOCALIZATION OF THE RESPIRATORY CENTRE IN THE SKATE. By /da
Hl. Hyde@ 6 6 8 we Se a ee
ON THE LOCAL APPLICATION -OF SOLUTIONS OF SALINE PURGATIVES
TO THE PERITONEAL SURFACES OF THE INTESTINE. By John Bruce
MacCallum . eee wt a we re
No. VI, FEBRUARY I, 1904.
A STUDY OF THE VARIATIONS IN THE COURSE OF THE NITROGEN,
SULPHATE, AND PHOSPHATE EXCRETION, AS OBSERVED IN SHORT
PERIODS FOLLOWING A SMALL INCREASE IN THE PROTEID INGESTED.
By P. B. Hawk and Joseph S. Chamberlain. . . . » 2 = = 2ee
THE RELATION BETWEEN SOLUTION-TENSION, ATOMIC VOLUME, AND
THE PHYSIOLOGICAL ACTION OF THE ELEMENTS. By Albert P.
Mathews 2. 2 6 6 ee ee on a en
ON THE PRODUCTION OF CONTACT IRRITABILITY WITHOUT THE PRECIP-
ITATION OF CALCIUM SALTS. By W. D. Zoethout . | eee
EFFECT OF IONS ON THE DECOMPOSITION OF HYDROGEN PEROXIDE,
AND THE HYDROLYSIS OF BuTYRIC ETHER BY A WATERY EXTRACT
OF PANCREAS. Sy C. Hugh Neilson and Orville H. Brown . . . 335
DorEs AN ANTAGONISM EXIST BETWEEN ALKALOIDS AND SALTS? By
Martin ff. Fischer. 0 6 oe ew 8 rr
Contents. Vil
THE SIMULTANEOUS ACTION OF PILOCARPINE AND ATROPINE ON THE
DEVELOPING EMBRYOS OF THE SEA-URCHIN AND STARFISH. —A
CONTRIBUTION TO THE STUDY OF THE ANTAGONISTIC ACTION OF
POISONS. “By @orald Sollmann eos =. 2s shes 8 fe 8 we 6352
THE EFFECT OF DIURETICS ON THE URINE, WITH A DIET POOR IN
Peete i hs). Pisces as kat 8 et -cun ita se sas 2 =) 302
No. VII, Marcu 1, 1904.
SOME PHENOMENA OF ANIMAL PIGMENTATION. By R. C. Schiedt . . 365
FURTHER EXPERIMENTS ON THE INFLUENCE OF VARIOUS ELECTROLYTES
ON THE TONE OF SKELETAL MuscLes. By W. D. Zoethout . .« 373
EFFECTS OF CERTAIN SALTS ON KIDNEY EXCRETION, WITH SPECIAL
REFERENCE TO GiycosurIA. By Orville Harry Brown . . . . 378
ON THE MoRPHOLOGICAL CHANGES IN THE BLOOD AFTER MUSCULAR
EReRGISE.. ills Be Tae “e) Giie Soba emer a eB ol ee 384
THE RATE OF THE NERVOUS IMPULSE IN THE SPINAL CORD AND IN
THE VAGUS AND THE HYPOGLOSSAL NERVES OF THE CALIFORNIA
HaGFISH (BDELLOSTOMA DoMBEY!). By A. J. Carlson . . . ~- 401
THE RELATION OF IONS TO CILIARY MOVEMENT. By Ralph S. Lillie. 419
THE RELATION BETWEEN THE DECOMPOSITION-TENSION OF SALTS AND
THEIR ANTIFERMENTATIVE PROPERTIES. By Hugh McGuigan . . 444
THE ALLoxuRIC BASES IN ASEPTIC FEVERS. By Arthur R. Mandel . 452
PROCEEDINGS OF THE AMERICAN PHYSIOLOGICAL SOCIETY . . . . ix-xliv
Ie Ls acs Sia ck MM Say ee aS a Sh, gh Ser Hal aah! ARO
PROCEEDINGS OF THE AMERICAN PHYSIO-
BOGICAL (SOCIETY,
SIXTEENTH ANNUAL MEETING.
PHILADELPHIA, PA., DECEMBER 29 and 30, 1903.
EEOCEE DINGS OF THE AMERICAN PHYSIOLOGICAL
SOCIETY:
THE SURVIVAL OF IRRITABILITY IN MAMMALIAN NERVES
AFTER REMOVAL FROM THE BODY.
By W. D. CUTTER anp P. K. GILMAN.
MAKING use of the fact noted by other observers (Bernard, Schiff,
Israel, Greene), that the mammalian nerve retains its irritability for
some time after removal from the body, the authors attempted to
determine the duration of this survival, the variations in irritability
during the period of survival, and, lastly, the effect of prolonged
anesthesia upon the phenomenon. Irritability was determined by
measuring the action current of the nerve when stimulated by a
series of induction shocks. The experiments were made upon dogs,
and the sciatics of both legs were taken for observation. One sciatic
was removed as soon as the animal was aneesthetized sufficiently for
the operation. The nerve was placed at once in the moist chamber,
and its action current was determined at intervals of half an hour, as
long as a response could be obtained to stimulation. With the
values of these action currents as ordinates, a curve was constructed,
showing the duration and variations of irritability in the “ unanzs-
thetized nerve” during the period of observation. The other sciatic
was left in the animal for a period of four to six hours, and during
this time the animal was kept completely anesthetized by morphia
and ether. At the end of this period, there was a considerable fall in
rectal temperature (30°-31° C.). The “ anzsthetized” nerve was
then removed, and galvanometric observations were made similar to
those just described. The results obtained show that the nerve
removed from the anesthetized (and cooled) animal survives for a
longer period than that taken from the animal at the beginning of
the period of anzesthesia, the difference in time of survival being as
Xl
X11 Sixteenth Annual Meeting.
much as four or five hours. A more marked difference, however, is
that the “‘ anzesthetized”’ nerve exhibits throughout a much greater
irritability. The curves obtained were irregular; but that for the
‘“ unanesthetized”? nerve shows a small increase in irritability occur-
ring shortly after the excision, and soon followed by a steady decline
to zero; while that for the ‘“‘ anesthetized” nerve exhibits, as its most .
marked feature, a large and sudden increase in irritability coming on
some hours after the excision, and followed by a more rapid fall to
zero.
THE CONDITION OF THE VASOCONSTRICTOR NEURONS
IN SHOCK” *
By W. T. PORTER anv W. C. QUINBY.
THE normal fall of blood-pressure produced by stimuli of uniform
intensity applied to the central end of the depressor nerve was
measured in the rabbit and the cat. In the same animals shock
was then brought on, and the measurements repeated. Following
are abbreviated protocols:
July 7, 1903.— Cat, anesthetized with ether. 9.25 a.m.: Carotid blood
pressure, 105 mm. Hg. 10.20 am.: Stimulated central end of left
depressor nerve ; blood-pressure fell from 80 to 45 mm. (44%). 10.23
A.M.: Section of spinal cord in dorsal region; blood-pressure 36 mm.
10.50 to 11.10 a.mM.: Injected 75 c.c. 0.6% sodium chloride solution in
jugular vein ; blood-pressure 80 mm. //./0 A.M.: Stimulated depressor ;
blood-pressure fell from 80 to 43 mm. (46%). //.35 a.m.: Blood-
pressure 30 mm.; injected 70 c.c. sodium chloride solution; blood-
pressure rose to 77 mm.; stimulated depressor ; blood-pressure fell from
77 mm. to 43 mm. (44%). 11.50'a.M.: Blood-pressure 30 mm.
September 22, 1903, — Rabbit, anesthetized with ether; blood-pressure in
left carotid, 87mm.Hg. 9.20 a.m.: Blood-pressure 70 mm. ; stimulated
central end of left depressor three times ; blood-pressure fell to 50 mm.
(297), 55 mm. (21%), and 52 mm. (26%) respectively ; burned intestines
and part of parietal peritoneum with nitric acid. 9.25 a.m.: Blood-
pressure 40 mm.; rectal temperature 37.6°. 9.35 am.: Blood-pressure
* A note of clinical interest regarding this investigation was published in the
Boston medical and surgical journal, 1903, cxlix, pp. 455-456.
Proceedings of the American Physiological Society. xiii
rises to 60 mm., probably from stimulation of sensory nerves by the
nitric acid. 17.50 4.m.: Rectal temperature 31°; blood-pressure 30 mm. ;
stimulated depressor; blood-pressure fell from 30 to 21 mm. (30%).
12 M.: Heart feeble; injected glycerine extract of suprarenal gland ;
blood-pressure rose to go mm., but sank thereafter. /2./0 p.m.: Rectal
temperature 30° ; blood-pressure 33 mm. ; stimulated depressor ; blood-
pressure fell from 33 to 25 mm. (24%). 12.15 p.M.: Repeated injection
of suprarenal extract; blood-pressure rose temporarily to go mm. 12.50
p.M.: Blood-pressure 72 mm. ; stimulated depressor ; blood-pressure fell
to 55 mm. (24%); blood-pressure then rose to 72 mm.}; repeated the
stimulation ; blood-pressure fell from 65 mm. to 48 mm. (26%).
September 24, 1903, 9 a.M.— Rabbit, aneesthetized with ether ; carotid blood-
pressure 80 mm. Hg. 9.7/0 a.m.: Both vagi cut. 9.75 a.m.: Rectal
temperature 38°; blood-pressure 67 mm.; stimulated central end of
depressor nerve; blood-pressure fell to 36 mm. (46%). 9.20 a.M.:
Exposed intestines; ligated mesenteric artery; applied nitric acid to
intestines ; blood-pressure rises. 9.40 A.M.: Rectal temperature, 36.6° ;
electrical stimulation of nerves near mesenteric artery causes the blood-
pressure to fall from 70 mm. to 55 mm. Hg. 3.25 p.m.: Rectal tempera-
ture 26° ; no anesthetic has been necessary for many hours; blood-pressure
53 mm.; stimulated depressor; blood-pressure fell to 30 mm. (43%).
4 ~.M.: Blood-pressure has for some time been about 30 mm. Hg. 4.50
p.M.: Blood-pressure 40 mm. ; stimulated depressor ; blood-pressure fell
to 22 mm. (45%). 4.16 p.m. (8 hours, 16 minutes after the beginning
of the experiment): Rectal temperature 25° ; blood-pressure 35 mm. ;
stimulated depressor ; blood-pressure fell to 23 mm.(34%).
In all cases the pressures given are diastolic, recorded by a slightly
damped Hiirthle membrane manometer. After stimulation of the de-
pressor, the blood-pressure regained its former level.
From these protocols it is clear (1) that the normal percentage fall
in blood-pressure may be obtained by stimulating the depressor
nerve during shock; (2) if during shock the blood-pressure be raised
to normal values by the injection of suprarenal extract or normal
saline solution, and the depressor nerve be stimulated while the
pressure is still high, the absolute fall in blood-pressure may be
as great as it was in the same animal before shock began.
Exhaustion in the vasoconstrictor neurons cannot therefore be the
essential cause of the symptoms termed shock.
xiv Sixteenth Annual Meeting.
A STUDY OF THE ERRORS INVOLVED IN THE DETERMINA—
TION OF THE BLOOD-PRESSURES IN MAN, TOGETHER
WITH A DEMONSTRATION OF THE IMPROVEMENTS IN
THE SPHYGMOMANOMETER THEREBY SUGGESTED.
By JOSEPH ERLANGER.
THE experiments herein recorded were performed with the object
of determining the errors involved in the estimation of the maximum
and minimum pressures with the sphygmomanometer devised by the
author (This journal, 1902, vi, p. xxii), the particular problem being
the determination of the effect of increasing the length of artery
compressed.
In the first series of experiments performed, use was made of an
artificial circulation scheme. These experiments showed that, as a
result of putting the walls of the tubes upon the stretch, the deter-
mination made with the short tubes were higher than those made
with long tubes; this effect disappeared when the length of the tube
compressed exceeded 4 cm.
These experiments were repeated upon animals, the pressure
being applied directly to the intact artery. The length of artery
compressed (3-6 cm.) had but very little effect upon the results.
These experiments indicate that the maximum pressure, as deter-
mined with the sphygmomanometer, is the maximum lateral pressure
of the artery of which the one explored is a branch. As the mini-
mum pressure is practically the same in all of the larger arteries
(Dawson), the sphygmomanometer, when applied to the brachial
artery of man, determines approximately the lateral pressures —
maximum and minimum — in the aorta.
In order to be able to study the effect of the tissues upon pressure
determinations, estimations of pressures in dog’s thighs were made
with cuffs of different widths. With narrow cuffs (3.5 cm.) the
resistance offered by the tissues may produce an error of 50 mm. Hg.
But with broad cuffs (g cm.) this error, under normal conditions, is
probably never greater than 10 mm. Hg. The error resulting from
the resistance offered by the tissues is the same for both the maxi-
mum and minimum pressures. The error that results from using the
sense of touch as an indicator of the return of the pulse wave may dy
chance be balanced by the error resulting from the resistance offered
by the tissues. Therefore, if an accurate method of determining the
Proceedings of the American Phystological Society. xv
return of the pulse wave could be adopted, the ‘ pulse pressure”
(difference between the maximum and minimum pressures) could
be determined with almost absolute accuracy.
As a result of this work a broad cuff (12 cm.) has been substituted
for the narrow one, and a method of determining the maximum pres-
sure by letting the cuff “feel” the return of the pulse has been
adopted. The mechanism of the instrument has been so simplified
that but a single stopcock is now required.
THE RELATION BETWEEN BLOOD-PRESSURE, PULSE-PRES-
SURE, AND THE VELOCITY OF BLOOD-FLOW IN MAN.
By JOSEPH ERLANGER anp DONALD R. HOOKER.
EXPERIMENTS performed with the sphygmomanometer show that,
upon changing from the recumbent to the standing posture, the
minimum pressure is increased and the pulse-pressure (difference
between maximum and minimum pressures) is diminished. Upon
assuming the sitting posture, these pressures almost return to the
values they had had while recumbent. Experiments performed with
the v. Kries tachygraph show that, in agreement with the changes
in pulse-pressure, the acceleration of the blood-flow per heart-beat
is greatest while recumbent, and smallest while standing. Further-
more, there is an inverse relation between the pulse-rate on the one
hand, and the pulse-pressure and the velocity of flow, on the other.
Under perfectly normal conditions, the product of the pulse-pressure
by the pulse-rate, therefore, tends to remain constant, as does the
product of the acceleration by the pulse-rate. Furthermore, in any
two postures, the pulse-pressures are to one another as the accelera-
tions per pulse wave. This is in accordance with the law governing
the flow of fluids through large elastic tubes.
Experiments tend to indicate that the explanation of the pressure
changes that accompany changes in posture is to be found chiefly in
changed hydrostatic conditions.
XVl Sixteenth Annual Meeting.
THE RELATION OF BLOOD-PRESSURE AND PULSE-PRESSURE
TO THE SECRETION OF URINE AND TO THE SECRETION
OF ALBUMIN IN A CASE OF SO-CALLED PHYSIOLOGICAL
ALBUMINURIA.
By JOSEPH ERLANGER anp DONALD R. HOOKER, °
EXPERIMENTS described in the preceding report have served the
purpose of studying the relation between the ‘“‘ pressures” and the
secretion of urine, the composition of the urine, and the secretion of
albumin in a case of physiological albuminuria. In all the experi-
ments performed, including the effects of posture, of baths in the
erect posture, of muscular exertion, of meals and of daily routine, a
distinct relation has been found between the magnitude of the pulse-
pressure and the amounts of urine and of albumin, an increase in the
pulse-pressure accompanying an increase in the amount of urine and
a diminution in the amount of albumin. The rate of secretion of
the urine and of albumin is apparently independent of pulse-rate
and of the minimum and maximum blood-pressures; and no relation
can be proved between the secretion of urine and of albumin, and the
product of the pulse-rate by the pulse-pressure.
Furthermore, experiments show that posture, and hence, presum-
ably, pulse-pressure, has a marked influence upon the composition
of the urine. The amounts of the chlorides, of the phosphates, and
of the total nitrogen, are much smaller in the standing than in the
recumbent posture, the variations in the amounts of the phosphates
and of the total nitrogen being smaller than the variations of the
chlorides.
ON CO-ORDINATION OF THE VENTRICLES OF THE HEART
By W. T. PORTER, ror C. FROTHINGHAM, Jr., anp W. E. LADD.
IN an investigation published in this journal, 1899, vol. ii, pp. 127—
136, W.T. Porter found “ That any portion of the ventricle will
beat synchronously with any other portion, so long as the two are
connected by muscle tissue, but that synchronism immediately fails
when the muscle bridge is broken, in spite of the fact that both por-
tions may retain their normal connection with the uninjured auri-
Proceedings of the American Phystological Soctety. xvii
cles.’ In 1899 von Vintschgau! observed that when a complete
physiological division of the frog’s ventricle is made by crushing the
ventricle lengthwise with forceps until the physiological continuity of
the two halves is destroyed, both halves, united only by auricular
tissue, beat synchronously. This discrepancy makes it desirable
(1) that the batrachian ventricle be subjected to the same method
of division used by Professor Porter on the mammalian ventricle,
namely, longitudinal section; (2) that Professor Porter’s investi-
gation be repeated, for his conclusion, though well supported by
numerous experiments, was after all negative; and (3) that similar
experiments be performed upon hearts in which the muscular con-
nection between auricle and ventricle is less general than in the
frog and tortoise, and at the same time more definite than in the
mammalian heart, so that the relation of co-ordination to the muscular
fibres uniting auricle and ventricle may be determined.
The present note deals with the first of these studies.
When the ventricle of the bull-frog or the tortoise heart is com-
pletely divided by an incision passing into the auricle, the separated
portions may still be observed to contract synchronously, although
their only muscular connection is through the auricle. Division was
made in different planes, but the results were the same in all cases.
THE PASSAGE OF DIFFERENT FOOD-STUFFS FROM THE
STOMACH.
By W. B. CANNON.
X-ray shadows cast by various foods (mixed with bismuth sub-
nitrate) afford a method of estimating the relative amount of food in
the intestines at different times and in different animals. As the
diameter of the intestines varies only slightly, the aggregate length
of the shadows may be taken to indicate the amount of food. The
faults of the method, due to absorption, to the intestinal loops not
being parallel with the fluorescent screen, and to variations in the
thickness of the food masses, are slight compared with the great
differences in the amounts of the different food-stuffs in the intes-
tine in the early stages of digestion.
In all cases the animals (cats) were given 25 c.c. of food, and the
1 Von VINTSCHGAU: Archiv fiir die gesammte Physiologie, 1899, Ixxvi, p. 59.
XVIll Sixteenth Annual Meeting.
different foods were as nearly as possible of the same consistency.
Tracings of the shadows on tissue paper were made at regular inter-
vals after feeding.
The following average figures illustrate the characteristic differ-
ences in the aggregate length (in centimetres) of the intestinal
content with the various food-stuffs :
Hours after feeding .
Fats (15 cases)
Proteids (10 cases)
Carbohydrates (14 cases) .
The remarkable difference between carbohydrate and _proteid
figures in the early stages can be accounted for by assuming that
free acid is the stimulus opening the pylorus. Both carbohydrates
and proteids provoke abundant gastric secretion (Pawlow). With
carbohydrate food there is free acid immediately; with proteid food
there is no free acid so long as acid unites with proteid. Thus the
discharge of proteids would be delayed. Feeding acid proteid and
also crackers wet with one per cent sodium bicarbonate solution gave
further evidence favoring this theory.
Hours after feeding .
Acid proteid
Alkaline carbohydrate .
Note that acid proteid leaves the stomach quite as rapidly as
carbohydrate food, and that alkaline carbohydrate, like ordinary pro-
teid, passes out slowly.
The addition of 0.4 per cent hydrochloric acid to carbohydrate food
does not increase the rapidity of the discharge from the stomach.
Hirsch and Serdjukow have noted that acid in the duodenum checks
the emptying of the stomach. Observation shows no cessation of
gastric peristalsis after food has passed the pylorus. The acid must
therefore act on the pylorus. Tying the bile and pancreatic ducts
greatly decreases the rate of discharge of carbohydrate food.
Evidence thus points to free acid in the stomach opening the
pylorus, and in the duodenum closing the pylorus. But acid in
- Proceedings of the American Physiological Soctety. xix
the duodenum stimulates alkaline secretions, and the acid is thereby
neutralized; whereupon the acid in the stomach again opens the
pylorus to allow more food to pass out. Thus proteids would be
retained in the stomach until acted upon by gastric juice, and thus
automatically the intestine would be guarded from being overwhelmed
with food and with secretions interfering with the intestinal ferments.
THE EMPTYING OF THE HUMAN” STOMACH.
By W. B. CANNON.
THE usual idea of the shape and position of the stomach is taken from
the figures of His and of Luschka, now copied in many text-books of
anatomy. The greater curvature in these figures reaches a point
considerably lower than the pylorus. Surgeons use this conception
in making the gastro-enterostomy opening, for purposes of “ drain-
age,” at “the most dependent part.” Conceptions of the shape and
position of the stomach based on appearances in the cadaver, or in a
living person relaxed in anesthesia, may not be true for the function-
ing organ.
Observations with the X-rays on a normal human stomach con-
taining food mixed with bismuth subnitrate show that while diges-
tion is proceeding, and the stomach is emptying itself, it shortens,
just as if the longitudinal and oblique fibres passing over the surfaces
to the greater curvature lifted the organ up toward the one fixed
point of the contracting fibres,— the cardia. Since the pylorus is
also more or less fixed, it does not rise with the rest of the stomach.
The consequence is that in the late stages of digestion, when the
gastric contents are more fluid than in the earlier stages, the pylorus
becomes the lowest point in the stomach, and the contents do not
therefore have to be lifted in order to be passed out.
SUPRARENAL GRAFTING IN THE KIDNEYS OF RABBITS WITH
SURVIVAL OF AN ANIMAL AFTER SUBSEQUENT REMOVAL
OF THE REMAINING SUPRARENAL.
By F. C. BUSCH anp C. VAN BERGEN.
Tue authors have made suprarenal grafts in a series of rabbits,
transplanting a section of the gland into the animal’s own kidney.
The experiment was successful, histologically, in several animals, and
Xx Szxteenth Annual Meeting.
physiologically, in one. Rabbits, where total ablation of the supra-
renals has been made, have invariably died from eight to ten hours
after the operation. One rabbit, from whom the remaining supra-
renal was removed eighty-three days after the introduction of a
graft, died ten hours after the operation. The graft was found to
have become entirely necrotic, and was surrounded by a thick layer
of connective tissue.
In three other cases, where the animals died from ten days to
two months after the introduction of the graft, either through an
anesthetic or other accident, a partial preservation of the supra-
renal was found upon histological examination. In contradistinction
to the results of other investigators, the authors found a survival of
medullary cells and a necrosis of the cortex. This may possibly have
been due to the manner in which the grafts were made.
In the single physiologically successful case (Rabbit X), at the
first operation, the left suprarenal was removed 77 foto. Longi-
section of the gland was made on each side, and the middle half of
the gland was introduced into an opening made in the cortex of the
lower border of the left kidney, a piece of the kidney cortex of cor-
responding size having been previously removed. The graft was
secured by silk ligatures. Eighty-six days later the remaining right
suprarenal was removed. The rabbit made an uninterrupted re-
covery and was apparently normal in all respects.
Twenty-one days after the removal of the remaining suprarenal
the rabbit was killed, in order to examine the condition of the graft.
It was found that both original suprarenals had been entirely re-
moved, without leaving any stump that might have carried: on the
function of the gland. No accessory suprarenals were found. The
remains of the graft, upon histological examination, were found in
the cortex of the right kidney just under the capsule, the estimated
thickness of the surviving graft being about 0.8 mm. The cells of
the graft appear to belong to the medullary portion of the supra-
renal. The rest of the original graft seems to have been replaced
by connective tissue. The vascular supply of the surviving cells
is good. .
Proceedings of the American Physiological Society. xxi
ON THE ABSENCE OF A CANE-SUGAR INVERTING ENZYME
IN THE GASTRIC JUICE.
By GRAHAM LUSKR,
In a former paper by Ferris and Lusk! the cane-sugar inverting
power of the gastric juice was attributed to free hydrochloric acid,
which was shown adequate to effect the results observed. The
experiments of Miura were accepted as indicating the absence of
inverting enzymes. Recently Widdicombe? has said that: gastric
juice contains an inverting enzyme. In consequence of this state-
ment, the following experiments were instituted : —
Dog I.— 1. Dog had fasted 48 hours. 1 c.c. of fresh gastric juice (stimulus
largely psychical, less than 5 grams of meat fed) was collected in drops
from a gastric fistula 40 minutes after the stimulus. Free acid present
indicated by tropzeolin oo. Mixed with 1 c.c. of a 5 per cent cane-sugar
solution, and kept at 37° C. Inversion in one hour indicated by the
reduction of Fehling’s solution.
2. 5 c.c. of the same gastric juice is permitted to digest fibrin until
there is no reaction for free acid, then 1 c.c. of a 5 per cent cane-sugar
solution is added, and the mixture put in a thermostat at 37° for 23 hours.
The proteid was precipitated by the alcohol method, and after neutraliza-
tion of the filtrate, and evaporation of the alcohol, no reaction for invert
sugar could be obtained.
Dog II.— 3. Fasting dog, anaesthesia by morphine and ether. Pylorus tied.
Gastric stimulus through 25 c.c. of a 25 per cent solution of alcohol
injected into the duodenum (method of Wallace and Jackson). One
hour later cardia ligated, stomach extirpated and put in thermostat 4}
hours. 15 c.c. of gastric contents found, no free acid present, but acid
tomitmuss Filtered. 3 c.c. of the filtrate, with 1-¢.c./of-a 5, per cent
solution of cane-sugar, after standing in the thermostat for 24 hours,
showed no inversion.
4. The gastric mucosa of the same dog, with about 10 c.c. of the above-
mentioned gastric contents, were digested for 24 hours with 200 c.c. of a
0.4 per cent hydrochlori¢ acid solution. Reaction shows free acid.
Fibrin is added, strong proteolysis takes place, and free acid disappears.
3 c.c. of this filtered digest, and 1 c.c. of a 5 per cent solution of cane-
sugar, showed no inversion after standing in a thermostat for 24 hours.
1 Ferris and Lusk: This journal, 1898, i, p. 277.
2 WIDDICOMBE: Journal of physiology, 1902, xxviii, p. 175.
XXII Sixteenth Annual Meeting.
Pig. — 5. A pig’s gastric mucosa was treated as above with rooo c.c. of 0.4
per cent hydrochloric acid. After 24 hours, no reaction for free acid.
15 c.c. of the filtered digest, to which was added 0.5 gram of cane-sugar,
showed no inversion after 24 hours in the thermostat.
é
The above experiments demonstrate that no cane-sugar inverting
enzyme exists in the gastric secretion, and confirm the belief that
such inversion of cane-sugar as takes place in the stomach is alone
due to the presence of free hydrochloric acid.
IMPROVED CAGE AND DIET FOR USE IN METABOLISM
EXPERIMENTS ON DOGS.
By WILLIAM J. GIES.
Cage. — After using cages of several types of construction during
the past few years, the author has improved a simple form by adding
various mechanical devices which ensure quantitative accuracy as
well as comparative convenience in the collection of excreta (urine,
feeces, hair).
Diet. —The improved diet consists of hashed meat, cracker meal,
lard and water, with a quantity of dove ash equivalent to that from
a moderate amount of bone. Bone ash is a very desirable addition
to the diet of dogs in metabolism experiments, and has already been
used in the experiments reported by Taltavall and Gies at the previ-
ous meeting of the society, and in the experiments reported by Hawk
and Gies at this meeting.
The addition of bone ash to the diet of dogs increases the bulk of
the fecal matter and makes its discharge more frequent and regular.
The faeces have the typical consistency and appearance of the fecal
matter eliminated from dogs subsisting on a diet containing bone.
Bone ash does not introduce into the diet anything that is injurious
to the dog, either mechanically or chemically. The fondness of dogs
for bones is well known. There is never any diarrhoea as a result of
the presence of the bone ash in the food. The faces are eliminated
in lumps that do not adhere to the cage, but are very easily and
completely removable from it. They dry very readily on the water
bath 7 a few minutes and may be quickly and easily ground to a
etal
Proceedings of the American Phystological Soctety. xxiii
fine, fluffy powder. Charcoal marks off period-faeces very distinctly
when bone ash is fed, because of the sharp color contrasts. Ten
grams of bone ash to a medium-sized dog in the average daily diet
suffices to produce the effects desired.
That the bone ash has little effect on the taste of the food or on
the dog itself, is evident from the fact that I have succeeded in
feeding large amounts of the material to several animals: one dog
weighing 17 kilos took daily as much as 100 grams of bone ash ad-
mixed with 250 grams of hashed meat, 70 grams of cracker meal,
30 grams of lard, and 500 c.c. water, without showing any observable
effects whatever, except more frequent and abundant defecation.
Although such a mixture, containing an excess of the bone ash,
looks much like powdered chalk and water, the dog ate it as readily
as it did the hash without it. The animal referred to was a well
nourished animal to begin with. Moderate amounts of bone ash do
not appear to interfere with digestion or absorption. I have not
yet determined the effect on excretion of earthy salts in the urine.
Nearly all of these substances in the bone ash seem to appear in the
faeces, however.
DEMONSTRATION OF WORKING MODELS OF THE
CIRCULATION.
By YANDELL HENDERSON.
In the first model, valves of sheet rubber anatomically similar to the
mitral and semilunar (together with a reservoir supplying water, a
rubber bulb serving as pump, and an air-chamber representing arte-
rial elasticity) are so arranged that the movements of the valves can
be observed under varied conditions.
The second model is a mechanical device consisting of a pump,
air-chamber, stop-cocks, by-passes, etc., designed to determine the
relative systolic volume, arterial elasticity, peripheral resistance, and
the factors incident to conditions of stenosis and regurgitation, when
the model is arranged to give pressure curves similar to intraventric-
ular and pulse tracings.
XX1V Szrcteenth Annual Meeting.
DEMONSTRATION OF RABBIT’S NERVES, SHOWING THE
EFFECT OF LIGATION UPON VITAL STAINING.
By S. J. MELTZER.
A SINGLE ligation of a nerve has no influence upon the staining of
the nerve on either side of the ligature. When, however, two liga-
tures are applied, the section of the nerve between the ligatures
remains free of color, while both ends are stained. This is the case,
even if the section between the ligatures comprises nearly the entire
length of the nerve.
ON THE ENZYME OF THE THYMUS.
; By WALTER JONES.
A xkILoGRAM of finely divided thymus gland was suspended in twice
its weight of chloroform water and allowed to remain for five days, at
the body temperature. The product was then treated with a few drops
of acetic acid, heated to boiling for the coagulation of the proteids,
and the filtered solution made strongly alkaline with ammonia, when
the usual precipitate of magnesium ammonium phosphate occurred.
This was filtered off, and the solution treated with silver nitrate
in ammonia. The profuse gelatinous silver precipitate thus pro-
duced was submitted to the scheme proposed by Kriiger and Solo-
mon for the separation of xanthine bases, and there were finally
obtained 2} grams of xanthine nitrate and 110 milligrams of hypo-
xanthine nitrate. As the hydrolytic products of thymus nucleic
acid which belong to this group are guanine and adenine, the follow-
ing experiment was undertaken to show that the xanthine in ques-
tion results from a decomposition of the nucleoproteid and not from
some hitherto unrecognized constituent of the gland. A kilogram
of prepared gland was extracted with water and the nucleoproteid
precipitated with acetic acid. This was purified by alternate solution
in sodium carbonate and precipitation with acetic acid until a neutral
opalescent solution was finally obtained, free from all constituents
of the gland except the nucleoproteid, and the enzyme which adheres
to it. This material was treated with chloroform and allowed to
digest at the body temperature. At the end of sixteen hours the
presence of xanthine bases could be shown. After a week, the pro-
Proceedings of the American Physiological Society. xxv
ducts of the digestion were examined by the method stated above.
1.7 grams of xanthine nitrate and 70 milligrams of hypoxanthine
nitrate were finally obtained. The production of phosphoric acid
was also shown.
ON THE ENZYME OF THE SUPRARENAL GLAND.
By WALTER JONES.
THE prepared gland was treated with four times its weight of chloro-
form water, and maintained at the body temperature for a week.
The product was then treated with acetic acid, heated to coagulate
the proteids, and the filtered solution evaporated to a small volume
under diminished pressure. The deposited sediment was found to
consist principally of xanthine, but contained also a small quantity
of hypoxanthine. The absence of other xanthine bases was definitely
shown. The nucleoproteid was then prepared by extracting the
gland with water and precipitating with acetic acid. By alternate
solution in sodium carbonate and precipitation with acetic acid
the material was freed from soluble constituents of the gland, and
finally placed in the thermostat in neutral solution with chloro-
form. At the end of a week the product was examined by the
method described in connection with the thymus gland, and with the
same results.
It is thus shown that there is present in the thymus, and also in
the suprarenal gland, an enzyme which exerts its activity on the
nucleoproteids of the glands giving rise to phosphoric acid and
xanthine bases, and that the xanthine bases formed under the influ-
ence of the enzyme are not the same as those formed when the
nucleoproteids are boiled with dilute acid.
THE PHYSICAL FACTORS CONCERNED IN. URINE
FORMATION.
By TORALD SOLLMANN (witH THE COLLABORATION OF R. A. HATCHER).
I. The excretion of chlorides by excised and perfused kidneys:
1. If saline solutions are perfused, the chloride-con'ent of the per-
fusing fluid and of the ureter filtrate are identical.
2. If mixtures of defibrinated blood and sodium sulphate are cir-
XXV1 Sixteenth Annual Meeting.
culated, the ureter filtrate contains almost as much chlorides as the
serum. The slight retention which occurs is different in kind as
well as in degree from the retention which occurs in the living ani-
mal. This “living retention” has been lost.
3. This is not due to changes in the chlorides of the serum pro-
duced in defibrination; for defibrination has no effect on the chloride
retention of living animals. It must be due to injury to the kidney
produced by very brief interruption of the renal circulation.
II. Changes occurring in the kidney during perfusion with one per
cent sodium chloride under uniform conditions.
These are shown to be mechanical, due to kinking of the renal
vessels. A period of two hours exists when the vein and ureter so
are almost precisely constant.
Ill. The influence of the injection pressure.
The volume of the kidney, and the vein and ureter flow, are par-
allel to the arterial pressure.
IV. The effect of compressing the renal vein.
Occlusion of the vein practically abolishes the ureter flow. This
begins quite suddenly when the vein-pressure reaches 50 cm. of water.
The vein flow stops suddenly when the vein-pressure reaches 70 cm.
of water. A kinking of the ureter and of the vessels must occur at
these pressures.
V. Results of occluding the ureter.
A sudden relative decrease of ureter flow occurs when the ureter-
pressure reaches 60 cm. of water. The urinary tubules must be partly
occluded by kinking at this pressure. The vein flow shows a compar-
atively small diminution, and the oncometer a small increase. The
maximal ureter-pressure is always less than the injection pressure.
VI. When the injection is made through the vein, but little solution
can be forced through the vessels.
VII. The vein and ureter Tow. are diminished by zuzcrease of vis-
cidity of the solution.
VIII. Concentration of the injection fluid. .
The flow from vein and ureter are parallel to the concentration of
the injection fluid. The effect is very great. The oncometer shows
definite oscillation; the final volume of the kidney is not greatly
altered.
IX. J/sotonic solutions, compared with one per cent sodium chloride.
The following produce definite changes, which are not, however,
very great: cane sugar, glucose, potassium, ammonium.
iw»
Proceedings of the American Physiological Society. xxvii
The following cause a very considerable diminution in vein and
ureter flow: urea, calcium, barium, acid, alkalies, carbonate. Mag-
nesium causes an increase. Urea also causes a decrease of kidney
volume. After flushing with salt solution, the vein flow recovers
with most of the solutions, but not the ureter flow.
X. Various substances added to two per cent sodium chloride.
Sodium fluoride and mercuric chloride (1 : 1000) diminish both
ureter and vein flow very greatly. No recovery occurs when pure
saline is circulated. Sodium arseniate and formaldehyde (1 : 1000)
produce absolutely no effect. A number of other substances which
were tried, had little, if any, action.
EXPERIMENTS ON THE PRECURSORS OF URINARY INDICAN,
FRANK P. UNDERHILL.
THE recent experimental observations of Ellinger and his coworkers
have made it probable that the indican of the urine does not arise
from nitrogenous products of tissue decomposition (as Blumenthal
and others have assumed), but rather owes its origin to the activity
of bacteria which form indol in the intestine. They have also
demonstrated that tryptophan may be a precursor of indol in putre-
faction. The discovery of the constitution of tryptophan as skatol-
amido-acetic acid by Hopkins and Cole, and their announcement that
the Adamkiewicz (glyoxylic acid) reaction of proteids is attributable
to the tryptophan group in the latter, has afforded the occasion for
the present experiments. Various proteid substances yield the Adam-
kiewicz reaction with different degrees of intensity, —in some cases
with entirely negative outcome. Of the substances which fail to give
the test, gelatin is the most familiar. Feeding experiments on dogs
have indicated, in conformity with theoretical considerations, a marked
decrease in the excretion of indican when gelatin is the chief nitro-
genous constituent of the diet. The relationship between the guadity
(as well as the quantity) of the nitrogenous foods ingested and the
indican output in the urine has been demonstrated; and thus a new
factor has been introduced into the consideration of the physiological
significance of indican formation. The details of the investigation
will soon be published.
XXVIl1 Sixteenth Annual Meeting.
THE INFLUENCE OF HEMORRHAGE ON PROTEID
CATABOLISM.
By P. B. HAWK anv WILLIAM J. GIES.
Tue authors have recently completed some experiments in this con-
nection on dogs brought to a condition of nitrogenous equilibrium.
Blood was withdrawn from the femoral artery, or one of its branches,
while the animals were under ether-chloroform anesthesia. The
influence of anzesthesia and of our usual operation, with or without
ligaturing the artery, was determined on each dog in order to check
the results of blood-letting under anesthesia. The operations were
conducted under aseptic conditions. The diet was always eagerly
taken and was the same as that referred to in another report at this
meeting by Dr. Gies. The periods of recuperation were from one to
two weeks in length. In one experiment the animal was under
observation continuously for eighty-five days, and was subjected to
five losses of blood equal each time to from 2.5 per cent to 3.5 per cent
of the body weight.
In spite of losses of blood-nitrogen varying between 8 and 15
grams, hemorrhages of about 3 per cent of the body weight caused,
among other effects, (1) diminished secretion and decreased specific
gravity of the urine at first, the reverse after twenty-four to forty-
eight hours; (2) a slight temporary increase in the amount of nitro-
gen and sulphur in the urine, and a decrease in the quantity of
excreted phosphorus.
The amount and consistency of the feces were unaffected. The
changes in the amounts of excreted nitrogen, sulphur, and phos-
phorus were registered solely in the urine.
In the experiments in which the effect of anesthesia with operation
was determined defore hemorrhage, the catabolic effects above re-
ferred to were almost as great. Anzesthesia with operation after
hemorrhage caused a reverse effect — decrease instead of increase.
Repeated hemorrhages from the same animal resulted in (1)
cumulative quantitative catabolic effects in harmony with those after
single losses of about 3 per cent of the body weight, and were fol-
lowed by (2) steady decline in body weight, and (3) gradual increase
in daily volume of urine, even when the animal ate the same amount
and kind of food as at the beginning of the experiment. (4) Nitro-
genous equilibrium seemed to be repeatedly established in a few days
canna wee
Proceedings of the American Physiological Society. xxix
after each hemorrhage, on successively lower planes, by the original
amount of food.
After successive hemorrhages at intervals of a few days the con-
tent of nitrogen and sulphur in the blood, as well as specific gravity
and number of erythrocytes, gradually diminished, whereas the leu-
cocytes steadily increased in number. The phosphorus content
remained about the same.
Experiments on the flow of urine directly from the kidney con-
firmed previous statements to the effect that a hemorrhage of
3 per cent of the body weight entirely stops urinary formation for
about a half-hour.
Our data confirm the general metabolic results obtained years ago
by Bauer and others, and disagree with the contrary conclusions
lately announced by Ascoli and Draghi.
NITROGENOUS METABOLISM AFTER SPLENECTOMY.
By LAFAYETTE B. MENDEL (wituH R. B. GIBSON).
AN opportunity to carry out a series of experiments on metabolism
in man after splenectomy has enabled the writers to control the
observations on splenectomized animals reported to the Society in
1899 by Mendel and Jackson. It was demonstrated by them, con-
trary to current opinions expressed in physiological literature, that
the spleen is by no means the chief organ involved in uric acid pro-
duction in the living body, if, indeed, it normally plays any part
whatever in this process. In the present case, a study extending
over many days was made of the composition of the urine under
known conditions of diet. The chief points of interest indicated by
the detailed protocols presented are: the normal character of the
curves of postprandial hourly excretion of uric acid and other nitroge-
nous constituents of the urine; the retention of chlorides during
febrile conditions; the undiminished capacity of the organism to
form uric acid from its precursors (purin bodies of the food); the
relatively large output of uric acid of “endogenous” origin on a
purin-free diet; the pronounced elimination of urobilin at times.
The writers are inclined to attribute some of these features to
impaired hepatic functions, of which the clinical data (ascites, cir-
XXX Szxteenth Annual Meeting.
rhosis, urobilinuria) give evidence. Should further studies likewise
indicate a high “endogenous” uric acid output in hepatic disease,
the quantitative determination of the uric acid of the urine after a
strict purin-free diet may prove of value in clinical diagnosis. A
complete record of the investigation will be published.
COEFFICIENTS OF DIGESTIBILITY AND AVAILABILITY OF
THE NUTRIENTS OF HOOD,
By W. ©. ATWATER.
Four years ago, an attempt was made to find coefficients which would
express the proportions of the nutrients of common food materials
which are digested and made available to the body when eaten in
ordinary mixed diet by people in good health.!. The data were found
in the results of ninety-three digestion experiments with men, in
which the amounts of protein, fats, and carbohydrates in the food
were compared with the corresponding materials in the intestinal
excreta. The latter include both the undigested residues of the food
and the residues of metabolic products. The coefficients of digesti-
bility, properly speaking, would be found by subtracting the un-
digested residues from the food. The coefficients of availability are
found by subtracting the total excreta from the total food. In
accordance with common usage, these coefficients of availability are
also designated as coefficients of digestibility. In the article referred
to, coefficients were given for different classes of food materials, as
meat, fish, and dairy products, making together one class, cereals a
second, dried legumes a third, sugars and starches a fourth, and so
on. From the data thus observed, factors were assumed for the total
food of ordinary mixed diet. Lately, the results of four hundred and
eleven such experiments, made during the past nine years, have been
collated and the coefficients of availability calculated? The factors
actually found in the average of all these experiments agree very
‘ ATWATER and BRYANT: Report of the Storrs (Conn.) Experiment Station,
1899, p- 73-
* These experiments. were made in a number of different laboratories, and
belong to an inquiry regarding the nutriticn of man, which is being carried out
in different parts of the United States under authority of the U. S. Department
of Agriculture.
Proceedings of the American Physiological Soctety. XXxX1
closely with those assumed from the smaller number, as appears
from the following comparison : —
COEFFICIENTS OF AVAILABILITY OF NUTRIENTS.
|
: ; Carbo-
| Protein. | Fat. hydrates.
per cent per cent per cent
Pemased factors; . . i 1s.» + « | 920 | 95.0 97.0
Factors found in average of 411 experiments | 91.1 | 94.8 | 96.8
The kinds of food used in these experiments were such that the
proportions of vegetable food materials were, on the whole, larger
than in what might be called ‘ordinary mixed diet” in the United
States. The coefficients for protein are smaller in the vegetable
than in the animal foods. The difference between the proposed
factor for protein, 92 per cent, and that found in these experiments,
QI.I per cent, may be easily accounted for by this difference in pro-
portions of food materials.
The inference is that the proposed coefficients represent very
nearly the actual average availability or digestibility of the nutrients
of ordinary mixed diet.
THE INFLUENCE OF HEMORRHAGE ON THE FORMATION
AND COMPOSITION OF LYMPH.
By E. R. POSNER anp WILLIAM J. GIES.
AFTER hemorrhage, as is now well known, fluid passes from the
tissue spaces into the blood-vessels, and the volume of the blood is
soon restored. At first the percentage content of water in the regen-
erating blood is higher than usual, at the same time its percentage
content of organic solids is subnormal. We have attempted to ascer-
tain the facts of post-hemorrhagic lymph flow and composition in
the region drained by the thoracic duct, having been led to this
research by the contradictory results obtained a few years since by
Tscherewkow and Hoche.
We have had four classes of experiments thus far, with controls:
the influence of hemorrhage (1) on the production of lymph in
XXX1l Sixteenth Annual Meeting.
(a) recently fed and in (2) fasting animals, and (2) on the effects
induced by lymphagogues, like (c) sodium chloride and (@) leech
extract.
Thus far after hemorrhage we have always observed diminished
flow of lymph from the thoracic duct, in recently fed as well as fast-
ing dogs. After a 2.5 per cent hemorrhage, the injection of 22 grams
of sodium chloride (dissolved in 75 c.c. H,O) into a dog weighing
28 kilos gave lymphagogic effects as decided within a few minutes,
as in a normal animal. The same effect was noted after a hemor-
rhage of 3 per cent of the body weight, followed by the injection of-
leech extract (27 c.c. of 0.8 per cent NaCl extract of eleven leech
heads into a dog weighing 19 kilos). In the latter cases the in-
creased production of lymph was noted after the hemorrhage had
previously much diminished its flow.
We have not yet observed any regularity in the proportion of
organic solids eliminated in the lymph under these conditions, except
after injection of lymphagogues. Without lymphagogues, hemorrhage
was followed in some instances by decrease of organic solids, in others
by increase, but in each of these cases the inorganic matter was the
same in amount throughout, and equal to the proportion of ash in
the blood.
The hemorrhages have amounted usually to about 3 per cent of
the weight of the animal. In some cases the effects of smaller and
repeated losses of blood have been ascertained.
Additional experiments are in progress, in which a study is being
made of the content of proteid, fat, and other constituents of the
lymph collected under conditions similar to those indicated above.
REPORT OF AN EXPEDITION TO: CRIPPLE CREEK a
PIKE'S PEAK-TO STUDY THE EFER@i@e
ALTITUDE ON THE BEOOD:
By GEORGE T. KEMP.
THE party was composed of Professor Kemp, Messrs. O. O. Stanley
and E. R. Hayhurst, Assistants in Physiology, C. E. Harris, Fellow,
Miss Henrietta Calhoun and Mr. E. L. Draper, graduate students.
Each had had at least a year’s experience in making the observations
assigned them, each was in good health, and each was the subject of
Proceedings of the American Physiological Society. xxxiil
a daily experiment, involving the following points: count of the red
corpuscles, count of the leucocytes, determination of the numerical
ratio of the blood-plates to the red corpuscles, determination of the
hzemoglobin, determination of the specific gravity, special microscop-
ical examination with one-twelfth oil immersion lenses of any inter-
esting structure which presented itself.
The party first took records at Champaign (700 ft.), then went
direct to Cripple Creek (9,400 ft.), thence to Pike’s Peak (14,200 ft.),
thence returned to Champaign.
The results given are from a composite curve made by averaging
the daily records on the blood of each of the six in the party. Where
fluctuations occur in the curve they represent fluctuations in the curves
of the majority of the party, not excessive variations in the curves of
one or two of the individuals.
Red corpuscles. — Upon going from Champaign to Cripple Creek
there was no change in the number of red corpuscles on the after-
noon of the day of arrival.
In two days the count rose from 5,100,000 to 5,900,000, and main-
tained this level for three weeks, fluctuating between 6,100,000 and
5,600,000.
At Pike’s Peak there was a rise of 400,000 the first day, then a
slight fall, and the level was maintained at about 6,000,000 for
eight days, —the duration of our stay.
From Pike’s Peak we returned to Champaign, and six days after
leaving the Peak the count had fallen to 5,400,000, and was still
falling.
It is a known fact that morning counts of red corpuscles run
slightly higher than afternoon counts. On Pike’s Peak these dif-
ferences were regularly enormous, amounting in individual cases to
1,000,000. It is important to remember this in counting the blood-
corpuscles of patients treated at high altitudes.
From the end of the first week at Cripple Creek microcytes were .
present in sufficient number to attract attention; later they became
numerous.
Nucleated red corpuscles were observed in isolated instances; they
were only exceptionally present, and always too few to count.
The changes in the blood-plates are more marked than those in the
red corpuscles. This is true both qualitatively and quantitatively.
The change in the ratio of the blood-plates to the red corpuscles is
more striking than any other change in the blood; it is greater in
XXXIV Sixteenth Annual Meeting.
degree, more regular in progression, and of longer duration than the
change in the number of the red corpuscles.
This was observed, without exception, in the blood of six healthy
individuals, so it practically represents a law.
It is fair to assume that this may be taken as an index to the
changes normally produced in the blood by altitude, and may be
used to determine whether the blood of anzmics treated at high
altitudes is following normal lines of improvement.
At high altitudes the fluctuations in the count of the red corpuscles
at different times of the day is enormous. The ratio of the blood-
plates to the red corpuscles is not affected by these fluctuations in
any marked degree.
The ratio of the blood-plates to the red corpuscles may be deter-
mined more easily and with smaller error than the count of the red
corpuscles by the hemocytometer. This ratio cannot be determined
in dry preparations, owing to the adhesion of the plates.
The first change indicating a connection between the blood-plates
and the red corpuscles was the presence of. a considerable number of
large plates, some of them closely approximating the size of red cor-
puscles. They were mostly oval in form, delicate (thin) in structure,
and colorless. At about the same time the number of small red cor-
puscles became so large as to attract attention. Later (about ten
days after reaching Cripple Creek), colored blood-plates began to
appear, and were more or less in evidence during our stay at high
altitudes, but were most numerous during the second and third week.
In the same drop of blood could be seen all intermediate stages be-
tween the plates and the reds.
The blood-plates were tested from time to time by Deetjen’s
method, with sodium hexaphosphate on agar. They were seen to
throw out the processes which Deetjen called amoeboid. This was
also true of the delicate plates containing haemoglobin, but was not
true of the thicker red corpuscles, either young or old.
Altitude appeared to have no effect on the leucocytes, either as to
number or kind. This would indicate that the plates are not related
to the leucocytes.
The amount of hemoglobin in the blood is a very unsafe index for
the number of red corpuscles.
In the long run, and in a general way, the haemoglobin may be said
to go with the red corpuscles, but the daily variations were often in
the opposite direction. At Cripple Creek the rise in the amount of
Proceedings of the American Phystological Society. Xxxv
haemoglobin was fully twenty-four hours in advance of the increase
in the number of the red corpuscles.
The daily fluctuations in the hemoglobin were much less than the
daily fluctuations in the red corpuscles, showing that when fewer red
corpuscles were circulating each carried an increased per cent of
hemoglobin. This point, taken in connection with the morning and
afternoon fluctuations in the number of red corpuscles, suggests a
new function for the spleen. It seems incredible that 1,000,000 cor-
puscles per cubic millimetre of blood should be built up and broken
down daily. It is far more likely that they are temporarily withdrawn
from the circulation. It is an old observation that many faintly
colored red corpuscles are often found in the spleen. Our observa-
tions show over and over again that the amount of haemoglobin per
corpuscle varies in the same blood from day to day; hence we infer
that it is lost by one corpuscle and taken up by another, and we
suggest that this transfer may take place in the spleen.
Parallel observations on the specific gravity, haemoglobin, and red
corpuscles show that the increase, per cubic millimetre, of the latter
cannot be accounted for by different degrees of concentration of the
blood.
It is a disputed point as to whether the specific gravity can be
taken as an index to the number of red corpuscles. Our observations
show repeatedly that it cannot, especially where such variations occur
as are observed at high altitudes.
EFFECT OF INTRAVENOUS INFUSION OF SODIUM BICAR-
BONATE AFTER SEVERE HEMORRHAGE.
By PERCY M. DAWSON.
THE fluids infused contained sodium chloride and sodium bicarbonate
in varying amounts (0.5 to 0.8 per cent and o.1 to 1.0 per cent re-
spectively). During anesthesia with morphia and ether, the animals
(dogs) were bled through the carotid artery and then infused through
the jugular vein.
In recording the systolic and diastolic pressures, the exclusive use
of the valved or of the Hiirthle manometer was found to be unsatis-
factory. The former, though accurate, is slow, and cannot be used
for obtaining pulse-curves, while the latter (under the conditions
XXXV1 Sixteenth Annual Meeting.
present in these experiments) is often quite unreliable. The author
therefore connected the “ Hiirthle” with one femoral artery and
the valved manometer with the other, and both instruments with
the same pressure bottle, so that at any time the readings of the two
might be compared.
From the data obtained, it is seen that solutions containing sodium
bicarbonate bring about an increase in cardiac action, thereby restor-
ing the blood-pressures (systolic and diastolic) to a higher level than
when sodium chloride alone is employed. When a solution contain-
ing 0.8 per cent NaCl and 0.25 per cent NaHCO, is infused slowly,
the beneficial action persists for a considerable time.
RICIN.
By T. B. OSBORNE anv L. B. MENDEL.
THE proteins of the castor bean are an albumin, a globulin, and a
proteose. The two latter, when completely separated from the al-
bumin, are without toxic action. These two bodies constitute much
the greater part of the protein matter of this seed, although the
amount of albumin is not inconsiderable. The globulin was sepa-
rated from the saline extracts of the seed by dialysis, the albumin by
saturation with magnesium sulphate (some albumins of vegetable
origin are precipitated by saturating their solutions with magnesium
sulphate), and the proteose by precipitation with alcohol, after free-
ing the solution from salts by dialysis.
The toxicity of the purified albumin was very great, 0.002 milli-
gram per kilo being sufficient to cause the death of rabbits when
subcutaneously injected, or ,!; the minimal quantity required in the
intravenous introduction of Cushny’s preparation. The post-mortem
appearance of the tissues was characteristic in every case. The ricin
preparation also possessed very marked agglutinating and sediment-
ing properties towards mammalian red corpuscles.
Our preparation of ricin was completely soluble in distilled water,
coagulated slowly between 70° and 80’, gave all the characteristic
reactions of proteins in a very pronounced manner, contained C 49.02
per cent, H 6.80, N 14.54, and throughout behaved exactly like a
protein body.
Proceedings of the American Phystological Society. xxxvil
These results in the main confirm those obtained by Cushny; and
considered in connection with the minimal toxic dose, they lend little
support to the recently advanced view that the toxic substance is a
non-proteid body. It is our immediate intention to prepare a much
larger quantity of this most powerful poison, if possible in a purer
condition, in order to obtain more positive evidence respecting its
true nature.
THE EFFECTS OF A SUBCUTANEOUS INJECTION OF ADREN-
ALIN ON THE EYES OF CATS WHOSE SYMPATHETIC
NERVE IS CUT, OR WHOSE SUPERIOR CERVICAL GANG-
LION IS REMOVED.
By s> J. MELEOZER:
WHEN the sympathetic is cut, a subcutaneous injection of adrenalin
causes a retraction of the nictitant membrane, and no change is seen
in the size of the pupil or the width of the palpebral fissure. When,
however, the superior cervical ganglion is removed, an injection
causes a strong dilatation of the pupil, a considerable widening of
the palpebral fissure, and a retraction of the nictitant membrane.
ON THE INFLUENCE OF ETHER ANAESTHESIA.
By P. B. HAWK.
THREE phases of the subject have been studied as follows: (1)
Glycosuria following ether anzsthesia; (2) Changes in the blood
produced by ether anesthesia; (3) Influence of ether anzsthesia
upon urine flow, and the excretion of nitrogen and chlorine.
The experiments were all made upon dogs, the length of the ex-
periments varying from twelve to thirty-nine days. On normal ani-
mals the plan was to bring the organism to nitrogen equilibrium by
means of a suitable diet, then induce ether anzsthesia for periods
of various lengths. The periods thus far investigated have ranged
in length from thirty minutes to four and a half hours.
A slight diuresis invariably followed, and accompanying this diu-
resis was a small increased nitrogen output and a large increase in
XXXVill Stxteenth Annual Meeteng.
the chlorine excretion. Glycosuria always occurred after ether
anzesthesia.
In general, the changes noted in the blood after anesthesia were
an increase in the number of red corpuscles, and a lesser increase
in the number of leucocytes; the haemoglobin was more irregular.
In the course of a few hours, the red corpuscles became normal in
number, whereas the leucocytes increased rapidly and produced an
extensive leucocytosis in from three to five hours.
Cannulas inserted in the ureters of normal dogs showed a very
slow urine flow during anzsthesia periods of from six and one-half
hours to eleven and one-half hours. In one case the flow entirely
ceased in three and one-half hours. The urines voided in these
experiments showed a very high content of nitrogen and sugar.
The red corpuscles were increased from 30 per cent to 50 per cent
in these experiments, and the leucocytes were greatly decreased in
number. The body temperature of a dog at the end of an anzsthesia
period of eleven and one-half hours was 23° C.
Experiments have been made on animals subjected to various
periods of fasting. In the case of a dog etherized, on the twenty-
second day of fasting a strong diuresis was noted, but, contrary
to the conditions obtaining in normal animals, this diuresis was
unaccompanied by any signs of glycosuria. The changes produced
by ether anzsthesia in the blood of dogs during periods of fasting
of from one to twenty-two days, are similar to the changes produced
in the blood of normal animals, z.¢., an increase in red corpuscles
and the accompanying leucocytosis.
THE END-PRODUCTS OF SELF-DIGESTION OF ANIMAL
GLANDS.
By SR? Ay -EEVENE.
THE results of the analysis of the crystalline end-products of self-
digestion of the pancreas gland and of the liver are presented in this
communication. Thus far only the amino acids were analyzed. The
pancreas was subject to self-digestion in a 0.5 per cent solution of
sodium carbonate, the liver in 0.2 per cent solution of acetic acid.
Alanin, amino-valerianic acid, leucin, glutamic and aspartic acids,
tyrosin, and phenylalanin could be identified. The presence of
a-pyrolidin-carbonic acid could not be established with certainty.
Proceedings of the American Physiological Society. xxxix
THE END-PRODUCTS OF TRYPTIC DIGESTION OF
GELATINE.
By PP. AL LEVENE:
THE object of the investigation was to compare the composition
of peptone with that of native proteid and with that of proteoses. It
was found that the molecule of gelatose contained more glycocal
than that of gelatine. Gelatine peptone contained less glycocal than
gelatose. In order to explain these observations, the crystalline pro-
ducts of digestion were studied.
There were found glycocal (in very large quantities), leucin, glu-
tamic acid (in smaller quantities), phenylalanin and a substance of
the composition of inactive pyralidin-carbonic acid :
Calculated for Found
(C49H,gN.0,)CuH,O
ET ON er aes Sie nh eee OOO te 8 ar as, creel
For the dry substance:
(Che t+ eae ileaeys Dene AWG) s 2 22's 40166
Le eek oe MCT CORE, once or oe ok emcee ee 7A0)
IN eee pees oe. 9 Reet OO2M me aout eee OU),
Oh ee eae ty areeape 2/4 lel (6) eee, ai bey) 0)
The copper salt differed in appearance from that of the a-pyralidin-
carbonic acid,
AN IMPROVED KYMOGRAPH.'
By W. T. PORTER.
THE improved kymograph (Fig. 1) consists of a drum revolved by
clockwork and also arranged to be more rapidly revolved or “ spun”
by hand.
The drum is of aluminium, cast in one piece, turned true in the
lathe to a circumference of 50 cm. The height-is 15.5 cm. The
weight is about 600 grams. In each head of the drum is placed a
thin steel plate pierced to admit a steel shaft about which the drum
revolves and on which it may be held at any desired height by a
spring clip. The steel shaft passes through both the heavy plates
1 I am indebted to Mr. C. E. Roy, foreman of the machine-shop, for valuable
assistance in the improvement of the kymograph.
xl Stxteenth Annual Meeting.
containing the clockwork and is securely bolted to the bottom plate.
The motion of the clockwork is communicated to the drum by a
brass sleeve surrounding the lower part of the steel shaft and
fastened to the intermediate or sleeve-gearwheel shown in Fig. I.
eer,
FIGURE 1.
The sleeve is crowned with a
disk upon which rests a brass
block, carrying a steel rod
which passes through an
opening in each head of the
drum, and finally is bent over
until its blunt steel point
rests in a shallow cup on the
head of the steel shaft. The
lower end of the steel rod is
held fast in the brass block
by a horizontal screw. The
brass block is held fast to the
disk of the revolving sleeve
by a vertical screw, the head
of which presses against the
under surface of the disk.
When these screws are set,
the revolving sleeve, the brass
block, the steel rod, and the
drum form one rigid piece,
the entire weight of which is
suspended on the point of the
steel rod where it bears on
the top of the steel shaft.
Friction on the steel shaft
occurs only at the point of
the steel rod, at the thin
steel plate in each head of
the drum, and at the bearing
surfaces of the sleeve, which
are made as small as possible.
When the vertical screw is given several full turns from right to
left so that its head no longer touches the disk of the revolving
sleeve, and the side screw is given a half turn from right to left, the
brass block may be raised until it no longer rests on the sleeve-disk ;
Proceedings of the American Physiological Society. xii
it may be held in its new position by a half turn of the side screw
from left to right. The brass block, steel rod, and drum will now be
free of the sleeve, and will rest suspended on the point of the steel
rod. The drum may now be “spun” by hand. A single impulse
will cause the drum to revolve more than a minute, making more
than one hundred revolutions. The speed in any one revolution,
except at the beginning and the end of the series, will be practically
uniform.
The clockwork consists of a stout spring about six metres in
length, driving the chain of gears shown in Fig. 1. A slow and a
fast set of speeds are provided. The slow speed is obtained by lower-
ing the sleeve until its collar is flush with the plate covering the
clockwork, lowering the brass block until it rests on the sleeve-disk,
fastening the block to the steel rod by a half turn of the horizontal
screw, and turning the vertical screw until its head presses against
the sleeve-disk. The pinion of the gear shown on the extreme right
of Fig. 1 now engages with the gear of the sleeve and transfers to it
the motion of the spring-gear. The slow set of speeds gives place to
the fast set, when the horizontal screw is released from the steel rod
by a half turn of the screw from right to left, the block with the
attached sleeve moved upward as far as possible on the steel shaft,
and the sleeve and block secured in their new position by a half turn
of the horizontal screw from left to right. The pinion of the gear
on the right now no longer engages with the sleeve-gear, but runs
“idle,” and the sleeve-gear engages with the spring-gear directly.
To pass from the fast to the slow speeds the horizontal screw is
loosened, the block and sleeve dropped as far as possible, and the
screw tightened again.
These operations are easily and rapidly performed, though, as in all
gear mechanism, an instant’s pause is sometimes required to enable
the gear teeth to engage. The clockwork should be in motion, with-
out the fan, when the adjustments are being made.
With both fast and slow gearing four fans of different areas may
be used. They are slipped upon an extension of the last pinion shaft
in the chain. Five slow and five fast speeds (exclusive of spinning)
are thus obtained. An additional slow speed (50 cm. per hour) may
be obtained with a very large fan. All speeds are regulated by a
governor consisting of two heavy metal wings fastened to the same
shaft that carries the fan. With one winding, the drum will revolve
from about one to about seven hours, or longer, depending on the fan
employed.
xlii Srxteenth Annual Meeting.
RESPIRATION SCHEME.’
By W. I. PORTER.
Tue glass cylinder (Fig. 2) represents the thorax. The surface of
the water in the glass cylinder represents the diaphragm and movable
chest walls; its level may be changed by raising or lowering the
large rubber tube, in the free end of which is placed a second glass
cylinder, not shown in Fig. 2. The interior of the cylinder above
the water represents the thoracic
cavity, and the rubber balloon
the lungs. The paraffined cork
is pierced by a pleural and a
tracheal tube. The upper end of
the pleural tube enters a rubber
tube in the wall of which is a
small hole, closed by a short
glass rod. Through this hole
the pleural cavity may be opened
to the atmospheric ain ie
tracheal tube opens below into
the lung, above into a rubber
tube, in the wall of which is a
small opening, which represents
the glottis, and which may be
partly or wholly closed by a glass
rod. The left manometer shows
the intra-thoracic pressure, the
right manometer the intra-pul-
monary pressure. The normal relations between intra-thoracic and
intra-pulmonary respiration may be reproduced with this apparatus.
The pressure changes in forced respiration, obstructed air passages,
asphyxia, coughing, sneezing, hiccough, and perforation of the pleura
may also be studied.
FIGURE 2.
1 IT am indebted to Mr. Frederick Haven Pratt for assistance in the details of
this design.
Proceedings of the American Physiological Society. x\iii
“MUSCLE WARMER.”
By W. T. PORTER.
A DISK, supported by a rod, bears three pins (Fig. 3). One of the
three pins is prolonged and bent at a right angle near its lower end.
To the bend is fastened one end of the muscle under experimentation.
About the other end is tied a fine copper wire
which passes through a hole in the disk to reach
a muscle lever. A second opening in the disk
is provided with a short metal tube, in which a
thermometer is held by a piece of rubber. The
bulb of the thermometer may be placed on a
level with the belly of the muscle. When these
adjustments are complete, a glass cylinder is I
brought against the under surface of the disk,
where it is held in position by the ‘‘ spring” of \h)
the three pins. A beaker or other vessel con-
taining water is now placed beneath the cylinder \1
and raised until the cylinder is sufficiently im- =
mersed. The temperature of the muscle is
altered by heating or cooling this water. . Direct electrical stimula-
tion of the muscle may be made by connecting one electrode with
the metal parts of the apparatus and the other with the copper wire
attached to the upper end of the muscle.
FIGURE_3.
THE DELINEATION OF THE Moror CORTEX IN THE Doc. By H. CusHINe.
THE SIMULTANEOUS ACTION OF PILOCARPINE AND ATROPINE ON THE DEVEL-
OPING EMBRYOS OF THE SEA-URCHIN AND STARFISH. By T. SOLLMANN.
This journal, 1904, x, pp. 352-361
A METHOD OF DEMONSTRATING THE LOCALIZATION OF POTASSIUM IN ANIMAL
AND VEGETABLE CELLS. By A. B. MACALLUM.
A New Heap HOo.Lper For Raspirs. By FREDERIC S. LEE.
DEMONSTRATION OF EXPRESSIVE MOTIONS IN A DECEREBRATE ANIMAL. By
R. S. WoopworTH.
xliv Sixteenth Annual Meeting.
DEMONSTRATION OF THE EFFECTS OF SUBCUTANEOUS INJECTION OR SUBCON-
JUNCTIVAL INSTILLATION OF ADRENALIN UPON THE PUPILS OF RABBITS
WHOSE CORRESPONDING SUPERIOR CERVICAL GANGLIA ARE REMOVED.
By S. J. MELTzer. |
ON THE NUCLEOPROTEIDS OF THE Brain. By P. A. Levene and L. B.
STOOKEY.
FURTHER CONTRIBUTIONS TO MuscLe Tonus. By T. A. Srorey and W. T.
PORTER. :
To be published shortly in this journal.
‘
THE
American Journal of Physiology.
VOL. X. SEPTEMBER 1, 1903. NO. I.
NOTES ON THE HEART ACTION OF MOLGULA
MANHATTENSIS (VERRILL).
By GEORGE WILLIAM HUNTER, JR.
NVESTIGATION undertaken by the writer on the nervous system
of Molgula manhattensis (Verrill) shows conclusively the exist-
ence of a set of connective fibres between the groups of ganglion cells
situated on or near the distal ends of the heart tube and the central
nervous system.! The course of these fibres will be treated at length
in a later paper.
The purpose of this paper is to indicate some of the observed phe-
nomena which point toward the physiological connection of the heart
and central nervous system in Molgula manhattensis (Verrill).
HISTORICAL.
It is not the place of this article to treat exhaustively the subject
of cardiac activity. Recent investigation on both invertebrate and
vertebrate material seems to demonstrate the myogenic activity of
heart-muscle.
The presence of regulative nerves in the vertebrates, both accel-
erators and depressors, is too well known to need comment. With the
exception of one of the lower fishes,” these nerves seem to be present
throughout the vertebrate group. Accelerator and depressor fibres
have also been demonstrated either anatomically or physiologically,
1 HUNTER, G. W., Jr.: Anatomiseher Anzeiger, 1902, xxi, p. 241.
2 GREENE, C. W.: This journal, 1go2, vi, p. 318.
2 George Wilhiam Hunter, Jr.
in several groups of the invertebrates, proofs being especially strong
in the Crustacz and Mollusca.1
The actual presence of a regulative apparatus for the heart in the
Tunicata has never been proved, although the physiological work of
Kruckenburg,? Lahille,? Lingle, Loeb® and others seem to indicate
the existence of such an apparatus. These researches, together with
that of Schultze,® will be treated when under direct reference.
Tne NormMAL HEART-BEAT OF MoLGuLa MANHATTENSIS.
It was the endeavor of the writer first to obtain some light on the
normal heart-beat of Molgula. Four factors must here be taken
into consideration: first, the normal rate of the heart-beat in an ad-
visceral direction 7; second, the number of heart-beats in an abvisce-
ral direction; third, the number of pulsations in either given direction
before reversal; and, fourth, the duration of the sest period just
previous to a reversal of the heart.
The normal ab- and advisceral rate of beat. — The following data
were obtained by counting the heart-beat in each individual examined,
for a period of time at least ten minutes in duration. An average
1 DoGIEL, JEAN: Archiv fiir mikroskopische Anatomie, 1877 xiv, p. 59;
YunG, E.: Archives de zoologie expérimentale, 1881, ix, p. 421; CONANT and
CLARK: Journal of experimental medicine, 1896, i, p. 341; YUNG, E.: Archives
de zoologie expérimentale, 1878, vii; DoGiEL: Comptes rendus, 1876, Ixxxii,
pp- 1117, 1160; Ransom, W. B.: Journal of physiology, 1884, v, p. 261;
FosTER, M., and DEw-Smiru, A. G.: Proceedings of the Royal Society, London,
1875, p- 318; F. Borrazzi and P. ENRIQUES: Archives italiennes de biologie,
1900, Xxxiv, p. 111; R. A. BuDINGTON: Unpublished work on Mollusca.
* KRUCKENBURG, C. Fr. W.: Vergleichend-physiologische Studien, zu Tunis,
Mentone, und Palermo, 3 Abt., Heidelburg, 1880.
® LAHILLE, F.: Contributions a l’étude anatomique et taxonomique des Tuni-
ciers, Dissertation, Paris, 1890.
* LINGLE: wide LOEB, J.: Comparative Physiology of the Brain and Com-
parative Psychology, Igoo.
° LoEB, JACQUES: Einteilung in die vergleichende Gehirnphysiologie und
vergleichende Psychologie mit besonder Berucksichtigung der wirbellosen Tiere,
Leipzig, 1899; Comparative Physiology of the Brainand Comparative Psychology,
1900.
® SCHULTZE, L. S.: Zeitschrift fiir Naturwissenschaften, Jena, 1901, xxviii,
pr zer
7 The use of the terms “ad- and ab- visceral” is taken from SCHULTZE. By
an advisceral contraction, we mean that the wave of contraction in passing over the
heart moves in the direction of the viscera; an abvisceral contraction moves in
the opposite direction. The terms endostylar and rapheal might be substituted for
the above-given terms.
On the Heart Action of Molgula Manhattensis. z
mean pulsation rate was thus obtained for each individual. Three
lots of Molgulz were examined, between twenty and thirty specimens
in each lot, at intervals of time separated by two weeks. Labora-
tory conditions were kept as nearly alike as possible for all animals.
The water temperature was between 21° and 22° C. For purposes
of comparison, the Molgulz were divided into three groups marked
large, medium, and small, respectively.
ABVISCERAL.
Time per 100 seconds.
ADVISCERAL.
Time per 100 seconds.
Size of
speci-
Slowest
observed
pulsation
rate.
Fastest
observed
pulsation
rate.
59 : 100
47 : 100
98 : 100
Average
normal
pulsation
rate.
i522 100
105 : 100
Slowest
observed
pulsation
rate.
Fastest
observed
pulsation
rate.
Average
normal
pulsation
rate.
54 : 100
99 : 100
74: 100
72: 100
52: 100
82 : 100
67 : 100
50: 100
90 : 100
G22 MOO
43:2 + 1m.
Average for all
Average for all
specimens
specimens.
The average mean normal rate of pulsation obtained from these
results (together with all other specimens examined) gives the fol-
lowing for all examined Molgulz: 43.2 per minute in an advisceral
direction, and 43.2 per minute in an abvisceral direction, with a rest
period of two seconds between the pulsation periods..
An unexpected feature of the above table is that it shows the
smaller individuals to have a slower pulsation rate than those of
larger size, and of presumably greater age. This is contrary to
Schultze’s findings regarding the rate of the heart-beat in species of
Salpze where the body size is a noticeable factor. I personally have
not investigated this point in the species of Tunicata available at
Woods Hole, where my investigations were carried on. For our pur-
pose, such results may be disregarded, and only the general average
obtained from some eighty specimens need to be used.
It is seen, however, that very great individual variations are the
4 George Wilham fLunter, Jr.
rule, and such results as are tabulated above may be of little value in
comparison.
The general agreement of the above results with those of Schultze
may be found in the following statement (page 233): ‘‘The result
of my observations was that the frequency of the abvisceral] and ad-
visceral pulsations is in general the same.” Kruckenburg, however,
finds a difference between the frequency of the ab- and advisceral
pulse in certain of the Salpz.
What is the normal number of heart-beats that take place ina given
direction before a reversal, or during a single pulsation period? Cana
normal average of the number of pulsations be established, and, when
once established, is this number constant for any length of time ?—In
answer to the latter question, it must be said that constant change is
the rule. Many factors, the discussion of which would not be in
place here, cause a frequent, and, ofttimes, erratic change in the
time (duration) of the pulsation periods in either the ab- or ad-
visceral directions. In making observations with the view of obtain-
ing the average length of time occupied in a given pulsation period
in either direction, time enough must be allowed in the series of
observations so that the above-mentioned changes can be averaged
fora general mean. In making the observations noted in the fol-
lowing tables, each animal was watched, and the heart-beats counted,
for at least fifteen minutes in succession, more frequently for from
half an hour to an hour; the rate and number of heart-beats were
entered in tabular form, and an average made from the total number
of ab- and advisceral beats, respectively. Frequently the same heart
was counted at a later time, and the results then obtained were com-
pared and averaged with the first record. In this way, an estimate
could be formed of the average duration of a pulsation period. The
following table, based on the results obtained from the comparison of
forty-three Molgulz, is self-explanatory.
TABLE SHOWING NUMBER OF PULSATIONS BETWEEN REVERSALS IN THE NORMAL
HEART OF MOLGULA MANHATTENSIS.
Over 200.|/200to150.}}150 to 100.|| 100 to 50. ||50and under.
Ab. | Ad.|| Ab.| Ad.|] Ab: | Ad.
No. of specimens 2
Percentage. . . | 4. ; 44.2) | 44.2) 25.6) | Zoe
On the Fleart Action of Molgula Manhattensts. 5
A second lot of thirty-eight Molgulz gave the following results:
Over 200.|/200to150.|/150 to 100.|; 100 to 50. |'5Oand under.
Ay ii Ades i Atb:) || Aid:
Ab. | Ad.
No. of specimens | 1 Ws ee 4 g, 9 13 13 1] 11
34.2 | 34.2 || 28.8 | 28.8
Percentage.
Observations by Kruckenburg and Schultze along this line point out
(as do the tables given above) the great variability in the length of
the pulsation period. Examples of the heart beating in one given
direction much longer than in the opposite direction are noted by the
above-mentioned writers. . This lack of balance in the duration of
the alternating pulsation periods has been noted for many cases in
Molgula.
It is the exception, rather than the rule, to have a pulsation
period in an abvisceral direction exactly balance the next succeeding
pulsation period in an advisceral direction. It is only when a large
number of heart-reversals are taken into consideration that we find
the number of beats in the two series approximating each other in
duration and in the number of alternating beats.
What is the duration of the rest period between reversals of the heart
in Molgula ?— The rest period is also a matter of great irregularity,
as is shown by the following table:
Rest period, || Rest period, || Rest period, || Rest period,
under 1 sec. 1-2 secs. 3-5 secs. over 5 secs.
Abv. | Adv. || Abv. | Adv. |} Abv. | Adv. ee Adv.
No. of specimens 18 18
Refcentage. . 7. 40 45
Forty-two and five-tenths per cent of all observed specimens had a
rest period of between one and two seconds duration. The writer
believes this condition to represent a normal condition; while the
rest period of very long or of very short duration is a pathological
indication. A long, and in some cases, at least, a short rest period,
is seen in animals that have suffered from the effects of long captivity
or from unfavorable laboratory conditions.
6 George Wilham Hunter, Jr.
The findings of Schultze are in accord with mine regarding the
length of the rest period. He, too, finds exceeding irregularity in
the length of the rest period.
Is the rhythmical activity of the heart of Molgula myogenic im
origin 2— The experiments of Lingle, quoted by Loeb,’ seemed to
show that the heart of Molgula, when cut into two pieces, would
beat continuously from the uncut ends. If a piece was cut out of
the middle of the heart, the ends would continue to beat, but the
middle piece would remain passive. These results, obtained over
ten years ago, have since been verified several times by the pupils
_in Loeb’s physiology classes at Woods Hole, and by myself in the
summer of 1900.
Schultze, however, after a series of careful experiments, came to
the conclusion that when the heart of certain of the Salpze, and that
of Ciona intestinalis, was cut into small pieces, these pieces would
continue to contract rhythmically.
These findings led the writer to make a more careful series
of observations on the heart of Molgula, with a view of deter-
mining this point. The heart, with the surrounding tissue, was
removed from the body and left in sea-water in covered glass
dishes until after the shock effect had worn off. After beating was
completely re-established, the two ends of the heart were cut away
and the middle piece left in the dish. Frequent observation of
these middle pieces revealed no immediate beating in any of the
specimens. In a small percentage of cases, however, pulsations of
a more or less rhythmical character were observed. Furthermore, if
the middle piece of the heart was cut up into smaller pieces, each
piece less than 2 mm. square, and proper precautions taken, the
smaller bits of heart-tissue were observed to beat. In all the observed
cases, the rate of pulsation was much less than the normal rate, or
that of the cut ends. When the hearts were cut into small pieces,
these pieces were usually from one to two millimetres in width, and
extended completely around the heart tube, thus forming small seg-
ments of that organ. In the smallest of these pieces the pulsations
were usually of a very irregular character, and occurred at infrequent
intervals, sometimes from one to two minutes elapsing between pul-
sations. Beating of the fragments of the above-mentioned material
has been noticed five hours after operation; and in one specimen,
* Logs, J.: Comparative Physiology of the Brain and Comparative Psychology,
1900.
On the FHleart Action of Molgula Manhattensts. 7
a small piece of heart-tissue three millimetres long, taken from the
centre of a heart, was observed to pulsate at the rate of thirty-three
beats to the minute.
So far as could be seen, when the heart was divided into two
nearly equal parts, these pieces uniformly continued to pulsate from
the uncut end toward the cut end. In hearts that were so cut as to
leave only one-third of the heart intact, beating would still take place
from the uncut end. Ina very few cases, where a minute fragment
was removed from one end, irregular pulsations were observed to
take their origin from the cut end of the heart. This latter phenom-
enon, however, was never observed immediately after the operation
upon the heart, and was rare. After the removal of a small strip of
tissue from such a specimen as has been mentioned above, the heart
would go on beating from the uncut end, and in no observed case did
beating again take place from the cut end.
It is the opinion of the writer that with the aid of a Ziegler’s?
compressorium, such as was used by Schultze, it would be possible
to diminish the error still more, and to obtain pulsations in a
larger percentage of hearts than by means of the methods described
above.
THE PHYSIOLOGICAL EVIDENCE FOR THE CONNECTION OF THE HEART
OF MOLGULA WITH THE CENTRAL NERVOUS SYSTEM.
Does the isolated heart of Molgula beat with a normal rhythm ? —
It is a well-known fact that the heart of a tunicate, when removed
from the body of the animal, and left in normal salt solution, or in
salt-water, will continue to pulsate rhythmically. This fact has been
noted by all of the earlier workers on the Tunicata, and especially
in the Salpze, as early as 1827 by Chamisso. The recent work of
Schultze shows that the isolated heart of Salpa will beat rhyth-
mically for long periods of time, but always with a slower rhythm
than the normal rate. Lack of co-ordination is not particularly
noticed by Schultze, although he notes changes in the time of dura-
tion of the pulsation period.
In the series of experiments performed upon Molgula by the
writer, the heart, with the tissues immediately surrounding it, was
completely isolated, and left in covered dishes for a few minutes to
1 ZIEGLER, H. E.: Zeitschrift fiir wissenschaftlichen Mikroscopie, 1897, p. 145.
8 George Wilham Hunter, Jr.
allow for the shock (inhibitive) effect to wear off. The heart so
treated very soon establishes a series of rhythmical contractions, the
waves of which originate as in the unoperated heart. The rate of
these contractions is usually much slower than those of the normal
heart, rarely exceeding thirty-two pulsations per minute, against a
normal rate of over “forty-three per minute. Instead of the normal
rest period, and the abrupt change from the ab- to the advisceral
pulse and vice-versa, as seen in the living normal animal, the change
is in this case usually effected by a series of beats from one end of
the heart gradually overcoming the established rhythm, and starting
an opposition series in the other direction. This method of reversal
in the heart of Molgula has been observed by Loeb’s pupils at the
Marine Biological Laboratory at Woods Hole as the normal method
of reversal. It was found by the writer, however, that only very
fresh specimens of Molgulz could be depended upon for results, and
as the available supply of animals for this work had to be brought
from New Bedford, a point twelve miles from the laboratory, it was
often found impractical to use them for careful physiological work.
These specimens were used in the laboratory for class work, and it is
by no means unlikely that the recorded observations were made on
such animals. Furthermore, in the operated hearts, careful observa-
tion shows that in a larger percentage of cases both ends of the
heart are beating at the same time, although one end may display a
markedly stronger rhythm. Irregularity of the rhythm, lack of co-
ordination between the ends, and long pulsation periods only in one
direction, the last named phenomenon one of the so-called ‘“ death-
signs” of Schultze, are all exhibited by the isolated heart.
Does the removal of the ganglion or“ brain” of Molgula affect the
heart rhythm ?— The findings of Schultze are here of great interest.
He notes that immediately after the removal of the ganglion from the
body a marked depression in the rate of the heart takes place. This
depression, however, can also be produced by cutting off a small piece
of the tissues surrounding the hyperbranchial groove, or even by
removing a small piece of tissue from any part of the animal. In
other words, the depression is not dependent upon the particular
tissue removed, but upon the amount of tissue removed and the
amount of blood lost from the blood-canals. Hence he comes to the
conclusion that the heart in the Salpe is not connected with
the central nervous system.
While the loss of blood and “ body-fluid”’ does, undoubtedly, lower
On the Heart Action of Molgula Manhattensts. 9
the blood-pressure in the body sinuses and in the heart, and thus
lower the rate of the heart-beat, still it is possible to reduce the loss
of blood to a minimum by the use of a cauterizing apparatus! A
comprehensive series of over fifty experiments with Molgule which
had been cauterized in the ganglionic region showed the following
results. (The effects of cauterization are decidedly more marked
when the posterior end of the ganglion and the dorsal nerve chain
are destroyed.)
1. In all experiments the immediate result of cauterization is the
complete inhibition of the heart in diastole for a shorter or longer
period, this period rarely lasting more than a few seconds. After
the heart begins to beat again, the rate is usually much slower than
the normal. The rhythm, however, increases slowly until it ulti-
mately becomes nearly stationary again, but still at a rate much
below the normal. The normal rate of heart-beat for all animals
examined (averaged) was 43 + per minute for the ab- and advisceral
pulsations. In thirty operated Molgulez in which the shock effect
had worn off, the rate was 31.2 per minute. This rate varied in the
individual animals from as low as 18 per minute to as high as 59
per minute.
2. Great irregularity in the heart-rhythm is noticeable. This
irregularity may take the form of alternating long with short beats,
fluttering beats, almost fibrillar in character, irregular pauses
between beats, and pulsation periods of widely differing duration
in time.
3. The heart may beat from both directions at once, either with
or without a co-ordinating rhythm. Series of rhythmical contractions
may be established by both ends of the heart, this state of affairs
continuing until one end of the heart seemingly gains control, and
asserts itself more strongly, thus causing the contractions to take
‘their origin from that end. Both ends, however, may continue to
contract; but one end, giving rise to a series of stronger pulsations,
gains the mastery of the blood-current. Often a “return beat” is
established by the end of the heart which is beating less strongly, and
a rebuff of at least part of the blood contained in the organ takes
place before it has left the heart. This latter phase is met with in
a large percentage of cauterized animals, or in those in which the
1 Thanks are due to Prof. C. B. SUMNER, of the College of the City of
New York, for the privilege of using a cauterizing apparatus (electrical) devised
and used by him in experimental embryological work.
10 George Wilham Hunter, Jr.
ganglion was carefully removed. In all the above-mentioned cases
the origin of the contraction moving in either direction is from the
extreme distal ends of the heart tube. In cases where the animal
may have lost much blood, and where the heart is not completely
filled in diastole, the wave of contraction in a given direction (A)
seems to end at a point near the middle of the heart tube, and a
second wave of contraction continues the beat to the end of the
heart. This second wave arises at the very instant that the next
succeeding wave starts from the given end (A). It is possible,
however, that this seeming double wave may be only an appearance
due to the flaccidity of the heart wall, and consequent wrinkling or
folding of the muscle at the middle of the heart.
The following experiment shows some of the above-mentioned
points:
EXPERIMENT XXIV.
August 25, 1902. — Ganglion cauterized at 9.15 a.m. Time of observation, 11.15 A. M.
to 11.38 A.M.
ABVISCERAL. ADVISCERAL. ABVISCERAL. ADVISCERAL
Beats. Time. Beats. Time. Beats. Time. Beats. Time.
min. sec. 5 min. sec. min. sec.
0 15 501 25 Abe SU) 8! 0 40
Se ]. 21 6 0 29
1 Beating irregular in rhythm.
2 Beating from both ends of the heart at once.
Next observation made at 3 P. M., when the heart rhade 200 beats
between 3.00 and 3.08 P.M., both ends of the heart beating at the same
time and with a like rhythm. The stroke in the advisceral direction is
stronger. After each advisceral beat a secondary return pulsation in the
opposite direction occurs.
3:10 P.M. Pulsations still in both directions at once (25 to the
minute), but stronger in the abvisceral direction.
3-34 P. M. Pulsations are stronger in the abvisceral direction, but now
show a return stroke after each beat in the opposite direction. A small
On the Fleart Action of Molgula Manhattensts. II
bit of débris in the heart was carried by the blood-current, time after time,
to the centre of the heart, and was then returned to the advisceral end as
if from a vortex, to have the next abvisceral beat repeat the operation.
This bit was eventually forced out of the heart on an abvisceral beat.
The rate of the heart at this time was 26 to the minnte.
7-52 P.M. Heart beating 16 to 18 per minute in an abvisceral
direction.
10.02 P.M. Heart beating at the rate of 7 per minute in an abvisceral
direction.
7.55 A.M. Heart pulsations occur at the rate of from 20 to 22 per
minute in an advisceral direction with a return beat in abvisceral direction
after each pulsation. Specimen lively ; reflexes not recovered in region
of siphons.
12 Noon. Animal not lively, heart pulsating faintly ; very irregular.
2.30 P.M. Animal dead.
Most of the other experiments show all or most of the above-
mentioned irregularities, and the above-quoted case is a fair repre-
sentative of some thirty to forty others.
4. Rhythmical pulsations take place in one direction only, for
abnormally long periods of time. As has been previously shown,
nearly 43 per cent of normal Molgulz examined had a pulsa-
tion period ranging from fifty to one hundred beats between
reversals; over 26 per cent had a pulsation period of less than
fifty beats between reversals, and only 3 per cent showed a
pulsation period of two hundred beats or over. In the cauterized
animals a very different condition exists. The heart, having once
established a rhythm in a given direction, will frequently continue
to beat in that direction without reversal for as long a period as two
to three hours. Several uncounted cases beat for over three hours
before reversing ; one case showed an actual time of two hours thirty-
three minutes, with a total of forty-two hundred pulsations; another
specimen gave thirty-three hundred and ninety-five beats in an ad-
visceral direction without a break, the time occupied being one hour
and thirty-one minutes. Numerous cases of over five hundred beats
were observed. Schultze gives such cases of abnormally long pulsa-
tion periods as a sign of approaching death. This does not seem to
hold true in Molgula, as in many of the recorded cases the animals
showing the above abnormality in the pulsation periods lived for
several (two to six) hours after the observations.
5. Especially after the cauterization of the posterior end of the
ganglion and the anterior region of the dorsal nerve cord, a series of
PS; vig George Wilham Hunter, Jr.
double beats, quicker in time than the normal beat and seemingly
interfering with it, may be established. The two beats follow each
other (in the same direction) so quickly that at first sight it would
be said to be a single pulsation instead of two. Close observation,
however, shows a second contraction immediately after the first, the
two pulsations following down the length of the heart tube only a
few millimetres apart.
The effects of electrical stimulation. — The earlier experiments of
Dew-Smith and Ransom and the more recent research of Schultze
have failed to give any definite information regarding the connection
of the ganglion and the heart.
Dew-Smith did get a slight lengthening of the pulsation period
in Salpa after stimulation of the ganglionic region; but other writers
have tried in vain for results. The difficulties which stand in the
way of accurate results are great, and a repetition of some of the
experiments of the above-mentioned writers resulted in negative
findings. The experiments were, however, far from what they should
have been, because of inadequate apparatus, and I hope to be able
to repeat them at a future date.
The results obtained with chemical stimuli.— Much previous in-
vestigation has been done on the invertebrata by means of chemical
stimuli, with especial reference to the connection of the heart and
the central nervous system. On the Crustacea, the work of Yung,!
Dogiel,? and others; on the Mollusca, that of Yung,? Foster and
Dew-Smith,* Ransom,”° and Bottazzi and Enriques;® on the Tunicata,
that of Kruckenburg,’ Lahille,® Lingle,? and Schultze is most note-
1 YUNG, E.: Archives de zoologie expérimentale, 1878, vii.
2 DoGIEL : Comptes rendus, 1876, lxxxii, pp: 1117, 1kGa:
a MONG, I. 206. cit; 1881 ,A%, Dp. AZT:
* Foster, M., and Dew-Smiru, A. G.: Proceedings of the Royal Society,
London, 1875, p. 318.
* Ransom, W. B.: Journal of physiology, 1884, v, p. 261.
* Borrazzi, F., and P. Enriques: Archives italiennes de biologie, 1900, xxxiv,
pare.
" KRUCKENBURG, C. FR. W.: Vergleichend-physiologische Studien, zu Tunis,
Mentone, und Palermo, 3 Abt., Hemelbas. 1880.
8 LAHILLE, F. = Conmalarone: a l'étude anatomique et taxonomique des Tuni-
ciers, Dceeietion. Paris, 18go.
* LINGLE: vide LoEB, J.: Comparative Physiology of the Brain and Cannar
ative Psychology, 1goo.
SCHULTZE, L. S.: Zeitschrift fiir Naturwissenschaften, Jena, IQOI, xxviii,
p. 145.
On the Heart Action of Molgula Manhattensis. 13
worthy. Such results as bear directly on my observations will be
noted when reference to such work is made.
For experimental purposes the following substances were used in
solutions varying in intensity from 0.01 to 0.00001. Wherever
possible, the solutions were made in normal salt. The substances
used were alcohol, atropin, caffein, curari, digatalin, hellebore,
muscarin, nicotin, and strychnin. I shall call attention only to such
of the obtained results as bear directly on the question of the heart
inhibition and acceleration.
For the obtaining of the results which follow, the normal Molgula
was placed in a chemically clean glass dish with sea-water, and left
undisturbed until the heart-beat became normal. Then the sea-
water was carefully drawn off and a solution of the poison in sea-
water put in its place, or the poison was added directly to the water,
at first contained in the dish, such water having previously been
carefully measured. The heart-beat was carefully taken for a few
pulsation periods just before the addition of the poison to the water,
and again immediately after it had been added. Later observations
on the animals were made at short intervals ; thus the immediate
effects of the drug, as well as the after effects, were noted. For the
sake of comparison, wherever it was possible, check experiments
were made on animals from which the ganglion had been removed.
Inasmuch as such animals sometimes live for days, this comparison
was readily possible.
Heart tracings were not obtained because of the lack of apparatus.
It is hoped that graphic results may be shown in a future paper.
Owing to the recurrent action of the heart, it is manifestly im-
possible to plot a curve, showing inhibition or acceleration, that
would compare graphically with such a curve as is obtainable in any
other of the invertebrates or vertebrates. A graphic representation
of heart depression or acceleration can, however, be shown by plot-
ting the numerical result obtained. Such results are seen in the
figures following. In all the cases here shown the abscissas give
the time marked in periods of one hundred seconds; each one
hundred second period having been taken from a single, successive,
pulsation period. The ordinates show the number of heart pulsa-
tions taking place in a given period of one hundred seconds. The
mean, made for the sake of comparison, is obtained by taking the
mean of a number of pulsation periods counted just before the poison
was added. These diagrams are in no sense to be taken as curves
14 George William Flunter, Jr.
obtained in the usual manner, but merely as figures which show
graphically obtained results and abbreviate explanation.
Experiments with alcohol. — The heart of a normal Molgula is not
easily susceptible to the presence of alcohol. Using a solution of
I part absolute alcohol to 100 parts of sea-water the heart-beat
appeared to be neither accelerated nor depressed to any great
degree. Depression was noted in some cases. The duration of the
pulsation period was not noticeably varied. There was, however, a
marked irregularity of rhythm. Long stops between strokes were
noticed in some specimens, with a subsequent recovery and an
acceleration of the heart rhythm. Animals thus treated with 1 per
cent alcohol often lived for
rTP several hours after immer-
“4 ay se EE it aac sion in the alcohol. The
an heart of an animal in
Z TIN +7 =6which the ganglion was
tient 1 A-AH ch removed stopped beating
Seana Se BEeee! within five minutes after
aa mee HH -_-EEEPE_PE, immersion in a solution
i : lat |_| 4 of 1 per cent alcohol. In
i 7 Biel ine Ey such an experiment, the
z+} | H Co EF -t~» alcohol undoubtedly
dl ESR: | reaches the heart tissues
FIGURE 1.— The upper curve is from a normal directly by way of the cut
animal; mean, at 60; the lower from an ani- area. With a solution of
mal with ganglion removed; mean, about 51; 1 to 1000 of alcohol the
* marks | : 1000 alcohol. :
normal heart is acceler-
ated so as to beat from four to eight strokes per minute faster than
the normal. In specimens in which the ganglion was removed, the
same strength of alcohol gave an immediate depression in the rate of
beating. Irregularity and a shortening of the pulsation period also
occurred in some of the specimens.
Normal animals, when placed in a1 to 10,000 or I to 100,000
solution, failed to show any marked symptoms. In one or two cases
a slight acceleration was noticed where animals were placed in a I—
10,000 solution; but as this soon became normal, the change of
rhythm might be attributed to the shock caused by the changing of
the fluids in the container. No results were obtained with operated
specimens, the weak strengths of alcohol seemingly having no effect
on them.
On the Hleart Action of Molgula Manhattensis. 15
The present unsettled views as to the real nature of the influence
exerted by alcohol, both. on nerve and muscle tissue, makes the
above notes of some interest. These experiments seem to fit in
with the most recent evidence taken from the higher vertebrates,
and with some of the findings in the invertebrate groups. (See
Cushney,! Herter,? Kraepelin,? and others.) Fig. 1 shows graphi-
cally the effects of certain strengths of alcohol on normal and gan-
glionless animals.
Muscarin. — The well-known effects of muscarin sulphate, and the
previous results obtained with it by Kruckenburg on Salpa, and by
Lingle on Molgula, render these experiments hardly worthy of re-
petition. In the experi-
ments of both of the
above-mentioned investi-
gators, muscarin had the
characteristic depressant
effect on the heart.
Only a few experiments
could be made by the
writer, owing to the diffi-
culty of obtaining pure
muscarin. Experiments
with one set of solutions
of muscarin turned out FIGURE 2.—The upper curve is from an animal
successfully; all the with ganglion removed; mean, about 83; the
others failed. Either be- lower curve is from anormal animal; mean, 75 ;
* marks 1 : 1000 muscarin.
cause of the poor quality
of the drug obtained, or ieeatee of the extremely small quantity of
muscarin present, solutions of 1-100,000 strength of muscarin sul-
phate had no effect, either on normal or ganglionless animals.
With a strength of 1-10,000 and 1-1000, the effects were immediate
and marked. These effects were: (1) depression of the heart-beat ;
(2) increase in the length of the rest period between the pulsation
periods ; (3) increased irregularity in the heart rhythm. The ulti-
mate effect, especially and immediately noticeable after the use of a
I-1000 solution, was the death of the animal. Frequently death
was immediate, or within fifteen minutes after the addition of the
1 CUSHNEY: Pharmacology, 1goo.
? HERTER: Chemical pathology, 1902.
3 KRAEPELIN : vide HERTER’S Chemical pathology, 1902.
16 George Witham Hunter, Jr.
poison. More frequently, especially with a solution of 1-10,000, life
continued for four, five, or even nine hours after the addition of the
drug to the sea-water.
EXPERIMENT X®.
August 25, 1902.— Specimen size, 16 X 18 mm. Fresh. Temperature of water, 21°.
ABVISCERAL. ADVISCERAL.
Time of obs.|_ Beats. Time. I INGSE: Beats. Time. Rest.
min. sec. min. sec.
10.20 A.M. 67 i 47 2 59 ] 35 1
oH) 1h 33 é 67 1 46 2
63 eee 2 55 1922 2
S44 28) | 1 44 1 10 3
Added 1.1000 muscarin sulphate at 10.40 a.m.
48 TG I 35 i lane! ay
41 I 7 1 40 1 10 7
41 IF 33 1 36 i eee 4 6
43 i= tks 2 37 ib Ue 6
41 Lg. 2 38 Ee, 5
tat 1 20 1 46 1 21 8
43 20 1 Aa 22: 8
43 MA? 1 43 1) az if
45 123 0 41 L220 $
41 18 2 4) 1 =20 10
11.13 A.M. 39 Wie! 1 49 TiS 10
12.19 p.m. 39 ae Sure fia 52 1 20 3
43 hg) 2 48 1321 4
36 1 16 1 60 Tsocel 3
1 Trregular.
Fig. 2 shows graphically the depressant effect of muscarin sul-
phate on the normal heart of Molgula manhattensis. It is noticeable
Ox the Hleart Action of Molgula Manhattensts. 17
that in some cases, at least, the advisceral pulsation appears to be
depressed to a.much greater degree than is the abvisceral beat.
In one case, however, the opposite result was noted, z.¢. the greater
depression of the abvisceral beat.
The net results of several experiments show a depression which
may be immediate, but which has a maximum several hours after
the addition of the poison. One example shows an immediate de-
pression of from six to eight beats per minute, and a lengthening of
the rest period of the abvisceral pulse. This experiment is given in
the table on page 16. |
Another experiment shows a depression from the normal of fifty-
five beats per one hundred seconds, fifteen minutes after the appli-
cation of muscarin of 1-1000 strength. Other cases give a depression
of from thirty to fifty-five beats per one hundred seconds, two and
one-half to three hours after putting the animals in a solution of
I-1000 strength. In one case (I-10,000 strength) there is a depres-
sion of twenty-five beats per one hundred seconds, two and three-
fourths hours after immersion in the poison. In nearly every specimen
under the influence of muscarin, an irregularity amounting almost
to a fluttering pulse is seen. This irregularity also makes its appear-
ance in the heart rhythm, as can be seen in the irregularly longer
rest periods between single beats in a given pulsation period.
A comparison of the results thus obtained with those reached by
the immersion of the ganglionless animal in weak solutions of mus-
carin are instructive. In the case of extirpation of the ganglion or
of the removal of the heart from the body, the influence of the drug
appears to be slight. Instead of an inhibition we find an actual slight
acceleration in some cases. The heart is very irregular. In no
observed instance was death immediate. A typical experiment is
the following:
Extirpated ganglion at 8.05 p.M., Sept. 1, 1902. 8.15 Pp. M., heart beating at
the following rate per minute: 24, 26, 31, 31, 32, 36, 36, heart very ir-
regular, beating from both ab- and advisceral ends at once. Added
I-1000 strength solution of muscarin sulphate at 8.23 P.M. At 8.25 P.M.
heart beating very irregularly, mostly in an abvisceral direction, at the
following rate: 38, 37, 37, 36, 38, 33, 37, 30, (reversal) ; 34. During the
above beating the heart appeared to be accelerated, making the strokes
very quickly ; but with long stops between the series of beats in a given
direction. 9.01-g.09 P. M., heart beating irregularly, mostly in advisceral
direction, at the following rate: 24, 27, 30, 32, 31, 35, 37-_ In the above
18 George William Hunter, Jr.
series every twenty-fifth to thirtieth stroke there appeared a stronger beat
which interrupted the advisceral beat, but which was quickly overcome by
it, after which beating in the advisceral direction was resumed, until inter-
rupted again about a minute later by this one strong beat in the abvisceral
direction.
Comparison of these results with those of the investigators men-
tioned previously as workers on the Tunicata, with those of Ransom:
and Yung on the Mollusca, with those of Dogiel and Yung on the
Crustacea, and finally with the many experiments on vertebrate
material quoted by Gaskell,! shows the generally depressant effect of
the drug. (Yung, however, finds acceleration preceding depression
in certain of the Mollusca.) There has existed a difference of opin-
ion as to the exact action of the drug in its depressant action on the
heart. Ransom and some of the older writers favored action on
the heart muscle; the later writers believe the drug acts through the
nervous system. The fact that muscarin is ineffectual on the em-
bryonic vertebrate heart, in which the nerve centres have not yet
appeared, seems conclusive proof of the latter view. If the drug
acts on the preganglionic fibre at the point of its connection with the
nerve cell, as Gaskell believes is true for the Vertebrates, then the
above-mentioned results might be taken to indicate the same state of
affairs in Molgula. As heart ganglia and endings on heart-muscle
have been found, together with connective fibres, the anatomical
connection of which have not yet been fully worked out, it looks as
if the connection between the heart and the central nervous system
did exist in the Molgule.
Nicotin. — Experiments have been performed on the Salpz by
Kruckenburg, Lahille, and Schultze. According to Kruckenburg,
poisoning with hellebore and nicotin affects only the duration of the
advisceral pulsation period, and consequently influences only the hypo-
branchial end of the heart. He believes that the change in direction
of the contractions is brought about by connection with the ganglion.
‘The results of poisoning with hellebore and nicotin . . . appear to
me to indicate that the reversal of contractions is reflex and is brought
about through the ganglion.” He was, however, unable to prove the
existence of ganglia on the heart, or of connective fibres between the
central nervous system and the heart. He finds that the particular
effect of nicotin is to reduce the number of advisceral pulsations,
! Vide GASKELL’S article in Textbook of physiology, edited by SCHAFER,
London, 1goo.
On the Heart Action of Molgula Manhattensis. 19
while hellebore increases the number of advisceral pulsations.
Schultze took up the investigation to verify Kruckenburg’s results, and
obtained different findings. He used solutions of 1-10,000, I—25,000,
and 1-100,000 of nicotin, and worked on the large Salpa africana-
maxima. With a 1-10,000 solution he finds an almost immediate and
strong reduction in both ab- and advisceral pulsation periods. The
animal dies after one-half an hour’s immersion in the poison. With
I-25,000 strength, practically the same result is obtained except that
the animal lives for one and one-half hours. In both of the above
experiments Schultze gets a decided depression of the heart-beat and
great irregularity. With 1-100,000 strength his figures show a slight
increase in the length of the pulsation period, together with a slight
depression in an abvisceral and acceleration in the advisceral direc-
tion. After a little 1-25,000 solution is added to the above, the
heart almost immediately shows the effect by greater irregularity,
depression, and a shortening of the pulsation period.
In my own experiments three solutions were used: 1I-1000,
I-10,000, and 1-100,000. With the first-named strength, an almost
instantaneous effect was obtained. Upon addition of the nicotin, the
heart was seen to change its direction at once; then, after from one
to two minutes beating, which was characterized by great irregularity
of rhythm and depression, death would ensue.
The following are typical experiments:
Specimen, size 18 X 20mm. Fresh. Temperature of water, 21°. Time of
observation, 11.30 A. mM. Normal animal in sea-water.
ABVISCERAL. ADVISCERAL.
Time. Time.
in. sec. min. sec,
42 20 1 32
46 38 be 2
58 45 2 5) 1 42
11.40 a.m. Added 1.1000 nicotin. Heart beating in advisceral direction.
After 10 to 12 beats, it reversed.
26 1 32 20
1 ay
No more beating observed.
20 George Wilham Flunter, Jr.
Specimen, 12 X 14. Fresh. Temperature of water, 21°.
ABVISCERAL. ADVISCERAL.
Beats. Time. Time.
min. sec.
3°28
5 ee
Zz 49 1
Added 1.1000 nicotin. Heart-beat both directions at once for 10 seconds.
1 OZ 20
1.10 i
No more beating.
With a strength of 1-10,000, death usually occurred within half an
hour, frequently more quickly. The characteristic changes caused
by the stronger solution were observed here, but in a less marked
degree. It is interesting
— r a= 2 | 7
e0 ||. to note that in both of the
4 | . above series of experiments
7 [tl the pulsation periods are
i | shortened greatly, as ob-
| Pineietaph 771° served by Kruckenburg
| . and Schultze, the abvis-
—r-} ceral pulse being affected
{||| the more decidedly. With
aE a I-100,000 strength an
|| {| \, immediate but very slight
HEE EHH depression of the heart,
FIGURE 3.—The upper -curve is from a normal and a marked irregularity
animal; mean, about 51; the lower from anani- of the rhythm, occurred.
la a le removed; mean, 30; * marks The length of the pulsation
was seemingly unaffected.
Fig. 3 following gives a graphic view of the heart action when
under the influence of 1-1000 and 1-10,0c0 nicotin. A comparison
of the above results with those obtained with the use of Molgulz
in which the ganglion was extirpated is of interest. In the case
of the strong solution (1-1000), the effect is somewhat like
Ox the Heart Action of Molgula Manhattensis. a1
that obtained in the unoperated Molgule with a solution of the
same strength.. The heart in all observed cases stopped beating
within twenty-five minutes. The zmmediate depressant effect of the
poison was not seen as in the normal animals, although present toa
marked degree. The shortening of the pulsation period was not so
marked as in the ganglionated animals. The characteristic abnormal
irregularity of the heart rhythm, however, was very noticeable. In
solutions of 1-10,000 and 1-100,000 strength, the depressant effect
of nicotin is not observable, many cases showing an actual accelera-
tion. Irregularity of rhythm is here characteristic, as in the other
cited cases. A long rest period, irregular in time of appearance, is
found between single beats. The example given below illustrates
many of the experiments:
Ganglion removed g.25 A.M., Sept. 3, 1902.
10.27 A.M. Heart beating in advisceral direction at the following rate
per minute: 29, 29, 29, 29, 28, 29, 28.
10.35 A.M. Added 1-1000 nicotin.
10.36 A.M. Heart beating in advisceral direction at following rate:
Bon aly 314.30, 30; 31:
10.52 A.M. Heart beating irregularly in both directions at once.
Pulsations very firm. One or two stops of 3 to 5 seconds each. Rate:
Soest, 40, 30, 30, 30:
11.30 A.M. Beating in advisceral direction ; interrupted by a few
beats in the opposite direction; pulsations then taking place in both
directions at the same time. Rate: 32, 32, 34, 32, 34-
These experiments are of interest when considered in comparison
with the well-known experiments of Langley on vertebrate material.
If nicotin acts on the junction of the inhibitory fibre and the ganglion
cells in the heart, we should expect to obtain the very results that
were obtained in the above experiments on the operated Molgule,
providing such an apparatus exists in the latter animal.
Strychnin. — According to Reid Hunt! the drug probably acts on
certain parts of the nervous system (the vaso-motor centre especially),
rather than on the heart itself in the vertebrates. In the frog, the
heart is stimulated by small amounts of the poison, while large
amounts weaken and retard it. In the invertebrates there seems to
exist a divergence of opinion as to the action of the drug, and its
meaning. Strychnin sulphate, according to Cushney, dissolves in a
1 Hunt: Reference Handbook of the Medical Sciences, 1901, pp. 687-703.
22 George Wiliam Hunter, Jr.
little over 8000 parts of water. My experiments were made with
solutions of 1-10,000, and 1-100,000. With a saturated solution in
sea-water the effect was immediate. The heart was strongly de-
pressed, there was loss of co-ordination, a shortening of the pulsation
periods in both directions, and death within twenty-five to thirty
minutes.
The following experiment is typical:
Specimen size, 12 X 16. Three days in aquarium. Temperature of water, 21°+.
ABVISCERAL. ADVISCERAL,
Time of obs.| Beats. Time. Beats. Time.
23 43
13 38
22 50
30
31
34 6
in sulphate at 8.30 P.M.
8.40-9.00 P.M. Stor stots rerere | 1
43 1
1
1
0
0 0
0 noge 0
Beating at this time starts at one end of the heart and returns slowly.
9.02 P.M. Slight contraction at abvisceral end of heart.
9.04 P.M. Slight fibrillar contractions. No more movement observed.
The results obtained with a 1-10,000 solution were practically
the same as those recorded above, except that the effect is not so
immediate or so marked.
Ganglionless Molgulz, when put in a I-10,000 solution, did, not
show an immediate depression of heart-beat, nor did death follow
On the Heart Action of Molgula Manhattensis. 23
immediately. (Unfortunately I was prevented, through lack of
material, from trying the saturated solution on operated Molgulz.)
With a 1-100,000 strength solution, the characteristic strychnin
acceleration was obtained when working with normal Molgule. This
acceleration was immediate and from seven to twelve beats per
minute above the normal rate. In some cases, after ten to fifteen
minutes had elapsed, irregularity in rhythm set in. This irregularity
seemed especially marked in the advisceral beat, causing a decided
depression. After one to two hours the rate of beating would be-
come lowered so that it was below the normal. Along with this
depression would appear great irregularity, both in the ab- and ad-
visceral directions, but es- ‘
pecially marked in the ae i | |
advisceral beat. In gen- | Leer Fr
eral, the advisceral end of 4 St
the heart seemed more HALLE [ ie
profoundly affected by the ~ ee CI
drug than the opposite eal
end. aaa al
The tables on pp. 24 and |
25 show the effect of strych-
nin sulphate, 1—100,000
solution, on a normal
Molgula.
f= beet
ThEP
=I
FIGURE 4.— The upper curve is from a normal
In Molgulz in which the animal; mean, 80; the lower from an animal
ganglion was removed the with ganglion removed; mean, about 43;
ks 1: trychnin.
effect of a I-100,000 solu- * marks 1: 1000 strychnin
tion was very slight, if, indeed, any effect could be noticed. In a
few experiments a very slight depression was obtained, followed by
a rise to the normal rate within a few minutes. This same effect
might be produced by pouring water over the animals, and may be
due to such an action. The graphic records showing the action
of strychnin do not show the extreme depression and reduction in
the advisceral direction observed in some animals. The results are,
however, inserted for the sake of comparison.
Although results were obtained with the use of atropin, curari,
and hellebore, it is not thought that these results have any direct
bearing on the subject of acceleration or depression through the
central nervous system, and so allusion to them, more than to say that,
in a general way, they conform with experiments made on the verte-
brates, will be omitted in this paper.
24 George Wilham Hunter, Jr.
ADVISCERAL. ABVISCERAL.
Time of obs. Beats. Time. Time.
min, sec.
2.05-2.20 P.M. 47 10 1 36
51 24
48 22
46 19926 21
50 12s |
Added 1.100,000 strychnin sulphate at 2.30 P.
65
65
55
non oe WwW WOW YO WHO HT LD W
— —_
—
Oo
- KF KF DY DY NH HBS Ff WO KF WO NYS FP FW WS KY fF WS KY FS
CO. Co Or SS Tt) | bo
KF OF FF FF Fe Fe Ee Ee
1 Trregular.
On the Heart Action of Molgula Manhattensts. 25
ADVISCERAL.
Time of obs. Beats.
45
45
41
Time.
min. sec.
1 20
24
13
15
13
15
ABVISCERAL.
Time.
min. sec,
1
0
0
il
1
0
0
0
§.06-8.12 P.M.
1 Trregular.
2 Sensory reflexes acute.
8 A quick beat is rebuffed by another in the opposite direction (abvisceral)
almost immediately.
26 George William Hunter, Jr.
Caffein and digitalin, drugs which are believed to act more or less
directly on heart muscle, were also used in the series of experiments
made on Molgula. The results obtained, although incomplete, will
be given in part.
Caffein.— According to Bock,! caffein acts, in the vertebrates at
least, chiefly on muscle. Acceleration takes place in the isolated
heart. In the normal animal, however, inhibition may occur from
the stimulation of the cardio-inhibitory centre.
A solution of 1-1000 strength of caffein was tried on the isolated
heart of Molgula with the result that there was a slight but noticeable
acceleration of the heart rhythm, and an increased regularity of the
rhythm. The heart-beat seemed stronger and fuller than it was
before the addition of the drug. Normal animals, when placed
in a I-1000 solution of caffein, showed an immediate depression of
the heart-beat, which appeared within a few minutes (five to ten)
after the addition of the drug, but which seemed less marked after
perhaps half an hour. The general irritability of the muscle was
much increased, and a decided irregularity of rhythm appeared,
especially in the case of animals that had been some time in the
solution. Results have not yet been obtained with solutions of less
strength.
Digitalin.— According to Cushney,? who has made the most ex-
tensive researches with vertebrate material, the drug acts in two
ways: 7.é., it first depresses the heart by acting on the inhibitory
apparatus, and later accelerates the heart by its action on the heart
muscle. This action he obtained with small doses of digitalin.
My experiments here are very incomplete, and I will only cite one
or two. In animals deprived of the ganglion, and put in a 1-1000
solution of digitalin, the heart was accelerated and the rhythm ren-
dered more regular.
The normal animal, when treated with the same strength solution,
also showed a slight acceleration, but with accompanying irregularity
of rhythm. There seems no doubt of the effect of the drug on the
muscle in both of these experiments, because of a peculiar double
heart-beat which appears and which seems to be characteristic of the
digitalin stimulation of the muscle fibres.
* Bock: Archiv fiir experimentelle Pathologie und Pharmakologie, 1900, xliii,
P- 397.
? CUSHNEY: Journal of experimental medicine, 1897, ii, p. 254.
On the Heart Action of Molgula Manhattensts. 27
CONCLUSIONS.
The above results may be summarized as follows:
1. The normal heart beat of Molgula manhattensis varies greatly
in different individuals as to rapidity of rhythm, duration of pulsa-
tion period, and rest period.
2. The average number of heart-beats in a given direction, either
ab- or advisceral, was 43.2 per minute.
3. The length of the average rest period was two seconds.
4- In approximately 70 per cent of the animals examined the aver-
age number of heart-beats in a pulsation period in either direction
did not exceed one hundred; in 30 per cent of the animals the pulsa-
tion period in a given direction exceeded one hundred beats, but
rarely more than three hundred and fifty beats. Animals having a
pulsation period of over three hundred and fifty to four hundred beats
may safely be said to be abnormal in that respect.
5. In some observed cases the middle piece of an isolated heart
of Molgula will beat when placed in sea-water; but in no cases did
these pieces beat immediately after operation.
6. The removal of the posterior part of the ganglion (brain), or
of the anterior end of the visceral nerve cord, affects the heart-beat of
Molgula in the following respects: depression of the heart; irregu-
larity of the heart rhythm (long pauses between beats, double beats,
etc.); loss of co-ordination between the two ends of the heart; beating
from both ends of the heart at the same time, either with or without
co-ordinative rhythm; lengthening of the pulsation period in one
direction to an abnormal degree, death not immediately following.
7. After treatment with certain specific nerve poisons, the heart
of normal Molgulz reacted in a different manner from those in
which the ganglion had been removed.
8. Some specific muscle poisons affect the heart of normal and
ganglionless animals in an almost identical manner.
In conclusion, the writer wishes to express his thanks to Prof. C. O.
Whitman and Prof. F. R. Lillie for the privileges of an investigator’s
room at the Marine Biological Laboratory, Woods Hole, and to Prof.
Jacques Loeb for many helpful suggestions. My thanks are also
due to the members of the staff of the biological department of
Columbia University for the courtesy shown me there.
CONTRIBUTIONS FROM THE ZOOLOGICAL LABORATORY OF THE
MUSEUM OF COMPARATIVE ZOOLOGY AT HARVARD COLLEGE.
E. L. MARK, Director. No. 143.
THE SKIN AND THE EYES AS RECEPTIVE ORGANS
IN THE REACTIONS OF FROGS, TO GiGHe:
By «G, H. PARKER:
INTRODUCTION.
LTHOUGH the frog has probably served as the subject of more
laboratory investigations than any other animal, its photo-
tropism seems never to have excited more than passing comment.
Graber (’84, p. 121) observed that when specimens of Rana esculenta
were put in a box one half of which was illuminated and the other
half in shadow, the animals were found about three times in the dark
to twice in the light. This result might be interpreted to indicate
that frogs are xegatively phototropic, as, in fact, Loeb (go, p. 89)
subsequently intimated. Plateau (89, p. 82), however, found that
when specimens of Rana temporaria were liberated in a dark-box illu-
minated only by a pair of windows at one end, they jumped toward the
windows. Thus R. temporaria seems to be fosztvely phototropic.
In view of these somewhat contradictory statements, I attempted to
determine whether our common leopard frog, Rana pipiens Schreber,
is negatively or positively phototropic, and what parts are concerned
as receptive organs in its reactions to light.
NORMAL FROGS.
Frogs upon which no operations had been performed were tested
with Nernst lamps in adark-room with blackened walls. These
lamps possess the advantage of requiring no glass protection for
their filaments, and thus reflections such as often cause great incon-
venience in ordinary incandescent lamps are avoided. Moreover, their
light in quality is much more nearly like daylight than that from
incandescent lamps or the are light. The Nernst lamps used were
a “single glower” lamp heated by a current of 110 volts and a “ six
glower” lamp ona current of 220 volts. Heat was eliminated by
28
Reactions of Frogs to Light. 29
passing the light through a layer of seven centimetres of distilled
water contained in a glass vessel with flat sides. With this heat screen
attached, the one glower lamp had an apparent intensity of 24 to 25
candles. The six glower lamp under similar conditions gave a light of
about 320 candle-power. These two lamps with their heat screens
were the sources of light for all my experiments except where other-
wise stated.
When frogs were put on a moist plate so that their long axes were
at right angles to the direction of the rays of light, they sooner or
later turned toward the source of light, and generally jumped in that
direction. This occurred irrespective of the side of the frog that
was exposed to the light, and was observed in animals that were
5 metres from the 25 candle-power lamp and consequently were in
light of 1 candle-metre intensity, as well as with those that were 12.5
centimetres from the 320 candle-power lamp and were in light of an
intensity of 20,480 candle-metres. Many intermediate intensities were
tried and always with the same general results, namely, the frogs
turned toward the source of light and usually jumped in that
direction.
With the lower intensities the animals often did not react for from
five to ten minutes or even longer, and the jumping response was
frequently omitted ; but their orientation was finally always with their
heads toward the source of light, that is, positive. In some instance
after a frog had remained ten minutes or more without changing its
original position, it was induced to jump by being touched from
behind, and, when this was done, the animal almost invariably turned
first and then jumped toward the source of light. With high intensi-
ties, it was remarkable how persistently the frog would face the source
of light and jump in that direction, even when the light was unbear-
ably strong to the human eye.
From these experiments, I conclude that between the intensities of
I and 20,480 candle-metres Rana pipiens is positively phototropic.
THE EVYEs.
It is natural to suppose that the positive phototropism of R. pipiens
is dependent on the eyes; but it is conceivable that this phenomenon
may depend upon the skin, or, since the tissues of the frog are more
or less permeable to light, upon the direct stimulation of internal
organs such as the brain and spinal cord.
30 G. H. Parker.
To test the efficiency of the eyes in this respect, I attempted to
eliminate the possible action of the skin and deeper parts by cover-
ing the animal, excepting the eyes, with opaque material. In my first
trial I made a suit of clothes in one piece from soft light-proof cloth.
This was slipped on the frog and held in place by a thread passed
through the mouth like a horse’s bit. The frog when thus covered
remained motionless both on land and in water, its limbs taking un-
usual and often unsymmetrical positions, as though it weredead. On
removing its covering, however, it immediately assumed a normal
position and began jumping about. Apparently the cloth covering
so stimulated it as to inhibit ordinary locomotion, and therefore I
abandoned this form of experiment.
Frogs, even when closely crowded together, move continually, thus
showing that the contact of the skin of one frog with that of
another has no such inhibitory influence as the cloth exerted. I
therefore killed a large dark-colored frog and removed its skin in one
piece. This was turned inside out, thoroughly washed, and slipped
over a somewhat smaller frog, on which it was held by a bit-like thread,
as in the former experiment. The frog thus covered moved about
with almost normal agility. The only portions of the living frog that
were exposed were the front and hind feet, the snout, and the eyes.
Four such animals were tested in light having an intensity of 50
candle-metres, and I found that these animals turned toward the light
and jumped toward it much as normal frogs do. The experiment
was made first with the covered frogs alone, but afterwards, for the
sake of comparison, I introduced a normal frog into the receptacle
each time I tested a covered one. In most instances the normal
frog responded more quickly than the covered one, but the difference
was not so great that it might not have been due to the purely
mechanical interference of the covering skin.
It might be assumed, since the dead skin with which the frog was
covered was not perfectly opaque, that even in these experiments the
reactions really depended upon the influence of light on the skin;
but this assumption is not warranted, for when the dead skin was
drawn up over the eyes all evidence of phototropism disappeared. I
therefore believe that I am entirely safe in concluding that Rana
pipiens is positively phototropic to light stimuli received through
the eyes.
Reactions of Frogs to Light. 31
THE SKIN.
Having found that the eyes were concerned in the phototropism of
the frog, it remained to ascertain whether other parts acted as recep-
tive organs for phototropic reflexes. To test the skin in this respect,
I operated on frogs in the following way. By a single, vertical,
transverse cut just behind the eyes, these organs and the cerebral
hemispheres were removed with the snout of the animal. It is well
known that frogs in this condition may with a little care be kept
alive many weeks, and that the chief difference between these and
normal frogs is the great reduction in spontaneous movements shown
by the former. Frogs without cerebral hemispheres move, as a rule,
only when stimulated by some obvious means.
I prepared in this way eleven frogs, and tested them in light of
50 candle-metres intensity. Of these, two never showed clear reac-
tions to light, but the other nine were unmistakably phototropic.
The following record of frog No. 5 will give a fair idea of the nature
of these responses.
Frog No. 5.— Cerebral hemispheres and eyes removed May 1. Tested May 8.
4.35 P.M. ‘The frog was placed with its /ef¢ side toward the source of
light. It soon began turning, a little at a time, directly toward the light,
till at
4.48 P. M. it was facing the light.
4.50 P.M. ‘The frog was placed with its 7igh¢ side toward the light.
It soon began turning toward the light and at
4.54 P. M. it was facing the light.
4.55 P. M. It was again placed with its ft side toward the light. By
5-02 P. M. it was facing the light.
5.03 Pp. M. It was placed with the head directly away from the light.
It began turning to the right and at :
5-16 P. M. it was facing the light.
May 9. At 1.35 P. M. the frog was placed with its head away from the
light. By
1.46 P. M. it was facing the light and continued in this position till
2.38 P. M., when it jumped toward the light.
These records are fair samples of those obtained from the nine
responsive frogs, and my observations on these have shown that the
animals will turn by the shortest course either to the right or to the left
toward a source of light, and, having obtained the position of positive
32 G. H. Parker.
orientation, they will remain facing the light for a considerable period,
usually terminated by a jump toward the light. In other words,
eyeless frogs, like those with eyes, are posztzvely phototropic.
These reactions are observable not only in artificial but also in
natural light. Thus frog No. 5,o0n being placed with its left side
toward a window through which bright diffuse daylight was entering,
turned repeatedly, in from eight to twenty minutes, so that it faced the
window. When, late in the afternoon, it was placed sidewise in sun-
light it turned with every trial almost immediately toward the sun.
Thus positive phototropism is observable in natural as well as in
artificial light.
It is strange that the positive phototropism of eyeless frogs has not
already been recorded, for in the majority of individuals it was found
strikingly characteristic. Moleschott and Fubini (79), in their numer-
ous experiments on the influence of light on the excretion of carbon
dioxide from frogs with and without eyes, must frequently have had
animals under observation that should have shown this phenomenon,
and yet they make no mention of it. Possibly the European species
may differ in this respect from the American; but however this may
be, there can be no doubt that positive phototropism is a characteris-
tic not only of the normal Rana pipiens, but also of its eyeless condi-
tions. In this respect R. pipiens resembles certain planarians,
which, as Loeb (94, p. 225) first observed, and Parker and Burnett
(:00) subsequently worked out in detail, are phototropic both with
and without eyes.
Having found that eyeless frogs were usually positively photo-
tropic, it remained to ascertain what were the receptive organs in this
reaction. Since in my first experiments I worked with frogs in which
the optic lobes were somewhat exposed to light, it might be suspected
that these organs received the stimulus directly. 1 therefore prepared
other frogs in such a way that after the removal of the eyes, etc., a
fold of skin was left to cover the exposed portion of the brain, and
thus protect it from light. With this precaution, however, the frogs
still continued positively phototropic. But it might still be supposed
that the small amount of light which penetrated the tissues of the
frog might reach the central nervous organs and act as an orienting
stimulus. ‘Yo test this possibility, I observed the effects of illuminat-
ing only a part of the frog’s body. When an eyeless frog is placed
in light of 50 candle-metres intensity, and about half of the skin on
the exposed side is kept in shadow, the positive reactions often fail
Reactions of frogs to Light. 33
to appear. If the obstruction is gradually removed, there comes a
time before complete illumination when the animal will orient posi-
tively. By placing a screen in an appropriate position, it was possi-
ble to throw a shadow on that part of the frog’s body which contained
the central nervous organs, and still leave the greater part of the skin
illuminated. Under such conditions the frogs in the great majority
of cases turned toward the light, showing that the nerve endings
in the skin were stimulated. Since no response was obtained from
frogs that were covered with the skin of another frog so cut as to
admit light to the region next the brain and spinal cord, I believe that
these deep-seated organs are not only not essential as receptive organs
in phototropism, but that they are not stimulated by such light as
may reach them. I therefore conclude that the positive phototropism
of eyeless frogs depends upon the capacity of the nervous structures
of the frog’s skin to be stimulated by light.
That the skins of some animals are normally open to stimulation
by light has long been known. Willem (’91, p. 338), who twelve
years ago prepared a résumé of this subject, enumerated some thirty-
five species of metazoa in which this had been demonstrated, and
Nagel (’96) subsequently added considerably to this number.
Among the animals enumerated by Willem there is only a single
representative of the vertebrates, Triton cristatus. This animal
was studied by Graber (’84, p. 96), who prepared young individuals
for experimentation by removing their eyes and then covering their
heads with a layer of black wax. Animals thus prepared were put
in a chamber part of which was illuminated and part in shade. In
a total of 2102 observations the animals were found 674 times in
the light and 1428 times in the shade. Graber, therefore, concluded
that the skin of this newt could be stimulated by light.
Recently Beer (:o1, p. 30) has called attention to a second species
of vertebrate in which like conditions occur. This is Proteus
anguineus, which, according to Configliachi and Rusconi ('19),
becomes restless on sudden illumination and retreats eventually to
the darkest situation it can find. This sensitiveness, which was
also noticed by Semper (’81, p. 79), was attributed by Configliachi
and Rusconi not so much to the stimulating effect of the light on
the eyes, as to its influence on the skin. But the eyes of Proteus
though very rudimentary, have been shown by Kohl (’95, p. 207) to
possess all the essential parts of a functional organ of vision, and the
reactions observed by Configliachi and Rusconi, so far as the evidence
34 G. H. Parker.
advanced by them is concerned, may be explained entirely on the
assumption that the rudimentary eyes are the receptive organs.
Dubois (’90, p. 358), however, has experimented on Proteus with
much more conclusive results, for he has shown that it will respond
to a small beam of light thrown on its tail. Moreover, when the
eyes are covered with gelatine and lampblack, the animal will still
respond to light. There is then good reason to suppose that the
skin of Proteus is a receptive organ for light.
Finally Koranyi (’93) has shown that, under conditions of ex-
ceptional excitability, a beam of strong light, when thrown on a frog’s
back, will induce reflex movements in the legs. It is thus evident
that several amphibians, including the frog, possess skins with end-
organs, sensitive to light.
The observations of Eigenmann (:00, p. 113) on the reactions to
light of the blind-fishes Chologastre and Amblyopsis indicate that
the nerve-endings in the skins of fishes also may be stimulated by
light. I know of no evidence, however, that the skins of air-
inhabiting vertebrates are ever thus normally stimulated. It is of
course well known that when strong light is concentrated by a lens
it may become a powerful stimulus for the nerve terminals in the
human skin, but the organs that are affected by this are temperature
organs and not organs of sight. Nevertheless it must be borne in
mind that, as Koranyi (’93, p. 7) has demonstrated, the skin of a
frog is stimulated both by radiant heat and by light, and that these
two influences, distinct as they seem to our senses, are members of
one physical series in that they are both ether vibrations, varying
only in wave lengths. It is therefore conceivable that in the lower
vertebrates, like the frog, the end-organs in the skin are stimulated
by radiant energy of a wide range, including what is for us both
radiant heat and light, and that the descendants of these organs in
the skins of the higher vertebrates are more restricted in function
and are ordinarily sensitive only to radiant heat and its effects. It
is thus possible that the temperature sense organs in the skins of
the higher vertebrates may be specialized derivatives of radiant
energy organs such as presumably occur in the skin of amphibians.
Reactions of Frogs to Light. 35
SUMMARY.
I. Rana pipiens is positively phototropic to light varying in intensity
from 1 to 20,480 candle-metres. This applies always to the orienta-
tion and usually to the locomotion of the frog.
2. Individuals with the skin covered and the eyes exposed are
positively phototropic.
3. Individuals with the eyes removed and the skin exposed are
also as a rule positively phototropic.
4. The receptive organs in the phototropism of the frog are the
eyes and the skin, but not the central nervous organs.
BIBLIOGRAPHY.
BEER; 1.
:o1. Ueber primitive Sehorgane. [Separate from Wiener klinische Wochen-
schrift, 1901, Nr. 11-13. ]
CONFIGLIACHI, P., e Rusconi, M.
"19. Del Proteo Anguino di Laurenti Monographia, Pavia. [This work is
known to me only through the citation given by BEER (: 01, p. 30), and
the extended abstract published by EL.is (’21). ]
DuBsols, R.
90. Sur la perception des radiations lumineuses par la peau, chez les Protées
aveugles des grottes de la Carniole. Comptes rendus de l’Académie
des Sciences, Paris, cx, pp. 358-361.
EIGENMANN, C. H.
700. The Blind-fishes. Biological Lectures; Marine Biological Laboratory at
Woods Holl, 1899, pp. 113-126.
GIS. 2):
21. Observations on the Natural History and Structure of the Proteus
Anguinus by Sig. CONFIGLIACHI and Dr. Rusconi. Edinburgh philo-
sophical journal, iv, pp. 398-406; v, pp. 84-112.
GRABER, V.
83. Fundamentalversuche tiber die Helligkeits- und Farbenempfindlichkeit
augenloser und geblendeter Thiere. Sitzungsberichte der kaiser-
liche Akademie der Wissenschaften, Wien. Math.-naturw. Cl., lxxxvii,
Abt. I, pp. 201-236.
GRABER, V.
84. Grundlinien zur Erforschung des Helligkeits- und Farbensinnes der Tiere,
322 pp.
KOHL, C.
95. Rudimentare Wirbelthieraugen. Dritter Theil. Bibliotheca Zoologica,
Heft 14, pp. 181-274.
36 G. fl Parker.
KoRANYI, A. v.
by
93- Ueber die Reizbarkeit der Froschhaut gegen Licht und Warme. Central-
blatt fiir Physiologie, vi, pp. 6-8.
LoEB, J.
‘90. Der Heliotropismus der Thiere und seine Uebereinstimmung mit dem
Heliotropismus der Pflanzen. Wiirzburg, 118 pp.
LOEB, J.
‘94. Beitrage zur Gehirnphysiologie der Wiirmer. Archiv fur die gesammte
Physiologie, lvi, pp. 247-269.
MOLESCHOTT, J., et FUBINI, S.
’79. Sull influenza della luce mista e chromatica nell’ esalazione di acido
carbonico per l organismo animale. Atti dell’ Academia delle Scienze,
Torino, xv, pp. 55-219.
NAGEL, W. A.
‘96. Der Lichtsinn augenloser Tiere. Jena, 120 pp.
PARKER, G. H., and BURNETT, F. L.
700. The Reactions of Planarians, with and without Eyes, to Light. This
journal, iv, pp. 373-385.
PLATEAU, F.
89. Recherches expérimentales sur la vision chez les arthropodes. Mémoires
couronnés de |’Académie royale des sciences, des lettres et des beaux-
arts, de Belgique, xliii, 91 pp.
SEMPER, C. ;
81. Animal Life as affected by the Natural Conditions of Existence. New
York, 12°, xvi 472 pp:
WILLEM, V.
‘gt. Sur les perceptions dermatoptiques. Bulletin scientifique de la France et
de la Belgique, xxiii, pp. 329-346.
INFLUENCE OF RENNIN UPON THE DIGESTION OF
THE PROTEID CONSTITUENTS, OF MILK.
By P.. B. HAWK.
[From the Sheffield Laboratory of Physiological Chemistry, Vale University.]
CONTENTS.
Page
eM LECHLUICEONV OR etn a) ee) a) eeiog, Scmect. s, gie ite eee Ge a ka” Pals 37
Il. Experimental Tew cyl eats Pts. Ik Sa al eh 38
1. Influence of rennin upon the gastric digestion of milk proteids. . . 38
2. Influence of rennin upon the pancreatic digestion of milk proteids . 43
3. Comparative tests upon fluidegg-albumen . . ....... =. 45
MIRREN USIOUS Mls cs) en We will
return to the discussion of this problem later.
The medium ratios. — The four medium ratios between 3.10 and
3-40 can be explained by a partial retention of the sugar or by a
failing kidney.
The influence of feeding fat.— Loewi® points out that the varia-
1 JACKSON: Proceedings of the American Physiological Society, 1902. This
journal, 1903, viii, p. xxxi.
* Loew!: Archiv fiir experimentelle Pathologie und Pharmakologie, 1go1,
xlvii, p. 56.
8 Lusk: Zeitschrift fiir Biologie, rgor, xlii, p. 33.
* Loewr: Archiv fiir experimentelle Pathologie und Pharmakologie, 1902,
xlvill, p. 410.
® Pavy, Bropig, and SIAv, in the Journal of physiology, 1903, xxix, p. 467,
claim for Pavy the priority of this idea, referring to an article in the Lancet, 1902,
p. 816. But the wording of the Lancet article gives a different impression. It
reads as follows: ‘In both pancreatic and phlorhizin glycosuria there was a
parallelism between sugar and nitrogen eliminations, showing that the sugar was
derived from proteid catabolism, but various facts pointed to the sugar formation
occurring in the latter case solely in the kidney, and in the former in the general
system. It seemed probable that under the influence of phlorhizin the renal cells
acquired the power of catabolizing proteid with the liberation of dextrose.”
° Loew!: Archiv fiir experimentelle ‘Pathologie und Pharmakologie, 1901, xlvii,
p. 68.
ee
On the Action of Phlorhizin. 71
tions in the ratio are not due to feeding fat, which accords with
Lusk’s ! previous statements.
High ratios, after feeding very large quantities of fat, have indeed
been obtained by Hartogh and Schumn,,? but in this case there may
have been a large fatty acid excretion in the faces, and the balance
of glycerin belonging to the fatty acid may have been synthesized
to dextrose, for Cremer® has shown such a synthesis after feeding
glycerin in phlorhizin diabetes.
As a result of this evidence, we can claim that the usual D: N ratio,
in dogs living on proteid and fat, varies between 3.40 and 3.90: 1, or
a sugar production from proteid of between 62 and 54 percent. In
the following paragraph another cause which may producea variability
in the ratio will be shown.
THE PROTECTION FROM COMBUSTION OF SUGAR FORMED
FROM PROTEID IN THE ORGANISM.
We have already made the statement that there is a total phlorhizin
diabetes within certain limits, but these limits must not be over-
stepped by flooding the organism with sugar. This is true as regards
proteid sugar as well as sugar directly fed. An example of this
occurs in Loewi’s * work. A dog was fed with 250 gms. of meat and
received 2.5 gms. of phlorhizin every eight hours; the average ratio
was D:N::3.6:1. During the next 24 hours, 500 gms. meat and
5 gms. of phlorhizin were given every eight hours; the ratio fell to
2.6:1. The following day the feeding was reduced to that first
mentioned, and the ratio rose to 3.7. On the day 15co gms. of meat
were fed, 136 gms. of sugar were excreted; whereas 161 gms. were
called for, as indicated by the proteid metabolism. Probably a part
was burned, since the kidney could not effect its removal with suffi-
cient rapidity. Loewi suggests that more phlorhizin might have
brought about a complete elimination of the sugar on the day when
1500 gms. of meat were fed. A part of Loewi’s experiment, repre-
senting eight hour periods, is here reproduced (page 72).
Two comments upon this experiment occur to us. In the first
1 Lusk: Zeitschrift fiir Biologie, 1901, xlii, p. 34.
2 HARTOGH und ScHuMm: Archiv fiir experimentelle Pathologie und Pharma_
kologie, 1900, xlv, p. II.
3 CREMER: Miinchener medicinische Wochenschrift, 1902, p. 944.
4 Loewr: Archiv fiir experimentelle Pathologie und Pharmakologie, 1901, xlvii,
47; P- 54-
72 Percy G. Stiles and Graham Lusk.
place, the nitrogen of Period 1 on the third day is less than in Period
3 of the first day, although more meat was fed on the third day
period, and the influence of the larger meat feeding of the second
day must have been active eight hours after the last portion of 500
gms. of meat! Perhaps some burning of proteid sugar took place,
sparing some of the proteid from combustion. .
The second comment is on the evenness of the ratios following the
feeding of meat in excess. Reilly, Nolan, and Lusk found that if
500 gms. of meat were fed to a phlorhizinized dog (Dog Va), the
sugar and nitrogen can be eliminated twelve hours after feeding.
LOEWYS EXPERIMENT.
But on repeating this experiment four days later (Dog V b), there was
a lag in the sugar excretion indicating decreased kidney function.
In another dog a similar lag took place after feeding $70 gms. at one
meal. But this sugar which was retained was not burned, but was
eliminated in the after periods. This is shown in the table of ratios
given on the following page.
Within limits, therefore, proteid sugar seems to be protected from
combustion while awaiting removal by the kidney. The power of
the phlorhizinized organism to retain proteid sugar unburned is also
illustrated by the following experiment. In some of our work it was
inconvenient to give phlorhizin later than 11 or I 1.30 P.M., or earlier
than 8.30 or 9 A.M. Consequently there was a night period of nine
to ten hours during which it seemed possible that the phlorhizin effect
1 REILLY, NOLAN, and Lusk: Loc. cit., p. 402.
On the Action of Phlorhizin. 7a
might begin to wear off and the sugar be retained to some extent.
The supposition was strikingly confirmed. In the table of our work
on Dog II (p. 77), the preliminary period of the first day is given as
fifteen hours. The urine of this period was collected in two portions.
REILLY, NOLAN, AND LUSK’S EXPERIMENTS.
Fasting, 12 hours
Meat, 12 hours .
Following 12 hours
Following 6 hours .
Following 24 hours
Total, subsequent to meat
The first represented the twelve hours from 9.10 P.M., June 10, to
9.10 A.M., June 11. The phlorhizin (2 gms.) was given at II P.M.,
and hence the last hour of this period was the tenth since the injec-
tion. The second portion of the urine was collected from 9.10 A.M.
to 12.10 noon, three hours immediately following an injection of
phlorhizin. The analyses showed the following:
I, 12 hours
Il, 3 hours
Total
The interpretation is clear. During the latter part of the first
period the sugar elimination lagged, and there was an accumulation
in the system. Under the influence of a second dose of phlorhizin
this accumulated sugar was swept out in addition to the dextrose
properly belonging to the period. Consequently the sugar of the
first period is too low and that of the second much too high. Com-
74 Percy G. Stiles and Graham Lusk.
bining the two we have results for fifteen hours which there is every
reason to regard as trustworthy.
Ten hours is then a longer time than should be allowed to inter-
vene between injections, when it is desired to collect the urine for
short periods. It will be noticed that the nitrogen excretion remained
practically constant, instead of falling and rising with that of the
sugar. We cannot suppose that the dextrose which was held back
was burned to any appreciable extent; it seems to have appeared
almost if not quite quantitatively in the second period.
At the close of this experiment (Dog II) we made another trial
on the following plan. The usual dose of phlorhizin, 2 gms., was
given at the beginning of a twelve hour period. This period was
subdivided into periods of eleven hours and one hour, with the ex-
pectation that the urine of the last hour, the twelfth since the in-
jection, would show a considerable lessening in sugar content. The
findings follow:
Period.
11 hours
] hour
Period.
3 hours
An inspection of these figures shows that our expectations were not
met. The dextrose in the twelfth hour after the first injection was
equal to or slightly above the average for the fifteen hours. The fall
of the ratio in this hour was due to a slight rise of the nitrogen above
the mean. As to the reason why the phlorhizin effect was distinctly
impaired on June 11, when ¢ez hours had elapsed since the injection,
while on June 13 it was not diminished in ¢we/ve hours, it can only
be suggested that the longer time a dog has been receiving phlorhizin
the more susceptible it becomes to its action, so that the effect of
each successive dose tends to be more prolonged. The duration of
the action of phiorhizin therefore varies even in the same dog.
On the Action of Phlorhizin. 75
It is clear from the foregoing evidence that the phlorhizinized dog
has some power of retaining proteid sugar unburned in his organism
if the kidney is not in a position to eliminate it. Some irregularities
in the ratio may be explained by the lack of maximal phlorhizin action
on the kidney, during the later hours after injection. The duration
of this maximal activity varies with the individual.
THE PROTECTION FROM COMBUSTION OF DEXTROSE ADMINISTERED
SUBCUTANEOUSLY.
If the phlorhizinized organism has the power of retaining for a
time a certain quantity of proteid dextrose without burning it, then
the subcutaneous injection of small amounts of dextrose in a fasting
diabetic dog should result in the quantitative protection of such dex-
trose and its elimination in the urine. Perhaps there is a limited
quantity of some body which, combining with dextrose, renders it non-
combustible, —a compound which can be split by the phlorhizinized
kidney.
Two out of three dogs showed the quantitative elimination of
dextrose subcutaneotisly administered. We quote the successful
experiments. The unsuccessful result in one dog may have been
due to imperfect kidneys.
‘ The general plan of the method adopted was as follows. The
animal was given phlorhizin subcutaneously (in 1.2 per cent sodium
carbonate solution) three times daily until —by the third or fourth
day —the diabetic condition was well established and the elimination
of nitrogen and dextrose was proceeding evenly. After one or more
preliminary periods, during which the urine was collected and an-
alyzed, the subcutaneous injection of sugar was made. The dextrose
used was a pure preparation (Kahlbaum’s). A o.5 per cent solution
of this dextrose, made up by weight, gave a value of 0.515 per cent,
as determined by the Allihn method.
After the injection the urine was collected for periods of three or
six hours, and the amount of extra sugar eliminated, as compared
with that before the injection, could be noted. Six hours after the
introduction of the dextrose, the values had reached practically the
initial level.
This experiment shows, (1) an unusually high ratio elsewhere com-
mented upon. (2) The maintenance of this high ratio during a
76 Percy G. Stiles and Graham Lusk.
twelve-hour period, when 150 gms. of meat had been fed and only
0.5 gms. of phlorhizin. The smaller dose of phlorhizin likewise
DOG I.
Fasted three days; then 3 gms. phlorhizin every eight hours for two days.
Weight = 18 kg.
Phlor- ave n. ID:N Extra
“aN: Remarks.
hizin. sugar. 5s ;
8.40 A.M.
11.40 A.M. .O) |
>| 3.18-6.18 P.M.
9.30 P.M.
8.45 a.m.| 0.5 | 9.19-12.19 44|.... | 1230-1235. 5
gms. D sub-
12.15 P.M. B 12.19-3.19 : oo or | +4264; cut. in 50 c.c.
water.
| 3.19-6.19 85| 1.96 | 2.2} 122
9.45p.m.| 0.5 | 6.19P.M. to et. ae ... [6.20 pm. 150
= gms. meat.
8.45 A.M.
sufficed for the elimination of the sugar ME utaneously injected.
(3) The apparent elimination of extra sugar to the amount of 5.86
gms. six hours after the subcutaneous injection of 5 gms. of dex-
trose. The extra sugar is obtained by multiplying the nitrogen for
the period by the prevailing D:N ratio, which in this case is 4.39
(average of 4.44 and 4.34) and subtracting this proteid sugar from
the total as determined in the urine.
A second experiment on similar lines ran the following course.
(See Table, Dog IT.)
This experiment is a type of the total phlorhizin diabetes. The
average D:N ratio (excluding the sugar-injection periods) i
3.60:1. On June 11, after injecting 5 gms. of dextrose sucht
neously, extra sugar amounting to 5.47 gms. were found in the urine.
On June 12, after injection of 4 gms., 3.43 gms. of extra sugar were
eliminated through the kidney in six hours. The figures here and
in the previous experiment demonstrate the quantitative elimination
of dextrose administered in small quantities. We have seen that
dextrose in excess is burned, but in small quantity it apparently
enters into a chemical combination which is not combustible, and
which may be split by the phlorhizinized kidney.
On the Action of Phlorhizin. 77
DOG Il.
Fasted three days; then three days with 2 gms. phlorhizin every eight hours.
Weight = 18 kg.
Remarks.
I, II, 15 hours 39:22
III, 6 hours 20.28 nie 5 gms. dextrose in 50
c.c. water, subcuta-
IV, 3 hours 8.01 sabe neously.
V, 12 hours! 28.78 The)
VI, 3 hours 7.53 2.06
VII, 6 hours 17.90 4.02 aes : 4 gms. dextrose, sub-
cutaneously.
VIII, 3 hours 7.07 1.93 | 3.66
IX, 11 hours 25.58 7.06 | 3.62
1 5 gms. phlorhizin at 11 P.M.
This experiment has been used before in this article. It will be
noticed that Periods I and II are added together, because the even-
ing injection of phlorhizin did not cause the complete elimination of
HOURLY EXCRETION, DOG II.
Period. D. per hour. N. per hour.
ee Whours 2 2). 2.61 0.735
TU Gyhours: (3. 59s 3; sane 0.720
BV Sthours! hi Ooh 5. Bs Seta 0.683
Wet2phours)) 405. °°. 0.666
Wales Hours... -. -s 0.687
WalleGyhours. < = « earls 0.670
MELE S hours) .... 1 sai: 0.643
IX Mbhourse.7 sth “< 0.642
all the sugar, during the night, but this all came out in the three
hours following the morning injection. Five gms. at II P.M. on
June 12 caused a maintenance of the ratio, and during the night of
78 Percy G. Stiles and Graham Lusk.
June 13 2 gms. sufficed for eleven hours — something not possible
two days before. Arteaga! found that o.1 gm. doses become in-
creasingly effective in the cat. It has been a recent custom in our
laboratory to give 5 gm. doses for a day or two, and then 2 gms.
of phlorhizin. The ratio is then usually established on the second
day.
The evenness of the elimination of nitrogen and dextrose is re-
markable. In the experiment on Dog II the hourly excretion of
sugar and nitrogen is presented in a table on p. 77. The nitrogen
and dextrose constantly and slowly fall together, the ratio remaining
the same.
It must be noted in such experiments as these that after withdraw-
ing the urine by a catheter, the bladder should be washed three
times with warm water, and this operation should be finished and the
catheter quickly withdrawn within half a minute of the appointed
time. This is easily accomplished.
CONCLUSION.
It may be of advantage to state concisely what this discussion in-
dicates concerning the nature of phlorhizin diabetes. If phlorhizin be
administered in 2 gm. doses every eight hours to fasting dogs, there
is a preliminary sweeping out of the body’s sugar, and thereafter there
is established a ratio between nitrogen and dextrose in the urine of
3-75 : I (or 3.60 to 3.70, perhaps, in the light of more numerous ex-
periments than were at first at hand). If the phlorhizin action on
the kidney be complete, this ratio remains constant even in three-hour
periods. After feeding meat, gelatin, or casein, the ratio is un-
changed in the aggregate, although the sugar tends to be eliminated
before the nitrogen belonging to the proteid fed.?
Dextrose fed in small quantities per os or injected subcutaneously
is not burned, but eliminated quantitatively. Diminished action of
phlorhizin may lower the ratio for a time, but quick renewal of the
phlorhizin will bring about a ratio higher than normal, on account of
the fact that sugar from proteid has accumulated within the body
without being burned. The aggregate of two such periods shows
the normal ratio. Phlorhizin diabetes is therefore a total diabetes.
Dextrose within limits cannot be burned. It seems possible to ac-
1 ARTEAGA: Loc. cit.
2 REILLY, NOLAN, and Lusk: Loc. ciz¢.
On the Action of Phlorhizin. 79
cept Loewi’s hypothesis of a blood-sugar combination, with the addi-
tional hypothesis that the sugar while in this combination cannot be
burned. Phlorhizin will decompose it and permit the elimination of
the sugar in the kidney. Any free dextrose unites with the com-
bining radical and is protected. If the quantity of sugar rises above
this combining power, immunity from destruction is lost and the sugar
burns.
Vosburg and Richards! have shown that the blood-sugar increases
in that form of diabetes which is produced by the effect of intra-
peritoneal injections of adrenalin upon the pancreas. After such
injection, diabetes usually ensues within a few minutes. In one ex-
periment by Herter and Richards,” a fasting dog was treated with
phlorhizin and the urine tested until it was sugar-free. Intraperi-
toneal injection of adrenalin failed to produce glycosuria within four
hours, but during the following four hours a small amount of sugar
was eliminated. This can be explained if this dog’s organism, freed
from dextrose through phlorhizin, possessed at first a sugar-com-
bining power (in Loewi’s sense, with colloid) which, when satisfied,
was followed by that extra accumulation of free blood-sugar which
Loewi claims to be necessary for sugar elimination through the
kidney.
If phlorhizin diabetes is to be produced, animals with sound kidneys
are essential. Our experience has taught us never to use the same
animal at different dates, for the first experiment may have done vio-
lence to the kidneys and the 2.8 ratio is likely to be obtained.. In this
case the kidney has lost the power of splitting a dextrose combination
formed from a definite percentage of the proteid sugar, a compound
which is always burned in animals having the lower ratio.
1 VospurG and RicHarps: This journal: 1903, 1x, p. 35.
? HERTER and RICHARDS: Medical news, 1902, Ixxx, p. 201.
EXPERIMENTS ON THE DIGESTIBILITY OF
VEGETABLES.
By AUP) BRYAN TE AnD EK Diy MILNER:
[From the Chemical Laboratory of Wesleyan University.] °
CONTENTS.
Pag
OCONEE tea te ees eis sgn ia, eso. he er ay welds Sepeue® topes 81
eR EVICISME OC KD eRe win Wa 5 is Aes l Taw hss opieen far ts oo ok det JG, Aol ee Wale Bes $2
PeeDeRnVentS@hCherreporteduns 4 - (lie meatie gt ip te ay caus +) fa) cme 83
EpenulenfalpmethOdss eamis0 j=) clr otasees mies ect 22 3. Moms aac eter ee. c 8+
MreapmMentwOrtaeCeSvs.. Sp ome ee Se eh ck sr o's, TS, aul uee se sc 86
Sanapinmsof LOOdematenialswen oe ey te co) “ae ) fobcli len Ginsh 7) oa 86
IDEALS. OF WHET ep oe ay ke ont Poem ©e | peee eo Loo mmOn Ss SET 87
PepewmMents WithnoUpject, D.) etal suis t0iiry if Jaume) =) )pseen 87
BE pemimentSawithme Subj eGte lia.) sells oy ose ree ch et Soa d-y ore ce yo cmes 89
EE@pemmnentSmwiths SUD} ECUMVa js) Go Ware reins ) \ Ga" a) a eau 90
WesuirstOmtnecnexPenMentsies tt) v3) vu es nieee oe Mec) es) 2.) dae eh eps anes 90
IBferEROfesIZGy Ofte prablOnt striae eee ees) 028 ceed 3, Pe une ce 91
Waniatonsuin, individuals: tevsass 74 Cs eneee-” | Baeiralls, Sar) ae Caenee 91
pier eiecstibility. Qf Cabbage sao [Uhl ke is seen i os Ger ca ee | hee 95
suheacdioestibilityOtspOtatOes. fe. memmsrt ito. Weal Pit Mitr yates ames 96
Siemdiceshibilitveotubects; tea ainis +) =>,
Experiments on the Digestibility of Vegetables. 95
TABLE II — (concluded).
Protein
hydrates.
Experiment No. 16.
grams. | grams.
Basalerationy te sie Yc. eee ©. BK 276.6 | 251.0
(EEGCHrCOLM Lf.) a shee es ee ee : 85.2| 24.3
ECE SG mia Non ete ee co cual eh Cee Gil 53:61) 27.9
Digestibility of total ration, % . .|.... 90.7} 89.9 3 | 76.6
Estimated digestibility of corn, % .| .... | 83.9] 41.2} 966) 59.3
three subjects. Subject B. appeared to get the least, and Subject M.
the most, from the material consumed, though later in the series the
latter subject was unable to continue with the diet, and there was a
distinct decrease in the efficiency of digestion, as seen in the results
of Experiment No. 10. In all cases, however, the differences in the
proportions of carbohydrates digested by the different subjects were
small; in fact, the uniformity was unusually satisfactory.
The digestibility of cabbage.— Of the vegetables studied in these
experiments the results with cabbage were lowest. According to the
estimates for the vegetable alone, the digestibility of protein was par-
ticularly low. Considering that cabbage contains so little protein,
and that at least a half of it is non-proteid, the results for protein
in cabbage are of little importance. The ether extract of cabbage,
designated fat, probably consists of chlorophyll and other matters sol-
uble in ether with little or no food value. On the average for the
three subjects, 82 per cent of the carbohydrates was digested and
utilized by the body. Of the total energy of the cabbage only 60 per
cent was contained in the digested material, and when allowance is
made for the energy of the incompletely oxidized portion excreted in
the urine, not much over 57 per cent of the energy of the cabbage
was actually available to the body.
In the experiment reported by Rubner, in which cabbage was the
only article of the diet, the proportions digested were: protein, 81.5
per cent; ether extract, 93.9 per cent; carbohydrates, 84.6 per cent;
and mineral matters, 80.7 per cent.
96 A. P. Bryant and R. D. Milner.
The digestibility of potatoes. — Considering the average of the
three experiments with potatoes, about 73 per cent of the protein and
99 per cent of the carbohydrates appeared to be digested, while the
energy of the digested material was 94 per cent of the total amount
in the potatoes consumed. Correction for the energy lost insthe un-
oxidized protein would indicate that about 91 per cent of the total
energy of the potatoes was available to the body.
In Rubner’s experiment, in which potato was eaten with butter,
the digestibility of the nutrients was: for protein, 67.8 per cent; for ,
ether extract, 96. 3 per cent; for carbohydrates, 92.4 per cent; and for
mineral matter, 84.2 per cent. In one experiment by Constantinidi,
in which suet and gluten were eaten with potatoes, the digestibility of
the diet was: protein, 93.6 per cent; ether extract, 97.5 per cent; car-
bohydrates, 99.6 per cent; and mineral matters, 83.5 per cent. Ina
parallel experiment with the gluten omitted from the diet, the factors
for the different nutrients were: protein, 80.5 percent; ether extract,
98.8 per cent; carbohydrates, 99.3 per cent; and mineral matters, 87.7
per cent.
The digestibility of beets.— Although the beets were not particu-
larly relished by either of the subjects, their digestibility was high in
the experiments with all three, the average being: for protein, 72 per
cent, and for carbohydrates, 97 per cent; while go per cent of the total
energy of the beets was contained in the digested material. With
allowance for the energy of incompletely oxidized material, at least
87 per cent of the total energy of the beets was actually available to
the body. If the experiment with Subject M. were disregarded,
because of the indisposition of the subject, the results would be still
higher. .
There is no other experiment with beets with which to compare the
results from these. In Rubner’s experiment with carrots, which
closely resemble beets in composition, the factors for digestibility
were: protein, 61 per cent; ether extract, 93.6 per cent; carbohy-
drates, 81.8 per cent; and mineral matters, 66.2 per cent.
The digestibility of apple sauce. — The factors for the digestibility of
protein in the apple sauce were low for both subjects; but the quantity
of protein present was too small to be considered. Some fat, how-
ever, was present, as the sauce was made of baked apples to which
butter had been added, and the digestibility of the fat, as estimated
for the apple sauce alone, averaged 98 per cent for the two experi-
ments. The high factor for carbohydrates, averaging 99 per cent for
Ramer yey ee a
Experiments on the Digestibility of Vegetables. 97
the two subjects, was no doubt favorably influenced by the sugar
added in making the apple sauce. The digestible nutrients contained
99 per cent of the total energy of the apple sauce, nearly all of which
was actually available to the body, because so little of it was derived
from protein. In thirty experiments with fruits and nuts in various
combinations Jaffa found the digestibility to be on the average: pro-
tein, 75, fat, 86, carbohydrates, 95, and crude fibre, 74 per cent.
The digestibility of fibre. — On the whole, the results for the diges-
tibility of fibre were rather higher than might have been expected,
though the materials were all eaten before they had fully ripened, and
consequently the cellulose may have been in a more tender condition
and more readily acted upon by the digestive juices. The highest
factor was that for apple sauce, 95 per cent, and the lowest that for
green corn, 60 per cent. The latter, however, was with only one sub-
ject, and if single experiments are considered, the results with Subject
B. with potatoes and cabbage were both a trifle lower than that of
corn. It is noticeable that the average for beets, 84 per cent, was
above that for potatoes, 74 per cent; so also was that for cabbage, 77
per cent. In Constantinidi’s experiments with potatoes, the digesti-
bility of fibre was 78 per cent in one case, and 79 in the other.
Weiske ! made two experiments to determine the digestibility of fibre
in a diet of celery, cabbage, and carrots, in one of which he found 63
per cent digested, and in the other 47 per cent.
Income and outgo of nitrogen. — In order to calculate the balance of
income and outgo of nitrogen in these experiments, urine was collected
in all but three of them, and its nitrogen content determined. | Instead
of the urine for the whole period of each experiment, however, only
that for twenty-four hours, beginning at six o’clock on the morning of
the last day of the study, was taken, as it was believed that by that time
the body would have reached a stable condition as regards nitrogen
assimilation and excretion for the given diet. Table III shows the
elimination of urine for the day on which it was collected, and the
percentage and amount of nitrogen in it, together with the gain or
loss of nitrogen in the body as computed from the quantities of nitro-
gen in the food, feces, and urine, that in the food and fzeces being the
average per day for the experimental period.
Experiments Nos. 1-6 were with Subject B. In the first one, with
the large basal ration, there was a slight loss of nitrogen; in the
1 WEISKE: Zeitschrift fiir Biologie, 1870, vi, p. 456.
98 A. P. Bryant and Rk. D. Milner.
second, with the large basal reduced one-third, the loss was consider-
able. In the next study, when cabbage was added to the reduced
basal ration, the loss was somewhat less, but stilllarge. In the fourth
experiment, with potatoes added to the basal ration, there was aslight
gain of nitrogen, but in the next two, with beets and apple sauce, the
TABLE III.
URINE DaTA, AND INCOME AND OUTGO OF NITROGEN.
Nitrogen
gained (+)
or lost (—).
Experi-
ment
No.
Amount | Sp. gr. | Nitrogen | Nitrogen | Nitrogen | Nitrogen
of urine. | of urine. | in urine. | in urine. | in faeces. in foud.
per cent.
1.83
1.95
1.60
difference between income and outgo of nitrogen was practically nil.
In the two studies with Subject M. in which urine was collected, Nos.
8 and 9, with cabbage and potatoes respectively, there was almost the
same gain of nitrogen in both. In the studies with Subject W. there
was also a gain, which was small in the experiments with cabbage
and apple sauce, but large in the others. On the whole, except in
the study of cabbage with Subject B., the diet containing the vege-
tables seemed to supply the bodily needs of the different subjects for
. protein.
Experiments on the Digestibtlity of Vegetables. 99
CONCLUSION.
So far as sources of protein or fat are concerned, the vegetables in-
cluded in these studies may be considered as of little value. They
do, however, contain carbohydrates, which the results of these and
other experiments indicate to be quite well digested and absorbed;
and they may, therefore, be considered as of value as sources of energy,
a large proportion of which appears to be available to the body.
The chief value of many vegetables, however, is perhaps aside from
the nutrients or energy they furnish; they add a pleasing variety and
palatability to the diet, supply organic acids and mineral salts, and
give the food a bulkiness that seems to be of importance in its
mechanical action in maintaining a healthy activity of the alimentary
tract. Possibly the result of these conditions is a favorable influence
upon the digestion of other food eaten with the vegetable; at least
such an effect was suggested by the results of some of these experi-
ments. For instance, in the studies with Subject B., with potatoes
and with apple sauce added to the basal ration, the digestibility of the
total ration, including such material, was noticeably higher than that
of the basal ration alone.
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ON THE ACTION OF SALINE PURGATIVES IN
RABBITS AND THE COUNTERACTION OF
THEIR EPrFECE BY YCALCIUM
By JOHN BRUCE MacCALLUM.
[From the R. Spreckels Physiological Laboratory of the University of California. |
I. THe MECHANISM OF THE ACTION OF SALINE PURGATIVES.
ew attempting to explain the action of saline purgatives, Schmiede-
berg? states that these salts are absorbed with difficulty, and
hence reach the lower parts of the intestine unchanged. In the large
intestine, according to his theory, the salts prevent the faeces from
becoming compact by inhibiting the absorption of fluids from the
lumen. This hypothesis is supported by Wallace and Cushny,’ who
add that the absorption of fluids from the intestine is retarded
especially by the salts of those acids which tend to form insoluble
salts with calcium. Hofmeister‘ found that gelatin plates absorb
less water when soaked in solutions of sodium sulphate, tartrate,
citrate, etc., than they do when soaked in chlorides or bromides.
Schmiedeberg’s theory of the action of purgatives has been widely
accepted. The older idea of Liebig that the salt solution attracts
fluid from the blood into the lumen of the intestine on account of its
osmotic pressure has long been abandoned. The purgative action
of the salt solution does not increase with an increase in its con-
centration (Buchheim). Loeb,® while not denying the possibility of
the inhibiting action of the saline purgatives on absorption, states
1 A preliminary report of the results of these experiments was published in the
University of California Publications, Physiology, May 25, 1903, Vol.i, No. 2, p. 5.
2 SCHMIEDEBERG: Arzneimittellehre, Leipzig, 1883.
8 WALLACE and CusHNy: This journal, 1898, i, p. 411.
4 HOFMEISTER: Archiv fiir experimentelle Pathologie und Pharmakologie,
1888, xxv, p. I.
6 Loes: Decennial Publications of the University of Chicago, 1902, p. Io.
IOL
102 John Bruce MacCallum.
that these salts are identical with those which produce contact irri-
tability, muscular twitchings, and hypersensitiveness of the nervous
system. Further, he suggests that the increased peristalsis may be
due to an increase in the irritability of the nerves and muscles of the
intestine.
In order to decide this question, and to render more clear the
action of saline purgatives, I have made a series of experiments on
rabbits, testing a number of salts, including sodium citrate, sulphate,
tartrate, oxalate, phosphate, and fluoride, barium chloride, and mag-
nesium sulphate. It was found that the effects of the drugs could
be most satisfactorily studied by exposing the intestines and observ-
ing them directly in animals under the influence of morphine, Sub-
cutaneous injection of 5 c.c., I per cent solution of morphine proved
sufficient for surgical anzesthesia, without affecting the intestinal
movements in the rabbits which were used. These weighed on an
average twelve hundred grams. In addition to this method, many
experiments were made in which animals were kept in separate
cages, and the amount and character of the feeces observed during
several hours after the administration of the salts.
It was found that a// those salts which act as purgatives when
introduced into the stomach or intestine, have the same action
when injected subcutaneously or intravenously. The injection of
1-2 c.c.% sodium citrate solution? into the jugular vein of a rab-
bit causes a striking increase in the peristaltic movements. This
activity begins in from one to two minutes after the injection.
Loops of the intestine which before the injection lay quiet and
collapsed, are set in active motion. They become rounded and
prominent, and seem to occupy a greater volume. The fact that
the animal is under the influence of morphine does not interfere
to any extent with this action. Claude Bernard? has stated that
sodium sulphate introduced into the veins acts as a purgative;
and that the same action is obtained by subcutaneous injection of
magnesium sulphate.
When introduced into the intestine or stomach a much larger quan-
tity of the salt ts required to produce an equal effect, and the action
1 The salt solutions were made up with distiiled water as fractions of molecular
solutions. Thus % sodium citrate solution means } molecular weight of the salt
in grams (including water of crystallization) dissolved in tooo c.c. water.
* CLAUDE BERNARD: Lecons sur les effets des substances toxiques et médi-
camenteuses, Paris, 1857.
On the Action of Saline Purgatives in Rabbits. 103
takes place only after an interval of from ten to fifteen minutes. By
piercing the wall of the intestine or stomach with a hypodermic
needle, and forcing 5-10 c.c. % sodium citrate solution into the
lumen, a similar increase in peristalsis is brought about. The
movements begin about ten to fifteen minutes after the injec-
tion, not only in the loops of intestine which contain the solution,
or the loops near the stomach, but simultaneously in all parts of the
intestine.
When given subcutaneously, these salts do not act immediately, but
only after an interval of ten to fifteen minutes. By this method usu-
ally a fairly large quantity, ¢.g., 10 c.c. 4% sodium citrate solution
is necessary to produce increased peristaltic movements. When
the actual passage of feeces is observed, relatively large doses also
are necessary. No appreciable effect is obtained with less than
10 c.c. % sodium citrate solution given subcutaneously. A number
of control animals were kept in each case, and the average weight
and character of the normal faeces carefully determined. During the
first six hours following the injection of the salt, the faeces were
collected and weighed. The purgative effect usually takes place
during the first hour. Considerable variation in the action is to
be observed in different rabbits; but with subcutaneous injection
of the salt, there is constantly a marked increase in the weight. of
the fzces varying from two to six times the average normal
weight. The feces are sometimes of a semi-fluid character, a fact
which is the more striking because the normal feces of rabbits
are drier and more definitely formed than the dejecta of most
other animals.
Schmiedeberg’s theory was an effort to explain this increase in
the fluid contained in the feces following the administration of a
saline purgative. That the prevention of the absorption of fluids
from the intestine is not the main cause of the production of faeces
containing much fluid is shown by the following facts: In addition
to the increase in peristalsis caused by the administration of the
saline purgatives (whether subcutaneous, intravenous, or intraintes-
tinal) there zs to be observed also a more or less marked increase in the
secretion of fluid into the intestine. A short time (twenty to thirty
minutes) after the salt is given there is usually found in the intes-
tine a considerable quantity of a clear yellow fluid. This does not
resemble bile. The collapsed loops of intestine, in addition to being
set in active squirming motion, become gradually filled with fluid after
104 John Bruce MacCallum.
the purgative is administered.1. The actual passage of faeces takes
place usually within an hour after the intravenous injectiom of the
purgative. The exact nature of the fluid secreted into the intestine
has not been studied, and the details of its secretion remain to be
investigated. It was noticed also in a number of. cases that an
increased flow of saliva and urine occurred when one of these salts
was introduced into the body. These problems of secretion are still
to be studied. After the subcutaneous administration of sodium
fluoride solution, the salivation in,two or three cases was so marked
that the saliva fell in drops to the floor. In overdoses of sodium
citrate, the same phenomenon was observed in several cases; while
it is well known that barium chloride causes salivation. Repeated
urination is a fairly constant accompaniment of the subcutaneous
and intravenous injections of all of these salts.
In connection with the production by these purgative salts of an
increased flow of fluid into the intestine, together with salivation
and increased urination, it is interesting to note the subcutaneous
and intravenous administration of pilocarpine and physostigmin as
purgatives by veterinarians. These drugs act primarily upon the
glands of the body, and no doubt owe their purgative action to the
production by them of increased secretion of fluids into the intestine.
The action of the various salts which act as purgatives when ad-
ministered subcutaneously or intravenously varies in intensity; but
it is difficult to make a definite list in which they can be placed in
order of increasing strength. Barium chloride is the most powerful
of all. The action is rapid, and with very small doses marked purga-
tive effects are obtained. It is well known among veterinarians as a
powerful purgative, and is commonly administered by them intrave-
nously. An intravenous injection of 0.75-1 gram is sufficient to
purge a horse weighing one thousand pounds. The intensity of
action of sodium citrate, fluoride, sulphate, tartrate, oxalate, and
phosphate decreases approximately in the order named. Magnesium
sulphate, when given subcutaneously, is no less active than sodium sul-
phate, but has a somewhat more poisonous effect than sodium sulphate
or sodium citrate. Large subcutaneous doses often cause death in
a very short time in rabbits, while equal doses of sodium sulphate
seem to be harmless. As a subcutaneous purgative, it is not to be
1 As quoted by SCHMIEDEBERG (Loc. cit.), BUNGE has stated that sodium
phosphate in rabbits causes an increase in the fluids contained in the alimentary
canal. I have been unable to find BUNGE’s original statement.
Ox the Action of Saline Purgatives in Rabbits. 105
recommended. Barium chloride also produces unfavorable symp-
toms. Rabbits often do not recover from a relatively small dose.
Sodium salts, however, such as the sulphate, citrate, tartrate, etc.,
may apparently be given subcutaneously with impunity. With re-
gard to the common use of barium chloride by veterinarians, atten-
tion must be called to its dangerous nature, and to the possibility of
the occurrence of serious after-results.
In addition to the purgative effects of these salts, there is produced
by the injection of sodium citrate, tartrate, fluoride, oxalate, and
phosphate a condition of extreme hypersensitiveness of the muscles
and the nervous system, which is entirely analogous to that de-
scribed by Loeb! in isolated muscles, motor nerves, and the skin of
frogs. The muscular twitchings and increase in nervous excitability
which Loeb caused in isolated muscles and in living frogs by means
of these salts, I have been able to produce to a marked degree in
rabbits by the subcutaneous injection of the same solutions. Sub-
cutaneous injection of Io c.c. 4% sodium citrate solution produces
within half an hour well-marked twitchings of the muscles in all
parts of the body, more noticeable in those of the gluteal region,
These are accompanied by tetanic contractions of the limbs, and
in some cases by general convulsions of varying intensity. There
is always a hypersensitiveness of the skin, and all the reflexes are
much exaggerated. One of the first effects to be noticed is an inco-
ordination of the movements of the hind limbs. If the animal is
held up by the ears, the feet tremble, and, if touched, the hind limbs
jerk away violently, or become rigid. The muscular twitchings
appear immediately at the place of injection, but only after twenty
to twenty-five minutes on the opposite side of the body.
In animals to which repeated small doses of sodium citrate are
given, a chronic state of hypersensitiveness may be produced. Daily
doses of 5 c.c. % sodium citrate solution were given to a rabbit
throughout one month, and then discontinued. -A condition of
marked hypersensitiveness persisted for four weeks after the last dose
was given. In such an animal muscular twitchings may be produced
on the side of the body opposite to the injection in as short a time as
two minutes, while in a normal animal it takes twenty to twenty-five
minutes.
To recapitulate, then, those salts which produce muscular twitch-
1 Lors: Loc. cit.
106 John Bruce MacCallum.
ings and hypersensitiveness of the nervous system, produce also
increased peristalsis, and increased secretion of fluid into the intestine.
They produce this purgative effect not only when introduced into the
intestine, but also when injected subcutaneously or intravenously.
The presence of the saltin the lumen of the intestine ts therefore not
necessary for tts cathartic action. When introduced into the intes-
tine, it takes much longer to act, and requires a much larger dose
than is needed when injected into the blood. This seems to indi-
cate that che salt must first be absorbed into the blood before it can
act on the intestine. The essential feature in the action of saline pur-
gatives is not their presence in the lumen of the intestine, but their
absorption into the blood, and the production by them of a con-
dition of hypersensitiveness of the nervous system controlling the
intestine. It is difficult to say which part of the nervous system
is specially affected. It is possible that the muscle and gland
cells themselves are influenced; and it seems justifiable to sug-
gest that the action of these salts is not a specific one, but that
they cause a more general hypersensitiveness. It is difficult to
disprove Schmiedeberg’s theory, that the absorption of fluids
from the intestine is retarded by these salts; but it is certain
that the main factor in the production of fluid or less solid feces is
the increased secretion of fluid intotheintestine. Since an increased
secretion is caused by the absorption of the salt, the methods com-
monly employed in determining the rate of absorption of fluids from
the intestine must be defective because they consist in measuring the
fluid contained in a’certain loop before and after the introduction of a
measured quantity of salt solution. This quantity would .remain
relatively large, not because of a retardation of the absorption, but on
account of the increased flow of fluid into the intestine caused by the
absorption of the salt.
II. THe Errecr oF CALCIUM IN OPPOSING THE ACTION OF
SALINE PURGATIVES.
It has already been shown by Loeb that the muscular twitching
and hypersensitiveness of the nervous system produced by sodium
citrate, etc., may be inhibited by calcium salts. I have found that a
complete analogy exists between this action and the production and
inhibition of peristaltic movements in the intestine. Active peristalsis
produced by the tntravenous injection of a minimal dose of any purga-
iste og
On the Action of Saline Purgatives tn Rabbits. 107
tive sodium salt can be almost entirely suppressed by the subsequent
injection of an equal quantity of % calcium chloride solution. This
counteracting effect takes place within one to two minutes after an
intravenous injection, and in ten to twenty minutes after a subcutane-
‘ous injection, or the introduction of the solution into the intestine or
stomach. A much larger quantity is required when introduced sub-
cutaneously or into the alimentary canal. To illustrate this effect, one
of a large number of experiments may be cited. To two similar
rabbits,’equal doses of morphine were administered. The abdominal
cavities were opened, and in each case 1} c.c. % sodium citrate solu-
tion were injected into the jugular vein. Active peristalsis set in in
both animals. After five minutes 14 c.c. % calcium chloride solution
were injected into the jugular vein of one animal, while the other was
left without further treatment. Almost immediately the peristaltic
movements of the intestine of the animal which received the calcium
chloride ceased. Those of the other animal continued. Usually after
the injection of calcium the intestines remain motionless for one-half
to one hour, while in some cases slight peristaltic movements begin
after five to ten minutes. This depends largely upon the relative
amounts of citrate and calcium chloride solutions which have been
administered. It is possible after the movements have been entirely
inhibited by calcium chloride to make them active again by a second
injection of one of the purgative sodium salts. It is unimportant
whether the intestines are set in motion by sodium citrate, sulphate,
tartrate, phosphate, or oxalate. The calcium chloride has the same
inhibiting effect in all cases. The peristaltic movements produced by
barium chloride are usually not stopped by the administration of
calcium.
As suggested by Dr. Loeb, the salts which produce rhythmical con-
tractions in voluntary muscle are those which are liable to decrease
the concentration of the free calcium ions. The same is true of the
salts which act as purgatives; and the hypothesis that their action is
due to the removal of calcium from the tissues, or from the fluids
bathing them, is a plausible one. It is certain that the subsequent
addition of calcium to the tissues restores them to the condition in
which they existed before the injection of the purgative salt, and en-
tirely counteracts the purgative effect. It is only on these grounds,
and from the fact that these salts in some cases precipitate calcium,
that it is possible to state that the purgative action is caused by the
removal of calcium from the tissues.
108 John Bruce MacCallum.
It is not possible to make the general statement that the anion
stimulates and the kation inhibits the muscular contractions. In a
number of salts it is clearly the anion which causes the purgative
action and the muscular contractions, ¢. g., sodium citrate, sulphate,
tartrate, fluoride, etc.; while in other cases it is equally clear that the
kation has the same stimulating action, ¢e. g., barium chloride. The
kation of calcium chloride on the other hand has an inhibiting effect.
That this action also is not dependent upon the valency of the ions is
shown by the fact that barium chloride, sodium fluoride, and sodium
citrate all have the same effect, while barium chloride and calcium
chloride have effects which are directly opposed to one another.
Moreover, the effects produced by magnesium and sodium sulphate
on the intestine are similar.
Little can be said at present concerning the practical application of
these facts in the treatment of disturbances of the human intestine.
The administration of calcium, however, seems rational in cases of
persistent diarrhea, especially in those cases accompanying hysteria or
any disturbance of the nervous system. And since calcium itself has
an irritating action when given in large doses, the treatment would
seem to be contraindicated in cases in which the diarrhoea is caused
by an inflammation of the mucous membrane of the intestine. The
empirical use of calcium, usually in the form of chalk, is commonly re-
sorted to in cases of diarrhoea, its action being explained by the state-
ment that it reduces the acidity of the intestinal contents. But it is
evident that it can only act through the calcium, since calcium chlo-
ride is more efficacious even than chalk.
In addition to this, the fact that a chronic hypersensitive condition
of the muscular and nervous tissues may be brought about in rabbits
by the continued administration of salts which tend to diminish the
concentration of calcium ions, suggests that the addition of calcium to
the tissues might be of service in the treatment of similar conditions
in human beings. Those conditions which most nearly resemble that
produced in rabbits are hysteria, neurasthenia, and the allied states.
Whether there is in reality any analogy, and whether calcium will
prove of benefit in such cases, must be the subject of clinical investi-
gation.
The question arises also as to the possibility of administering pur-
gatives to human beings subcutaneously or intravenously. Although
in general such methods would be contraindicated, certain cases might
arise in which it would be of distinct advantage. The present experi-
On the Action of Saline Purgaiives in Rabbits. 109
ments seem to indicate that subcutaneous or intravenous administra-
tion of some of the salts, especially sodium citrate, might be safely
resorted to. Neither barium chloride nor magnesium sulphate should
be given in this way.
III. CoNncLusIONs.
1. In general, the saline purgatives act not only when introduced
into the intestine, but also when injected subcutaneously or intra-
venously.
2. The intensity of their action is greatest with barium chloride
and decreases approximately in the following order: barium chloride,
sodium citrate, fluoride, sulphate, tartrate, oxalate, and phosphate.
3. The purgative action of these salts is caused, first, by an in-
crease in peristalsis, and, second, by increased secretion of fluid into
the intestine, both of which can be directly observed.
4. Although I have not specially studied the secretion of other
glands, I have noticed that in a number of cases an increased flow of
saliva and urine occurred when these salts were introduced into the
body.
5. Intravenous injection of 1-2 c.c. % solution of these salts causes
increased peristalsis within one minute. When introduced into the
intestine, it takes ten to fifteen minutes, and five times the amount to
produce an equal effect.
6. This seems to indicate that even when these salts are intro-
duced into the intestine, they must be absorbed into the blood before
they can produce their purgative effect, and that they affect the in-
testine by increasing the irritability of the nerves and muscles, as
Loeb has suggested. Their action in producing less solid feces is
-not due to the prevention of the absorption of fluids from the intes-
tine, but to the production of an increased secretion of fluid into the
intestine.
7. By the continued administration of small doses of sodtum cit-
rate, a chronic condition of hypersensitiveness of the nervous system
may be brought about in rabbits, which persists for a considerable
time after the drug is discontinued.
8. By the injection of solutions of calcitim chloride, the peristalsis
caused by these salts can be entirely inhibited.
g. There is a perfect analogy between these actions and the pro-
IIO John Bruce MacCallum.
duction and suppression of muscular twitchings and nervous hyper-
sensitiveness.
10. The administration of calcium is, therefore, rational, especially
in those cases of diarrhoea in human beings which accompany
hysteria or nervous excitability of any sort.
The study of this subject was suggested to me by Professor Loeb,
and it is a pleasure to thank him for the interest which he has taken
in the experiments, and for many helpful suggestions.
fie CH REBRO-SPINAL FLUID IN HYDROCEPHALUS:
By [sADOK H. CORTAT:
[From the Chemical Laboratory of the Worcester Insane Hospital.]
eS of the cerebro-spinal fluid in hydrocephalus have
been rather infrequent, because of the comparative rarity of
this condition, and of the greater attention given to the anatomical
rather than to the chemical findings. During the last few years the
cerebro-spinal fluid has assumed great importance, from both a
normal and pathological standpoint. Much remains to be cleared
up, however, especially in regard to the form of proteid present, the
nature of the reducing body, and the molecular concentration as
established by the freezing point and its relation to the chloride
content, before we shall know what analogy, if any, exists between
this fluid and blood-serum. Unfortunately, the large amounts of
fluid required for an investigation of this nature can only be obtained
in hydrocephalic cases. So far as known, the fluid in this disease
does not differ in its composition from that found in normal con-
ditions. I have been able to discover only seventeen analyses of
hydrocephalic fluid. The latest are two by Panzer! and one by
Salkowski.2 In these three the fluid was obtained at an early
period of life. In Panzer’s case, 455 c.c. and 180 c.c. respectively
were obtained from two hydrocephalic foetuses; in Salkowski’s case,
that of a young child, there was 1050 c.c. of fluid.
My case was that of a congenital hydrocephalic imbecile, forty-two
years of age, who died suddenly, without any previous illness.
The fluid was obtained an hour and a half after death. The
pressure was considerable, for, following an accidental slight rupture
of the hemispheres during the removal of the brain, a stream of fluid
escaped to the distance of almost six feet. In spite of this, how-
ever, 750 c.c. was obtained, although at least 150 c.c. was lost.
1 PANZER, T.: Miinchener medicinische Wochenschrift, August, 1899, p. 805.
2 SALKOWSKI, E.: Festschrift fiir M. Jaffé, 1go1, p. 263.
DEE
112 Isador Hl. Cortat.
There was no spontaneous coagulation. The color was clear straw.
After precipitation of the proteid by alcohol, the alcoholic extract
was of a distinct amber color, and in the spectroscope there could be
detected a broad band of absorption extending from F in green to
beyond G. This coloring matter could not be extracted by shaking
with amyl alcohol. The specific gravity of the fluid was 1012, and
the reaction very slightly acid. A trace of lactic acid was present.
There was a marked reduction with Fehling’s solution. After re-
moval of the proteid by heat coagulation, this reducing body was
submitted to further study, with the following results. Fehling’s
test, positive. Nylander’s test, positive. Molisch’s reaction, deep
purple ring. Trommer’s test, positive. Sodium hydrate (with warm-
ing), deep yellowish-brown color. Fermentation test, positive after
twelve hours. A control test with the same sample of yeast and dis-
tilled water gave no reaction after the same length of time. Phenyl-
hydrazin test: 50 c.c. was heated in the water bath for an hour, with
I gram phenylhydrazin and 2 grams sodium acetate, and allowed to
cool slowly. There was obtained an abundance of long yellow
needles, arranged in sheafs, resembling phenylglucosazon and having
a melting point of 205° C. Ferric chloride, no reaction. Hydro-
chloric acid — phloroglucin— negative. Orcin, negative. Ammoni-
acal silver nitrate, strong reduction to metallic silver. The amount
of this reducing body (in terms of dextrose) was 0.917 gram per
1000 c.c.
Pyrocatechin, cholesterin, and cholin were absent. There was a
trace of fat. Urea was present, the amount being 0.750 gram per
1000 c.c. The freezing point was —o.65° C., and the relation —
per cent was 0.97. The total proteid was 1.180 grams per 1000 c.c.
Fibrin, nucleoproteid, albumose, peptone, mucin, serum albumin, and
fibrinogen (no coagulation at 56° C.) were absent, the entire proteid
content present being in the form of serum globulin. It coagulated
at 75° C., and was completely precipitated by saturating the fluid
with magnesium sulphate in substance.
The ash on elementary analysis was found to consist of phosphorus,
potassium, and sodium; calcium and magnesium were present in
traces, while iron was absent. The nitrogen of the proteid and urea
amounted to 0.5312 gram per 1000 c.c. The relation KCl: NaCl
Was 1) 1029-- 0f KO Na, Ouwas: 148.5.
Tests for a proteolytic ferment showed no digestion of fibrin after
The Cerebro—Spinal Fluid in Hydrocephalus. 113
twenty-four hours, either with or without the addition of hydrochloric
acid. A diastatic ferment, however, was present, but it was inactive
in a neutral or acid medium, and only became active after the fluid
was made slightly alkaline with sodium carbonate. It carried starch
digestion through the various dextrines to the final production of
maltose. The tabulated results of the quantitative analysis follow:
Substance. ok ea Substance.
Parts per
1000 c.c.
Vi 2iSith A 983.8 Potassium chloride. . . 0.6559
ixedisolids’= . .- . . |): 16.2 Potassium oxide... . 0.4143
Volatile substances. . . 10.6 Sodium oxide... . . 3227
ANS) “gs an se ee 5.6 Reducing body — ae. = - 0.917
sotalsproteid... =.) . ; 1.180 Motalinitrogen) ~ 7.) 1.260
Extractives and salts . . 15.020 inca antes Bee et acto: 0.750
hlonides; = 1. 6. 2) on Phosphoric acid. =. -- 0.090
Sodium chloride .
The coloring matter was a lipochrome, resembling serum lutein
(lutein of Kiihne), but differed from it in not being soluble in amyl
alcohol, and in presenting before the spectroscope one broad band
of absorption, instead of two narrow ones, although occupying the
same relative position in regard to the lines. The reducing sub-
stance was of the hexose group, as it failed to give the characteristic
reaction for pentoses with orcin or hydrochloric acid — phloroglucin.
In all probability it was dextrose, as it responded to all the usual
tests, reducing the salts of the various metals in alkaline solution,
producing fermentation with yeast, and the osazon, in both crystalline
form and melting point, exactly resembled phenylglucosazon. Panzer’s
second case contained glucose, and Salkowski in his last analysis
speaks of the fluid as containing a fermentative sugar. This is in
direct opposition to Halliburton’s claim, that the reducing substance
in cerebro-spinal fluid is pyrocatechin, for a careful testing of the
fluid in my case for this substance, using the method recommended
by Halliburton, failed utterly to detect it. The characteristics on
which he bases his claim, namely, the non-production of alcoholic
fermentation and the failure to reduce bismuth salts, were certainly
114 Isador F1, Cortat.
absent in this analysis. In large amounts of fluid obtained from cases
of general paralysis, I also found this reducing body responding to
the tests detailed above. At one time I believed that it was closely
related to the nitrogenous glucosides (cerebrins), but further investi-
gation along this line, using the method for isolation of the cerebrins
recommended by Koch, has shown this to be untenable. The amount
of dextrose present was greater than that contained in blood. The
chloride content is greater than in normal serum, and the larger part
of it is in combination with the univalent radicals, sodium and potas-
sium. The molecular concentration, as shown by the depression of
the freezing point, is also greater than in normal serum or in defibri-
nated blood, for Hamburger has shown the freezing point of the
latter to be that of blood-serum. This lower freezing point is due to
the greater quantity of dissociated salts, especially the chlorides.
A proteolytic ferment was absent, but, as in the blood, there was
present a ferment capable of hydrolyzing starch into sugar (maltose).
The proteid consisted entirely of globulin. Its coagulation point was
the same as that of serum globulin, and furthermore, it was com-
pletely precipitated by magnesium sulphate in substance. There
was no cholin, thus proving the absence of any recent active degen-
eration in the central nervous system.
ON THE TIME RELATIONS OF PROTEID METABOLISM!
By, Bae Ba EEAWie
[From the Chemical Laboratory of Wesleyan University.]
CONTENTS.
Page
Inligiomcallsiniiineiay sake Eke: TGiRGn es To) Soe nascnaon pa temeremrin Un meny ao toe eno ellis)
Description MTR sy se Se ate! oy SMe a sy cokes 119
Purpose and plan of the experiments here reported. Diet. Subjects. Daily
schedule. Preparation of samples, methods of analysis, etc.
Discussion of general results . Oe Sh ee ee tet ee em ee
Urine volume. Nitrogen excretion by Subject H. Nitrogen excretion by
Subject R. Sulphur excretion by Subject H. Sulphur excretion by Sub-
ject R. Phosphorus excretion by Subject H. Phosphorus excretion by
Subject R. Income and outgo of nitrogen (Subject H.). Income and
outgo of nitrogen (Subject R.). Income and outgo of sulphur (Subjects
Hi. and R.). Income and outgo of phosphorus (Subjects H. and R.).
Ratio of nitrogen to the heat of combustion of the unoxidized material in
the urine.
(SOROMOANS: 74” io ME eh Bd Onecare come ORE te ett Nema prem bores of iaceten tris
124
HISTORICAL SUMMARY.
NVESTIGATIONS! upon the time relations of proteid meta-
bolism seem to have been inaugurated about 1855 by Becher.”
About this date Lehmann? writes, “It is noteworthy that very soon
after the ingestion of food rich in nitrogen an increase in the urea
excretion occurs, and five-sixths of the nitrogen contained in the
food is often eliminated in twenty-four hours, as urea.” Lehmann
does not appear to have made accurate observations upon the in-
i It is not the author’s purpose, however, to give a complete review of this sub-
ject, but rather to report the results of his experiments, with only such references
to other work as seem called for in this connection.
2 BECHER: Zeitschrift. fiir rationelle Medicin, 1855, vi, p. 249.
3 LEHMANN: Lehrbuch der physiologische Chemie, second edition, i, p. 163.
115
116 P. B. Hawk.
crease in the urea output from hour to hour, neither does he draw
any conclusions from his observations.
Karl Voit,! in 1857, described experiments made with dogs fed on
different quantities of lean meat. He found that the urea excretion
was increaséd the first hour after the meal, reaching its maximum in
the seventh hour. His figures indicate that in twenty-four hours
after ingestion, an amount of nitrogen equivalent to that contained
in the food was excreted mainly as urea.
Winternitz? in a series of experiments carried out upon himself
reached the conclusion that the maximum urea excretion occurred
sometimes in the third, and sometimes in the fourth hour after
normal feeding. In one experiment this investigator took 40 c.c.
“rum” with an ordinary meal, determined the urea content of the
urine in hourly periods thereafter, and found the maximum excretion
during the first hour (8-9 a.M.). The food was not analyzed, and
there were no periods with normal diet for comparison.
J. W. Paton? in a series of investigations with himself and Gamgee
as subjects, observed, among other things, the effect of severe mental
work upon metabolism. He found that with mental work the amount
of urine and its nitrogen content increased, and the phosphoric acid
content slightly decreased. On returning to comparative rest after
the severe mental work, the nitrogen excretion was greatly diminished
in every case, while the phosphoric acid excretion was slightly in-
creased with one subject and unchanged with the other. The author
holds that urea has no relation to mental work except in so far as the
latter influences the excretion of water. Under this “ perverted
nervous action” a general wash-out takes place, and the urea is there-
fore increased.
Forster,’ in an experiment of twenty-four hours’ duration, upon a
man, collected the urine in six four-hour periods. The experiment
began with a breakfast of meat supplying eighteen grams of nitrogen ;
no other food was taken. The largest excretion of nitrogen was during
the second, and of phosphoric acid during the first, four-hour period.
The author shows that when Voit’s results are tabulated in four-hour
periods, the maximum excretion of nitrogen is also in the second
four-hour period, or from five to eight hours after the ingestion.
1 Voir: Physiologische chemische Untersuchungen, Augsburg, 1857, p. 42.
WINTERNITZ: Wiener medizinische Jahrbiicher, 1864, xx, p. 3.
PATON, J. W.: Journal of anatomy and physiology, 1871, v, p. 285.
3
4 Forster: Zeitschrift fiir Biologie, 1873, ix, p. 383.
On the Time Relations of Proted Metabolism. 117
Panum! and Carl Philip Falck? made valuable experiments with
dogs. Those of Panum give interesting data concerning the influ-
ence of fat upon the excretion of urea. Upon a diet of five hundred
grams of beef the dog excreted its maximum of urea in from two and
one-half to five hours after the ingestion, whereas when thirty grams
of pork fat were added to a like amount of beef, the maximum urea
excretion did not occur until the sixth or eighth hour thereafter.
When one hundred and fifty grams of rye bread were added to the
latter ration, the maximum urea excretion occurred after only one and
one-half to four hours. Unfortunately the periods of collection of
urine were irregular, and the results are consequently less definite
than might be desired. Falck fed his dogs quantities of beef, ranging
from one-half to one and one-half kilograms, and made hourly collec-
tions of urine thereafter by use of a catheter. The maximum excre-
tion of nitrogen occurred during the seventh to twelfth hour after the
ingestion of the food, being in general later as the quantity of meat
was larger.
Oppenheim? in experiments upon himself found the maximum of
urea generally from four to seven hours after somewhat irregular
meals.
Feder? conducted an extensive series of experiments upon the
time relation of proteid metabolism with dogs. In each case one-
half kilogram or one kilogram of meat was taken at the beginning
of the experiment. The maximum nitrogen excretion was reached
in four to six hours with one-half kilogram and in six to eight hours
with one kilogram of meat. The excretion of sulphur as sulphate in
the urine reached its maximum in one experiment in four to six hours
and in another in two to four hours. In every case the excretion of
phosphorus as phosphate rose more rapidly than that of nitrogen
and reached its maximum earlier (in two to four hours); a rapid
decline soon followed. The author found that when five grams of
sodium chloride were added to the meat diet the excretion of nitro-
gen was accelerated and increased. Feder also made a series of
experiments with a diet containing fat, supplied in 150-200 grams of
1 Panum: Nordiskt medicinskt Arkiv, 1874, Jahresbericht fiir Thierchemie,
1874, iv, p. 361.
2 Farck, C. P.: Beitrage zur Physiologie, Hygiene, Pharmacologie, und Toxi-
cologie, i, Stuttgart, 1875.
3 OPPENHEIM : Archiv fiir die gesammte Physiologie, 1880, xxiii, p. 446.
4 FEDER: Zeitschrift fiir Biologie, 1881, xvii, p. 531.
118 P. B. Hawk.
bacon, added to 400-500 grams of beef. The maximum nitrogen
excretion occurred in two to six, that of sulphur in one to two, and
that of phosphorus in one to four hours after taking the food. There
were no analyses of the food and no central periods.
The results of such comparatively recent investigations as those of
Rosemann,! Tschlenoff,? Riazantseff,? Veraguth,* Roeske,® and Graf-
fenberger ® have already been sufficiently reviewed in this journal.’
In an inquiry regarding the excretion of uric acid, Frangois Marés ®
gives data regarding the elimination of nitrogen. In nearly all of his
experiments the urine was collected hourly and the nitrogen content
determined. The subjects were boys and men ranging in age from
thirteen to forty-five years. They received amounts of beef varying
from one-half to one and one-half kilograms at a single meal. The
maximum nitrogen excretion appeared in six to nine hours after the
ingestion. The author concludes that uric acid excretion is a “ func-
tion of age and individuality,’ and that the nitrogen excretion is
variable, depending principally upon the quantity of nitrogenous food
ingested, and is independent of the age and individuality of the
subject.
Gley and Richet® in experiments with themselves on uniform
diet found the maximum urea excretion three to four hours after the
ingestion of the food. The conditions were, however, somewhat irreg-
ular and the data by no means complete.
Sondén and Tigerstedt in the discussion of the results of an
inquiry on “Die Respiration und der Gesammtstoffwechsel des
Menschen,” refer to the nitrogen excretion in the urine during dif-
ferent hours of the day, but the point of maximum excretion is diffi-
cult to determine, as the different days are not divided alike.
1 ROSEMANN: Archiv fiir die gesammte Physiologie, 1896, lxv, pp. 343-392.
2 TSCHLENOFF: Correspondenz-Blatt fiir Schweizer Aerzte, 1896, xxvi, p. 65.
8 RIAZANTSEFF: Archives des sciences biologiques, 1895, iv, pp. 895-896.
4 VERAGUTH: Journal of physiology, 1897, xxi, p. 112.
® ROESKE: Ueber den Verlauf der Phosphorséure-Ausscheidung beim Men-
schen. Dissertation, Greifswald, 1897.
5 GRAFFENBERGER : Zeitschrift fiir Biologie, 1892, xxviii, pp. 318-344.
7 SHERMAN and Hawk: This journal, 1900, iv, p. 26.
8 MaREs: Archives slaves de biologie, 1887, iii.
® GLEY et RicHeET: Comptes rendus de la société de biologie, Paris, 1887,
Ie, FS Sh/7/-
0 SONDEN und TIGERSTEDT: Skandinavisches Archiv fiir Physiologie, 1895,
Mi, ila
On the Time Relations of Proted Metabolism. 119
Hopkins and Hope; in experiments with men, found the maximum
nitrogen excretion in general during the third or fourth hour after
the ingestion of nitrogenous food.
Sherman and Hawk,? in experiments upon themselves in this
laboratory, observed the effects of sudden increase of nitrogen in
the food from eating extra amounts of lean meat at breakfast. The
maximum rate of nitrogen excretion occurred in six to nine hours
after the extra proteid was ingested; the rise and fall of the sulphate.
excretion was nearly parallel to that of nitrogen. The course of the
phosphate excretion was entirely different from that of either nitrogen
or sulphate.
Herfeldt,? Johansson,‘ Bert,® Kaupp,® and Van Noorden? have also
conducted investigations upon nitrogen metabolism which are closely
related to those of the present paper.
METHODS.
Purpose and plan of the experiments here reported. — The aim of the
present investigation has been to determine: (1) The length of
time elapsing between the ingestion of large amounts of proteid food
and the excretion of increased amounts of nitrogen, sulphur, and phos-
phorus in the urine. (2) The balance of income and outgo of nitro-
gen, sulphur, and phosphorus. (3) The relation between the nitrogen
content of the urine and the heat of combustion of its water-free
substance.
A preliminary period of four days (Period I) was passed on a diet
containing 14.86 grams of nitrogen and about 2900 calories of energy.
On the fifth day (Period II) at breakfast, a portion of the normal
diet having a nitrogen content of 2.46 grams was replaced by a pro-
teid food containing 12.60 grams of nitrogen. In other words, the
1 Hopkins and Hope: Journal of physiology, 1898, xxiii, p. 271.
2 SHERMAN and Hawk: Loc. cit.
8 HERFELDT: Mittheilung aus der Wiirzburger medicinische Klinik., 1885, i,
p. 61, Centralblatt fiir die medicinische Wissenschaften, 1885, xxiii, p. 515.
4 JOHANSSON; Skandinavisches Archiv fiir Physiologie, Leipzig, 1898, viii,
pp. 85-142.
> BERT: Jahresbericht ftir Thierchemie, 1879, ix, p. 291.
® Kaupp: Archiv fiir physiologische Heilkunde, 1856, p. 554.
™ Van NoorDEN: Pathologie des Stoffwechsels, 1893, Physiologische Theil-
ung, p- 45.
120 P. B. Hawk.
amount of nitrogen ingested at the meal was increased by 10.14
grams, During the remainder of that day, and for the four days
following (Period III), the constant diet of the preliminary period
was.again maintained. Urine was passed every three hours during
the day, beginning at 6.30 A.M., and in a nine-hour period at night,
beginning at 9.30 P. M.
The total nitrogen, sulphur, and phosphorus were determined in
foods and feces. The total nitrogen, the sulphur as SO,, and the
phosphorus as P,O, were determined in the urine.
Diet. — The foods used in the constant diet were soda crackers,
butter and whole milk (see Table I). The extra proteid of the fifth
TABLE LI.
DIET: AMOUNT AND COMPOSITION.
Amt. of food
Heat | eaten per day.
of
com-
Dry : : : . | P2O;.| bus-
matter. tion
F per
gram.
proteid
Day of extra
ingestion.
Beef (par- per cent. per cent. .| per cent. | p. cent. | p. cent. | sm. cal.
tially dried) 96.40 7.70 14.00 | 2.24 | 1:87 || 5565
Crackers 90.53 6.20 1.84 | 0.28 | 0.24 | 4216
Butter 91.44 Ee e (1.2% | 212... | gos
casein)
Milk (par- a ‘
tially dried) | 9°52 34.09 441 | 1. 5986
Babcock = 4.3
Milk (fresh) | .... | Ether extract | ...- 0.56 ne 782 | 1650 | 1375
= 422
Beef (partially dried) = 40%. Milk (partially dried) = 12.71%.
day was furnished by 225 grams of a specially prepared and very
lean beef. Three meals were eaten each day: the first one at
6.30 A.M., the second at 12.30, and the third at 6.,0 p.m. With one
exception, explained below, each meal consisted of 550 grams of whole
milk, 20 grams of butter, and 100 grams of soda crackers. Upon the
morning when the extra proteid was to be taken, 275 grams of whole
milk and 50 grams of crackers were replaced by 225 grams of beef.
This substitution increased the ingested nitrogen for the whole day
by 10.14 grams, though the energy of the diet was increased by only
————
On the Time Relations of Protetd Metabolism. 121
80 calories, or from 3027 calories to 3107 calories. The particular
feature in this case was that the extra proteid material was ingested
all at one meal, without any further interruption of the usual condi-
tions of the experiment.
The constant diet became at no time distasteful, and seemed emi-
nently fitted for the needs of the subjects. Feelings of hunger were
entirely absent, and at the same time no unpleasant sensation, such
as excess of food sometimes causes, was experienced. When the
added beef was taken, however, Subject R. found a slight difficulty in
consuming the entire two hundred and twenty-five grams. This was
due, no doubt, mainly to the fact that the only method feasible for
preparing the beef before eating, was to heat it upon a water-bath.
No frying-pan or spider could be used, for with either of these a loss
of material would have occurred; and the quantity of butter in the
diet was too small to grease the pan. The prepared beef was quite
dry and tasteless, and the subjects found the quantity of saliva
secreted rather inadequate for the consumption of the large amount
of unmoistened beef.
Subjects. — The subjects of the experiments here reported were two
men twenty-five years old in normal health. Both were wholly unac-
customed to the use of alcoholic beverages. One was accustomed to
use moderate amounts of coffee and tobacco, but during the time of
the experiments and for a number of days previous he abstained
from both. The weights of the men, without clothing, at the begin-
ning of the experiment were: H. 56.2 kg. and R. 60 kg., and at
the end they were: H. 56.4 kg. and R. 59.2 kg. Thus H. had gained
0.2 kg. during the nine days, whereas R. had lost 0.8 kg. The weights
were taken on the first and last days, about 6.30 a.m., before any food
was eaten, and immediately following micturition and defzcation.
The attempt was made to have the contents of the alimentary canal
and bladder as small and nearly alike as practicable at the times of
weighing.
Daily schedule. — The subjects rose at 5.30 A.M., and as they
lived near the laboratory they were ready for work at 6 A.M. The
time up to 6.30 A.M. was passed in preparing breakfast and in begin-
ning the analytical work of the day. Immediately after breakfast the
urine samples completed at 9.30 the previous evening, and at break-
fast time, were weighed, their specific gravity taken, and the urine ali-
quoited for composite samples, one-fifth of the urine of each subject
being used for this composite.
122 P. B. Hawk.
The milk for the day was next thoroughly mixed, sampled, both
for individual sample and for composite, and the fat determined by
the Babcock test.
At 9.30 A.M. each subject drank 300 c.c. of water. Regular ana-
lytical work connected with the experiment proper or with investiga-
tions with the respiration calorimeter occupied the subjects from that
hour until about 12.15 p.m. The midday meal was then prepared,
and at 12.30 was eaten. From this hour until 3.30 Pp. M., regular
analytical work and computation and tabulation of results occupied
the subjects. At 3.30 P.M., 300 c.c. of water was again taken by
each.
The time between 5 and 6 p.m. was devoted to active exercise,
either brisk walking out of doors or, when the weather was unfavor-
able, light exercise in the university gymnasium near by. The sub-
jects returned to the laboratory at about 6 pP.M., prepared and ate
their evening meal, and passed the remaining time to about 9.30
in the routine operations of weighing and analyzing samples, and cal-
culating and putting into tabular form the results obtained from pre-
vious analytical work. At 9.30 P.M. each subject drank 300 c.c. of
water and at about 10.15 P.M. went to bed.
All meals were taken in the laboratory building, thus rendering it
unnecessary to leave the laboratory except for the usual afternoon
exercise. In order to obviate any possible influence of nervous ex-
citement upon metabolism, the subjects aimed to maintain absolutely
normal conditions at all times and to eliminate as far as possible
excitement of any kind.
Preparation of samples, methods of analysis, ete. The crackers
were purchased at a local grocery, and during the experiment were
kept in a large tin can in order to insure constancy in the moisture
content. Enough for the whole experiment were secured at one
time, and sampled by taking crackers at random throughout the
lot.
The butter was the best product of a neighboring creamery. A
quantity sufficient for the whole experiment was obtained at one
time, and the amount needed for each subject for each meal of the
experimental period was accurately weighed and placed in a small
ointment pot, before the experiment proper began. During this
process small portions were taken occasionally for the sample.
The milk used in the experiments was a portion of the product of
five cows, isolated from a large herd for this special purpose. The
Ox the Time Relations of Proteid Metabolism. 123
beef was specially prepared and as free from fat as possible. In
preparing the foods for analysis the usual method was followed."
The faeces were dried at 100° C., then weighed, and ground, and
the nitrogen, heat of combustion, etc., determined as with the foods
(Table II). Separation of faeces was made by taking, at the be-
ginning of the first meal of each period, two gelatin capsules each
containing about 0.2 grams of powdered charcoal. Defaecations were
normal throughout.
TABLE II.
AMOUNT AND COMPOSITION OF FACES.
Fat. | Ash. | Nitrogen. , POs.
Dry matter.
combustion
grams. | percent. | percent. | per cent. | p. cent. |} grams. - | percent. | grams.
77.8 | 97.24 | 15.23 | 3202 | 3.89 | 3.02 10.63 | 8.27
33:5 || 94:86 | 16.02 | 28:08 | 4.24 | 1.42 8.97 | 3.01
72.0 | 96.83 | 14.82 | 31.86 | 4.31 | 3.10 10.63 | 7.66
I | 118.2 | 94.28 | 15.14 | 29.62 | 3.67 | 4.34 oF 6.10
TS 59:0) 94-93) 1 14-908) 29:25) 4h Sel 1636 93:04 | ALLO SiS6n ro45
PS heol-y| 96.65) 1) 12-67: | 31.97 3:95.) 3:23 4:25 4 3:48) |) 9.24. | 7-56
1 Period I = 4 days. Period II = 1 day. Period III = 4 days.
The fusion method ? was used for the determination of total sulphur
and phosphorus in the foods and faces. The casein content of the
butter was determined by dissolving out the fat with ether, weighing
the residue, igniting the latter, and subtracting the weight of the
ash, as recommended by the Association of Official Agricultural
Chemists. The other methods have already been described in this
journal.?
All analyses of the foods, as well as the urine and feces, were made
in duplicate. The daily analyses of milk and the analyses of urine
1 See Bulletin 44, Office of Experiment Stations, United States Department of
Agriculture.
2 See HAwK and GIEs, This journal, 1gol, v, p. 493-
8 SIMERMAN and HAWK: Loe. cit.
124 PBs Taw:
for different periods of the day were controlled by analyses of com-
posite samples. All specific gravity determinations, as well as all
weighings of food, feces, and urine, and all burette readings when
convenient, were checked by two men in order to eliminate as nearly
as possible any error which might arise from inaccurate readings. .
TeABIEH elie
ANALYSIS OF COMPOSITE URINE SAMPLES.
Subject. Amount. Nitrogen. SO3.
grams. grams. grams,
H. 1414.2 13.040 1.856
669.2 12.250 1.776
726.6 13.007 | 1.870
702.2 12.780 1.950
1080.3 19.120 2.930
740.3 14.435 2.080
723.4 13.100 1.895
768.6 13.260 2.047
680.6 13.480 2.032
1664.6 14.450 2.086
1041.2 4212 2.085
1553.6 15.536 2.280
1428.8 14.430 2.285
1599.6 Zier oo 3.432
1081.0 15.566 2.265
1269.4 = ERAS 2.288
1218.7 14.260 yap 15)
1655.3 13.800 1.997
DISCUSSION OF GENERAL RESULTS.
The data relating to the diet will be found on page 120 (Table I).
The amounts and composition of the faeces of the two subjects are
given on page 123 (Table II). In Table III, page 124, will be found
het ena:
On the Time Relations of Protetcd Metabolism. 125
the results of the analysis of the various composite samples of urine.
Table IV, page 126, shows the volumes of urine for the various
periods of the different days. For more convenient reference, the
amounts of the nitrogen excretion for the various periods of the
several days are condensed in Table V, page 128. Similar data for
the excretion of SO, and P,O; may be found in Tables VI and VII,
on pages 134 and 137, respectively. The data of income and outgo
of nitrogen may be found in Table VIII, page 139. Similar data for
sulphur and phosphorus are given in Tables IX and X, on pages 141
and 142, respectively. Table XI, on page 143, summarizes the ratios
of the nitrogen and heat of combustion of the composite sample of
the urine of each day. The hourly excretions of nitrogen, sulphur,
and phosphorus by the two subjects during the several experimental
days are shown by curves in Figs. 1-4, pp. 127, 130, 133, 138.
Urine volume. — When we consider the fact that the daily con-
sumption of liquid (water and milk) was the same for each subject,
we are struck with the marked variation in the volume of the urine
excreted by the two men. It is commonly stated that the total
volume of the urine excreted during any definite period varies in a
fairly regular degree according to the amount of liquid taken into the
body during that period. In the case under consideration, we had
two men of approximately the same age, weight and lung capacity,
maintaining the same constant diet and drinking precisely the same
amounts of water and milk daily; yet Subject R. excreted as an
average 1400 grams of urine per day, while Subject H. excreted as
an average but 850 grams (Table IV, page 126). The time being
mid-winter, the subjects did not perspire very freely, and as far as
could be judged the rate of sensible perspiration was about the same
for each subject. It is difficult to explain why Subject H. should
excrete but 60 per cent as much urine as Subject R. when the two
men were engaged in exactly the same occupation, took the same
amount of exercise at the same hour of the day, and worked side by
side in the same room, thus securing like conditions as regards tem-
perature, etc. As has been said (page 121), Subject H. gained 200
grams in body weight during the experiment, while Subject R. lost
800 grams in the same period, but this would account for only a very
small part of the discrepancy between the urine volumes of the two
men for the entire experimental period. Insensible perspiration may
account for a portion of the variation in urine volumes, but it does
not seem probable that Subject H. could have lost, in that way, 550
grams per day more than Subject R.
126 P. B. Hawk.
A similar phenomenon is observed in comparing by periods, the
urine excretion of the two subjects. On every day except January 14,
the volume of the urine excreted by Subject R. during the fourth
period of the day (3.30 P.M. to 6.30 P.M.) was greatly in excess of
that for any other period of the day for this subject, and also far
greater than that excreted by Subject H. during this same period.
TABLE IV.
URINE VOLUME.
Jan. 14.} Jan. 15. .| Jan. 18. | Jan. 19.| Jan. 20. | Jan. 21.
Subject.
grams. grams. 5 grams. grams. grams. grams.
90.7 | 112.4 102.3 | 108.1 81.2 88.3
102.8 | 114.0 119.7 | 127.2 97.6 83.5
77.2 82.1 OS 82.3 92.0 83.5
19:9 86.1 82.9 76.9 Midendk 78.9
105.9 78.5 82.0 72.4 99.4 | 1075
227) \\Be253:5 262.1 | 256.5 238.9
=| 1414.2 |. 669:2' | 726:6 740.3 | 723.4 ; 680.6
1A GIES ls372 130.4 | 1426
AS a ES AS WSS 117.2 | Uses
190.5: |) 173-5" | T5633 85.7 | 109.0
129-4 |) WOLi6s | 399:0 214.1) | A akG
153225 | 1124 |S 1Or0 NTIS: ASS.
268.5 | 308.6 | 501.6 AZT || 319g
Tot.| 1664.6 | 1041.2 | 1553.6 1081.0 | 1269.4
The average excretion by Subject R. for this fourth period was 393.6
grams; while the average excretion by subject H. was but 126 grams,
or only 32 per cent as great as the excretion by Subject R. Further-
more, examination of the tabulated data shows the volume of Subject
R.’s urine for the fourth period to have been greater than the com-
bined volumes for the third and fifth periods, on every day except
January 14 and 20. On six of the nine experimental days the urine
0 ar ling le
On the Time Relations of Proteid Metabolism. 127
volume of Subject R. for this fourth period was from 25 per cent to
43 per cent of the total twenty-four hours’ urine. At 3.30 P.M., each
subject drank 300 c.c. of water, as has been stated, and following this,
at 5 P.M. came the customary exercise. Why these factors should
so differently influence the urine volume of the two subjects, how-
ever, is not entirely clear.
Nitrogen excretion by Subject H.— The graphic representation of
the nitrogen excretion by Subject H. will be found in Fig. 1,
below, while the actual amounts of nitrogen excreted are stated in
FiGurRE 1.— The nitrogen excretion by Subject H. The ordinates show the average
hourly excretion in grams and the abscissz the time in days.
Table V, page 128. The curve for this subject shows a general
tendency for the nitrogen to be excreted in such a manner as to
form two very well-defined maxima. This does not hold true for
the day upon which the large amount of extra proteid food .was
ingested, nor for the two days immediately succeeding, but as soon
as the normal level for the nitrogen excretion was again obtained
(January 20) the two maxima reappear.
In every instance there was a very decided fall in the rate of ex-
cretion during the night period, and this was succeeded, in general,
by a very perceptible rise during the first period of the day following,
z.é., just after the morning meal. This rise was without doubt due,
at least in part, to the influence of the ingested food.
Taking into account the days which show the two maxima, it is
noticed that in every case except one (January 15) the highest point
reached by the day’s excretion was in the fifth period or from 6.30
P.M. to 9.30 P.M. It will be remembered that just preceding this
period (5 p.m.) the subjects were accustomed to take their daily
128 P. B. Hawk.
exercise. This light exercise can hardly be considered an important
influencing factor, however, as it is very generally believed that any
increase in the nitrogen excretion due to muscular work does not
appear before the following day.1 The evening meal, at 6.30 P.M.,
very likely had an influence upon the excretion of nitrogen similar
to that attributed to the morning meal. The taking of the 300 c.c.
TABLE V.
NITROGEN EXCRETION BY PERIODS.
.| Jan. 14. | Jan. 15. | Jan. 16. | Jan. 17. | Jan. 18.| Jan. 19. | Jan. 20. | Jan. 21.
Subject.
grams. grams. grams. grams. grams. grams. grams. grams.
1.392 | 1.692 | 1.645 | 1.983 | 1.908 | 1-811 | Dssieiisan
—
al
1.646 | 1.568 | 1.636 | 1.657 | 2826 | 2.053 | L657) ei
1697 |} 1.560 | 1.580] 1610} 2.956! 1.849] 16621 1.762) I766
1.965 | 1.578 | 1.670 | ° 1.530 | 2.599} 1.808 | 1.638 |" USsse ore
2.072 | 1.880 | 1.650 | 1.778 |. 2.551 |. 1.775. |. 15890) eee
4.196 | 4.212 | 4.766 | 4.693 | 6305 | 4.967 | 4.720 | 4.666] 4.910
13.108 | 12.190 | 12.994 | 12.853 | 19.220 | 14.360 | 13.277 | 13.162 | 13.659
1.890 | 1.873 | 1.918] 1.643 | 1.803 | 2.200 | 1.8474) U6e27 yess
1.835 | 1.716 |. 2.361 | 2.134 | 2.803 | 2.028 | 2.124) “6645 aZioGs
Pallas ||. oyasteye|| PAAOINS || ieresispall « skarlle: : 1.919 | 1.965 | 2.225
2.199 ; 1.910); 2.334, 2.582 | 3.928 | 2.912 | 2466) 2272002
1.731 | 1.753 | 1.670 | 1.578 | 2.858. | 1.975 | 1.8069) oye meee
4.484 | 4.691 | 5.016 |. 4.555 | 6.653, | 4.8071] 4.626) 94:92GN meee
Mots W273 199) oss 14.277 21.458 | 15.652 | 14.788 | 14.405 | 13.871
of water at 3.30 P.M., followed by the customary ration of milk at
supper, making in all very nearly a litre of liquid, may possibly have
accelerated the washing out of the urea formed and thus assisted
in the attainment of the point of greatest excretion at this hour.
These results when considered with the fact that the subjects gen-
1 Various references cited by SHERMAN and HAwk: Loc. c@d.
On the Time Relations of Protetd Metabolism. 129
erally did the most fatiguing work of the day between 6.30 P. M. and
9.30 P.M., lead the author to believe that some or all of these factors
had at least a slight modifying influence upon the appearance of the
high rate of nitrogen excretion at this point.
There was apparently a very well-established tendency to reach
the minimum excretion for the day during the long nine-hour night
period. :
As was to be expected, the taking of the large amount of extra
proteid upon January 17 very materially altered the normal curve for
the nitrogen excretion. In the first place, instead of showing two
well-defined maxima, the excretion of this day showed but one. This
maximum occurred with Subject H. during the third three-hour
period (12.30 P.M. to 3.30 P.M.) or from six to nine hours after the
extra proteid food was ingested. In this respect my results agree
with those obtained by Sherman and Hawk.! It will be observed
that this maximum came at a different hour of the day than either
of the daily maxima normally observed preceding the ingestion.
The rise in the rate of excretion after the ingestion of the beef at
breakfast was very rapid and began immediately. This sudden rise
was followed at once by a fall, somewhat less abrupt than the rise,
which continued unchecked even over that portion of the day at
which, under normal conditions, the point of most rapid excretion ap-
peared, and extended into the first period of the day following. The
most important characteristics of the curves for the two days follow-
ing the day of the “chief maximum” are the same as those of the
curve of the 17th, z. ¢., one maximum followed by a gradual fall. On
the 20th and 21st the normal excretion curve with two maxima (the
highest point of excretion being in the fifth period) was again
attained. The failure of the excretion of the 17th, 18th, and roth to
show the normal maximum of the fifth period was evidently due in
part at least to the fact that a large portion of the extra nitrogen,
ingested January 17 at 6.30 A.M., had been excreted before this time
of day (6.30 P.M.), and the rate even then being far above the nor-
mal, those influences which upon other days were apparently such
potent factors in producing a maximum at that point were not able to
check this seemingly well-defined tendency of the excretion to reach
the normal level of former days.
Taking 12.8 grams as the normal level for the nitrogen excretion
1 SHERMAN and Hawk: Loc. cit.
130 PBA rawe:
on the days preceding the beef ingestion, it will be seen that even
upon the day of the actual ingestion 6.4 grams or 63 per cent of the
10.1 grams of extra nitrogen was excreted. The total elimination of
the extra nitrogen for the forty-eight hours following the ingestion
of the beef was 8 grams or 80 per cent, while the last day of the ex-
periment showed 97 per cent of this extra nitrogen eliminated, and
the rate of excretion at that time slightly above the level of the
normal days mentioned.
Nitrogen excretion by Subject R.— The data for the nitrogen ex-
cretion of this subject will be found in Table V, page 128, and the
graphic interpretation in Fig. 2.
FicuRE 2.— The nitrogen excretion by Subject R. The ordinate shows the average
hourly excretion and the abscissa the time.
Subject R. maintained during the preliminary period a normal
level very appreciably above the level maintained by Subject H. In
general, the curve representing the excretion by Subject R. was simi-
lar to that showing the excretion by Subject H., except that the
points of maximum excretion were somewhat more accentuated, and
also, as has been said, the elimination as a whole was on a higher
level. As in the case of Subject H. there was the same very rapid.
fall in the rate of excretion during the night, and also the accom-
panying rise. ‘There was a fairly well-defined tendency for the maxima
on normal days to occur somewhat earlier than the maxima in the
case of Subject H., the point of highest excretion being reached on
On the Time Relations of Protecd Metabolism. 131
normal days generally in the fourth period, instead of the fifth, as with
Subject H. By consulting Table IV, page 126, it will be seen that
this fourth period was the time of the copious discharge of urine by
Subject R., and the author is inclined to think that the passage of
such large amounts of water through the system may have tended,
in a measure, to cause this occurrence of the maximum excretion at
an earlier hour.
Upon the day when the extra proteid was ingested at breakfast,
there was the same immediate rise in the rate of excretion as was
observed in the graphic representation of the excretion by. Subject H.
In the present instance, however, instead of reaching its highest
point during the third period, as with Subject H., the excretion con-
tinued its rapid rise, and only attained its maximum during the fourth
period of the day (3.30 P.M. to 6.30 P.M.), nine to twelve hours after
the proteid had been ingested, and three hours after the maximum
was reached by Subject H. upon the same increased ingestion. Even
during the third period, where Subject H. reached the maximum, the
excretion of Subject R. was far above that of Subject H., owing to his
elimination being at a higher level normally; but when the custom-
ary maximum discharge of urine occurred during the next period, it
seemed that the excretion by Subject R. followed its normal course
and attained its highest point at that time. This fact would seem to
indicate, as has already been mentioned, that the large amount of
water eliminated during this period had a tendency to give the
maximum a somewhat more advanced position than it would have
had if the amount of water passed had been no larger than in the
contiguous periods. The urea which would have come normally
somewhat later, and thus assisted in the formation of the fifth period
maximum, shown so plainly in the excretion by Subject H., may,
through the agency of this large volume of water, have been re-
moved at an earlier hour, thus causing the second daily maximum
for Subject R. to fall in the fourth period.
When we compare Periods I-IV inclusive, of the first four days
with the analogous periods of the fifth day, we find that Subject H.
eliminated 3.9 grams or 38 per cent of the extra nitrogen during that
time, while Subject R. eliminated 3.8 grams or 37 per cent, showing
in this respect very marked agreement. Taking into consideration
the whole of the day upon which the beef ingestion occurred, we see
that Subject R. excreted 7 grams or 70 per cent of the 10.1 grams of
extra nitrogen, and showed in this respect an increase of 0.6 grams
12 P. Be tlawk:
or 7 per cent over Subject H. However, during forty-eight hours
Subject R. eliminated 8.1 grams or 80 per cent of the extra proteid ;
this again agreeing with the data for Subject H.’s excretion. During
the final day of the experiment Subject R. showed a falling off in the
rate of excretion, due no doubt to a storage of nitrogen, and hence
no time relation between the consumption of the extra proteid food
and the ultimate total elimination of its nitrogen content can be
determined.
The days immediately following the day of the “chief maximum ”
seem to differ from those in the case of Subject H. in the fact that
with Subject R. the point of greatest excretion continued to occur
at the same hour as on the day of the beef ingestion. This may have
been due to the fact that just preceding this period the excretion
was at a level, lower in some instances even than the level for the
excretion of Subject H. at the same time. Remembering that the
normal level for Subject R. was something over one and one-half
grams per day above that for Subject H., it is easy to imagine a very
natural attempt to at least regain the level, and this attempt being
made just at the time that the great surplus of water was eliminated,
it is not difficult to see how the maximum may have fallen very
naturally at the point indicated.
Sulphur excretion by Subject H.—In general the course of the
sulphur excretion followed that of the nitrogen. A notable difference,
however, was the greater regularity in the position of the points of
maximum excretion in the case of the sulphur. Upon every day of
the experiment, including the day when the beef was ingested, there
were two well-marked maxima in the excretion of this substance
(Fig. 3, page 133). An added regularity was noted in this connec-
tion, inasmuch as the first maximum occurred, in every instance,
during the third period, while the second maximum fell in the
fifth period. Thus on the day of the increased ingestion of proteid
following the uniform diet, the “chief maximum” occurred, as
in the case of nitrogen, six to nine hours after the beef was
eaten.
By reference to Fig. 3, it will be seen that the rate of sulphur
excretion was low during the nine-hour night period, but that instead
of beginning a sudden rise during the first period of the day follow-
ing , as was customary with the nitrogen excretion, the course of the
sulphur excretion was lower during this period than at any other
time in the twenty-four hours. Hence the rise following the ingestion
On the Time Relations of Proteid Metabolism. 133
of the beef began, not immediately after the food had been taken,
but only after a lapse of three hours. The rise from this hour was
rapid, and the maximum point was reached in the third period. The
excretion then returned rapidly to its normal level, which was practi-
cally reached by the next morning, and maintained throughout the
remainder of the experimental period. In increasing its rate of
excretion somewhat tardily after the ingestion of the extra proteid,
and in regaining the normal rate at an earlier hour, the excretion of
sulphur differed very markedly from that of nitrogen.
As was previously shown by Sherman and Hawk,! the ratio
between nitrogen and SO, was lower on the day following the in-
gestion of the extra proteid than on any of the other experimental
FIGURE 3.— The SOg3 excretion by both subjects. The ordinate shows the average
hourly excretion and the abscissathe time. The curve for Subject R. is a broken and
that for Subject H. a solid line. ,
days, due to the fact that the SO, reached the normal level more
quickly than the nitrogen (see Table VI, page 134).
Sulphur excretion by Subject R.— The curve representing the course
of the sulphur excretion of Subject R., while possessing many of
the features common to the curve for the excretion of Subject H.,
differed from the latter in a few notable directions. We fail to find,
in our examination of this curve, any such regularity in the occur-
rence of the maxima as was so plainly exhibited in the excretion of
Subject H. Instead of showing two daily maxima, falling in the third
and fifth periods respectively, many of the days show but one maxi-
mum, and its occurrence seems to be unguided by any well-regulated
tendency. There appears, however, to be a somewhat masked tend-
ency toward the formation of maxima in the third and fifth periods,
as was so plainly set forth in the graphic representation of the ex-
cretion of Subject H. This was shown by the fact that on five days
there was a maximum occurring in the fifth period, and on four days
in the third period. As was true of Subject H., the rate of excretion
1 SHERMAN and Hawk: Loc. ciz.
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134
TA WIAVL
On the Time Relations of Protetcd Metabolism. 135
during the night was low; but opposed to the conditions which
obtained in the excretion of Subject H., the minimum rate of excre-
tion occurred in the nine-hour night period instead of in the first
period of the day.
The excretion of sulphur followed that of nitrogen by commencing
an immediate rise as soon as breakfast had been taken. Everything
considered, there was a fairly satisfactory agreement between the
courses of the nitrogen and sulphur excretions of Subject R., though
perhaps hardly as close as that shown in the curves for the nitrogen
and sulphur elimination of Subject H.
Upon the day when the extra proteid was given, there was the
customary rise immediately after the first meal had been taken. This
was followed, at the commencement of the second period, by a very
pronounced increase in the rate of excretion, which continued until
the third period, and formed the “chief maximum” in six to nine
hours after the extra proteid was ingested. In this it agrees with the
positions of the nitrogen and sulphur maxima for Subject H., but falls
three hours sooner than the corresponding maximum for Subject R.
As was true in the case of Subject H., the decline from the point of
maximum excretion was rapid, and the normal level of the prelimi-
nary period was fully regained during the early periods of the day
following.
Recalling now the positions occupied by the nitrogen and sulphur
maxima of the two subjects upon the day when the added proteid
was consumed, we note that three of the four maxima fall in the
third period. This may lend some force to the supposition that the
maximum for nitrogen in the excretion by Subject R. was carried
along to a later period simply through the agency of the unusually
large volume of urine passed during that period.
Phosphorus excretion by Subject H.— The course of the phos-
phorus excretion by Subject H. was somewhat more comparable to
the sulphur than to the nitrogen excretion of this subject. We note
the same evident tendency toward the formation of two maxima in the
third and fifth periods respectively. On some days, however, the
highest point was at the end of the night period. This phenomenon
formed a very marked contrast with the excretion of nitrogen and
sulphur, for in the latter cases the rate of excretion at that time was
very low. At the beginning of the first morning period, the rate of
the excretion of phosphorus underwent a very rapid fall, and reached
a decided minimum at the end of the period. This same character-
136 P.. BE TTAWE.
istic, in a much less accentuated form, was noted in the sulphur
excretion of Subject H. The minimum point in the phosphorus
excretion was followed by a rise, just as marked in nature as the fall,
and the level of the night period was not regained on normal days,
until the excretion had reached the third or fourth period.
On the day of the extra proteid ingestion there was the customary
fall immediately after the ingestion, followed by the usual rise. This
rise, however, was more prolonged and reached the maximum in the
second period, or three to six hours.after the meal. The stimulating
effects produced by the ingestion of this large amount of proteid food
may have been a factor in the formation of this maximum in the
second period rather than in the third, as was evidently the course of
the phosphorus excretion on the other days of the experiment. Fol-
lowing this “ chief maximum,” the second maximum occurred as usual
in the fifth period, after which the rate descended during the night
period and reached the normal level in the first period of the follow-
ing day.
The constant diet furnished normally 4.96 grams of P,O; (see
Table VII, page 137), and upon the day when the extra proteid was
ingested this amount was augmented by 0.86 grams. The average
total excretion of P,O, for the first five periods of the four days com-
prising the preliminary interval was 1.47 grams, and the total for the
similar periods of the fifth day was 2.09 grams. Thus 0.62 grams, or
72 per cent of the extra P,O, ingested, was eliminated by the urine
in fifteen hours after the food was ingested, or in twelve hours after
the rate of the P,O, excretion began to rise. In the twenty-four
hours following the ingestion of the extra proteid, 0.70 grams, or 81
per cent of the extra P,O; ingested at this time, was eliminated by
the urine and the rate of excretion was almost precisely at the normal
level maintained during the four preliminary days.
Phosphorus excretion by Subject R. — As was seen to be true of
this excretion in the case of Subject H., there was a somewhat closer
relationship to the sulphur excretion than to the nitrogen excretion.
There was here, however, a very evident inclination toward the for-
mation of but one maximum, which generally fell in the third or fourth
period. A falling off in the rate of excretion during the night was
noted, and formed in many cases a marked contrast to the rising tend-
ency of the phosphorus elimination of Subject H. The extremely
rapid decline in the rate during the first period of the day, which was
so plainly set forth in the graphical representation of Subject H.’s
On the Time Relations of Protercd Metabolism. 137
excretion, did not obtain to so great an extent here; but the excre-
tion followed somewhat more closely the moderate course taken by
the sulphur excretion of Subject H.
The point of mininum excretion occurred, as with Subject H., dur-
ing the first morning period, and the failure of the excretion to ex-
hibit the extremely rapid fall so characteristic of the phosphorus
TABLE VII.
PHosPHORUS (P,0O;) EXCRETION BY PERIODS.
Jan. 13. | Jan. 14. Jan. 16. | Jan. 17.
grams. grams. é grams. grams.
0.164 | 0.148 0.123 | 0.212
0.289 | 0.192 0.272 | 0.489
0235, | 0:37 0.322 | 0.481
0.417 | 0.381 0.293 .| 0.438
0.458 | 0.425 0.360 | 0.474
TESS | C49 iGo | HEIs)
Tot.| 2.701 | 2.570 25050) |) 32289
0.289 | 0.289 0.290 | 0.314
0311 | 0.326 0.368 | 0.505
0.380 | 0.491 0.491 | 0.636
0.578 | 0.460 0.589 | 0.547
0.376 | 0.505 gaay || (58%:
0.946 | 1.095 0.966 | 1.081
2.880 | 3.166 3.148 | 3.616
excretion of Subject H., was evidently due to the fact that the
single maximum in the case of Subject R.’s excretion occurred
either at 3.30 P.M. or 6.30 P.M., and was followed by a gradual fall
for a period of fifteen to eighteen hours. Under such conditions,
the effect of the extreme rapidity of the customary morning fall
would be somewhat neutralized by the falling off in the rate at so
early an hour. In many instances with Subject H.,as we have seen,
138 Po BD: tear:
the morning fall, following as it did the gradual rise of the previous
long nine-hour night period, caused the graphic representation of the
course of the excretion at this point to assume a somewhat more
striking appearance than would otherwise have obtained.
Following the ingestion of the beef on the fifth day, the excretion
rose rapidly after 9.30 A. M., and reached the “ chief maximum” in the
third period, or six to nine hours after the ingestion. In this it
Ficure 4.— The P,O; excretion by both subjects. The ordinate gives the average
hourly excretion and the abscissa the time. The curve for the excretion by Subject
R. is in a broken line and that for the excretion by Subject H. in a solid line.
agreed with the excretion of sulphur of both subjects, but occurred
three hours later than the “chief maximum” of the phosphorus ex-
cretion of Subject H. In descending from this point of maximum
excretion there was no formation of a second maximum as occurred
in the course of the phosphorus excretion of Subject H. At the end
of the first period of the day following the extra proteid ingestion, the
level was somewhat lower than the normal level of the preliminary
period, and continued practically unchanged to the end of the experi-
ment. But 0.42 grams or 50 per cent of the extra P,O, was excreted
during the fifteen hours following the ingestion, and but 0.47 grams
or 55 per cent during the whole twenty-four hours, differing in this
respect quite materially from the percentage excretion by Subject
H. during this time.
Income and outgo of nitrogen (Subject H.).— During the preliminary
portion of the experimental period, upon a constant diet affording
14.86 grams of nitrogen per day, Subject H. excreted on an average
12.8 grams, showing an evident storage in the body each day of
approximately two grams of the ingested nitrogen. After the extra
proteid had been ingested and 80 per cent of its nitrogen content
eliminated by the urine inside of forty-eight hours, the rate of
excretion of nitrogen again fell, and during the final days of the
experiment gave an average daily elimination of 13.37 grams, or
about 1.5 grams less than the ingested amount.
—
On the Time Relations of Proted Metabolism. 139
Taking into consideration the entire experimental nine days, Sub-
ject H. showed a gain of 11.51 gramsof nitrogen. We may rightfully
assume that the major part of this nitrogen was retained in the tissues
of the body. Nevertheless if we apply the figure advanced by Voit}
for the nitrogen content of fresh muscle (3.4 per cent), it is easily
demonstrated that this gain of 11.51 grams of nitrogen, if used en-
TABLE VIII.
INCOME AND OuTGo OF NITROGEN (NITROGEN BALANCE).
NITROGEN.
Exp. | Length of
period. | period.
Average
gain or loss
per day.
Gain or
In food. | In urine. | In feces. Tess.
grams. 2 grams.
59.44 +5.27
25.00 +4.36
59.44 +1.88
143.88 : 411.51
59.44 —2.96
25.00 ale
59.44 —2.50
143.88 . —3.55
tirely for the formation of muscular tissue, could in itself account for
more than the total gain of 200 grams body weight. This gain in
nitrogen was evidently accompanied by a loss of water from the
system, otherwise a greater increase in body weight would appear.
The average gain of nitrogen per day was 1.28 grams (see Table
VIII).
The greater amount of nitrogen in the faces on the day when the
extra proteid was taken, over that of the normal days of the milk diet,
seems to indicate, either that beef entails a larger amount of meta-
bolic products in its digestion than milk, or, that the normal diet
1 Voit: Zeitschrift fiir Biologie, 1865, 1, p. 97+
140 P. B:. Hawk.
gave an amount of proteid adequate to the needs of the body,
whereas when the extra proteid was taken, the body could not im-
mediately adapt itself to the new conditions, and the nitrogen content
of the faeces was consequently raised.
Income and outgo of nitrogen (Subject R.). — Upon an examination
of the nitrogen balance for this subject (Table VIII, page 139) we
note a marked change in conditions from those which obtained with
Subject H. Upon a daily ingestion of 14.86 grams of nitrogen dur-
ing the preliminary period, there was an average daily elimination of
14.5 grams by the urine. In thus approximating nitrogen equi-
librium, the excretion of Subject R. differed decidedly from that of
Subject H., which ran on an average level 1.7 grams per day lower.
However, during the forty-eight hours following the beef ingestion,
80 per cent of the extra nitrogen was eliminated by this subject,
thus showing perfect agreement with the percentage elimination of
Subject H. After the effects of the extra proteid had subsided, the
excretion again assumed practically the same level it held during the
preliminary period. It is thus seen that the normal level for Subject
R. during the entire experiment was about 1.7 grams higher than the
normal level for Subject H.
As we see from the balance of income and outgo, Subject R. suf-
fered a loss in nitrogen aggregating, during the experiment, 3.55
grams. The loss of 800 grams body weight shown by him at the
end of the experiment, may in a measure serve as a means for the
explanation of this discrepancy. However, a large part of this 800
grams loss was doubtless due to the abstraction of water from the
system. Some time previous to the commencement of this investiga-
tion, Subject R. had been upon a diet having a somewhat lower
proteid content than that of the constant diet of the experiment.
Hence the continuous ingestion of a larger amount of proteid food
during the experimental period may have stimulated proteid metab-
olism and thus served as the chief factor in the production of the
loss sustained by this subject.
Income and outgo of sulphur (Subjects H. and RD): — As is univer-
sally understood, the method for determining SO, in the urine by
precipitation as barium sulphate fails to reveal the total SO, present.
The exact percentage left undetermined is to a certain extent a
matter of opinion and speculation. Different writers present differ-
ent solutions of the problem, and as far as the author is informed,
there seems to have been no invariable factor presented as represent-
-"
On the Time Relations of Proterd Metabolism. 141
ing the relation existing between the total SO, and that obtained by
the precipitation method. It is the author’s opinion that no rigid
factor can be deduced, inasmuch as it seems to be the tendency of
the ratio to vary with the conditions. Circumstances were such dur-
ing the progress of this investigation as to exclude the determination
of the total SO, in the urine by the reliable fusion method. There-
fore rather than employ any assumed factor in calculating the total
TABLE IX.
INCOME AND OutTGo OF SO, (SULPHUR BALANCE).
Income. Outgo.
Average
Wene th Osim Gain or gain or loss
period. loss. arda
In food. | In urine. | In feces. P Me
grams.
+0.55
+0.78
+0.39
SO, from the SO, as determined, this somewhat low value for the
SO, content of the urine was used in the calculation of the income
and outgo of sulphur (Table IX). Recognizing the deficiency in
the outgo however, all discussion of income and outgo of sulphur
has been omitted.
Income and outgo of phosphorus (Subjects H. and R.).— The method
for the determination of P,O, in the urine by titration with uranium
acetate having been shown in this laboratory! to give results closely
comparable with those obtained by the fusion method, the author felt
1 SHERMAN, H. C.: Bulletin 121, Office of Experiment Stations, United States
Department of Agriculture.
142 P. B. Hawk.
he could safely rely upon the data thus secured in the preparation
of a phosphorus balance. The striking point about this balance is
the large percentage of P,O; eliminated by the faeces (Table II, page
123). In the case of Subject H. for instance, 18.94 grams or 44.2
per cent of the total P,O; elimination passed out through the intes-
tines, and with Subject R. the amount thus passed was the slightly
lower value of 17.11 grams or 38.5 per cent of the total elimination.
During the nine days of the experiment, Subject H. gained 2.69
grams of P,O,, whereas Subject R. gained but 0.90. This was a
TABLE X.
INCOME AND OuTGO OF P,O; (PHOSPHORUS BALANCE).
Income. Outgo.
Average
gain or loss
per day.
Length of
period.
In food. | In urine. | In feces.
grams. grams. grams. grams.
19.84 10.31 8.27 +0.32
5.82 3.28 3.01 —0.47
19.84 10.28 7.66 +0.47
45.50 23.87 18 94 +0.30
19.84 12.60 6.10 +0.29
5.82 3.62 3.45 1.25
19.84 els 7.56 +0.29
45.50 27.35 17.11 +0.10
daily gain of 0.3 grams by Subject H., and of 0.1 gram by Subject
R. (see Table X). There is thus one point of similarity between
the nitrogen balance and the phosphorus balance, in that in each
case the gain by Subject H. was far above the gain by Subject R.
With the sulphur balance (Table IX, page 141), the conditions were
practically identical in the two cases.
Relation between the nitrogen content of the urine and the heat of
combustion of its water-free substance. —It has been assumed from
work done in this laboratory and elsewhere, that the ratio between
the heat of conbustion of urine and its nitrogen content will remain
On the Time Relations of Protecd Metabolism. 143
fairly constant so long as a definite amount of some specific proteid
is daily ingested. The previous work seems to indicate that the
ratio will be somewhat higher upon a diet of crackers and milk than
upon a diet consisting principally of beef. The data from this inves-
tigation verify this conclusion. Upon a diet of crackers and milk
TABITE Xi
RELATION BETWEEN THE NITROGEN AND HEAT OF COMBUSTION OF URINE.
Heat of | Total heat
Date Weight | combus- | of com- | Nitrogen
(1900). of urine. | tion per | bustion of | excreted.
gram. urine.
Ratio of | Calories
nitrogen | In excess
tocalories.| of urea.”
small calories. | large calories.
Jan. 13 74.3 105.1 : 8.1
14 LS eZ 105.2 Gan
15 153.3 NIA: : 8.6
16 151.5 106.4 : 8.3
17 139.0 150.1 : 7.85
18 160.1 118.5 Rone
19 159.5 115.4 : 8.8
20 167.8 129.0
21 170.4 116.0
1 The urine of Subject R. decomposed before satisfactory analyses could be made.
2 See SHERMAN and Hawk: Lec. cit.
8 Paraffin was used for sealing sample bottles and a few small pieces were found.
in this composite urine sample. The heat of combustion of paraffin being about
9075 small calories per gram, the combustion of a very minute portion could easily
cause the variation in the ratio noted on this day (20th).
during the four preliminary days, an average ratio of 1:8.3 was main-
tained. Upon the fifth day, when the constant diet was altered so as
to secure a change in the ingestion from milk proteids to beef pro-
teids, the ratio fell immediately to 1: 7.85, but regained its former
level when the constant diet of crackers and milk was again taken
upon the following days.
As was shown by Sherman and Hawk! the “calories in excess
of urea” assume a fairly constant level for the whole experimental
1 SHERMAN and HAWK: Loc. ci¢.
144 Pi Bi Hawk:
period, and are to a great extent unaffected by the variations in
the excretion of nitrogen. This seems to indicate that the organic
matter less highly oxidized than urea was eliminated in fairly con-
stant amounts from day to day.
CONCLUSIONS.
The normal curve for the nitrogen excretion by each subject
showed two points of maximum excretion. On the day of the extra
proteid ingestion a single maximum was observed, and the return to
the normal condition of two maxima occurred on the second day
following with Subject R. and on the third day following with
Subject H.
After the ingestion of the extra proteid, the nitrogen excretion
began an immediate, rapid rise to the point of maximum excretion,
which was reached in six to nine hours with Subject H., and in nine
to twelve hours with Subject R. This point of maximum excretion
was followed at once by a very rapid fall which passed, in a few
hours, into a more gradual return to the normal rate of excretion.
The minimum rate of nitrogen excretion for each subject oc-
curred during the night period.
In general the course of the sulphur excretion followed that of the
nitrogen. With Subject H. the sulphur excretion of every day of
the experiment showed two well-defined maxima; the excretion by
Subject R. did not show this regularity.
The sulphur excretion of Subject H. was lowest during the first
period of the day; the corresponding excretion of Subject R. was
lowest during the night.
After the ingestion of the extra proteid, the sulphur excretion by
Subject H. began to rise after the lapse of three hours; the sulphur
excretion by Subject R. began to rise at once.
The maximum sulphur excretion occurred with each subject six to
nine hours after the ingestion of the extra proteid. The normal rate
of excretion was regained in twenty-four hours.
The course of the phosphorus excretion by Subject H. showed a
tendency toward the formation of two maxima daily; the phosphorus
excretion by Subject R. generally showed a single maximum.
The minimum phosphorus excretion by each subject occurred
during the first three-hour morning period.
After the ingestion of the extra proteid at breakfast, the customary
———
~~
On the Time Relations of Proteid Metabolism. 145
fall in the phosphorus occurred, and was followed by a rise which
caused the excretion to reach the maximum in three to six hours
after the ingestion with Subject H., and six to nine hours after the
ingestion with Subject R. The normal level was regained with each
subject at the first period of the following day.
The ratio of the nitrogen content of the urine to the heat of com-
bustion of its unoxidized material was somewhat lower on the day of
the extra proteid ingestion than on normal days.
The ratio between nitrogen and SO, was the lowest on the day
following the ingestion of the extra proteid, due to the fact that the
SO, reached the normal level more quickly than the nitrogen,
On aconstant diet accompanied by like water ingestion, one sub-
ject excreted but 60 per cent as much urine as the other.
The author wishes to express his gratitude to Prof. W. O. Atwater
and Dr. H. C. Sherman for many valuable suggestions and for their
continued interest in the investigation. His thanks are also due Mr.
A. E. Roberts, who acted as subject and assisted in the analytical
work, and also Messrs. E. Osterberg, W. R. Frazier, and E. M. Swett,
who were very helpful in carrying out the details. The author is
also very grateful to Mr. R. D. Milner for many suggestions during
the preparation of this paper.
ON THE DISTRIBUTION OF OSSEOMUCOID.
By CHRISTIAN SEIFERT anp WILLIAM J. GIES:
[From the Laboratory of Physiological Chemistry of Columbia University, at the College of
Physicians and Surgeons, New York.)
HE question whether or not bone contains glucoproteid seemed
to be settled in the negative in 1892, when Young, working
under Halliburton’s superintendence, failed to extract from bone,
with calcium hydroxide or barium hydroxide, any substance that
could be precipitated with acetic acid.1 Several years ago, however,
on investigating this matter, we were able to show in this laboratory
that the rib and femur of the ox contain an appreciable quantity of
mucoid.
In our first communications? on this subject we outlined the
method of separating this substance, which has been termed osseo-
mucoid, and we also pointed out the mechanical obstacles in the way
of success in the method employed by Young for its detection. Later,
our studies in this connection were devoted to perfecting the method
of separating osseomucoid, and to determining its composition,
reactions and heat of combustion.®
On failing to detect mucoid in the bone shavings and powder under
examination, Young concluded that ‘in the process of ossification
the connective tissue matrix is apparently completely calcified.” The
results of our own work proved, however, that in the ossification of at
least the femur and rib of the ox, the “connective tissue matrix” is
not entirely removed. Until osseomucoid could be shown to exist in
1 YounG: Journal of physiology, 1892, xiii, p. 213.
2 GiEs: Proceedings of the American Physiological Society, This journal,
1900, iii, p. vii; Proceedings of the American Association for the, Advancement
of Science, 1900, p. 131 ; Biochemical Researches, 1903, i (a and b), pp. 31-33-
8 Hawk and Giles: This journal, 19o!, v, p. 387; MEAD and Gigs: Proceed-
ings of the American Physiological Society, This journal, 1902, vi, p. xxvili;
Gres: Biochemical Researches, 1903, i (bb), p. 53, and reprints Nos. 2 and 3.
146
~~
On the Distribution of Osseomucord. 147
the bones of other animals it was impossible, of course, to say that
Young’s deduction as to complete substitution of the “connective
tissue matrix” during ossification, was not, perhaps, correct in the
main.
During the past two years we have been making a study of the
distribution of osseomucoid in the bones of various animals... Our
purpose thus far has been to ascertain merely the extent of its distri-
bution in animal species rather than in the various bones of individ-
uals. Our work in this connection has been wholly qualitative.
The bones under examination were, in practically all cases, the
larger osseous structures of the limbs. They were thoroughly freed
- of all extraneous matter and subjected to the treatment in acid de-
scribed for previous preparations.2 Large bones were converted into
shavings after treatment with dilute hydrochloric acid. Small bones
were merely softened in the dilute acid and when nearly all of the
inorganic matter was removed they were thoroughly minced in a
meat-chopper. The quantities of moist ossein in each experiment
varied from a few grams to several hundred, the amount in use de-
pending on the bulk of the available bony material. The lime water
extracts were treated with acid in excess, as usual, and the precipitates
were thoroughly tested to determine mucoid identity.
The results of our chemical examinations of the precipitates were
invariably as follows:
A. After thoroughly washing free from acid, each product thus obtained was
found to be acid to litmus.
Each precipitate gave proteid color-reactions.
Each substance was free of phosphorus.
On hydration in pure hydrochloric acid, solutions of each product were
formed which always contained :
a. An insoluble proteid portion.
4. Proteose and peptone.
¢. Sulphate.
@. Reducing substance.
&. Each precipitate readily dissolved in dilute solutions of alkalies and
of alkaline salts.
F. Each substance was insoluble in a moderate excess of cold mineral acid, as
well as organic acid.
Sad
1 Gies: Proceedings of the American Physiological Society, This journal, 1903,
viii, p- xiii ; Biochemical Researches, 1903, i (cc), p. 54.
ecrEes:: Loc. cit.
148 Christian Seifert and William J. Gres.
These results made it evident that all of our precipitates consisted
of mucoid.
By these methods, osseomucoid was detected in, and separated
from, the larger bones of each of the following animals :1
Mammats. Birds. Reptiles. Fish.
Man ‘Woodpecker Alligator Cod?
Rabbit Sea gull Turtle
Seal Partridge
Cat Chicken
Dog Turkey
Black bear Marsh hawk
Ox Blue heron
Sheep Surf snipe
Deer Flamingo
Caribou Ostrich
Pig
Tapir
Kangaroo
Our results make it seem very probable that osseomucoid is a
normal constituent of a// bones.
We take pleasure in acknowledging our obligations to Prof. Henry
F. Osborn, and to Director Wm. T. Hornaday of the New York
Zoological Park, for their interest in this research and for much of
the material under investigation. Weare alsoindebted to Mr. Alfred
Malik for assistance in procuring some of the animals.
1 All bones tested thus far have yielded positive results.
2 The bones of the head were the ones examined.
ON THE VARIATIONS OF BLOOD-PRESSURE DURING
é THE BREATHING OF “RAREFIED AIR.
By FREDERIC HH. BARTLETT.
[from the Physiological Institute (Hallerianum) of Bern University.]
1 ae the summer of 1903 it was my privilege to carry out the
following investigation under the direction of Professor Kron-
ecker. I wish to take this occasion of expressing my gratitude to him
for his constant suggestion and unflagging interest in the work.
Indeed the whole investigation belongs to him in its origin and plan.
The following study is one of a series which Professor Kronecker
‘has made in the course of his inquiry into the causes of mountain
sickness, and its results have contributed further confirmatory evi-
dence of his theory as stated in his “ Gutachten iiber die Frage: Ob
und unter welchen Bedingungen sowohl der Bau als der Betrieb einer
Eisenbahn auf die Jungfrau ohne ausnahmweise Gefahrdung von
Menschenleben (Gesundheit) mdoglich sei.”! He there maintains
that mountain sickness is caused by disturbances of the circulation,
saying that “the pulmonary vessels, exposed to the diminished air-
pressure found also in the lungs, dilate and thereby produce conges-
tion in the lesser circulation.” 2
1 KRONECKER: Beilagen zum Konsessions-Gesuch fiir eine Jungfraubahn,
Zurich, 1894.
2 KRONECKER: Jéid.,p. 47. ‘Wohl derart, dass unter dem auch in der Lunge
verminderten Luftdruck die Blutgefasse der Lunge aufschwellen und hierdurch
Stauungen in kleinen Kreislaufe entstehen.”
The views of Albrecht von Haller, the early Bernese investigator, are inter-
esting in this connection, as he also sought for an explanation of mountain
sickness in circulatory disturbances. Rarefied air, he maintained, is injurious
because it does not fully expand the lungs. Pressure is withdrawn from the
vessels of the whole body, which therefore offer less resistance to the heart and
are easily ruptured. Since denser air expands the lungs better, facilitating the
passage of blood through them, better supplying the left heart with blood, and
making it contract more strongly, we can with difficulty bear sudden changes in
air-pressure such as we have in high altitudes. Rarefied air expands the lungs
incompletely and thus withdraws the stimulus that excites the left heart to con-
traction. Elementa Physiologiae Corporis Humanii, iii, pp. 194-197.
149
150 Frederic 1. Bartlett.
I tested this theory by making rabbits breathe from a bottle
constantly supplied with rarefied air from Waldenberg’s apparatus.
This is constructed on the same principle as the gasometers in gas
factories. Its outer cylinder, 1 metre high and 30 centimetres in
diameter, contains water. The escape tubes being closed, the density
of the air is regulated by pressing down or drawing up the inner
cylinder by means of weights. The bottle from which the rabbit
breathed contained five litres, and was connected by a rubber tube
with the escape-tube of the cylinder. Two other tubes entered the
bottle through the same cork as the first. One opened into the out-
side air and constantly supplied the bottle with from one and a half
to five litres of fresh air a minute, about five times as muchas the
rabbit actually needed; the other ended in a quicksilver manometer
for measuring the air-pressure in the bottle. The rabbit breathed
through a tracheal cannula connected with a short tube which entered
the bottle near the bottom. The corks and tubes were sealed with
paraffin. The rabbit was first given an injection of morphine suffi-
ciently large to prevent pain, the reflex sensibility, however, being in-
creased. The blood-pressure was measured by means of a manometer
connected with the common carotid artery, and arranged to write on
a Ludwig Kymograph.
The tables which follow will show in what way the blood-pressure
and respiration changed with the variations in the pressure of the air.
1 WALDENBERG: Centralblatt fiir die medicinischen Wissenschaften, 1847,
p. 106.
Blood-Pressure During the Breathing of Rarefied Air. 151
TABLE I. July 14, 1903.
Dura- | Differ-
tion of | ence of
breath- |air-pres-
ing. sure.
Blood-| Pulse Respir-| Respir-
pres- fre- atory | atory
fre- excur-
sure. |quency. ;
quency.| sion.
Remarks.
mm. Hg | mm. Hg
0.0
—3.6
11 0.0 90 160
160 36
88 160 38 8 Several convulsive movements.
38
10 —5.1 84 164 62 6-8 | Noise in room—convulsive move-
ments. 34 litres fresh air per min.
0.0 86 160 44 U :
82 164+
62
10 0.0 85 158 34 8 | Noise in room—convulsive move-
: ments.
9} | -81 81 130 68 4-§ | Frequent convulsive movements at
end of period. 2 litres fresh air
per minute.
0.0 90 156 36 6
—8.8 86 Berets ...- | 8-10 |3 strong convulsive movements at
equal intervals during period.
0.0 90 Latrs eyelets ...- | Constant convulsive movements, re-
sult of noise in room and handling.
Blood-pressure at close, 90.
Wr SON ata he
Dyspnoea at close of period; four
convulsive movements at equal in-
tervals during period.
Several convulsive movements.
Several convulsive movements.
101
Rabbit extremely irritable throughout experiment.
152 Frederic H. Bartlett
TABLE II. July 15, 1903.
Respir-| Respir-
atory | atory
fre- | excur-
quency.| sion.
Pulse
fre-
quency.
Remarks.
In this experiment a water mano-
meter was used, in others a mercury
manometer.
Rabbit breathed air from flask with no
ventilation. No sign of dyspneea.
Two convulsive movements.
4 litres fresh air per minute.
Several convulsive movements.
Several convulsive movements.
Noise in room caused several convul-
sive movements.
Several convulsive movements. 4
litres fresh air per minute.
Several convulsive movements.
Several convulsive movements.
Writing on fresh paper. Several con-
vulsive movements.
Cet s | erie fee afo2h ei Wont eS foSy SS)
Expiration very active.
TABLE III. July 17, 1903.
Dura- | Differ-
tion of | ence of
breath-|air-pres-
ing. sure.
A.M.
min. sec.| mm. Hg | mm. Hg
15 15 0.0) 122
37 20} —20.0 Experiment to test ventilation of flask.
Except for slightly forced expira-
tion, no dyspncea. Three or four
convulsive movements.
Respir-| Respir-
atory | atory
fre- | excur-
quency.| sion.
Blood-| Pulse
pres- fre-
sure. |quency.
Remarks
0.0
—20.0 2-4 | No dyspneea.
Same rabbit. In the afternoon, —20 caused slight dyspnoea but no asphyxia.
After cutting vagus at —20, marked dyspnoea appeared, which was relieved by fresh
air. The blood-pressure thereupon fell. At —25 asphyxia ensued in a few seconds.
Blood-Pressure During the Breathing of Rarefied Air. 153
TABLE IV. July 21, 1903.
| Dura- | Differ- Respir- | Respir-
tion of |ence of | Blood- | Pulse atory | atory
breath- |air-pres- fre- | excur-
ing. | sure. quency.| sion.
pres- fre- Remarks.
sure. |quency.
min, sec.| mm. Hg mm. Hg
50 0 136 oad .... | 12-22 | Blood-pressure and respiratory ex-
cursion very irregular.
32 0 26 | 10-22 | During this period rabbit breathed
out of flask with no ventilation.
No dyspnea.
25 30 Rabbit moved two or three times in
middle of this period. Respiratory
excursion, 30.
30 S506 76 In middle of period blood-pressure
fell to 90, and then immediately
rose again to 104.
Several convulsive movements, but
no indication of dyspnea.
One convulsive movement.
Blood-pressure fell steadily to 92.
Series of convulsive movements.
Steady rise of pressure.
Blood-pressure varied greatly. Fig-
ures represent maximum and
minimum. Minimum occurred in
middle of period.
Dyspneea throughout, with constant
rise of pressure. Vagus irritation
at close.
Breathing and blood-pressure both
irregular.
At start, blood-pressure fell immedi-
ately to 88, then rose to 100, and
again fell steadily to 88 at close.
Blood-pressure rose to 106 and then
fell to 89.
Dyspneea at outset, followed by as-
phyxia and vagus irritation. Blood-
pressure showed characteristic rise.
Cut vagi at beginning of period.
Blood-pressure fell immediately to
68 and rose gradually to 92.
Blood-pressure rose rapidly to 116.
Marked dyspneea.
Continued dyspnea.
Continued dyspneea. Tried artificial
respiration, but could not restore
normal pressure and respiration
154 Frederic Hl. Bartlett.
TABLE V. July 22, 1903.
Respir-| Respir-
atory
excur-
quency.| sion.
Dura- | Differ-| pi o6q-
tion of | ence of
breath- |air-pres-
Pulse
pres- fre-
sure. | quency.
Remarks.
min. sec.| mm. Hg
3 58 0
4 30 —2
2 30
3 20 Dyspneea throughout.
6)
2
35
Dyspneea throughout. Forced ex-
piration.
CON tON IN 02 CN) eB IRS CN On
Dyspncea set in immediately at be-
ginning of period.
6
14-8 | Rabbit handled at beginning of period.
5 Breathing difficult, sterterous sound.
6 Cut vagiimmediately after this period.
Note extreme susceptibility to change of pressure shown by this rabbit. Autopsy
showed no particular lung lesion.
Blood-Pressure During the Breathing of Rarefied Air. 155
PABLE VI. “July 23, 1903:
Dura- | Differ- Respir- | Respir-
tion of | ence of aa rues atory | atory :
‘ pres- fre- Remarks.
breath- |air-pres- i fre- | excur-
ing. | sure. 48 y: quency.| sion.
mm.Hg | mm. Hg
7 50 0 97 180 28 2 4
Gmels = O3 162 24 2
3 12{| —20 88 123 48 6
4p Ges —3 92 156 28 2
7 52) —14 83 So o8 40 + Forced expiration.
elon 0 89 156 20 2
eet? 88 144 24 y
2 2) —14 86 4
Z 56| —18 84 136 39 5 Forced expiration. Noise at end
caused convulsive movements with
rise of blood-pressure.
Als, a 89 3 Several convulsive movements.
9 55 | —14 84 132 38 4
Suet | —3 86 152 24 2
tae S| — 10 84 156 32 7)
4 23 | —13 82 151 36 5 Forced expiration
S21 —18 83 4
2 46 | —18 84 4 Period began with strong convulsion
with marked rise of blood-pressure.
op 12) |) —18 90 2 Period began with strong convulsion.
S00) |) 23 79 4 Slight nose movement throughout.
3 92 1 Period began with convulsion.
2 6 Fresh drum. Rabbit disturbed in
change of drum.
0 6 Slight dyspnea.
8 Dyspneea throughout.
6 Less dyspnea. Gradual rise of blood-
pressure to 120 at close.
24 Dyspneea markedly less ; vagus irrita-
tion.
3} Noise in room raised pressure. Fell
to 101 at close.
156
Frederic Hl. Bartlett.
TABLE VI— (Continued).
Respir-| Respir-
atory | atory
_| fre- | excur-
quency+quency.| sion.
Pulse
fre-
Remarks.
Gradual rise in pressure till end of
period at 107.
This period began at 109 blood-pres-
sure and followed a period of
marked vagus irritation at —20.
Reading for —20 imperfect, so
omitted.
Respiration irregular; slight nose
movement.
Spasm after 30 seconds, with fall of
pressure.
Spasm after 35 seconds, with fall of
pressure.
Convulsion at close, with fall of pres-
sure.
One convulsion in middle of period
and several at close, with fall of
pressure.
Convulsions at close of period, with
fall of pressure.
Very slight nose movement, marked
vagus irritation.
Just before close of period, convul-
sions. At close vagi divided, fol-
lowed by dyspnoea and immediate
rise of pressure.
Uniform vagus irritation throughout
period.
Marked nose movements.
Vagus irritation.
In middle of period, vagus irritation
for 35 seconds.
Blood-Pressure During the Breathing of Rarefied Air. 157
TABLE VII. July 24, 1903.
Dura- | Differ-
tion of | ence of
breath- |air-pres-
ing. | sure.
Blood-| Pulse ea
pres- | fre- Siig |
: excur-
sure. |quency. an
Remarks.
min, sec. | mm. Hg | mm. Hg
55 0 104
ap) gall 104+ 1} litres fresh air per minute.
0
ie)
25 Hae aot Sade Movement of rabbit on being handled.
15
54
30 aries S060 Soc Several spasms. Rabbit in bad posi-
a tion.
55 Slight dyspneea.
45 50nc bode .... | Less dyspnoea than at — 24.
Dyspncea and spasms.
High pressure from spasmodic move-
ments.
1} litres of air per minute.
Expiration forced. Slight nose move-
ments.
Dyspncea, with rise of pressure in last
ten seconds.
Spasm at end.
Several spasmodic movements.
Blood-pressure taken at end of period.
0
7
0
1
1
1
1
i
0
1
4
1
3
1
2
0
0
0
2
0
1
—
Continued spasms.
Dura- | Differ-
tion of | ence of
breath- |air-pres-
ing. | sure.
min. ‘sec. | mm. Hg
6 40 0
6 §45: |" —20
220) 3
33 a) |) ales
230) —20
1 2 0
LON 22
1 10 0
I os) =F
I 30'| —20
3 15 | —29
3) IS || S22)
ORS ial 2o
Za 0
Z, 40 0
> 10) —20
Blood-
pres-
sure.
mm. Hg
110
107-96
110-104
Frederic HI. Bartlett.
TABLE VII.— (Continued.)
Respir-| Respir-
atory | atory
quency. fre- | excur-
quency.| sion.
Pulse
fre- Remarks.
164 105 4-6 | Breathing irregular. Expiration ex-
plosive. One spasm. Slight nose
movement.
6 One spasmodic movement.
175 5
4
7
6
4
2
3
126 156 4 Slight nose movements.
6-10 | Increasing dyspnoea Active nose .
movements,
18-40 | Asphyxia.
Vagi cut at end of last period. Spas-
modic movements continue. Blood-
pressure high.
Gradual rise of blood-pressure, in-
creasing dyspnoea, and finally con-
vulsions.
Blood-Pressure During the Breathing of Rarefied Air. 159
Dura-
ing.
a
bh = rnN KF Woe pe aS
—
Oo NH WwW
45
50
55
Differ-
tion of | ence of
breath-|air-pres-
sure.
Blood-
pres-
sure.
92
TABLE VIII.
Pulse
fre-
Respir-| Respir-
atory
fre-
atory
excur-
7 ohana quency.| sion.
176
206
214
160
240
240
204
150
54
16
28
52
14
12-50
July 27, 1903.
Remarks.
Noise in room caused spasmodic move-
ments of rabbit. Cannula final sepa-
rated from flask.
Quiet breathing, undisturbed.
Rabbit moved several times. Blood-
pressure much raised.
Noise caused movement. Blood-pres-
sure much raised.
Rabbit moved several times; ex-
tremely irritable.
Moved again.
Noise in room. Several movements.
Spasm, — rabbit handled.
At start blood-pressure rose, but fell
immediately.
Continual movements.
Rabbit moved once; duration of ef-
fect 15 secs. Blood-pressure and
respiratory excursion are for period
of quiet.
Rabbit moved at very beginning.
Movement, due to noise in room.
Nose movement became manifest and
expiration forced.
After preliminary fall of pressure,
dyspnoea and rise of pressure be-
gan at once. In 1 min. 15 secs.
spasms began; 45 secs. later con-
tinued convulsions.
160 fredevit Hl. Bartlett.
TABLE IX. July 30, 1903.
Differ- Respir-| Respir-
ence of | Blood-| Pulse
air-pres-
sure.
pres- fre- : Remarks.
sure, quency.
mm. Hg | mm. Hg
110 168
102 ote
102 sees Dyspnea.
iit
104 ae
108 Bfersie Dyspneea.
Asphyxia. Vagus irritation.
ae 3 oS MS
101 sinetc Marked depression of blood-pressure
after asphyxia.
108
104
102
Dyspneea followed by asphyxia.
Or ONM |
Marked depression of blood-pressure
after asphyxia.
1
5
1
1
2
2
2
4
After 2 min. 30secs. convulsive move-
ment — respiratory excursion 40—
after which blood-pressure fell to
93 to end of period.
At end of period a delay of several
minutes to remove blood clot in
cannula.
At close of period blood-pressure 100,
the average being about 107.
Dyspnea throughout. Blood-pressure
95 at close.
Dyspneea in 15 secs., followed in a few
seconds more by asphyxia.
Blood-Pressure During the Breathing of Rarefied Air. 161
The following figures are taken from the above tables to illustrate
more forcibly the relation between changes in air-pressure and
changes in blood-pressure.
Diff. of air-pressure.
0, —20, 0, —20, 0, —20, 0, —20, 0,—28, 0,—20, 0,—20, 0,—20, 0,—20.
Blood-pressure.
110, 102, 102, 90,96, 82, 92, 85,97, 80,104, 92,110, 92,97, 88,136, 90.
Diff. of air-pressure. —3, —20.
Blood-pressure. 122, 92.
These tables show clearly the general fact that rabbits react to
slight variations in atmospheric pressure. A rarefaction of the air
corresponding to 300 metres elevation caused in every case marked
difficulty of breathing and in some cases asphyxia.! The specific
results of the investigation may be stated as follows:
1. Rabbits breathing rarefied air show in the aortic system a fall
of blood-pressure. Up toa certain point the fall is greater with the
increase of rarefaction, the relation varying with different individuals.
The maximum fall of pressure was 46 mm. Hg and occurred with
rarefied air of —20.
2. Rapid rarefaction appears to lower the blood-pressure more than
does gradual rarefaction.
3. For the most part, rarefaction diminishes blood-pressure sud-
denly, but sometimes gradually.
4. When the pressure has been reduced from —20 to —32, the ani-
mal suffers from dyspnoea and asphyxia, and the blood-pressure rises.
Slowing of pulsation by vagus irritation arising from dyspnoea lowers
it again: spasms raise it.
5. The pulse frequency shows no clear relation to the rarefaction,
but in general decreases with the pressure.
6. The respiratory frequency increases with the rarefaction.
7. The respiratory excursions of the blood-pressure are weaker
when the rabbit is breathing rarefied air.
8. Rabbits with cut vagi experience dyspnoea even at normal air-
pressure. Their blood-pressure rises with slight rarefaction, and
remains high, even if they are allowed to breathe free air again...
The most important of these deductions is the first: the fact that
1 For example:
—10 mm. Hg = 105 metres rise of altitude.
—20 mm. Hg = 212 metres rise of altitude.
—30 mm. Hg = 321 metres rise of altitude.
162 Frederic H. Bartlett.
the blood-pressure in the aortic system sinks when the pressure of the
respired air falls. The point at which the blood-pressure falls varies
in each individual case, many rabbits resisting a pressure of —15,
others showing a fall in blood-pressure with —1o.}
The question now arises: What phenomena present themselves in
the pulmonary circulation at the same time with the fall of pressure
in the aorta system? According to Waidenburg’s theory, this fall of
pressure is accompanied by a reduction in the capacity of the
arteries.2_ The combined result of the diminished pressure and of the
reduced arterial capacity must be a swelling of the lung capillaries
and a lessening in the outflow of the blood from the lungs. Tiger-
stedt’s recent experiments on rabbits give further proof of this. He
has shown that the systolic blood-pressure in the pulmonary artery of
the rabbit is equivalent to 11—15—25 mm. of mercury, and that by
reducing the atmospheric pressure so that it is equal to the intrapleu-
ral pressure hardly any propelling force would remain and the com-
plex symptoms of lung swelling would result.2 Our experiments have
shown that by reducing air-pressure 15—25 mm., a marked fall in
blood-pressure is produced. The two sets of figures afford an inter-
esting comparison, and seem clearly to indicate that a reduction of
from fifteen to twenty-five millimetres of mercury is enough to bring
about that relation between intrapleural pressure and atmospheric
pressure which will cause the symptoms of lung swelling and of
storing of blood in the pulmonary vessels.
As to the nature of the lesion produced by stagnation of blood in
the lungs, I will cite but one reference. The experiments were
carried out by Welch and Cohnheim, and consisted in ligaturing the
branches of the aorta. They found out that “ Passive oedema first
occurs in the lungs when the obstruction which hinders the outflow
of the blood from the pulmonary veins can no longer be overcome by
the action of the right ventricle.” 4
1 It is clear that animals or men who are wholly within chambers of rarefied
air can endure a much lower pressure (400 mm. Hg or even lower) than those
who simply breathe rarefied air.
2 WALDENBURG, L.: Die Messung des Pulses und des Blutdrucks am Men-
schen, Berlin, 1880, pp. 140-141.
* TIGERSTEDT: Skandinavisches Archiv fiir Physiologie, 1903, xiv, p. 285.
* COHNHEIM: “Es in dem Lungen erst dann zum Stauungsédem kommt, wenn
das Hinderniss, welches dem Abfluss des Blutes aus den Lungenvenen entgegen-
steht, von der Action des rechten Ventrikels nicht mehr iiberwunden werden
kann.” Gesammelte Abhandlungen, Berlin, 1885, p. 594.
Blood-Pressure During the Breathing of Rarefied Air. 163
We may conclude, then, that there exists in mountain sickness an
increased amount of blood in the pulmonary vessels, due to an in-
crease in their capacity and to a stagnation of blood arising from an
equalization of the atmospheric and the intrathoracic pressures. The
lesions arising from this are comparable to those observed in the
experiments of Welch and Cohnheim. The symptoms are analogous to
those seen in dyspnoea and asphyxia, both of which invariably
appeared in our experiments, the stage of their appearance being
different with different rabbits. Finally, the symptoms of mountain
sickness are much aggravated in those who, in high altitudes, must
work. We found that by cutting the vagus of rabbits the effects of
rarefied air are much intensified.
y
REACTIONS TO TEMPERATURE CHANGES IN SPIRIL-
LUM, HYDRA, AND FRESH-WATER PLANARIANS.
; BY 53.0% MAST;
| From the Zoological Laboratory of the University of Michigan.]
HE general reactions of Spirillum, Hydra, and fresh-water Plan-
arians has recently been worked out; the reactions of Spirillum
by Jennings and Crosby! and Rothert,? of Hydra by Wagner,’ and
of Planarians by Pearl, and Parker and Burnett.> No work, however,
was done by any of these authors on reactions to heat and cold in
these organisms. My attention was called to this fact by Dr. H. S.
Jennings, and at his suggestion and under his direction this work
was taken up. I wish here to acknowledge my indebtedness to him
for valuable advice and criticism.
SPIRILLUM.
The Spirilla studied were found in cultures composed of decaying
hay and aquatic plants. There were probably several species, but
most of the Spirilla were thought to be S. volutans (Ehrenberg).
For a brief description of this organism, its movements, chemotaxis,
motor reflex, etc., see Jennings and Crosby.®
The thermotactic reaction of Paramecia‘ can be very clearly demon-
strated, (1) by mounting the Paramecia under a large cover glass,
and cooling or heating local areas with drops of cold or hot water
placed on the cover glass, and (2) by first cooling or heating the
slide containing the animals; then, if the slide was heated, putting
drops of cold water or small pieces of ice on the cover; if it was
JENNINGS and Crossy: This journal, 1go1, vii, pp. 31-37.
ROTHERT: Flora, 1901, Ixxxviii, pp. 371-421.
WAGNER: Not yet published.
PEARL: Quarterly journal of microscopical science, 1902, pp. 509-754.
PARKER and BuRNETY: This journal, 1900, iv, pp. 373-385. _
JENNINGS and Crossy: Loc. cit., pp. 33-35-
JENNINGS: Paper on “ Reactions to heat and light,” now in press.
165
~—
1
2
3
pei
5
6
i
166 S.C: Mast:
cooled, putting drops of warm water on the cover. The slide may
be conveniently heated or cooled under the microscope by placing a
flat bottle containing hot or cold water on the stage and laying the
slide upon it. Under the first condition the Paramecia leave the
area beneath the drops on the cover; under the second they collect
beneath the drops.
Experiments similar to those described above were performed,
using Spirillum in the place of Paramecium, but in no case did the
Spirilla collect or disperse under the drops. The only noticeable
effect of a change of temperature on Spirillum was an increase in
motion, both forward and rotary, when passing from regions of lower
to higher temperature; and a decrease in motion when passing from
higher to lower. Spirilla frequently reverse their direction of motion
(motor reflex ),! when coming in contact with solid particles or when
stimulated chemically, but such reactions could not be demonstrated
in the case of thermal stimuli. The organisms were subjected to
both sudden and gradual changes of temperature, varying from
nearly 0° to far above the ultramaximum, and indeed many were
exposed to temperatures so high as to prove fatal; yet no motor
reflex could be observed.
It is thus seen that while Spirilla react slightly to thermal stimuli
their reactions seem in no way purposive; 2.¢., they are not of
such a nature as to keep the organisms in regions of optimum
temperatures. Such non-purposive reactions were entirely unex-
pected, and since no similar reactions to thermal stimuli were found
recorded in the literature,” it was thought that further investigation
might prove them incorrect. In order then to carry on such further
investigations I used a modification of Mendelssohn’s apparatus for
the study of thermotaxis.*? By means of this apparatus temperature
changes can be more accurately regulated than by the method
described.
Mendelssohn’s apparatus as modified in these experiments consists
of two horizontal, parallel glass tubes of convenient length, with a
rubber tube attached to either end. The free end of one of these
rubber tubes is connected with a siphon in a jar of water situ-
1 JENNINGS and CrosBy: Loc. cit., p. 34.
2 JENNINGS personally stated later that he had obtained somewhat similar —
results in subjecting certain ciliate infusoria to different temperatures.
8 MENDELSSOHN: Journal de physiologie et de pathologie générale, 1902,
Pp. 409.
Reactions to Temperature Changes in Sptrillum. 167
ated considerably higher than the glass tubes; that of the opposite
rubber tube leads into a waste jar. The rate of flow of water is regu-
lated by means of adjustable pinch cocks. Thus by heating the
water in one of the outflow jars and cooling it in the other and
regulating the flow, the temperature of the glass tubes can be varied
at will. A slide or dish containing the organisms worked on is
placed on these tubes and thus subjected to different degrees of
temperature.!
Spirillum was mounted on a slide two inches wide and. covered
with a thin slide one inch wide supported by small pieces of wire
to prevent crushing. A little vaseline was smeared along the edges
to prevent evaporation. The slide was then laid on the glass tubes
in the apparatus described above, so that one of the tubes was near
either end of the slide. The temperature of one end of the slide was
then very gradually lowered almost to o°, by slowly passing water
containing ice through the tube under it and finally by placing small
pieces of ice on the slide over the tube; that of the other end was as
gradually raised to about 50° by slowly heating the water and allow-
ing it to flow through the glass tube under it. While these changes
in temperature, which required nearly two hours, were taking place,
the Spirilla remained, as nearly as could be judged, equally distributed
with reference to regions of different temperatures,” although all in
the region of highest temperature (50°) were killed before the
experiment was ended. During the course of the experiment many
of the organisms collected in dense groups, remained thus for a short
time, dispersed, and collected elsewhere. Several such groups were
formed, but these also were about equally distributed over the slide.
Soon after such a group is formed a small circular area may be
seen, in the centre of which there are but very few Spirilla. This
area gradually becomes larger and larger until the group disappears.
In the regions of comparatively high temperature the groups form
more quickly and disperse more quickly than in regions of lower
temperature. These aggregations are similar to those due, in the
case of Paramecium,’ to the excretion of carbon dioxide by the organ-
isms. If in the case of Spirillum they are also due to the excretion
of carbon dioxide, their more rapid formation when the temperature
1 This apparatus is figured in the unpublished paper of JENNINGS, already
cited.
2 A BrAus-DRUNER stereoscopic binocular was used in studying the reactions.
8 JENNINGS: Journal of physiology, 1897, xxi, pp. 258-322.
168 S. O. Mast.
is higher, is what would be expected, since an increase in tempera-
ture causes an increase in activity, and consequently would cause an
increase in excretion of carbon dioxide. Experiments with carbon
dioxide seemed to indicate, however, that this is not the cause of the
aggregations in the case of Spirillum.
A number of Paramecia which happened to be among the Spirilla
collected in a region about one centimetre wide, a little nearer the
cooler than the warmer end of the slide.
This experiment was performed repeatedly, and the same result
obtained in each case. The results confirm in every respect those
obtained by the first method. We may therefore conclude that
Spirillum volutans is not thermotactic in the true sense of the word.
This being true, a determination of the ultramaximum and ultra-
minimum temperatures will be of some interest.
The ultramaximum temperature, as above stated, was reached in the
region of highest temperature in the experiment with Mendelssohn’s
apparatus. It was then only necessary to measure this temperature.
This was done by laying on the slide bits of paraffin having different
melting points, and later determining the melting point of that which
melted over the region fatal to Spirillum. The ultramaximum tem-
perature was thus found to be between 49° and 50°
In order to determine the ultraminimum temperature, a solution
containing Spirillum was kept for twelve hours on ice, but without
freezing. This temperature did not prove fatal. Another solution
was gradually frozen and allowed to thaw very slowly while surrounded
by the freezing mixture, the mixture being uncovered and set in a
room at about 22°. After the ice thawed, the Spirilla were all found
to be dead. It is possible that the ice surrounding the Spirilla
reached a temperature slightly below o°, but it is not probable,
since as soon as the solution was frozen it was uncovered and
allowed to thaw. We may thus consider the ultraminimum tem-
perature of Spirillum volutans to be somewhat below o°. It is of
course probable that the physical changes which occur in freezing
or thawing are the real cause of death.
HypRA.
In the following work Hydra vulgaris was used exclusively. The
material was largely collected in the Huron River, where Hydras
were found in abundance attached to pond lily leaves and other
Reactions to Temperature Changes in Spirillum. 169
plants growing near the head of a mill pond in water having a very
slight current.
Two methods of applying thermal stimuli were used; one by in-
creasing or decreasing the temperature of the water in which the
animals were kept, the other by bringing an object of high or low
temperature near the animals. In the first method, Mendelssohn’s
apparatus was used with modifications; for the second method an
apparatus was constructed as follows: A small glass tube about
fifteen centimetres long was drawn out into a capillary tube in the
middle, and then bent on itself so as to form a long narrow U
tube. The two arms of the U tube were then passed through
holes in a cork, which served
to hold the tube and also to #& ‘a a
strengthen it. 22 Sa
A small rubber tube was slipped
over the end of each arm. One Ficure 1.— Apparatus used in applying
of these was connected with a pole pierre 4, cou byelass
siphon in a jar about one-half
metre above the table, the other opened into a waste jar. Thus the
water in the jar above the table could be siphoned through the U
tube, and by regulating the temperature of the water in the jar and
the rate of flow by means of a pinch cock, the temperature of the
U tube could be varied as desired, and kept nearly constant even
under water.
The results of applying stimulations according to the second
method will be discussed first. The Hydras were put into small
glass dishes containing water about seven millimetres in depth, and
left till they were well expanded. The U tube was then carefully put
into the water some distance from an animal, and slowly brought near
its tentacles, foot, or sides. If the tube is moderately warm (50°-
60°), the Hydras respond very readily to the stimulus by contracting,
whatever part of the animal stimulated by the tube. As to the
relation of the movement to the localization of the stimulus, the fol-
lowing may be noted. When the tube is brought near the foot, the
animal contracts of course toward the source of stimulation. (This
was found to be always true, no matter how great the stimulation.)
When the tube is brought near the sides, the animal contracts in a
line at right angles with the source of stimulation. In such contrac-
tions no lateral movements which would carry the animal away from
the source of stimulation could be detected. Only when the tube is
170 S. O. Mast.
brought near the tentacles from in front, do the animals contract so
as to get away from the source of stimulation. If the tube is kept
near the animal for some little time after it has contracted, it will
frequently expand again. The direction of such expansion, however,
bears no definite relation to the source of stimulation. The animal
is just as likely to move toward the source of stimulation as in any
other direction. Thus, on the whole, it is evident that the direc-
tion of the movements induced in Hydra by thermal stimuli has no
relation to the localization of the stimulus. If the temperature. of
the tube is increased sufficiently, the Hydras will no longer expand,
and may be killed without further movement. In no case was it
possible by this method to cause the animal to release its foothold
and to move away. Such movements were, however, produced by
another method. (See page 171.)
If water at O° is passed through the tube, and a Hydra treated
as described above, the animal reacts essentially as if hot water had
been used; but the reactions are much slower and less definite. In
fact, it is often impossible to tell whether such reactions are due to
a stimulus caused by a decrease in temperature, or whether they
are spontaneous reactions, since a Hydra, under normal conditions,
slowly contracts and expands once every two or three minutes.
The reactions of Hydras to thermal stimuli as described above
agree with their reactions to chemical and mechanical stimuli as
worked out by Wagner (/oc. cit.) with one exception. Wagner found
that if a Hydra is slightly stimulated mechanically for a long time, it
will release its foothold and move away; but the direction of such
movements, as in case of expansion after thermal stimulation, bears
no relation to the source of stimulation.
In reactions due to thermal stimuli, a Hydra does not always react
simultaneously throughout, z.¢., there may be local responses due to
local stimulations. One or more of the tentacles frequently contract
first, after which the rest of the animal may or may not contract.
Such reactions are especially prominent when Hydra is thermally
stimulated from the oral end. The body of an animal may also con-
tract without the tentacles, but this reaction is rare. If Hydra is
stimulated near the foot, the different parts of the body usually con-
tract simultaneously.
If a well-heated tube is lightly brought in contact with the side of
the body of an expanded Hydra, the animal will immediately bend at
the point of contact, toward the source of stimulation, until it nearly
ee ee
Reactions to Temperature Changes in Spirillum. 171
if not quite forms a right angle at that point. After thus bending, it
soon contracts; but when it re-expands an angle is again formed at
the same place as at first, though it is not soacute as the first. Thus
as the animal contracts and expands the angle gradually becomes
more nearly straight, until it finally disappears. Wagner (Joc. cit.)
obtained similar reactions to local chemical stimuli. He suggests
that these reactions may be traumatropic, 7z.¢., due to injuries ; and I
am inclined to believe they are, since they cannot be produced by
local mechanical stimulations or stimulations due to a decrease in
temperature. If the U tube through which water at 0° is flowing is
brought in contact with the body of a Hydra, it usually contracts
immediately, though rather slowly, but does not bend at the point of
stimulation.
The question as to the ability of Hydra to react in such a way as
to protect itself against unfavorable temperatures has already been re-
ferred to, but requires further consideration. It was found by exper-
iment with the U tube that the animals do not move away from the
source of stimulation by releasing their foothold or by contracting or
expanding in such a way as to avoid critical temperatures. It was
thought, however, that they might do so if temperature changes were
more carefully regulated. For this purpose, Mendelssohn’s! appa-
ratus, with modifications, was used. In place of.the slide, a tin box,
three centimetres deep, nine centimetres wide and twenty centi-
metres long was set on the two glass tubes so that one was near each
end of the box. The box hada cross partition, three centimetres from
one end, making a water-tight compartment. After filling the box
with water to the depth of about eight millimetres, several Hydras were
scattered over the bottom. The temperature of oneend of the box
was then gradually decreased to nearly o° by putting small pieces
of ice into the compartment above-mentioned, and that of the other
end was gradually increased to 38° by slowly passing hot water through
the tube under it. As the temperature increased, the rate of contrac-
tion and expansion of the Hydras also increased, and later the animals
began to release their foothold and to move away from the point at
which they had been attached. But the direction of such movements,
as well as the direction of expansion, had no definite relation to the
source of stimulation, z.e.,a Hydra was just as likely to move or to
expand toward the source of stimulation as away from it (cf. Fig. 2.),
so that when the ultramaximum temperature (34°) was reached the
1 Described under Spirillum, page 166.
FIGURE 2.— Diagram representing the reactions of Hydra to gradual increases of tem-
perature as seen in Mendelssohn’s modified apparatus. The diagram represents
one-half of the box used in connection with the apparatus. g, glass tube; 38°, 33°,
28°, 23°, temperatures at end of experiment. (The temperature of the right end of 4
the box, not represented, was nearly 0°.) a, 4, c, etc., Hydras; ¢, tentacles; #, body. :
The numbers, 1, 2, 3, etc., represent the successive positions taken by an animal as
the temperature increased; the highest number in each series shows the place where i
the animal died. «, between two numbers signifies that the animal changed posi-
tion by releasing its foothold. In nearly all other cases the animals changed position
by first contracting and then expanding in a different direction. Not all the contrac-
tions and expansions are represented. Occasionally, however, they changed position 4
by moving the oral end laterally. The animals represented by the diagrams without
numbers changed their position by contracting and expanding several times during
the experiment, as did also several others situated in regions of lower temperature in
that portion of the box not represented, but none of them moved by releasing their
foothold. It can be seen from this diagram that Hydras move at random, and are .
as likely to move toward the source of thermal stimulation as away from it; also A
that they move by releasing their foothold only when the temperature is increased to 4
about 31°, and not when it is decreased. ;
Sol
Reactions to Temperature Changes in Spiritlum. 173
animals died, although a movement of one or two centimetres in the
right direction would have carried them out of danger. Comparing
these results with those obtained by stimulating with a hot tube, it
will be seen that they agree in all respects save one. By stimulating
with a tube Hydra could not be forced to release its foothold, while
it could by stimulating as just described. This may be due to the
fact that with the U tube the foot could not be stimulated as much as
the rest of the animal, since as soon as the water was heated upward
currents were formed.
The Hydras in the region of lowest temperature (nearly 0°)
contracted and expanded very slowly and less frequently than
under normal conditions. None of these released their foothold
and moved away. In performing this experiment, the results ob-
tained are the same when the small compartment contains either
ice or water at about 22°. The experiments require one to one and
one half hours.
Considering the results above stated, it is difficult to see how the
reactions of Hydra due to changes in temperature can be of any
protective value to it, excepting in a rather accidental way. It is
hardly probable that the contractions and expansions of Hydra are
in any way protective against critical temperatures. The reaction
which consists in releasing the foothold and moving away from the
point of attachment in some random direction would, of course, occa-
sionally result in escaping critical temperatures, even if, as shown
above, the animal is as likely to move toward the source of stimula-
tion as away from it. But Hydra reacts by releasing its foothold
and moving away only in response to stimuli due to increase of tem-
perature and not to those due to decrease, and then only if the
temperature is increased to nearly 31°. Now Hydra in its natural
environment is seldom if ever subjected to a temperature of 31°, con-
sequently it is probable that few if any Hydras were ever under
natural conditions thermally stimulated in such a way as to cause
them to react by releasing their foothold and moving away. But
practically all of them will give this reaction if properly stimu-
lated; thus we are certain that all or almost all Hydras that
give this reaction in the laboratory in response to thermal stimuli
do so without previous experience. Are such reactions due to an
inherent property of protoplasm; or can they be explained on the
theory of ‘‘ natural selection ”?
It seems to me that the fact that the reaction to mechanical and
174 SO MaMast:
thermal stimuli are in all essential points the same, throws some
light on the question.
A mechanical stimulation produces a certain definite physiological
change in an animal, which causes it to respond with a definite reac-
tion. It is evident that if the same physiological change could be
produced in any other way, the animal would respond with the same
reaction. In other words, similar reactions in an organism are due
to similar physiological changes in the organism regardless of the
kind of stimuli that have caused the changes. Then, since the reac-
tions of Hydra to thermal and mechanical stimuli are similar, the
physiological changes which cause the reactions are probably similar.
If this is true, in order to explain the non-protective reactions of
Hydra to thermal stimuli, it is necessary only to explain the reactions
to mechanical stimuli. But the reactions to mechanical stimuli are
protective, and their development in Hydra may be explained by the
theory of “natural selection,” that is, through frequent mechanical
stimuli, which cause definite physiological changes, Hydra has, by
‘““ selection,” acquired the power to respond in a certain definite way.
When similar physiological changes are produced by thermal stimuli,
Hydra reacts in the same way, although its reactions may be of no value.
This question will recur in the account of the reactions of Planarians.
A number of experiments were performed in order to determine
the degree of sensitiveness of Hydra to temperature changes. In
carrying on such experiments, the animals were put into a beaker and
set in a shallow dish into which hot or cold water was siphoned, and
thus the temperature of the water in the beaker raised or lowered,
and the number of degrees of change necessary to cause contraction
noted. Conclusions derived from such experiments are subject to
two sources of error. In the first place, it was many times impossible
to tell whether the contractions of Hydra were due to temperature
changes or whether they were spontaneous. In the second place, it
was impossible to measure the temperature of the water next the
walls of the beaker; and it was the change in temperature in this
water, or even possibly the temperature of the walls of the beaker,
which gave the stimulations. The results of all these experiments
may be summed by saying that Hydras living in water at about
22° respond to a sudden increase in temperature of from I° to 2°.
If the temperature is first gradually increased to 28° or 29°, and then
suddenly increased, they respond to a change of from 3° to 1°.
Reactions due to a decrease in temperature were so varied and
Reactions to Temperature Changes in Spirillum. 175
uncertain that it was impossible to come to any definite conclusion,
but it was evident that a greater decrease than increase was necessary
to produce reactions. In fact, as will be shown later, the tempera-
ture can be easily decreased so slowly as not to cause any notice-
able modifications in the normal reactions.
The ultramaximum temperature for Hydra was roughly determined
in the experiments with Mendelssohn’s apparatus. To determine it
more accurately, and also to study more carefully the effect on Hydra
of gradually increasing the temperature, a beaker containing five
animals was set in a medium-sized bacteria dish full of water at 22°.
This water was then displaced, drop by drop, by conducting hot
water into the bottom of the bacteria dish. Thus the temperature
of the water in the beaker was very gradually raised; in fact, so
gradually that it required fifty-six minutes to raise it from 22° to 34°.
As the temperature increased, the rate of contraction and expansion
also increased slightly, until a temperature of about 28° was reached,
when the animals began to remain contracted longer, and did not
expand so fully as under normal conditions. At 31° they began to
release their foothold, and move away from the point of attachment.
Such movements continued from time to time until the temperature
reached 34°, when all motion ceased. It required twenty-three min-
utes to raise the temperature from 31° to 34°. During this time one
of the Hydras moved seven times, the rest from two to five times.
At the beginning two were attached to the sides of the beaker; these
moved down to the bottom. Those attached to the bottom were in
about the same position at the end of the experiment as they were
at the beginning. In moving they stretched out partially, turned
the free end down until it came in contact with the bottom so as to
form an arch, raised the foot, frequently remained supported on their
tentacles a minute or more, and then put the foot down near the
tentacles, sometimes on the same side upon which it had been before
it was raised, and sometimes on the opposite side, thus turning a
somersault. None were seen to crawl on their tentacles. Two of
the Hydras died while supported on their tentacles, and remained
thus after death. One hour after all motion ceased, all the animals
were nearly liquefied. We may thus conclude that the ultramaxi-
mum temperature for Hydra, living in a temperature of about 22°, is
about 34’.
The above experiment, in connection with those performed with
Mendelssohn’s apparatus, seems to me to lead to another conclusion
176 S. O. Mast.
worthy of mention. In the experiment just described, the animals
were stimulated equally on all sides; while in the experiment with
Mendelssohn’s apparatus, they were stimulated more strongly on one
side than on the other. Now the reactions under these two condi-
tions are found to be the same. If the reactions are due to a direct
effect on the motor organs, as Mendelssohn believes, in the case of
infusoria,! we should expect different reactions under these two differ-
ent conditions. And furthermore, according to the theory of a direct
effect on the motor organs, it is difficult to see why Hydra, when
stimulated equally on all sides, should release its foothold and move
away,—-a movement which requires unequal reaction in the muscle
fibres on two different sides of the body, which are subjected to
equal stimuli. These results seem to me to lead to the conclusion
that thermal stimuli act indirectly on the motor organs, and that the
direct cause of the reaction must be sought elsewhere.
If the temperature of the water in which Hydras are found is
gradually decreased from 22° to o° in about one hour, they do not
appear to be stimulated. They contract and expand from time to
time, as they do under normal conditions, but more slowly and less
frequently as the temperature approaches 0°. At low temperatures
Hydras also become much less susceptible to mechanical stimuli, and
more easily loosened from their attachment by the foot than under
normal conditions, so that those attached to the surface film, and
some of those attached to perpendicular surfaces, fall to the bottom,
frequently remaining expanded during such descent. This effect of
a decrease in temperature on the attachment of the foot may be due
either to a physical change in the mucus, or a physiological change
in the cells of the foot. The fact that the attachment is affected in
this manner by cold seems to be of some biological significance.
Hydras are usually found near the surface of the water in which they
live, and consequently, since a decrease in temperature does not cause
them to move, they are subject to freezing, which, as will be shown
later, is fatal. Now, since the attachment of the foot becomes much
weakened as the temperature decreases, the slightest jar will cause
them to fall to the bottom, and thus they may escape being frozen
and killed.
Under normal conditions the longitudinal axes of expanded Hydras
are usually nearly perpendicular to the surface to which they are
attached. As the decreasing temperature approaches 8°, the animals
1 MENDELSSOHN: Loc. cit., pp. 484-485.
Reactions to Temperature Changes tn Spirvitlum. 177
slowly bend near the foot so that the longitudinal axis, if not attached
to horizontal surfaces, becomes vertical. This bending seems to be
due to a decrease in the tonus of the muscle fibres near the foot, and
consequently an inability to hold the animal perpendicular with the
attached surface against gravity.
At 0° the activity of Hydra ceases almost entirely. At this tem-
perature some may be seen fully expanded lying on the bottom as if
dead; others remained contracted. But if the temperature is raised,
the animals recover at once. Several, thus kept at o° for twelve
hours without being frozen, recovered as soon as the temperature was
raised to normal. If the temperature of water is gradually lowered,
Hydras may be frozen in a partially expanded condition. Several
were thus frozen and examined in the ice. While some of them were
well contracted, the bodies of others were found to be from two to five
millimetres long, and their tentacles well expanded. Frozen Hydras
slowly thawed in a temperature of 22° do not recover. Their ultra-
minimum temperature is therefore below o°,— death being probably
due to the physical changes in freezing or thawing.
FRESH-WATER PLANARIANS.
As noted in the introduction to these studies, Pearl (/oc. czt.)
and Parker and Burnett (/oc. cit.) have carefully worked out the gen-
eral behavior of Fresh-water Planarians, excepting their reaction to
thermal stimuli.
A quotation from Pearl will show the extent of his work. ‘The
general natural history of the animal was studied as completely as
possible. All the normal movements were studied in detail. The
reactions to mechanical stimuli; the food reactions and reactions to
chemicals in general; electrotaxis; thigmotaxis ; rheataxis; the right-
ing reaction; the reaction of cut and regenerating pieces; and hydro-
taxis and the reaction during desiccation were investigated. No
work was done on the phototaxis or thermotaxis.” A study of photo-
taxis was made by Parker and Burnett (: 00).
Pearl in his study found that there are two principal, qualitatively
different reactions to stimuli, the positive and the negative reactions.
‘‘The negative reaction is given in response to strong unilateral
stimulation of the anterior portion of the body. It consists essen-
tially in a turning away of the head from the side stimulated.
“The positive reaction is given only in response to weak unilateral
178 ‘pd. AL ast.
stimulation of the anterior portion of the body. It is essentially a
turning of the head toward the source of stimulation. This reaction
is one of considerable precision, bringing the anterior end into such
a position that it points in most cases exactly toward the source of
stimulus.”! Pearl found that the positive reaction was given in
response to all weak mechanical, rheotactic, and chemical stimuli,
regardless of the substance used as a stimulus, and that the negative
reaction was given in response to all strong mechanical and chemi-
cal stimuli, regardless of the agent used as a stimulus with one
exception.2, The negative reaction could not be produced by rheo-
tactic stimulations. =
A brief review of some of Pearl’s results has been given because in
the following description frequent reference to them will be made and
some of his terms will be used. His experiments on mechanical
stimulations were repeated by me, and results obtained which agree
with his throughout.
Material. — Planaria dorotocephala was used almost exclusively in
the following experiments. The animals were collected in the Huron
River at Ann Arbor immediately below a dam, in a swift current,
where they were found on the under surface of rather large stones,
which had settled well into the substratum.
Reactions to local thermal stimulations. In giving local stimula-
tions, the U tube described under Hydra (Fig. 1) was used. Quan-
titative results are practically impossible in using this method of
stimulation. Some quantitative work, however, was done later.
If the tube, while hot water is flowing through it, is carefully
brought near the margin of a Planarian, anywhere in front of the
cesophagus, the Planarian turns towards the tube, —the source of
stimulation, —z.¢., it gives the positive reaction above referred to.
In thus turning the animal sometimes lifts its anterior end, and if the
temperature of the tube is not too high, it usually continues to move
towards the tube, and occasionally grasps it with the anterior end, as
is customary in food reactions. Then if the tube becomes too warm
on contact, as is usually the case, the animal turns its anterior end
away,—thus giving the negative reaction described above. Fre-
quently, however, in giving the positive reaction the animal does not
turn sharply enough to come in contact with the tube, merely turning
toward it slightly, and passing by. If the tube is very warm, the
animal usually raises its anterior end, brings it toward the source of
1 PEARL: Loc. cit., ps 700. 2406. CU we 5 ae 8. LotxGit De ses
Reactions to Temperature Changes in Spirillum. 179
stimulation, and then, as it reaches a region of comparatively high
temperature, suddenly throws its anterior end in the opposite direc-
tion until this frequently forms a right angle with the posterior half —
of the body; then gradually it swings its head toward the source of
stimulation again. A Planarian will thus not infrequently swing its
anterior end from side to side three or four times, as if in the act of
investigating matters, and then finally move away from the source
of stimulation, — giving the negative reaction. If the tube is very
warm, and is brought near the anterior end of the animal rather
suddenly, the negative reaction is induced, not preceded, by the
positive.
In the above description use has been made of the phrase ‘‘ turns
towards or away from the source of stimulation.” From this it must
not be understood that the animal orients itself and moves parallel
with the rays of radiation or convection, for this is only seldom true,
and then it is apparently accidental. The question of orientation
will be referred to again later (see page 184).
It is not always possible to cause a Planarian to give the posi-
tive reaction, whatever care is exercised in varying the degree
of thermal stimulation. Pearl found this to be true also with refer-
ence to mechanical stimuli, and showed that whether an animal can
be made to give the positive reaction or not depends upon its physi-
ological condition, ¢. ¢., animals in an excited condition or at rest will
not respond with the positive reaction!
It was found, however, that while this is also true with reference
to thermal stimuli, the positive reaction to thermal stimuli is more
definite than to mechanical stimuli, and can be more readily and more
frequently produced by the former than by the latter.
Thermal stimuli on any portion of the posterior half of a Planarian
cause, if weak, slight increase in gliding movement, and, if strong,
“crawling” movements. Crawling movements can be induced only
if the tube is Zot and brought very near the animal. They were not
induced by ventral stimulation. If the tube when very warm is held
over the anterior end of a Planarian near the so-called oral appen-
dages, the animal soon turns its anterior end to one side, but if the
tube is now moved over the oral appendages again and kept there,
the animal throws its anterior end up and twists it so that the ventral
surface of the head faces the tube. When the stimulation. becomes
too strong, due to the fact that the head is brought too near the hot
1 PEARL: Loc. cit., pp. 592-595.
180 SHG. Wass
tube, the worm suddenly withdraws the head, usually by contracting
‘the anterior portion of the body so as to get out from under the tube.
It then turns aside and moves on. Sometimes, however, instead of
thus contracting and backing out from under the tube, it glides for-
ward and escapes in this way.
Ventral stimulation. — Thermal stimulation can readily be applied
to Planarians gliding along the under side of the surface film with
their ventral surface up,! or by putting them in some water on a ¢hin
piece of glass and heating or cooling the glass from below.
If a hot iron is held above the anterior end of a Planarian gliding
along the surface film, it will throw its anterior end down (a negative
reaction) and swing it from side to side. If the stimulation is con-
tinued, the animal will leave the surface film and sink to the bottom.
A Planarian stimulated in the region of the head, from below, by
heating the stratum upon which it is gliding, will throw its anterior
end up, swing it from side to side, and if the stimulation is strong
enough, finally turn abruptly and move away. If the stimulation is
not very strong, the worm will pass over the heated region, after rais-
ing its head and swinging it from side to side a few (two to five)
times. In such cases the animal continues in about the same direc-
tion after stimulation as it did before.
Reactions to stimulation due to a decrease in temperature. — In
stimulating Planarians by decreasing the temperature, precisely the
same methods were used as in stimulating by increasing the tempera-
ture; and the reactions in response to such stimuli were essentially
like those in response to stimuli due to increase in temperature; 7. é.,
the anterior end was frequently raised and swung from side to side;
positive reactions were given to weak stimuli and negative reactions
to strong stimuli. The negative reactions to decrease in temperature
were, however, not so pronounced as those to increase in temperature ;
while the positive reactions appeared to be more pronounced, and the
head was also less frequently raised and swung from side to side in
response to cold than in response to heat.
The difference between reactions to stimuli due to decrease and those
due to increase in temperature is probably owing to the fact that as the
temperature decreases the animals become less susceptible to stimuli.
Then, too, it is possible to increase the temperature much more than
it is possible to decrease it; since if the temperature of the water in
which the animals live is 22°, it can be increased 78°, but decreased
2 PEARL: (Loc. 62. qin g3a:
AEE AP CNT Ah
es yy
Reactions to Temperature Changes in Spirillum. 181
only 22°. Thus one would be likely to give stronger stimuli in
increasing the temperature than he would in decreasing it.
Equal, simultaneous, thermal stimuli on all surfaces.— In order to
stimulate a Planarian equally on all sides at the same time, it is of
course necessary to have the temperature equal on all surfaces. At
first thought this appears to be a very simple matter, but in reality it
is not so simple as it appears. The chief difficulty lies in the fact
that Planarians move along the bottom or sides of the vessel in close
contact with its walls, which, under ordinary conditions, are warmer
or colder than the water in the vessel. This difficulty was at least
partially overcome in the following way. A Petri dish, about two
centimetres deep and twenty centimetres in diameter, without a cover,
was set on three small pieces of glass, one centimetre thick, in a large
bacteria dish, which was then filled with water at about 60°. Thus
the Petri dish was entirely surrounded by water, and its walls soon
became practically of the same temperature as the water within.
After the water in the bacteria dish had cooled to a desired tempera-
ture, a Planarian was carefully transferred from water at normal tem-
perature to the bottom of the Petri dish and the reactions studied.
A section lifter was found very’ convenient for transferring the
animals.
If the water in which the Planarians live is at 22°, and the water in
the Petri dish at 32°, and an animal is transferred from one to the
other, as soon as it reaches the bottom of the Petri dish it raises its
head slightly (less than when ventrally or laterally stimulated) and,
without forward motion, swings it from side to side five or six times
violently, so that it frequently touches the tail. Then the animal
starts to crawl, usually in a direction almost opposite that which it
faced when first put into the dish. It usually crawls only a short
distance, contracting two or three times, then glides rapidly, making
frequent curves in its course. These curves, however, soon disappear,
and the animal glides about in a perfectly normal way and soon comes
to rest. The time required to regain normal reactions after being
transferred varies with different animals from twenty-five to thirty-
five seconds. If a Planarian, on the other hand, is carefully trans-
ferred from a dish containing water at 22°, to a dish containing water
at the same temperature, it usually starts off at once in the direction
in which it faces when it reaches the bottom. Sometimes it crawls
a short distance, making two or three contractions, and then glides,
but it usually glides from the start, making but few curves in its =
course.
182 S. O. Mast.
If animals are transferred to the Petri dish from time to time, as
the temperature decreases, it is found that their characteristic reac-
tions, described above, become gradually less marked, until, at 25°, it
is questionable in many cases if there is any difference between their
reactions when transferred to the warmed Petri dish or to another
dish ,containing water at 22°. Thus it would seem that an increase
of 3° in temperature is necessary to produce a stimulus strong enough
to cause a response in Planarians, This matter will be considered
more in detail later (see page 188).
In analyzing the reactions of Planarians to equal simultaneous
thermal stimuli on all sides, one is led to consider the cause of such
reactions. Why should a Planarian swing its head from side to side
when the muscles and nerve endings of both sides are subjected to
the same temperature; and why make more frequent curves in its
course than under normal conditions? If the reactions to thermal
stimuli are due to a direct effect of the stimulating agent on the
motor organs, causing either a simple contraction or expansion, we
should expect neither of the above reactions. The fact, however, that
these reactions are given, leads to the conclusion we deduced in the
discussion on the reaction to equilateral simultaneous stimuli under
Hydra (see page 175), namely, the reactions to thermal stimuli
“cannot be due to a direct effect on the motor organs, nor to simple
single motor reflexes. Apparently the stimulus causes a change in
the physiological condition of the animal, and the movements are the
expression of this changed condition.
Optimum temperature. — In determining the optimum temperature
for Planarians, Mendelssohn’s apparatus was used, as modified in the
work on Hydra (see page 171).
If the temperature at one end of the box is reduced almost to 0°,
by keeping ice in the small compartment, and that at the other end
raised to about 32°, and then several Planarians are scattered in a
little water at the bottom of the box, the results are as follows: The
animals glide about the box for a short time, and then come to rest
scattered over a somewhat wide area near the middle of the box.
The lowest and highest temperatures at the two opposite limits of
sixteen such areas were taken. It was found that these varied con-
siderably. The temperature limits of the area at one extreme were
10° and 16°, while those at the other extreme were 20° and 29°.
The average lowest temperature of the sixteen areas was 17°, and the
average highest 26°. Thus we may consider the optimum tempera-
Reactions to Temperature Changes in Spirillum. 183
ture of Planaria living in a temperature of about 22° to be between
Banand 26°.
- Reaction by means of which Planarians reach regions of optimum tem-
perature. — If a Planarian in its wanderings about the box happens to
come near the warmer end (32°), it pauses a moment, raises its head,
swings it from side to side from one to five times, usually not moving
forward during this process, then moves on, after changing its course
FIGURE 3.
so as to form an approximate right angle with its original course;
z.¢., it gives the negative reaction. Its new course is then usually
more or less nearly parallel with the end of the box, and perpendicu-
lar to the rays of radiation and convection. The animal does not,
however, continue on a straight course, but gradually turns towards
the source of stimulation; 7z.¢., it gives the positive reaction. Soon,
however, it is brought to a region of comparatively high temperature,
and is strongly stimulated, when it again raises its head, swings it
184 S. O. Mast.
from side to side, gives the negative reaction, and starts off at an
approximate right angle with its old course. Its anterior end may
now be carried far enough away from the source of stimulation so
that it will not again be stimulated sufficiently to give the positive
reaction: the animal will consequently continue straight on its new
Ficures 3 and 4.— Diagrams of the reactions and typical courses of a Planarian coming
into and getting out of a region of comparatively high temperature, as seen in the
box used in connection with Mendelssohn’s apparatus. The temperature at one end
of the box was 32°, and that at the other end 0°. The dots represent places where
the animal stopped and raised its head. The small figures near the dots indicate the
number of times it swung its head from side to side before taking a new course.
The arrows indicate the direction of locomotion. The curves toward the warmer
region show that the animal responded with the positive reaction.
course, which may form almost any angle with the rays of radiation
and convection. It may, however, before leaving the region of high
temperature, alternately give the negative and positive reactions sev-
eral times, and in so doing move approximately parallel with the end
Reactions to Temperature Changes in Spirillum. 185
of the box, making several curves (see pages 183, 184, Figs. 3 and 4).
An animal after leaving the warmer may go straight toward the
colder end, and there give reactions similar to those described,
but not so pronounced. After this it may again reach the warmer
end, and give the reactions, thus continuing until it finally comes
to’ rest in any region where the conditions are such as not to
give an effective stimulation. It will be seen from this descrip-
tion and the accompanying figure that in reaching the optimum
temperature the animals do not orient themselves and move in any
definite relation to the source of stimulation or the rays of radiation
and convection.
Ultramaximum temperature and reactions to gradual increase in tempera-
ture. — The method used to determine the ultramaximum temperature
for Planarians was that already employed for Hydra (see page 175).
~ If a Planarian taken from a dish of water at 23°, is put into the
beaker and the temperature slowly raised, the animal gradually be-
comes more active, gliding about rapidly and turning frequently, but
it usually soon comes to rest. The reaction given is clearly that
that we-have described as the positive reaction. After the tempera-
ture has increased a few degrees, the animal begins to move again,
now raising its head frequently and swinging it from side to side,
as if investigating. « As the temperature still increases, the forward
motion decreases and the lateral movements increase. The negative
reaction is now very strongly marked. Finally both lateral and for-
ward movements cease, and the animal begins to make rapid, violent,
crawling contractions (crawling reaction); in this it moves forward
but little. Soon after this it begins to twist the body so that it
sometimes has two complete turns in it (righting reaction, Pearl).
Finally it turns the anterior and posterior end under, arching the
central part of the body upward. Usually the anterior end is turned
under farther than the posterior, and the animal goes over forward
onto its back (final reaction). Sometimes, however, it goes over
sidewise. After it gets on its back, the two ends may move forward
and backward a little, but the animal soon dies. -It may then be
concluded that the ultramaximum temperature for Planarians living
at a temperature of about 24°, is approximately 42°.
- We thus have, as the temperature rises and the stimulation in-
creases, the following reactions given consecutively: positive, nega-
tive, crawling, righting, and final. (All the reactions described by
Pearl (/oc. cit.), with the exception of some of the food reactions, and
186 SY 0O-Mast.
the “ final” reaction in addition.!) The positive reaction is conspicu-
ous between the temperatures of 23° and 26°, the negative between
26° and 38°, the crawling between 38° and 39°, the righting between
39° and 40+’, the final reaction between 40+° and 42°. These re-
actions were studied carefully in fourteen different animals, divided
into three groups, and were found to vary surprisingly little.
The reactions of Planaria in response to a gradual increase in tem-
perature bring up a number of interesting questions. How does it
happen that to a single stimulus, not localized, we obtain successively,
as it increases in intensity, such a varied series of reactions, — com-
prising indeed almost all those of which the animal is capable ? ~The
general impression given is that as the thermal stimulus increases,
the animal tries, in a sort of ‘hit or miss” way, every reaction which
it has at command, in order to get rid of the stimulation. The first
reaction (‘ positive”) is that which the organism gives when sub-
jected to any slight, non-injurious environmental change; it gives
the impression of “seeking.” The next reaction (‘‘negative”) is
that regularly induced by more intense environmental changes, which >
would in the long run be injurious; it tends under usual” circum-
stances (though not under those of the present experiment) to
remove the organism from the agent affecting it. The crawling
reaction is another method, perhaps still more effective, of producing
the same result. The “righting” and “final” reactions, to which
the organism has recourse when the stimulus becomes more intense
and the previous methods of response have proved ineffective, are
more difficult to interpret; under certain unusual conditions, how-
ever, they would be useful. It is remarkable that the organism gives
these reactions, when others have failed, even under conditions to
which they are not at all adapted.
The fact that we thus get such a varied series of responses to a
single method of stimulation again indicates that the results cannot
well be due to a direct action of the stimulating agent on the indi-
vidual motor organs. Under the conditions of the experiment the
animal is equally affected on both sides and ends of the body by
the stimulating agent. Yet it responds with “ positive,” “ negative,”
and other reactions, of exactly the sort that are produced under
other circumstances by stimulation at one side or one end. These
results reinforce our previous conclusion, that the direct result of
1 PEARL (@oc. cit.) describes reactions somewhat similar to the “final reac-
tions” in his study on reactions to desiccation.
Reactions to Temperature Changes in Spirillum. 187
stimulation is. to produce some change in the physiological state
of the organism, and that the reactions given are the results of the
changed physiological state. Under the influence of gradually in-
creasing heat, the Planarian undergoes a series of changes in physio-
logical condition, and gives the reactions corresponding to these.
The fact that practically all reactions which can be produced by
mechanical, chemical, rheotactic, and thigmotactic stimuli, varied in
strength and applied to different regions of the body, can also be
induced by thermal stimuli varying only in intensity and not vary-
ing in the regions of application, seems to show that reactions in
Planarians depend very largely on intensity, and but little upon the
location and kind or quality of stimulation. All the reactions to
thermal stimuli induced by gradual increase in temperature, with
the exception of the negative reaction, seem non-protective, under
the conditions of the experiment. Some of them are probably never
induced as reactions to heat, under the natural conditions of ex-
istence. The development of such reactions in Hydra has been dis-
cussed on pages 173 and 174. While the development of the positive,
negative, and crawling reactions to thermal stimuli in Planarians can
be explained by applying the same line of argument, greater diffi-
culties arise in thus attempting to explain the righting reaction and
the final reaction.
Ultraminimum temperature. —If Planarians are put into a beaker of
water and the temperature slowly decreased, they gradually become
less active and respond less readily to mechanical stimuli, until a
temperature of about 10° is reached. At this temperature they. no
longer respond to mechanical stimuli, and practically all motion has
ceased.— If the temperature is lowered still more, the animals turn
the two ends under, and usually roll over onto the back (“ final reac-
tion,” see page 185). They may remain an indefinite length of time
at o without harm, providing they do not freeze; but if they are
frozen, and thawed, they are killed.
Several Planarians were kept at 0° over night without freezing.
The following morning most of them were found lying on the back
with the two ends curled up. They were then transferred to water
at 22°, and almost immediately recovered.
Three Planarians were frozen and thawed in the same beaker with
several Hydras whose fate has been discussed (see page 177). The
freezing or thawing killed the Planarians. -We may consequently
consider O—° as their ultraminimum temperature.
188 SO. Mast.
Threshold of sensitiveness to thermal stimuli.— This question has
already been referred to (see page 182), but it may be well to collect
here all we have with reference to it.
Two methods were used in determining the degree of sensitiveness
of Planarians to temperature changes. In one the animals were
transferred from water in which they lived to water at various higher
temperatures, until a temperature was obtained in which (when the
animals were transferred to it) no difference could be seen between
the reactions and the reactions given when they were transferred to
water at the same temperature as that from which they were taken.
In ‘the other method, Mendelssohn’s apparatus, as modified in the
experiments under optimum temperature, was used and the tempera-
ture of the water over the tube increased until the animals would no
longer pass over it without giving definite reactions.
The results of a large number of experiments according to both of
these methods may be summarized by saying that Planarians respond
to stimuli caused by a rather sudden change in temperature of from
2° to 3°. ~That is, if the temperature of the water to which animals
are transferred is two or three degrees higher than that from which
they are taken, they respond by giving definite reactions. Likewise
when the water in Mendelssohn’s apparatus over the tube is from 2°
to 3° higher than that fifteen centimetres from it, the animals, on
reaching the region over the tube, give definite reactions.
Planarians then are susceptible to stimuli produced by an increase
in temperature of from 2° to 3°.
GENERAL SUMMARY.
I. SPIRILLUM.
Spirillum volutans is not thermotactic; that is, it does not react to
temperature changes in such a way as to collect in regions of opti-
mum temperature, or avoid regions of suboptimal or supraoptimal
temperatures. z
Increase in temperature causes increase in motion; decrease in
temperature, decrease in motion.
The ultramaximum temperature, when the temperature is gradually
increased, is between 49° and 50°.
The ultraminimum temperature is slightly below o°.
is
Reactions to Temperature Changes in Spirillum. 189
II. HYDRA.
Hydra has two methods of reacting to thermal stimuli, one by con-
tracting, the other by releasing its foothold and moving away from
the point of attachment. The second reaction is given only in re-
sponse to stimuli due to increase in temperature.
The reactions of Hydra to thermal stimuli are essentially the same
as its reactions to mechanical and chemical stimuli.
The reactions to thermal stimuli cannot be explained as due toa
direct effect of the stimulating agent on the motor organs.
The direction of movement in the reactions bears no definite rela-
tion to the source of increase or decrease in temperature, z.¢., Hydras
are as likely to move toward the source of stimulation as away from
it, and vice versa.
_ The reactions to thermal stimuli are not directly protective, though
they may be so at times in an accidental way.
Local contractions may take place in response to local stimuli.
Hydras respond at normal temperature to an increase in tempera-
ture of from 1° to 2°. They are less sensitive to a decrease in tem-
perature than to an increase.
The ultramaximum temperature of Hydra is about 34°, the ultra-
minimum is slightly below o°.
III. PLANARIANS.
Planarians respond to weak and strong thermal stimuli, just as they
do to weak and strong mechanical stimuli. That is, they turn to-
wards the source of a unilateral stimulus applied to the anterior
portion of the body, if it is weak, and away from it, if strong, and
they respond by crawling if the stimulus is applied to the posterior
portion of the body.
By gradually increasing thermal stimuli, applied equally on the
sides and ends of the body, Planarians can be made to give all the
different reactions given in response to mechanical, chemical, rheo-
tactic, and thigmotactic stimuli of different strength and applied to
different regions of the body.
The reactions of Planarians to thermal stimuli, depend, primarily,
upon the intensity of the stimulus, and, secondarily, upon the physio-
logical condition of the animal and the location of the stimulus.
— The nature of their reactions to stimuli in general bears little if
any relation to the quality of the stimulus.
190 S. O. Mast.
Planarians in their reactions do not orient themselves with refer-
ence to thermal rays of radiation or convection, z.¢., the path which
they follow is liable to form any angle with such rays.
- Their reactions are due, apparently, to a general physiological
change in the organisms, rather than to a direct effect on the motor
organs or a mere simple motor reflex.
Of all the reactions induced by thermal stimuli, the negative reac-
tion appears to be the only one that is under the conditions of the
experiments directly protective or useful.
- The optimum temperature, when the temperature is slowly in-
creased, is 42°, and the minimum below o°.
- Planarians at ordinary temperatures react to an increase in tem-
perature of from 2° to 3°.
EE ig a sty aimS
THE HYDROLYSIS AND SYNTHESIS OF FATS BY
PLATINUM “BLACK.
By HUGH NEILSON.
[From the Hull Physiological Laboratory of the University of Chicago.]
HE use of catalytic agents in accelerating chemical action has
long been known. Just how and why a catalyti¢ agent acts is
not well understood. Yet the similarity of catalytic action to that of
the so-called ferments or enzymes has led physiologists to teach that
enzyme action is a catalytic action. This relation has been especially
emphasized by Bredig in his classical work, Anorganische Fermente.
Among catalytic agents which are used for chemical purposes are
finely divided metals, as platinum, gold, silver, and palladium. To
this list may be added the colloidal solutions of platinum, gold, etc.,
prepared by Bredig. He finds in the action of these colloidal solu-
tions, especially of platinum, a similarity to the action of certain
enzymes in the splitting of hydrogen peroxide into water and atomic
oxygen. This action is a catalytic action and therefore shows a rela-
tion between the action of the platinum and the enzymes. The
similarity in their action is further shown by the following experi-
ments :-— |
1. The oxidation of alcohol to acetic acid is brought about by the
presence of Mycoderma aceti (Pasteur), or by the presence of finely
divided platinum (E. Davey).
2. According to O. Sulc! dilute oxalic acid is decomposed by
powdered palladium, platinum, etc., and also, according to Jorrissen,?
by the presence of certain fungi.
3. Laccase accelerates the oxidation of pyrogallol. This can also
be done with Bredig’s colloidal platinum, as K. Ikeda has shown.
4. Schénbein®? says that the oxidation of pyrogallol by hydrogen
peroxide can be accelerated by platinum black.
1 Succ: Zeitschrift fiir physikalische Chemie, 1899, xxvili, p. 719.
2 JORRISSEN: Chemisches Centralblatt, 1898, ii, p. 1084.
3 SCHONBEIN: Journal fiir praktische Chemie, 1863, Ixxxix, p. 24.
IOt
192 Hlugh Nevlson.
5. The inversion of cane sugar can be brought about by the action
of finely divided metals, according to Rayman and Sulc.1
6. The decomposition of hydrogen peroxide into water and atomic
oxygen can be very greatly accelerated by platinum, gold, silver, etc.
(Thenard #), as well as by many organic ferments (Schonbein ®).
On the one hand, oxydase splits up hydrogen peroxide into water
and atomic oxygen; invertase inverts cane sugar; zymase ferments
sugar. On the other hand, we find that platinum black will bring
about the same reactions. The only logical conclusion is that there
is a similarity in these actions; or as the action of the platinum is
catalytic, so also the action of the enzymes is the same or at least
similar, namely, catalytic.
In this series of experiments the similarity is carried still farther.
Platinum black is used instead of the fat-splitting enzyme, lipase.
In the experiments, attempts are made to answer the following
questions : —
1. Does platinum black accelerate the hydrolysis of a fat, as ethyl
butyrate ?
2. Is this action reversible, or will platinum black synthesize ethyl
butyrate from butyric acid and alcohol ?
3. Do poisons and antiseptics lessen its catalytic action?
4. Do these reactions obey the same laws of chemical kinetics as
lipase?
METHODS.
The experiments on the hydrolysis of ethyl] butyrate were carried
on in medium-sized test-tubes, while those on the synthesis of ethyl
butyrate from butyric acid and alcohol, were carried on in 250 c.c.
flasks. All glassware used in the experiments was carefully washed,
rinsed in distilled water, and then sterilized in a hot-air sterilizer.
The platinum black used was principally Merck’s preparation. Before
it was used in the experiment, it was washed with distilled water
until the wash-water was neutral to litmus or phenolphthalin. It
was then placed in a drying oven and afterwards heated to 100° C.
for an hour. After this it was carefully powdered, and kept in a
? RAYMAN and Sutc: Zeitschrift fiir physikalische Chemie, 1896, xxi, p. 481 ;
XXVill, p. 719.
* THENARD: Memoire de |’ Academie des Sciences, 1818, iii, p. 385.
* SCHONBEIN: Journal fiir praktische Chemie, 1863, Ixxxix, p. 24.
F[ydrolysis and Synthesis of Fats by Platinum Black. 193
desiccator. The ethyl butyrate and butyric acid were the ordinary
chemically pure articles.
The experiments, except those made to determine the effect of
different temperatures, were performed at a temperature of 38°-4o’ C.
The test-tubes, with the contents, were placed in the incubator for
thirty minutes, in order to get the required temperature, and then
the platinum black was weighed in the desired amounts and run into
the tubes through a paper funnel. These were then left in the incu-
bator the desired period of time, and afterward placed in ice-water
to stop the action as much as possible. After becoming cool, the
contents of each tube was poured into a medium-sized evaporating
dish. To this also was added the water in which these tubes were
afterwards carefully rinsed. This was then titrated with #4 NaOH
with phenolphthalin as the indicator. Owing to the platinum black
making the mixtures very dark, the phenolphthalin was used in pref-
erence to litmus as the color is brighter and the end point is thus
more accurately determined.
Knowing the amount of ethyl butyrate originally present, the per
cent of hydrolysis was easily calculated.
In each experiment a control was made to determine the amount
of hydrolysis (if any) without the platinum black. One was also
made to determine the acidity of the ethyl butyrate used. The
acidity of the control, due to both these factors, z. ¢., the small amount
of hydrolysis and the acidity of the ethyl butyrate, was deducted from
the acidity of the tube containing the platinum black. This differ-
ence was the amount of acid produced by the catalytic action of the
platinum black.
Owing to the difficulties of measuring such small amounts of ethyl
butyrate, and in order to avoid differences in the amount, a mixture
was made as follows: —
200 c.c. distilled water,
10.4 c.c. ethyl butyrate,
2 c.c. 1% solution of thymol, as an antiseptic.
Five cubic centimetres of the above mixture were placed in test-
tubes, together with 300 mgms. of platinum black. These tubes were
then placed in the incubator, and shaken at regular intervals. This
shaking was necessary, as the particles of platinum black are of com-
paratively large size and soon settle to the bottom of the tubes. If
left in this condition the catalytic action. of the platinum black was
194 Flugh Newtson.
much less than when the tubes were shaken. The shaking kept the
particles in suspension longer, and therefore the typical colloidal solu-
tion of platinum, as described by Bredig, was more nearly approached.
The same amount of shaking was given the control tubes as was
given those containing the platinum. An experiment was made to
determine whether the shaking had any effect on the hydrolysis
without platinum black. There was no increase of the acidity in the
tubes shaken over that of those not shaken. Therefore we are war-
ranted in asserting that the shaking merely keeps the particles in
suspension longer, and thereby increases the surface action of the
particles of the platinum.
I. Effect of time on the hydrolysis of the ethyl butyrate. — The temper-
ature was 40° C., the amount of platinum black 300 mgms., and the
quantity of ethyl butyrate 0.26 c.c. in 5 c.c. distilled water.
The results, with the time element taken as a variable, are shown
in the following table: —
EXPERIMENT 1. EXPERIMENT 2.
Time a Per cent Time x Per cent
c.c. gg NaOH. hydrolysis. fehours:| t|Cor a0 NaOH.
in hours.
hydrolysis.
0.9 2.4 24 4.10
1.55 4.0 48 11.25
1534) 8.9 72 14.80
4.39 11.0 96 19.40
6.55 16.5 27.00
23:3
Other experiments show approximately the same results, but with
slight differences in the amount of hydrolysis, owing perhaps to
differences in the amount of shaking, variations in temperature, etc.
One experiment was carried on one hundred and twenty hours, with
45.7 per cent hydrolysis. Another was carried on eight days, with
60 per cent hydrolysis. The per cent of hydrolysis calculated from
the increased acidity, is to some extent due to the acetic acid formed
from the alcohol produced in the hydrolysis. Therefore, the results
given are not entirely due to the butyric acid produced.
ed ee ee oe
ee EeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEeEOEOEOEeeeeeeeee ee ee
Fydrolysts and Synthesis of Fats by Platinum Black. 195
The liquid from a number of experiments was distilled over the
water bath. This distillate was again distilled. The second distillate
contained alcohol, as was shown by the iodoform test. The liquid
remaining from the second distillate had unmistakably the odor of
butyric acid. It also showed a small amount of acetic acid, when the
acetic ether test was used.
2. Effect of concentration of platinum black.— In this experiment the
concentration of platinum black is a variable, while the other factors
are constants.
Time, 32 hours; temperature, 40°; quantity of
ethyl butyrate, 0.26 c.c.
Platinum x Per cent
in milligrams. | © 26 byes hydrolysis.
25 0.35 0.8
50 0.63
100 Ny
Other experiments show the same or nearly the same results.
Here, evidently, the catalytic action is a function of the concentration
of platinum. But there is not a direct ratio, as the table clearly
shows. Especially is there a greater hydrolysis than we should
expect, between 250 mgms. and 300 mgms. Other experiments show
that this is not constant at this period; therefore it must be due to
some difference in the conditions.
3. Effect of temperature on catalytic action of platinum.— Jn this
experiment, the temperature is a variable, while the time, platinum,
and ethyl butyrate are constants.
In this experiment five temperatures were used: (1) 0 — 1° C,, by
a freezing mixture; (2) 10°, in a refrigerator; (3) 20°, by the tubes
being placed in running water; (4) 40°, in an incubator; (5) 60’, in
196 Flugh Newtson.
a second incubator. The time was twenty-four hours, the quantity
ethyl butyrate 0.26 c.c., and the amount of platinum black 300 mm.
The results are shown by the following table: —
Temperature. | c.c. 75 NaOH. spanice
0.3 0.75
1.00 25
2.05 5.1
5.00
6.9
The catalytic action of the platinum in this experiment increases
with the temperature. The increase from 40° to 60° is not as great
as might be expected. This increase is fairly constant in other
experiments and therefore cannot be a varying condition of this
experiment. Possibly this fact is due to the high temperature which
vaporizes the ethyl butyrate and thus removes it from the sphere of
action, or possibly it may be that the action of platinum is lessened
at 60°, as that of lipase is at 45°.
4. Effect of concentration of ethyl butyrate. — In this experiment the
concentration of the ethyl butyrate is a variable, while the other
factors are constants and the same as those of the other experiments.
Starting with 0.5 c.c. of ethyl butyrate to 5 c.c. of water, the amount
of ethyl butyrate is gradually decreased 0.0025 c.c. The results are
shown in the following table: —
Concentration of ethyl 1 Per cent
butyrate. SE eNO. hydrolysis.
5 c.c. HyO+
0.4 c.c. ethyl butyrate
OZ Cichamce
ONNCicy a
OlO5icicrmess
OOZ5ie-654
Hydrolysis and Synthesis of Fats by Platinum Black. 197
With 0.6 c.c. ethyl butyrate, the per cent of hydrolysis was about
two-thirds, as much as with 0.2 c.c. With 0.8 c.c, it was one-half as
much. This is probably due to the fact that the platinum is collected
in clumps by the drops of ethyl butyrate which have not gone into
solution, thereby decreasing the action. This experiment shows,
however, that the action of the platinum is independent of the con-
centration of ethyl butyrate.
5. Action of poisons on the catalytic action of platinum black. — Two
series of experiments were made.
a. In each test-tube 5 c.c. of 1-1000 solution of each poison were
placed. To this were added 300 mgms. of platinum black and
0.26 c.c. of ethyl butyrate.
6. 4 c.c. of a solution containing 200 c.c. distilled water and
10.4 c.c. of ethyl butyrate were placed in each test-tube. To this
was added 1 c.c. of a 1-1000 solution of each poison and 300 mgms.
of platinum black.
With the first series, the following results were obtained : —
A SERIES. B SERIES.
Time, 48 hours. Temperature, 40°. Time, 24 hours. Temperature, 42°.
n_ | Per cent| meh Per cent
Substance. Bie hydro- Substance. "29 | hydro-
Slee lysis: Hoes lysis.
2 ae : Shy Tiler eWater 0 -. coer, 25 13.2
Sodium fluoride . : 18.0 | Chloroform . . 11.2
Formaldehyde ., . 18.0 || Sodium fluoride . 95
Chloroform . . ; 14.5 UGMNENe Ne 5) ¢ 9.0
MOlWeEnG =. 11.0 Ehymolir. goues a 80
Mercuric chloride : 11.0 Salicylic acid . . as
Silver nitrate . . hil Formaldehyde . 6.6
Salicylic acid . . 5 5.8 Silver nitrate . . 5.6
LELNESNO eG oh wade : 4.3 Mercuric chloride | 50
Hydrocyanic acid ! 1E2, Pbenole ene 5.0
Potassium cyanide . 0.0 Hydrocyanic acid 1.6
Potassium cyanide 0.0
198 flugh Newtson.
In each of these series the results given are the averages of several
experiments. In Series A the lessened action of platinum black, in
the presence of chloroform and toluene, may in part be due to the
fact that the platinum black clumped together in little lumps which
would lessen its surface action. Each of these substances was used
in pure form. .
In ‘Series Bia Pinca solution of chloroform at 22° was used.
1 c.c. of pure toluene was added to the tube containing the 4 c.c. of
water. The lessened action of the platinum here may be due to the
fact that the toluene collecting at the surface gathered the platinum
with it. The action of the poisons was. not always the same as in
the order given in the above tables. But the order did not vary toa
large extent.
On comparing the actions of these poisons on the cardlyiele action
of platinum black, with their effect on lipase as given by Kastle and
Loewenhart, the following differences are noticed:
Sodium fluoride is one of the most destructive agents to the action
of lipase in their list. On platinum black its action is not so marked,
—in fact, being little greater than the action of chloroform.
Again, hydrocyanic acid, which is very destructive to the action of
platinum, is not so destructive to the action of lipase, — in fact, being
only about one third as destructive as sodium fluoride. The action
of other substances on platinum black approximates quite closely
their action on lipase.
O: Reversibility of the action of platinum black, as shown by the syn-
thesis of ethyl butyrate from alcohol and butyric acid. — The experiment
was made as follows : —
A mixture of 100 c.c. of j5 butyric acid, 40 c.c. of 20 per cent
alcohol, and a little thymol, was placed in each of two 250 c.c. flasks.
To one were added two grams of platinum black. This was thor-
oughly shaken, allowed to stand for a short time, and then the liquid
decanted off, leaving all the coarser particles of platinum black in the
flask. This mixture was placed in a flask the same size as the con-
trol flask and tightly sealed. A control flask was also prepared and
tightly sealed. Both flasks were then placed in an incubator regis-
tering 45°, and left for eight hours. Both were shaken at intervals,
and at the end of the eight hours, a distinct odor of ethyl butyrate
was noticed in the flask containing the platinum, while none was
noticed in the control. At the end of twenty-four hours, the odor
of ethyl butyrate in the flask containing the platinum had markedly
Flydrolysis and Synthesis of Fats by Platinum Black. 199
increased, while there was still no perceptible odor in the control
flask. At the end of forty-eight hours, the contents of each flask
was distilled over a water bath, and 10 c.c. of distillate collected from
each. This had a strong odor of ethyl butyrate in the distillate from
the flask containing the platinum, but there was no odor from the
distillate of the control flask. Each 10 c.c. was redistilled, and 4 c.c.
of distillate collected from each. These distillates were saponified
with sodium hydroxide, evaporated to dryness, and then sulphuric
acid was added. The odor of butyric acid was at once noticed in the
saponified distillate from the flask containing the platinum, while
none could be detected in the saponified distillate from the control
flask. When the second distillation was made, the distillate was
poured into 200 c.c. of water, and this then distilled; as with this
amount of water the ethyl butyrate comes over first, leaving the
butyric acid behind.
This experiment was repeated many times with varying amounts
and concentrations of butyric acid and alcohol, and always with posi-
tive results. No quantitative experiment was made to determine the
amount of synthesis, owing to the large amounts necessary for such
work, and the difficulty and expense of obtaining platinum black.
The synthesis is evidently not so pronounced as the hydrolysis.
This is also the case in Kastle and Loewenhart’s work.
CONCLUSIONS.
On comparing the catalytic action of platinum black with that of
lipase on ethyl butyrate, the following facts were observed : —
1. Platinum black accelerates the hydrolysis of ethyl butyrate as
lipase also does. But the action of the platinum is slower.
2. The action of the platinum increases with the increased concen-
tration of the platinum. This is also true of lipase.
3. The action increases with the temperature, reaching its maxi-
mum at 50, which is somewhat higher than the lipase.
4. The action of platinum is independent of the concentration of
the ethyl butyrate, which seems also to be true of the action of
lipase.
5- Poisons with the exception of sodium fluoride and hydrocyanic
acid, affect the catalytic action of platinum in a manner quite com-
parable to their action on lipase.
200 Hugh Netlson.
6. Platinum black synthesizes butyric acid and ethyl alcohol into
ethyl butyrate, as shown by increasing odor of ethyl butyrate and
saponification giving odor of butyric acid. This synthesis is also
brought about by lipase.
My thanks are due Professor Loeb for his valuable suggestions in
these experiments. |
THE STATIC FUNCTION IN GONIONEMUS.!
By LOUIS MURBACH.
INTRODUCTORY.
HE medusa Gonionemus, abundant during the whole summer
at Woods Holl,? displays more definite movements than other
medusz. Certain rotation experiments made several seasons ago
indicated that the animal has a sense of equilibrium that can be con-
fused. Accepting the view commonly held, I believed this to be the
function of the otocyst organs. The static function in lower animals
is naturally associated with the otocyst organs, which, as their name
indicates, were formerly held to be auditory in function, a view now
generally given up.
The otocyst organs make their first appearance in the free-swim-
ming Coelenterates, and are not generally present in sessile forms of
animals. On account of their structure and position in medusz, they
have been called ‘marginal vesicles,” and frequently “ marginal
bodies.”’ A large number of medusz and other free-moving inverte-
brates (most worms, many Crustacez, and all insects) do not have
these organs. As to their homology, it will suffice to say that their
origin differs in the groups of animals in which they are found. Even
in the craspedote medusz they originate in two ways, characterizing
the two orders. Although differing widely in structure in most
1 According to a recent note from Professor AGAssiz the genus name of
Gonionemus was derived from the words ydvv (knee) and véeyas (grove), making ,
the change of name recently used by some writers inapt. MURBACH: Science,
1903, Xviii, p. 373.
2 It gives me pleasure to acknowledge my indebtedness for working facilities
to the Director of the Marine Biological Laboratory.
8 In Ceelenterate literature, otocyst, statocyst, and lithocyst have been used
synonymously. In this paper otocyst organ is preferred, as it is more inclusive.
Many of the older authors used the name marginal bodies (Randk6rper) synony-
mous with otocysts. In rarer cases the subumbral or marginal papilla (Rand-
warzen) are also included. Otocyst organs also occur in worms, crystacee,
molluscs, and tunicates among the invertebrates.
201
202 Louis Murbach.
invertebrates, the otocyst organs agree in essentials: (1) A vesicle
or sac, the otocyst — containing sensory projections; (2) foreign or
secreted inorganic particles, —the otoliths. The more primitive
organs are those in which the otolith is formed within the sensory
basal portion of the otocyst; the next, those in which the otoliths are
attached to sensory projections; and the highest, those in which deli-
cate sensory projections alone are present.
HISTORICAL.
The true auditory function of the otocyst organs in Crustaceze was
doubted by Farre (according to Prentiss!) as early as 1843, but it
was not until 1887 that Delage? was convinced from some of his
experiments that they serve a static in addition to an auditory func-
tion; yet Bethe, in 1894, states that the experiments in the last half
of our century have indicated that the otocysts are organs of equilib-
rium. The strongest evidence that they are not auditory but static,
in Crustacez, has been given by Beer? and Prentiss.
In a review Lyon‘ holds that work done up to the time, except,
perhaps Kreidl’s, does not show that otocyst organs are necessarily
static in function. Ina later paper® he reiterates this opinion, sup-
ported by experiments.
Prentiss, especially in his excellent and exhaustive paper, combines
structural and functional study, successfully repeating Kreidl’s experi-
ments with iron otolith and a magnet. He shows that up to the
time of his work, three theories as to the function of the otocysts (in
Crustacez) were held: ‘1. That they are purely auditory organs.
2. That they are both auditory and static in function. 3. That they
are purely static in function, z.¢. organs of orientation.” -His experi-
_ments seem to leave little room for doubting the static function of
the otocysts in Decapods. Thus the best evidence that the otocyst
organs are static in nature has been gained from experimental work
in Crustacez, only a few investigators in Coelenterates having worked
1 PRENTISS: Bulletin of the Museum of Comparative Zoology, Harvard Uni-
versity, Igol. :
2 DELAGE: Archives de zoologie expérimentale et générale, 1887, v, p. I.
8 BEER: Archiv fiir die gesammte Physiologie, 1898, Ixxiii, p. 1; /d¢d., 1899,
Ixxiv, p. 364.
* Lyon: Journal of comparative neurology, 1898, viii, p. 238.
5 Lyon: This journal, 1899, iii, p. 86.
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:
mie EP PECTS OF VARIOUS "SALTS ON THE
PONTCITY "OF SKELETAL MUSCLES,
By Wi. D ZOE THOMET,
N an article! on the effects of potassium and calcium ions in the
striated muscles, I showed that potassium chloride increases the
tone of the gastrocnemius muscle of the frog, and that calcium chlo-
ride, and to a lesser extent sodium chloride, antagonize this action of
potassium chloride. Subsequently I found? that if a potassium salt
whose anion precipitates calcium is employed, the increase in tone is
much greater than when the chloride is used, and that the minimum
concentration of such a potassium salt necessary to cause increase of
tone is much less than that of potassium chloride.
This work I continued during the summer of 1902 at the Uni-
versity of Chicago, and found that several other salts, such as the
salts of ammonium, rubidium, and czsium, behave like potassium
salts, while the salts of strontium and magnesium are similar to
calcium.
Potassium salts. — Besides potassium chloride and those salts oi
potassium that precipitate calcium, the effects of which are discussed
in the above-referred-to articles, I tested the action of potassium
iodide and potassium sulphate. Concerning the power to cause
rhythmical contractions in the skeletal muscle, Loeb? found that
sodium iodide is far more efficient than sodium chloride. This led
me to surmise that a solution of potassium iodide would be more
effective in producing increase in tonicity than a solution of potas-
sium chloride of the same concentration. This I found to be true,
for whether the % KI be diluted with water, sodium chloride or
calcium chloride solution, the increase of tone produced was always
greater than that produced by the corresponding mixture of potas-
sium chloride. While the weakest solution of 4% KCl mixed with
ZOETHOUT: This journal, 1902, vii, p. 199.
ZOETHOUT: This journal, 1902, vii, p. 320.
Loes: Festschrift fiir Professor FICK, 1899, p. 104.
211
212 W. D. Zoethout.
% CaCl, capable of producing a slight increase in tone was found
to be 44 c.c. KCl + 54 c.c. CaCl,, a solution of 4 c.c. 4% KI + 6 c.c.
% CaCl, called forth a considerable increase in tone.
Potassium sulphate was also found to be more active than the
chloride, although this may perhaps be due to the partial precipita-
tion of the calcium in the muscle.
Ammonium salts. — If a muscle is placed in a 4% NH,Cl solution, the
tone of the muscle is increased, but instead of the immediate and
powerful increase produced by % KCl, the muscle findergoes a more
gradual and limited shortening which may be preceded by a latent
period varying from one-half to three minutes. Ammonium chloride
is, therefore, far less powerful in its action than potassium chloride,
as can also be readily seen from the following tables, which express
the minimum concentration of potassium chloride and ammonium
chloride necessary to cause increase in tonicity.
lec. 7 KC1+9c.c. HO 24 c.c. ¢ NH,Cl + 73 c.c. H2O
2cc. 7 KCl+ 8 cc. # NaCl 5 cc. ¥ NH,Cl +5 cc. % Na€l
44 cc. 7 KCl + 54. c.c. 7 CaCl, 9cc. F NH,Cl+ 1 cc. F CaCl,
If we express these facts in terms of the molecular concentration of
the potassium chloride and ammonium chloride, we obtain : —
KG NH,Cl
Tn EDO sn ca Fonteyo See eee et te
Inna Cla eaten oo. Bes ee
Ta CaCly: 4 18 ks ys dos aera ya) aes 3.68
By dividing the molecular concentration of the potassium chloride
by that of the ammonium chloride as given in the above table, we
obtain, for dilutions with water and sodium chloride, the quotient 23,
and for dilution with calcium chloride, the quotient 2; showing that
the power of potassium chloride to cause increase in the tonicity
is about twice as great as that of ammonium chloride.
From these tables it is also evident that sodium chloride and
calcium chloride antagonize this action of ammonium chloride; and
what is true for calcium chloride holds also for strontium chloride.
I determined the minimum concentration of “% NH,Cl when diluted
with 4% SrCl,, and found this to be 9 c.c. NH,Cl + 1 Ceiaeere
Magnesium and lithium chloride also have the power to abolish the
increase of tone caused by ammonium chloride.
Besides the chloride, I also tested the action of the nitrate, sul-
phate, succinate, and bromide. These salts are, as a rule, more effi-
Lifects of Salts on the Tontcity of Skeletal Muscles. 213
cient in producing tone than the chloride, but they are also more
liable to cause rhythmical contractions.
Czsium chloride. — If a motor nerve of a gastrocnemius muscle is
placed in a ¥ CaCl, solution, no contractions of the muscle take place,
nor is the tone increased. In this respect this salt resembles the
chlorides of potassium and ammonium; Mathews! found that these
salts do not stimulate motor nerves. When, however, the muscle
itself is placed in such a solution, the tone of the muscle begins
to increase after a latent period varying from one to two minutes.
This increase in tone is generally associated with small rhythmical
contractions, as was first noted by Loeb.2, However, at the time that
the tone has increased considerably, the rhythmical contractions are
still very small.
The minimum concentration which will induce an increase in
tonicity in from four to five minutes is represented by 3 c.c. 4%
CsCl + 7 c.c. ¥ NaCl or 9 cc. ¥ CsCl + 1 cc. ¥ CaCl, The in-
crease of tone brought about by this salt is also reversed by the
chlorides of sodium, lithium, magnesium, strontium, and calcium.
Rubidium chlorate.— A “% solution of rubidium chlorate has no
effect on the motor nerve. If, however, the muscle is placed in
such a solution, there is an immediate and powerful increase in ton-
icity. The power of rubidium chlorate to increase the tonicity is
of about the same magnitude as that of potassium chloride, as
can be seen from the following table, which gives the minimum
concentration.
1 c.c. | RbC1Oz + 9 cc. *¢ NaCl, or,
44 c.c. | RbC1Og + 54 c.c. F CaCle.
As pointed out by Loeb,’ rubidium salts give rise to rhythmical
contractions, but it should be noticed that the great increase in tone
which takes place immediately after the muscle is immersed in
rubidium chlorate is far more apparent and takes place much sooner
than the rhythmical twitches.
A few experiments were made with rubidium chloride which proved
that this salt has the same action as the chlorate. The tone induced
by rubidium chloride or chlorate is reversed by calcium chloride and
strontium chloride, and to some extent by sodium chloride and lithium
chloride.
1 MATHEWS, A. P.: Science, 1902, xv, p- 492.
2 Loe: Festschrift fiir Professor FICK, 1899, p. 104.
8 LoEB: Jézd.
214 W. D. Zoethout.
Sodium salts. — A ¥% solution of sodium chloride has no effect on
the tone of the gastrocnemius muscle.!_ In some of the experiments
the muscle was allowed to remain from three to five hours in the
solution without the slightest increase of tone being perceptible.
I have already shown that a sodium chloride solution abolishes the
increase in tonicity produced by potassium, czesium, ammonium, and
rubidium salts.
In a % Nal solution, the muscle may or may not increase its tone
to a very slight extent, with a latent period varying from ten to
|
nt ee ee |
‘
et a eee
a b
FicuRE ].— At a@the muscle is placed in % Nal. Time between a and 6 is
seven minutes.
thirty seconds. Sometimes this increase in tone is immediately ac-
companied by rhythmical twitchings, but in most cases the twitches
do not set in until the muscle has been immersed in the sodium
iodide from six to twenty minutes. In such cases the previous slight
increase of tone may disappear with the onset of the twitches, as
Fig. 1 illustrates. In this figure the muscle was placed in the sodium
iodide solution at a, and the tone is slightly increased; the rhythmi-
cal contractions began seven minutes later (at 4) and almost imme-
diately the increase of tone which had existed up till this time was
abolished. In some cases the sodium iodide did not produce any
increase in tone, although rhythmical contractions were present.
The slight increase in tone which may be developed by sodium
iodide is reversible by the chlorides of sodium, lithium, calcium, and
strontium. Even the tone induced by a % solution of sodium iodide
is almost instantly reversed by calcium chloride. If a muscle is first
treated with calcium or strontium chloride, the subsequent application
of sodium iodide never increases the tone nor produces rhythmical
contractions.
1 Cf. ZENNECK: Archiv fiir die gesammte Physiologie, 1899, Ixxvi, p. 21.
Effects of Salts on the Tonzcity of Skeletal Muscles. 215
As I stated in a former paper,! when a muscle is first treated with
a sodium salt whose anion precipitates calcium, the subsequent ap-
plication of potassium chloride causes a much greater increase of
tone than when the muscle is directly treated with potassium chloride.
I now found that this is to a certain extent also true for sodium
iodide. A muscle which had been subjected to the action of % Nal
for ten minutes, and which showed little or no increase of tone, was
at the expiration of that time placed in a bath of }c.c. % KCl+ 93
c.c. % NaCl, and a great increase of tonicity resulted. The strength
of the potassium chloride solution used in this instance is far below
the minimal strength required by the normal muscle. When potas-
sium chloride is used after the application of sodium iodide, the
twitches which may have been caused by the sodium iodide are
abolished by the potassium chloride at the same time that the tone
is increased.”
Sodium bromide acts upon the muscles in much the same manner
as the iodide, but its action is less pronounced. A muscle in ¥
NaBr generally increases its tone a very little almost as soon as it is
immersed. This increase in tone may or may not be accompanied
by fibrillar twitchings. After a period varying from five to fifteen
minutes, rhythmical contractions appear.
The tone produced by sodium bromide is reversed by calcium
chloride or strontium chloride, and to a lesser extent by sodium
a b
FIGURE 2.— At a the muscle is placed in 7 NaBr. At 6 this is replaced by % CaCl.
chloride. In those cases where the rhythmical contractions are
accompanied by tone, it was observed that generally the rhythmical
contractions ceased very speedily on the application of calcium
chloride, while the reversal of tone was a far slower process. This is
well illustrated in Fig. 2. At a the muscle was placed in a % NaBr
solution. After seventeen minutes (at 4) this was replaced by
calcium chloride. It will be noticed that while the twitches cease
instantaneously, the relaxation of tone proceeds but slowly. If a
1 ZoETHOUT: This journal, 1902, vii, p. 320.
2 Cf. Logs: Archiv fiir die gesammte Physiologie, 1902, Ixxxxi, p. 255.
216 W. D. Zoethout.
muscle is placed in sodium bromide for three or four minutes, its
irritability towards potassium chloride is increased.
Of all the sodium salts tested, the sulphate causes the greatest
increase in tone. If a muscle is placed in a 4% Na,SO, solution,
the tone is immediately increased. In some cases this increase
is considerable, in others it is moderate. The tone is generally
accompanied by twitchings, and is reversible by sodium chloride,
magnesium chloride, lithium chloride, and better by calcium chloride
and strontium chloride. .
Sodium nitrate has but little power to increase the tone of skele-
tal muscles. In about half the experiments, sodium nitrate induced
a very slight increase, in the others none at all. If tone is de-
veloped, it is abolished by sodium chloride and calcium chloride.
The irritability of the muscles towards potassium salts is increased
by sodium nitrate.
Sodium asparginate causes no increase in tone. In one experi-
ment the muscle was left in a 7% solution for 54 hours without any
shortening of the muscle being observed.
Neutral sodium butyrate and formate, having osmotic pressure
equivalent to a 0.7 per cent NaCl solution, have little or no effect on
the tonicity of skeletal muscles.
Lithium salts. — The action of % LiCl is very similar to that of
NaCl; it does not increase the tone of skeletal muscles. Even the
increase in tonicity produced by the action of 4 LiCl for five minutes
is exceedingly small. In two experiments with 7 LiCl there was no
shortening of the muscle in ten minutes.
From this it would appear that lithium chloride, like sodium chloride,
ought to counteract the action of potassium salts. This we found to
be so; in fact, the power of lithium chloride to abolish the effects of
potassium chloride is slightly greater than that of sodium chloride.
If one gastrocnemius muscle is treated with 2 c.c. % KCl + 8 c.c.
% NaCl, while the other is placed in 2 c.c. 4% KCl + 8 cc. ¥ LiCl,
the contraction caused by the former solution is greater and has
a shorter latent period than that caused by the latter solution. This
is also proved by placing one muscle in % LiCl for fifteen minutes,
while the other is placed for the same length of time in ¥ NaCl;
when the two solutions are exchanged for 2 c.c. KCl + 8 c.c. NaCl,
the contraction of the muscle previously treated with sodium chloride
is greater than that of the muscle treated with lithium chloride.
Lithium chloride also reverses the increase in tone produced by
Lfects of Salts on the Tonzcity of Skeletal Muscles. 217
sodium sulphate, caesium chloride, rubidium chlorate, ammonium
chloride, and other salts.
The bromide, iodide, and nitrate of lithium also act very similarly
to the corresponding sodium salts; their action, however, is less
powerful, and is preceded by a longer latent period. After a latent
period varying from twenty to fifty minutes, the fibrillar twitches set
in. They gradually increase in intensity, and are sometimes accom-
panied by a very slight increase in tonicity. This is removed by the
use of calcium and strontium chloride. Although these salts have
practically no tendency to increase the tone, yet, like the sodium
iodide and nitrate, they increase the contraction brought about by a
minimum concentration of potassium chloride.
This difference in the behavior of the salts of sodium and lithium
appears nowhere to better advantage than in the sulphates. While
sodium sulphate causes an immediate and distinct increase in the
tone, lithium sulphate has no effect till after a latent period of from
twenty to thirty minutes, when there is a gradual and small increase
in tonicity. In a few cases, it is true, a small increase took place
after a latent period of one or two minutes, but these are exceptions.
The twitches induced by lithium sulphate are also small compared
with those brought about by sodium sulphate. The chlorides of cal-
cium, strontium, magnesium, and sodium reverse the tone caused by
lithium sulphate.
Magnesium chloride. — This salt is very similar to calcium chloride
in its action on muscle tissue. A muscle placed in 4% MgCl, under-
goes no change in tone for a long time. After a period varying from
one-half to one hour, the tone is gradually increased, but in such a
solution the muscle soon loses its irritability.
Like calcium chloride, the magnesium chloride counteracts the
effect of potassium chloride, and its power in this respect is about
the same as that of calcium chloride, for the minimum concentration
of potassium chloride, when diluted with magnesium chloride, is the
same as when mixed with calcium chloride. Magnesium chloride
also abolishes the tone induced by salts of ammonium, czsium, and
rubidium, and it also stops the twitches produced by sodium sulphate,
sodium fluoride, lithium sulphate, etc.
Barium chloride. — Barium chloride in some respects resembles
calcium and strontium chloride, but in others it differs very greatly
from these salts. Like calcium and strontium chloride, the barium
chloride antagonizes the power of potassium chloride to increase the
218 W. D. Zoethout.
tonicity. A mixture of 7 c.c. 4 BaCl, + 3 c.c. @ KCl in two cases
produced no increase of tone, and in a third case a slight in-
crease after a latent period of twenty-six minutes. From this we
would expect that barium chloride itself had little or no power to
cause increase in tone. A slight tone may be developed in a %
BaCl, solution in ten or fifteen minutes, but, compared with potas-
sium chloride, or even ammonium chloride, this action of barium
chloride is very limited. In one experiment a % BaCl, solution pro-
duced no shortening of the muscle in ten minutes, and in the same
length of time a 4% solution caused but a small increase in tonicity.
It is needless to say that in such solutions the muscle loses its irrita-
bility very speedily.
As was first shown by Ringer,! and afterwards by Loeb, barium
chloride causes great rhythmical contractions. In some cases the
muscle in % BaCl, may exhibit powerful rhythmical contractions
without the tone of the muscle being increased, the lever descending
to the base line after each contraction. In the other cases the
rhythmical contractions may be accompanied by some increase in
tone. Not only is the rhythmical contraction continued, as Loeb
found, for a greater length of time in a 2% than in a more concen-
trated solution, but the twitches also appear sooner in the dilute
solution. Ringer also found that potassium chloride abolishes the
rhythmical contractions induced by barium chloride. This I fully
corroborated. A gastrocnemius muscle was placed in a solution com-
posed of 7 c.c. ¥ BaCl, + 3 c.c. # KCl, and for forty-one minutes no
rhythmical contractions appeared, although after thirty-one minutes
the tone was slightly increased. The control muscle was placed in
a bath composed of 7 c.c. ¥ BaCl, + 3 c.c. 4% NaCl, and in twelve
minutes this muscle was thrown into powerful rhythmical twitches
which were maintained to the end of the experiment.
Strontium chloride. — A % solution of strontium chloride does not
increase the tone of the muscle, except after the muscle has been in
the solution for one or two hours. As the muscle has lost all or
nearly all of its irritability when it has remained in the strontium
chloride for that length of time, the shortening of the muscle is no
doubt a phenomenon of rigor mortis. Even a ¥% SrCl, solution pro-
duced a comparatively small increase of tone in five minutes. In a
few cases this tone was reversed by the application of sodium chloride,
1 RINGER: Journal of physiology, 1886, vii, p. 291.
2 LoeB: Decennial Publications of the University of Chicago, 1902, x, p. 4.
Liffects of Salts on the Tonicity of Skeletal Muscles. 219
provided the strontium chloride had not acted for too long a
time.
Strontium chloride acts similarly to calcium chloride, in that it
prevents or removes the tone caused by the salts of potassium,
caesium, ammonium, and rubidium. The concentration of the stron-
tium chloride needed to overcome the action of these salts is about
the same as that of calcium chloride.
Calcium salts. — As already described under the effects of potas-
sium, ammonium, cesium, and other salts, calcium chloride has the
power to prevent or abolish the increase of tone induced by these
salts. A % solution of calcium chloride has very little or no ten-
dency to cause increase of tone. In fact, a muscle placed in such
a solution, as a rule, shows no change until after one or two hours,
and as the calcium chloride rapidly destroys the irritability of the
muscle, it is more than likely that such changes are due to mortifer-
ous processes. A ¥% solution causes a gradual shortening of the
muscle, which begins almost as soon as the muscle is placed in the
solution.
Calcium sulphate has a far greater tendency to increase tone. In
comparing #4 CaSO, with #4 NaCl, the increase in tone produced by
the calcium sulphate in fourteen minutes was very great, while that
produced by the sodium chloride for the same length of time was nil.
GENERAL CONSIDERATIONS.
From the above experiments, it is evident that the various salts
may be classified with reference to their ability to increase the
tonicity of the skeletal muscle, as follows:
KCl
RbCI NaCl CaCl,
Class I CsCl Class II 4 LiCl Class III | oes
om BaCl, MgCl,
In this table we have only included the chlorides. In Class I are
placed the salts which in % concentration produce an almost imme-
diate increase in the tone of the muscle. The power of potassium
chloride and rubidium chloride to increase tone is about twice
that of ammonium and czsium chloride.
In “ concentration, the salts in Class II do not increase the tone.
In fact, as we have seen, they counteract the effects of salts in Class
220 W. D. Zoethout.
I. The power of LiCl to antagonize the action of salts in Class I
is slightly greater than that of sodium chloride. The salts in Class
III have the same effect as those of Class II, but their antagonistic
action to the salts of Class I is far greater than that of sodium
chloride and lithium chloride. Another reason why I have not
grouped the chlorides of calcium, strontium, and magnesium with
Class II is because of the fact found by Loeb,! that while salts
of sodium and lithium produce rhythmical contractions, the salts
mentioned in Class III stop such’ activity. As to rhythmical con-
tractions, it may be repeated that Loeb found that rubidium chloride
and czsium chloride, grouped by us in Class I, also have this power.
The action of barium is somewhat peculiar. As already stated, its
power to counteract the effects of salts of Class I is about the same
as that of calcium, strontium, and magnesium; but, unlike these
salts, it does not abolish the rhythmical contractions induced by the
sodium and lithium salts; in fact, it augments these contractions.
We have, therefore: (1) salts which increase the tone, but do
not cause rhythmical contractions (potassium chloride); (2) Salts
which cause beth tone and rhythmical contractions (czesium chloride,
rubidium chloride); (3) Salts which do not cause tone, but which
do cause rhythmical contractions (sodium, lithium, and barium
chloride); and (4) Salts which neither increase tone nor cause
rhythmical contractions (the chlorides of calcium, strontium, and
magnesium ).
These facts would almost seem to indicate two different contractile
substances? in the skeletal muscle, as held by Bottazzi® for the
cardiac muscle. The stimulation of one of these, it would seem,
produces a tonic contracture, that of the other produces rhythmical
twitches. The first-named contractile substance, it may be assumed,
is readily affected by salts of Class I in such a manner that a tonic
shortening of the muscle takes place, and it is affected by salts of
Class II, and especially by those of Class III, in the opposite manner.
The other contractile substance is stimulated by salts of Class II,
and also by the chlorides of rubidium and cesium, and this action is
prevented or abolished by salts of Class III.
The supposition of the existence of two such contractile sub-
stances is still further strengthened by facts very similar to those
LorEB: Festschrift fiir Professor FICK, 1899, p. 104.
LoeEs: Archiv fiir die gesammte Physiologie, 1902, Ixxxxi, p. 258.
1
8 BorTazzi: Journal of physiology, 1897, xxi, p. I.
Effects of Salts on the Tonictty of Skeletal Muscles, 221
that led Bottazzi to hold this view in regard to the cardiac muscle.
I refer to those phenomena in which the muscles may at one time
exhibit tonicity without rhythmical activity, to be followed by a
period of ever-increasing rhythmical action and simultaneous de-
crease in tonicity (see Fig. 1).
Whether the effect of these different chlorides which possess such
varied and even antagonistic actions must be assigned to the anions
or the cations, I am hardly prepared to state. In regard to this
question, however, I may once more refer to the peculiar action
of barium chloride in conjunction with potassium chloride. As
already stated, while this salt, like the chloride of calcium and
strontium, inhibits or abolishes the increase in tonicity produced by
potassium chloride, yet its power to induce rhythmical contractions
is even greater than that of other members of Class II. And the
rhythmical contractions induced by barium chloride are, in turn,
inhibited by potassium chloride. As both these salts are chlorides,
as the one causes tonicity, while the other causes rhythmical contrac-
tions, and as both salts antagonize each other, it seems to me most
likely that we must ascribe these stimulating and inhibiting effects
to the cations, barium and potassium.!
SUMMARY.
1. The salts of potassium, czesium, ammonium, and rubidium in-
crease the tonicity of the skeletal muscles. The iodide, bromide,
and sulphate have a greater effect than the chloride.
2. The chlorides of sodium and lithium, and especially those of
calcium, strontium, and magnesium abolish this increase in tonicity.
3. Certain salts of sodium, such as the iodide, bromide, and sul-
phate, may increase the tone; but such action is generally exceed-
ingly slight compared with their power to cause rhythmical activity.
The action of the lithium salts is still less than that of the sodium
salts.
4. Barium chloride antagonizes the action of potassium chloride in
preventing tonicity, and potassium chloride antagonizes the action of
barium chloride in preventing rhythmical action.
1 Cf. Logs: Archiv fiir die gesammte Physiologie, 1902, 1xxxxi, p. 248, and
the Decennial Publications of the University of Chicago, 1902, x, p. Io.
SOME EFFECTS OF THE RONTGEN RAYS ON
THE DEVELOPMENT OF EMBRYOS.
By P. K. GILMAN anp F. H. BAETJER.
[From the Anatomical Laboratory of the Johns Hopkins University, Baltimore, Md.]
HE great interest aroused by the discovery of the marked alter-
ations produced in the human skin by exposure to R6éntgen
rays has given rise to a number of experiments on animals. This
work, however, has been confined in the main to a study of the
lesions produced in the skin of adult mammals! and has not been
much extended to the loweranimals and to embryos. F. Schaudinn,?
found that various protozoa were differently affected by exposure to
the Roéntgen rays. Those having a protoplasm very rich in water
seemed to be the ones most greatly affected. Recently Bohn® has
shown that radium causes deformities and arrest of development in
frog embryos.
In the following paper a brief account is given of the gross effects
noted in certain embryos exposed to the X-rays. A study of the
microscopical alterations produced in the tissues is at present under
way.
Eggs of Amblystoma were subjected to the influence of Rontgen
rays for fifteen minutes each day for varying periods. The eggs, just
covered with water, were exposed in a shallow glass dish, and were
disturbed as little as possible in transferring them to and from their
tank. Both exposed and control eggs were subjected to the same
conditions of temperature, daylight, and water-supply.
The first effect of the rays noted, on comparing exposed with
normal eggs, was an accelerated development for a short period.
This continued up to the tenth day in the case of some of the
1 A good summary of the literature on this subject may be found in the recent
work of PusEy and CALDWELL: “The Practical Application of the Réntgen Rays
in Therapeutics and Diagnosis,” Philadelphia, 1903.
2 SCHAUDINN, F.: Archiv fiir die gesammte Physiologie, 1899, Ixxvii, p. 29.
3 Boun: Comptes rendus de l’academie des sciences, 1903, CxXxxvi.
222
Liffects of Rontgen Rays on Development of Embryos. 223
embryos, though after the third or fourth day the development was
seen to be along abnormal lines. About the tenth or eleventh day
the normal embryos caught up in size with the exposed ones, and
from that time on continued to enlarge, while the abnormal ones
assumed grotesque shapes, and either grew no larger or became
smaller. The experiments were continued up to the twenty-second
day.
Animals exposed daily to the rays for four or five times, and then
allowed to develop undisturbed, showed a marked tendency to recover
and develop normally; but in less than half of these was complete
restitution of form effected. Animals exposed daily for twenty-two
or three days died soon after the last exposure.
Up to approximately the twelfth day the exposed animals were
noticeably larger than the control. After this period practically no
increase in size was noted. Although the shapes assumed by the
exposed embryos were not exactly the same, the following deformities
were noted in practically all of a large series, ranging from specimens
of the tenth to those of the twentieth day: (1) no external gills
developed on any of the embryos; (2) the body surface appeared
roughened and wrinkled in places, especially about the head portions,
which were slender, elongated, and more pointed than the normal
heads, and showed poorly developed eyes and distorted mouth parts;
(3) the bodies of the exposed animals all showed hemispherical
bulgings at the base of the neck on the ventral surface, over which
the body-wall appeared tense and of a lighter color; (4) the mem-
braneous portion of the tail was but slightly developed. .
Hens’ eggs were employed in another series of experiments. The
same care in regard to control material was observed. The eggs
were exposed each day for ten minutes, and the following points
observed.
The exposed eggs showed accelerated development during the
first thirty-six hours, after which the eggs of this series were retarded
by the rays. Specimens were preserved each day for histological
study.
The chicks showed the following features on exposure for four
days, the abnormalities becoming more marked on longer exposure
to the rays: (1) there were deformities of the occipital region, ac-
companied by hemorrhagic areas, which often extended along the
dorsal line; (2) the development of the eyes was retarded in many
of the specimens; (3) the membranes were generally quite firmly
224 P.K. Gilman and F. H. Baetzer.
adherent to the embryo, so that it was difficult or impossible to remove
them; (4) the limbs assumed grotesque positions, sticking out at
unnatural angles to the body. In older specimens, where feathers
had begun to appear, these were abnormally distributed in patches.
The bodies of the exposed chicks were distorted, and many of them
found hanging in abnormal positions within the shells.
In the experiments above described, the embryos were exposed to
very powerful rays. The apparatus used was a twenty centimetre
coil with an interrupter of the electrolytic type and a ten-inch coil
with a mechanical interrupter. Two styles of tubes were used, — a
Queen, self-regulating with heavy anode, and a Heinze with a medium
heavy anode. The vacuum of the tubes was so arranged that a light
of medium soft quality was obtained; z.¢., when the hand was held
before a fluroscope the flesh was very pale, and the bones were a
dark gray, standing out in a sharp contrast to the flesh.
With such a light a hen’s egg held eighteen inches from the tube
casts a gray shadow upon the fluroscope. By means of the self-regu-
lating tubes, the desired degree of penetration was readily maintained.
Sufficient current was sent through to keep the anode at a bright
red heat. The eggs were placed six inches from the anode, and were
arranged each day so that each egg received the same amount of
light.
Dr. Bardeen, at whose suggestion the work was undertaken, informs
us that in some preliminary exposures of hens’ eggs to the rays, he
found only those eggs affected which were exposed to rays of con-
siderable intensity. In a series of experiments carried on at the
Zoological Station at Naples, he was unable to obtain positive effects
on the eggs of certain sea-urchins and teleosts, when they were ex-
posed to rays of moderate intensity. It is, therefore, a matter of con-
siderable importance to determine the intensity of the rays, and the
length and frequency of exposure necessary to bring about marked
alterations in animal tissues.
1 Our thanks are due to Dr. Henry M. Hurd for the use of the X-Ray appa-
ratus belonging to the Johns Hopkins Hospital.
eS
fee EPFECTS OF IONS ON THE DECOMPOSITION
OF HYDROGEN PEROXIDE BY PLATINUM BLACK.
By C. HUGH NEILSON anp ORVILLE H. BROWN.
[From the Hull Physiological Laboratories of the University of Chicago.|
Rk. A. P. MATHEWS! has shown that certain salts have a stimu-
lating effect on nerves, while others in the same concentration
have a depressing effect. He ascribes the stimulating action to the
negative ion or anion, and the depressing action to the positive
ion or cation. When the anion of the salt solution used is more
powerful than the cation, stimulation takes place; when the cation
is more powerful, depression results. Bredig? has found that the
splitting of hydrogen dioxide into water and oxygen by his platinum
solution is accelerated by the hydroxyl ion, and retarded by hydro-
cyanic acid, cyanogen iodide, mercuric chloride, certain acids, anti-
septics, and some salts. Dr. A. C. Crofton,? working in this
laboratory, has shown that in the catalysis of hydrogen dioxide the
action of a nucleo-proteid from the liver was greatly accelerated
by the addition of certain substances, as salicylates and dilute
alkalies. Other substances, as nitrates and cyanides, greatly di-
minished the action. Kastle and Loevenhart* very recently pub-
lished papers on the decomposition of hydrogen peroxide by platinum
black and other substances. They also show that certain substances
inhibit, while others accelerate the catalysis. The above facts led us
to try a series of experiments on the action of salt solutions on the
rate of splitting hydrogen dioxide into water and oxygen by platinum
black, with the view of determining the effect of the anion and
cation respectively. We used the sodium salts of many inorganic
and organic acids to test the effect of the anion, and the chlorides
of the alkalies, alkaline earths, and some heavy metals to test the
effect of the cation on the rate of the decomposition of hydrogen
1 MATHEWS: Science, 1903, xvii, pp. 729-733.
? BREDIG: Anorganische Fermente, Leipzig, 1got.
3 CROFTAN: Medical record, 1903, lxiv, pp. 6-11.
4 KASTLE and LOEVENHART: American chemical joyrnal, 1903, xxix, pp. 397-
437 and 563-588.
225
226 C. Hugh Netlson and Orville H. Brown. -
dioxide by platinum black. The platinum black has the advantage
that it acts purely as a catalyzer, and thus all possibility of having
the reaction hidden by union of the salts with the proteid consti-
tuents in the enzyme is eliminated. The analogy between the
action of the enzymes and platinum black in splitting hydrogen
peroxide has been shown by many investigators, especially by Bredig;
hence the value of any results obtained in the catalysis of hydrogen
peroxide by platinum black in the presence of certain salts.
Methods. — To measure the amount of oxygen given off during any
interval of time, wide-mouthed bottles of 200 c.c. capacity were used.
TABLE I. —
= solution. ez solution. siz Solution. Control.
Salt used.
Cubic centimetres of oxygen set free in
i
wn A A © Ww FE
2 min. 1 min. 2 min. 1 min.
15 10 24 15
15 iy 32
20 10 25
17 11 25
21 15 30
20 28
18 24
21 27
15 20 |
Potassium chloride
—
Sal
Ammonium
Lithium
Calcium
Strontium
Barium
Magnesium
Cobalt
Aluminium
nm on 0 0 A © nM ME
The rubber stopper was fitted with a delivery tube leading to the
receiving vessel, which was an eudiometer tube of 50 c.c. capacity,
graduated in 51, c.c. In every experiment four bottles were used,
three containing 25 c.c. of the solution to be tested, and a fourth con-
taining 25 c.c. distilled water; the fourth served for control. In
each bottle was placed 5 c.c. hydrogen dioxide, which was kept in
ice-water. In order to get the same amount of platinum black
in each bottle, the platinum black was put into distilled water, which
was constantly stirred so that the platinum was suspended evenly.
While the stirring was continued, 5 c.c. of this mixture was placed
in each bottle. The corks were rapidly placed in the bottles, and
the amount of oxygen given off was read at intervals of one and two
Effects of Ions on Decomposition of Hydrogen Peroxide. 227
minutes. Three tubes were used, so that any variations in the
amount of substance taken could be seen in the different quantities
TABLE II.
% solution. | 7% solution. | sf, solution. Control.
Salt used.
Cubic centimetres of oxygen set free in
2 min. in. | 2min. | 1min. | 2min. | 1 min. | 2 min.
Sodium chloride . . . . 7) 6 Sas Yee rag!) Sas else
IDLOMIGE. seo es 3
nitrate .
hyposulphite
butyrate
chlorate
succinate .
acetate .
sulphite
lactate .
benzoate
acid carbonate .
tungstate .
fluoride
sulphate
tartrate
salicylate .
acid phosphate .
citrate .
valerianate
oxalate .
formate
of gas given off. The error in the reading due to the short interval
of time in fitting the stoppers was very small.
Effect of the cation. — The chlorides were selected to get the effect
of the positive ions. Three different concentrations of each solution
228 C. Hugh Nelson and Orville H. Brown.
were used, %, 74, and ;%5, respectively. A typical experiment is
recorded in Table I. In the column for controls, the general aver-
age is recorded as the controls were uniform.
From Table I, it is seen that the salt of the alkali, alkaline earths,
and the heavy metals used, uniformly cause a Gepression in the rate
of catalysis. As the strength of the solution diminishes, the rate of
catalysis is increased. Nearly normal action is allowed in the gz
solution.
Effect of the anion. — The sodium salts of many of the inorganic
and organic acids were selected to test the effect of the anion.
Three concentrations of solutions, %, ¢%, and sz, were employed.
All of the solutions used were practically neutral to litmus. The
results are shown in Table II.
It will be noticed that the salts of the inorganic monobasic acids,
with the exception of the fluoride, have a depressing effect on the
platinum black catalysis. The hyposulphite also has a slight hinder-
ing action. Sodium bromide is seen to depress the catalysis more
than any of the other sodium salts. It is at least an interesting coin-
cidence that bromides are recognized by pharmacologists as general
and typical depressants. The acid carbonate depresses slightly ;
the sodium sulphate has an accelerating effect in strong concentra-
tion; the acid phosphate and the salts of the organic acids accelerate
the catalysis. The formate, valerianate, oxalate, and citrate are the
most active. Sodium citrate is one of the most powerful acceler-
ators, and this may possibly be related to its power of stimulating
diuresis. There was no constant variation in the rate of catalysis
due to the different concentrations of solutions used. But asa rule
the 74 solutions were the most active. Papers will soon be pub-
lished by us containing the results of similar investigations on the
effect of ions on the action of a watery extract of the pancreas in
splitting hydrogen peroxide and ethyl butyrate.
We are deeply indebted to the criticism of Dr. Stewart and other
members of the staff of the Hull Physiological Laboratory.
CONCLUSIONS.
In the decomposition of hydrogen peroxide by platinum black, the
cation, in general, has an inhibiting or depressing effect, and the
anion has an accelerating effect.
_— pe met
CONCERNING THE FORMATION OF SUGAR
FROM LEUCIN.
By’ J. /T-SHALSEY:
[From the Pharmacological Laboratory in McGill University.]
NE of the questions which still remain unsolved in the physi-
ology of carbohydrate metabolism concerns the production of
sugar from proteid. From the work of various authors, especially of
Lusk,! we know that in certain conditions this production may amount
to as much as sixty per cent of the proteid involved. In explanation
of this phenomenon, two hypotheses have been formulated. One
hypothesis presupposes a large nitrogen-free portion of the proteid
molecule which is split off from the rest, and changed more or less
directly into dextrose; the other would explain the formation of sugar
from proteid by synthesis.”
The view that sugar is derived from proteid through synthetic
processes has recently received new support in the results of experi-
ments by Stiles and Lusk,? and by Knopf.4 The former have found
that dogs with phlorhizin glycosuria are able to convert a large
portion (about 40 per cent) of the end-products of proteid hydrolysis
into dextrose. The experiments of the latter showed that under like
conditions dogs can manufacture dextrose from asparagin.
Leucin, which forms so large a portion of the products resulting
from proteid hydrolysis, is naturally a constituent to which attention
would first be directed in this connection. Kossel® and Miiller® were
the first to point to this substance as a possible mother-substance for
1 Lusk: This journal, 1898, i, p. 395.
2 For discussion of these hypotheses, see CREMER and LANGSTEIN: Ergeb-
nisse der Physiologie, erster Jahrgang, Theil I, 1903, pp. 99 ff. and pp. 872 ff.
8 STILEs and Lusk: This journal, 1903, ix, p. 380.
4 Knorr: Archiv fiir experimentelle Pathologie und Pharmakologie, 1903,
mix, 123.
5 KossEL: Deutsche medicinische Wochenschrift, 1898, p. 58.
§ MULLER and SEEMAN: /dzd., 1899, p. 209.
22
230 J. T. Halsey.
sugar. Cohn! believed that his experiments with fasting rabbits
indicated that leucin was a “ glykogen-bilder.”
Halsey,” working with phlorhizin dogs, was unable to show that this
was the case.
A further investigation of this point seemed desirable, and in the
last two years experiments have been made in the Pharmacological
Laboratory of McGill University, in the endeavor to obtain additional
EXPERIMENT 1.
Mongrel bitch, weight about 15 kg. Has fasted 50 hours. Received 1.5 gms. phlorhizin
every six hours during the experiment.
Besvad: Dextrose in | Nitrogen in
gm. gm.
72.00 15.40 Fasting.
49.09 ‘S
42.72 : ae
57.67 : 100 gms. egg albumin.
53.93 95 gms. “ «“
34.86 ] Fasting.
51.72 5. 104 gms, nutrose.
46.48 100 gms. sty
14.64? Fasting.
14.642
15.002 ah ; 27 gms. leucin.
15.002
16.58 ; Fasting.
17.41 : : =
1 Urine fermented in bladder.
2 Analyzed together, and reduced to 12 hours basis.
light on this subject. The method of experimentation followed was
that elaborated by Lusk, and made familiar through his various papers
1 CoHN: Zeitschrift fiir physiologische Chemie, 1899, xxviii, p. 211.
2 HALsEY: Sitzungsberichte der Gesellschaft zur Beférderung der gesammten
Naturwissenschaften zu Marburg, 1899, p. 102.
Concerning the Formation of Sugar from Leucin. 231
on phlorhizin diabetes. Precautions were observed to avoid errors
through incorrect feeding, loss of urine, and other mishaps which
would tend to obscure results. The urine was drawn off regularly by
catheter; analyses were invariably made in duplicate. The sugar
was determined both by polarization and by reduction. Lehmann’s!
‘method was employed in place of the more usual Allihn method, it
having proved in the author’s experience more rapid and convenient,
and not less exact, than Allihn’s. The leucin was prepared from
digestion mixtures, according to usual methods, and was most care-
fully purified by repeated recrystallization. That used in experiment
I was prepared from the copper salt.
Five experiments were successfully completed; in these leucin
was fed six times. Twice (Exps. 2 and 3) the results pointed to a pos-
sible production of dextrose from the Jeucin. The conclusions were,
EXPERIMENT 2.
Mongrel bitch, weight about 15 kg. Has fasted 36 hours. Phlorhizin, 1.75 gms. every
6 hours.
Period. Dextrose. Nitrogen.
36 hrs. 56.21 8.96
12 hrs. 15.67 Ss)
12 hrs. 15.67 Belo
12 hrs.1 24.08 6.28 i Leucin, 25 gms.
12 hrs. 24.08 6.28
12 hrs. 19%5 oo Fasting.
12 hrs.1 10.51 2.42 3
12 hrs.
10.51 2.72 soe &
1 Analyzed together, and reduced to 12 hour basis.
however, rendered uncertain, in one case by a sudden alteration in
the metabolism (Exp. 1), in another (Exp. 3) by the development of
kidney lesions which made it difficult to draw positive conclusions.
In four experiments, the results indicate that no sugar was manufac-
tured from the leucin fed.
The ingestion of 27 gms. of leucin (Exp. 1) was followed by so
1 LEHMANN: Archiv fiir Hygiene, 1897, xxx, p. 267.
232 J. 1... fabsey:
slight an increase in excretion of dextrose that the results cannot be
held to show any production of sugar from leucin. The nitrogen
excretion, increased more than 3.0 gms. in Periods 11, 12, and 13,
indicates that the leucin was absorbed.
In Exp. 2 the ingestion of the leucin was followed by a sudden in-
crease in both dextrose and nitrogen excretion. If we subtract from
the nitrogen excreted in Periods 4, 5, and 6, 17.49 gms., 2.7 gms., as
leucin nitrogen, we have as remainder derived from body proteid
about 14.8 gms. nitrogen, which, multiplied by 4.00,! the D : N ratio,
gives us 59.2 gms. dextrose as the amount of dextrose corresponding
to 14.8 gms. nitrogen. 67.6 gms. were excreted, giving us an excess of
dextrose of about 8 gms., which could have been produced from the
leucin fed. The extraordinary increase in the metabolic processes
(Periods 4, 5, and 6), however, casts doubt upon such an interpreta-
tion as being the only tenable one.
EXPERIMENT 3.
Fox terrier bitch which has fasted 48 hours. Phlorhizin, 0.75 gm. every eight hours,
subcutaneously.
Day. | Dextrose. | Nitrogen.
grams grams
18.0 4.96 Fasting. |
28.8 6.74 : Ke
LOS : : 300 gms. meat.
25.5 : y f
31.0 : : 300 gms. meat, 11.5 gms.
leucin.
26.5 8.40 : 300 gms. meat.
21.0 7.18
1 Dog distinctly ill; vomits about half his food; urine contains albumin.
During the 5th day about 5.5 gms. more dextrose was excreted than
on the preceding day, but at the same time the nitrogen excretion
rose 1.20 gms. on this and the following day; whereas only 1.2 gms.
leucin nitrogen were fed. If we consider days 5 and 6 together, we
find 16.86 gms. nitrogen and 57.58 gms. dextrose excreted. Subtract-
ing from the 16.86 gms. nitrogen, 1.2 gms. contained in the leucin, we
1 Average of 4.20 and 3.86.
Senate Eaatiiomen “at
Concerning the Formation of Sugar from Leucin. 233
have left 15.66 gms. nitrogen, as nitrogen derived from body proteid.
This 15.66 gms. nitrogen, multiplied by the D: N ratio (3.6), gives
us 56.37 gms. dextrose, as the amount of dextrose which we would
expect to find in the urine derived from the proteid decomposed, as
indicated by the nitrogen excreted. The difference, about 1.2 gms.,
is so small as to be within the limit of analytical error.
The fact that on the 7th day the dog gave unmistakable evidence
of impaired kidney function prevents our judging whether or not this
interpretation be justifiable.
EXPERIMENT 4.
Irish terrier bitch, fasted 60 hours after receiving 2.5 gms. of phlorhizin by mouth.
During the experiment 0.5 gm. phlorhizin in alcoholic solution was administered, subcu-
taneously, each day at 6.30 A.M., and at 2.30 and 9.30 P.M.
Period. Dextrose. Nitrogen. : Diet.
grams grams
24 hrs. 50.35 Tell Fasting.
12 hrs. 19.00 4.845 s
12 hrs. 18.461 oonSe : a
3 hrs. 8.04 2.24 : Leucin, 9.5 gms.
3 hrs. 6.90 2.16 : Fasting.
3 hrs. 5.40 1.548
3 hrs. 7.54 2.504
12 hrs. 4 09%? Vai
1 During the night the dog had a loose movement of the bowels, but 115 c.c.
urine was obtained from the bladder at 6.30 A.M. In this second portion, dextrose
and nitrogen were determined quantitatively, and in the contaminated portion the
dextrose was determined polariscopically.
2 Dextrose and nitrogen determined in urine obtained from bladder, 150 c.c. An
earlier portion contaminated by feces.
During the two fasting periods of the second day, the dog excreted
19.0 gms. and 18.46 gms. of dextrose. In the first twelve hours of
that day, 4.845 gms. of nitrogen were excreted. In the twelve hours
following the feeding of leucin, 27.88 gms. dextrose and 8.452 gms.
nitrogen were excreted. Taking the figures of the preceding day
for comparison, these amounts represent a plus of dextrose of about
9 gms.; of nitrogen, about 3.6 gms. Of this surplus nitrogen not
234 J. TL tlasey:
more than 1.0 gm. can be looked upon as leucin nitrogen, leaving 2.6
gms. nitrogen to be considered as body nitrogen. ‘Taking 3.6 as the
D : N ratio, this nitrogen represents 9.36 gms. of dextrose, more than
enough to cover the surplus dextrose of the twelve hours. This cal-
culation would indicate that in this experiment no dextrose was
formed from the leucin fed.
EXPERIMENT 5.
Spaniel bitch, weight about 8.5 kg.' Fasted 140 hours. Phlorhizin given
subcutaneously in alcoholic solutions.
Phlorhizin. Diet.
0.5 gm. at 7 A.M. | Fasting.
and 6 P.M.
0.652% 1| 3.44 | 0.5 gm. at 9A.M.
and 2.30 and 9.30
P.M.
6-9 A.M., 3 hrs. : 1.50 3: 0.2 gm. at 6 A.M.
9-12 M., 3 hrs. 1.49 3: 0.2 gm. at9 A.M. | Leucin, 5 gms.
at 8.30, 5.7
gms. at 10.30.
12-3 p.M., 3 hrs. 1.47 0.2gm.at12™M. | Fasting.
3-6 P.M., 3 hrs. 1.48 : 0.2 gm. at 3 P.M.
0.2 gm. at 6 P.M.
535 ; 0.7 gm. at 9 P.M.
6 P.M.-6 A.M., 12 hrs.
6-10 A.o., 4 hrs. 3 : 0.4 gm. at 6 A.M.
10-2 A.M., 4 hrs. ; : 0.4 gm. at 10 A.M. | Leucin, 6 gms.
at 10.15. 7 gms.
at 12,12 gms.at
11.30.
2-6 P.M., 4 hrs. 5.02 ! 2.60 | 0.4 gm. at 2 P.M. | Fasting.
6-10 P.m., 4 hrs. 5.52 4 3.45 | 0.4 gm. at 6 P.M. of
10 P.M.-6 A.M., § hrs. |10.16 { 3.36 | 1.0 gm. at 10 P.M. se
1 Urine obtained from bladder used for analysis.
In this experiment the ingestion of 10.7 gms. leucin on the third
day, and of 25 gms. leucin on the fourth day, has produced few or no
noticeable effects on the excretion of sugar. The increase in sugar
in Period 2 of the fourth day is too small to be of significance when
compared with the amount of leucin fed, 25 gms. In this experiment
no diarrhoea occurred during or after the experiment. On the third
day after 10.7 gms. leucin were fed, 35.81 gms. dextrose and 11.29 gms.
nitrogen were excreted; on the fourth day after 25 gms. leucin, 33.32
gms. dextrose and 11.28 gms. nitrogen were excreted. The absence
Concerning the Formation of Sugar from Leucin. 235
of any evidence in our figures for the complete absorption of the
leucin is regrettable. The desirability of continuing the experiment
for another twenty-four hours was not recognized until too late.
Nevertheless the fact that the dextrose excretion did not rise after
the leucin feedings stands out prominently in the table.
Two other experiments in which leucin was fed, failed to show any
evidence of a production of sugar from the leucin, but the interpre-
tation of the results is made difficult by the occurrence in one case of
kidney disease, in the other of contamination of the urine by feces.
Although our experiments do not allow us to express too positive
an opinion, we believe that they indicate that leucin, when fed ina
pure form to phlorhizin dogs, is not changed into sugar.
There still remains the possibility that the leucin complex, as it
exists in proteid, may be concerned in the formation of sugar, or
that when leucin is fed with the other end-products of digestion, as
in the experiments of Lusk and Stiles, it, together with some other
substance or substances, plays a réle in the synthesis of sugar.
I take pleasure, at the close of this article, in thanking Messrs.
W. E. Ainley and J. C. Fyshe, of the class of 1904, and Messrs.
F. J. Tees, F. A. C. Scrimger, and E. T. F. Richards, of the class of
1905, of the McGill Medical College, for their assistance in carrying
out these experiments.
LOCALIZATION OF THE RESPIRATORY CENT gm
TRE SKAGE:
BY IDA TH -HYy DE.
[From the Physiological Laboratory of the University of Kansas.]
CONTENTS.
Page
Introduction’ ~ 000° 5%. 8 eye a ee ats ee te eto ctr sae rrr
Description of the medulla and the respiratory movements of the skate ... . 239
Experimental methods, .. Seen oc Meh ses, <0. oferta pete
Shockeffects 2. 2 6 «6 6% 6 is eo ge oa Ne fs. ote, com tel en rr
Sections anterior and) posterior to the medulla ~~...) a reese
Experiments on special nucler ©. 2.0.2 20, 0
Medisection of the medulla. 22. 6 a. won we we i te
Experiments on the lobes of thesmedulla. . <5 =< 2) sewn enroute To
Effect of transverse sections of the:medulla. . . = = . +) ce) mennennne nnn eannnmammnaEnE
Medisection followed! by hemisection’ 7 =. 5° 5. = a 1m tee ce
Conclusions: 3; 65. See we eee eras wes eta
INTRODUCTION.
ULPIAN' and Steiner? studied the respiratory centre in fishes.
They concluded that it is situated in the posterior limit of the
medulla, and that a lesion in this region produces an immediate arrest
of all respiratory action. When placed in water after such an opera-
tion, the fishes -remain immovable, respiratory movements are not
resumed, and soon all vital functions cease. ,
In my paper on the “Nervous Influences on the Respiratory
Mechanism in Limulus,” ® I have demonstrated that each ganglion is
the relay station for the sensory and motor nerves of the correspond-
ing segment. The ganglia of the cesophageal ring differ from those
of the abdominal cord, especially in that the motor nerves of the former
innervate muscles that cause definite movements of the mouth parts
of the corresponding segment, while those of the cord innervate
movements of the gills of the proximal segment. Any number of
' VupiIAN, A.: Physiologie générale et comparée du systéme nerveux, Paris,
1886.
* STEINER, J.: Die Functionen des Centralnervensystems und ihre Phylo-
genese, 2 Abt., die Fische, Braunschweig, 1888.
® Hype, I. H.: Journal of morphology, 1894, ix, p. 347.
236
>
»
5
a
4
th ha 1 Ce
Localization of the Respiratory Centre tn the Skate. 237
these ganglionic stations could be isolated, not only from each other,
but from the rest of the central nervous system, without losing their
identity in function. In short, the hypothesis of a superior centre
controlling the co-ordination of the parts that combine in the respira-
tory act in animals as low as Limulus was conclusively disproved.
The view that in such animals a part of the brain, corresponding to
the medulla of higher animals, governs the respiratory movements by
superior control, and thus regulates, through subsidiary guards, the
mechanism of respiration, must also be considered erroneous.
Investigations on respiration have been pursued in my laboratory
by several of my students whose work is about to be published. H. Z.
Ewing ! has experimented on the localization of the respiratory centre,
and O. H. Brown and L. W. Roller,? on the effects of gases and pres-
sures upon respiration in the Acridide, especially upon grasshoppers.
They have proved that the relative position of the respiratory centre
in the central nervous system of the Acrididz is practically the same
as in Limulus. They have also found that the respiratory activity of
one or more segments persisted, if the sensory and motor connection
between the ganglia and the muscles controlling the inspiratory and
expiratory movements of the segments was not injured. It was pos-
sible, therefore, to cut one of the Acrididz into several breathing
segments, each having a definite rhythm and force of action. More-
over, the movements of these segments are increased, decreased, or
stopped when exposed to the influences of different gases and pres-
sures, and they also respond to stimuli in the same manner as do the
segments in the intact animal, when it is placed under the same
conditions and subjected to the same stimulations. — .
Thus the evidence secured from a study of these two types,
Limulus and the Acrididz, lead to the conclusion that their respira-
tory movements are segmental processes, each ganglion controlling
the peripheral organs of its own segment.
In the higher vertebrates not only have whole systems of nerves
been lost and new ones developed, but the simple primitive relations
have been changed by distortion, due to shifting and overlapping.
Proximal ganglia have thus been separated, and through changes in
position and environment, their connections are difficult to trace, even
with the aid of morphological and neurological study. In the head
1 Ewina, H. Z.: Kansas University Science Bulletin, 1904, ii, No. 1.
2 Brown, O. H., and ROLLER, L. W.: Kansas University Science Bulletin,
1904, ili, No. I.
238 Ida Ff. Flyde.
for instance, certain segments have developed more than others;
predominating thus in size, they overlie more distant as well as
nearer related ones. Under these conditions it becomes difficult
to determine the exact ganglia which originally had close relations,
and which together controlled the respiratory mechanism.
The phrenic nuclei, regarded as the segmental ganglia of the dia-
phragm, belong to the ganglia that control a certain part of the
respiratory movements. They perform a function in some respects
similar to that of the ganglia which control, for instance, the move-
ments of the gill arches or spiracles in the skate. Any one of these
ganglia, as was proved in my work on the skate, can be isolated from
the other centres governing the respiratory movements, without sacri-
ficing its power of sustaining rhythmical contractions of the muscles
governing the activity of the segmentally related arches or spiracles.
But judging from Porter’s! researches on mammals, the phrenic nuclei,
unlike the more primitive segmental ganglia of invertebrates, or the
ganglia that control the movements either of the spiracles or gill
arches in the skate, represent in higher vertebrates subsidiary centres
that do not possess the power of discharging rhythmical contractions
of the diaphragm, excepting through impulses passing to them from a
superior respiratory centre in the medulla. Their functions were
possibly altered during the process of phylogenetic change, and they
may, therefore, have sacrificed some of the properties characteristic of
primitive segmental ganglia.
A survey of the results and conclusions reached by different physi-
ologists who have carried on investigations relating to the respiratory
centre in mammals, shows that the controlling power of the respira-
tory movements is believed to be collections of ganglia and nerve
‘fibres, connected chiefly with the root fibres of the fifth, seventh,
ninth, and tenth nerves.
Belief in the existence in vertebrates of a superior centre in
the medulla, controlling, through subsidiary centres, the rhythmical,
co-ordinating respiratory activity, seems, from the facts presented by
many experiments, justifiable, though there are physiologists who
still believe, with Brown-Séquard? and Langendorff,? that co-ordinated,
rhythmical respiratory movements executed by thoracic and abdomi-
1 PorTER, W. T.: Journal of physiology, 1895, xvii, p. 455; PORTER and
MUHLBERG: This journal, 1900, iv, p. 334.
* BROWN-SEQUARD: Journal de la physiologie, 1860, ii, p. 153.
8 LANGENDORFF, O.: Archiv fiir physiologie, 1880, p. 518, 1888, p. 283.
Localization of the Respiratory Centre in the Skate. 239
nal muscles are possible after severing the cord from the medulla,
and who therefore do not consider the respiratory centre to be wholly
confined to the medulla.
If we assume that the segmental control of the respiratory move-
ments prevails among invertebrate forms, and that the co-ordinated
rhythmical contractions of the inspiratory and expiratory muscles in
mammals are initiated in the brain, the question at issue is, might we
not find in the lowest vertebrate types a transitional stage of respira-
tory control, bridging the gap from the simple segmental to the com-
plex central apparatus existing in the brain? The respiratory centre
in the skate represents primitive segments that have during develop-
mental changes come to occupy definite areas in the medulla. The
relation of the ganglia to their segmental structures, although altered
is not in these and related forms so obscured by intrusion of neigh-
boring tissue as to make it difficult to trace their connections. It is
hoped, therefore, that a study of the respiratory centres in the skate
may contribute toward a better understanding of the innervation of
respiration in higher vertebrates.
DESCRIPTION OF THE MEDULLA AND THE RESPIRATORY MOVEMENTS
OF ‘THE SKATE.
The medulla and its nerves.—- The brain of the skate (Fig. 2),
lying in its cartilaginous cranium, is easily exposed without loss of
blood. It consists, as is well known, of large olfactory lobes, extend-
ing antero-laterally from the pros-encephal, a comparatively small
diencephal, a mesencephal with its optic lobes, and a large cere-
bellum that overhangs the fourth ventricle of the medulla.
By carefully removing the cerebellum, the medulla (Fig. 3), with
its dorsal lobes and nerves, is well exposed. At its antero-lateral
border is a convoluted ridge, the lobus linez lateralis, continuous
with the cerebellum and mid-brain. Posterior to, and partly covered
by the lobus linez lateralis, is a slight protuberance, the tuberculum
acusticum. It borders the posterior and lateral side of the fourth
ventricle. The floor of the fourth ventricle is marked by a distinct
median suture, at each side of which is a spindle-shaped ridge known
as the fasciculus longitudinalis posterior. Laterad to this, is the
lobus vagi, which bounds the lateral and posterior limit of the fourth
ventricle. It lies close to the tuberculum acusticum, and extends
anteriorly to about the middle of the lobus linez lateralis. In some
240 Ida FZ. fLlyde.
specimens, the lobus vagi of both sides meet in the median line,
forming thus a U-shaped ridge.
The large fifth and seventh nerves are seen at the anterior lateral
edge of the medulla, near the median plane of the linez lateralis. Ven-
tral to them is the eighth nerve, consisting of three branches. Lying
close to the lateral border of the tuberculum acusticum, may be seen
the ninth root, extending posteriorly to join the four root fibres of the
vagus. The last-named four roots emerge on a level with the pos-
terior end of the fourth ventricle. Each of these roots has its
special ganglion at its exit at the side of the medulla, as is well
shown in Herrick’s! illustration of the nerves in Menidia.
Electrical or mechanical stimulation of the peripheral cut end of
the fifth nerve causes movements of the mouth. Sectioning the
fifth interferes with the rhythmical activity of the mouth parts that
are seen during respiratory movements. Stimulation of the seventh
produces closure and opening of the spiracles, and also mouth move-
ments. Sectioning the seventh stops these movements. Stimulation
of the peripheral cut end of the ninth causes the dropping or expira-
tory position of the first gill arch, while lesion of this nerve abolishes
its activity. Stimulation of the first to the fourth vagus roots brings
about the characteristic inspiratory phases of the second to the fifth
gill arches, while cutting these respective roots stops their move-
ments. We see, therefore, that the motor nerves concerned in res-
piration are the fifth, seventh, ninth, and tenth nerves, of which the
fifth is the least important and the tenth the most important. This is
because the aeration of the blood is effected if only the last four gill
arches are active, all other parts ceasing. On the other hand, the
animal may live for an indefinite time, if the spiracle and first gill
arches continue their rhythm, all other parts of the respiratory ap-
paratus discontinuing. Attention. will be directed mainly to the
connections of the seventh, ninth, and tenth nerves in the medulla,
as these are the only ones that specially concern us.
From a study of Johnston’s? article on the brain of Acipenser, and
from the results of my experiments and dissections, I judge that the
description and histology of the lobes of the medulla of Acipenser
and those of the skate are not only homologous but also analogous.
Inasmuch as we possess no histological study of the brain of the
1 HERRICK, C. L.: Journal of comparative neurology, 1899.
2 JouNsTON, J. B.: Zoologische Jahrbiicher, Abtheilung fiir Anatomie und
Ontogenie, IgoI, xv, p. 59.
Localization of the Respiratory Centre in the Skate. 241
skate, and as it is desirable to know more of the structure of the
lobes of its medulla, I think it quite permissible to assume that
Johnston’s description of the lobes of the medulla of Acipenser may
answer also for those of the medulla in the skate. I believe that
future work will prove them very much alike in function.
The seventh nerve has extensive connections with the medulla.
First, through its dorso-lateral line roots which extend into the lobus
lineze lateralis, and second, through its ventral roots that enter the
tuberculum acusticum. Moreover, the anterior division of the lobus
vagi consists chiefly of root fibres for the dorsal seventh. Then too
the anterior portion of the fasciculus longitudinalis posterior gives off
motor fibres which leave laterally to form the motor seventh, at
about the level of the lobus linez lateralis. Therefore injury to
different areas of most of the lobes of the medulla, as will be shown,
easily stops or inhibits the movements of the spiracles. The sensory
nucleus of the ninth is situated in the lobus vagi, but the fasciculus
longitudinalis posterior gives origin to the motor or ventral ninth
root. The tenth nerve also has lateral line roots in the tuberculum
acusticum, while the posterior region of the lobus vagi receives its
sensory roots. In addition to these stations the fasciculus longitu-
dinalis posterior contains numerous vagus roots derived from the
motor vagus nuclei situated ventro-mesally. These make their exit,
as can be clearly seen, at the side of the medulla. The tenth nerve
covers thus a definite but wide area of the medulla. It is directly
and indirectly influenced by lesions especially in the posterior part of
the medulla and the lobus vagi. We learn from the above descrip-
tion that the lobus vagi embodies the sensory nuclei of the three prin-
cipal respiratory nerves; it is therefore of special interest to us.
The lobus vagi moreover gives rise to the secondary vagus tract, de-
scribed by Johnston. At the anterior limit of the medulla in contact
with the cephalo-ventral surface of the tuberculum acusticum, and at
the junction with the cerebellum, lies a group of nerve cells known as
the Rinden-Knoten, or the secondary vagus tract nucleus. At the
posterior limit of the medulla are two other groups of nerve cells, the
nucleus funiculi and the commissura-infima-Halleri. The secondary
vagus tract extends from the Rinden-Knoten posteriorly through the
tuberculum acusticum and commissura-infima-Halleri to reach the
lateral tract in the cord. The importance of this tract I believe lies
in the fact, that it acts as an intermediate agent between the vagus
and other centres.
242 Ida Ff. Hyde.
Respiratory movements. — The respiratory phases in the skate con-
sist of rhythmical co-ordinated movements for the passage of water
into and out of the gill chambers. During the inspiratory phase,
the spiracles (sf.
Fig. 2) and mouth
(7) open, the arches
(1a-5a) are elevated,
permitting the en-
trance of water, and
the gill slits (1s—5s,
Fig. 1)-elose the
openings to the gill
chambers. During
the expiratory phase
which follows, the
spiracles and mouth
close, the arches
FiGuRE ]1.— Ventral view of the skate, about half natural drop, and the gill
size. The outline of the first two gill arches are seen slits open for the
during the inspiratory, the last three in the expiratory exit of water. Oc-
phase. m, the mouth; la to 5a, outline of first to fifth
gill arch; 1s to 5s, first to fifth gill slits.
a =
casionally there is a
gasping movement
consisting of a forcible closure of the gill slits and dropping of the
gill arches. During this interval, water and waste matter are ex-
pelled through the mouth. Ina definite interval of time the respi-
ratory movements may greatly vary, from twenty to seventy or more
inspirations per minute. The variation is influenced by depth from
the surface, irritation, or internal conditions.
EXPERIMENTAL METHODS.
The skate was kept in position on a board by means of a piece
of fish netting, and sea-water was passed constantly from a reservoir
through a rubber tube into the mouth. The fish remained perfectly
quiet for hours, and even days. In this way, or by placing the
board with the tube and fish in an aquarium, artificial respiration
could be kept up for days. When ether was used, another tube
connected with a reservoir of seawater and ether was placed in the
mouth. By means of a T-tube joined to the rubber tube and the
ether reservoir, ether vapor, ether water, or both could be made
Localization of the Respiratory Centre in the Skate. 243
to enter the fish’s mouth. The respiratory movements can be
seen either from the fish’s dorsal (Fig. 2) or from its ventral
(Fig. 1) side. If the dorsal side is up, the opening and closing
of the spiracles, and the rising and falling of the gill arches can
be followed. With the ventral side up, not only the changes in
the movements of the mouth, gill arches, and gill slits are seen,
but also variations in the heart’s action can be studied. The skate,
therefore, is an excellent object for the study of changes in the
respiratory, as well
as the heart’s
movements. I
therefore employed
it in my investiga-
tion on the effects
of intravenous in-
jections of solu-
tions on the central
nervous system,
with special regard
to the heart and
respiratory move-
ments. This paper
is about to be pub-
: P : FicurE 2.— Dorsal view of the skate, showing the outline of
lished. The brain the first two gill arches in the expiratory, the last three in
was exposed either the inspiratory phase. The dorsal view of the brain is
from the dorsal side exposed, and the right spiracle represented closed, the
b tat i left open. 1a to Sa, first to fifth gill arches; 5, 7, 9, 10,
y fae Oe = corresponding cranial nerves; sf, spiracle; ¢, eye; Pros.,
overlying cartilage, cerebrum; mes, midbrain; met, cerebellum; #, medulla.
or from the ventral
side by cutting through the mucous lining and the roof of the
mouth, with scarcely any loss of blood. The effects on respiration,
of lesion of any part of the brain, are instantly noticed.
Electrical and mechanical stimulations, incisions with small or large
scalpels, punctures with needles, or destruction of tissue with red hot
needles were employed for different purposes. Stimulation with fine
electrodes and threshold value of induction shocks, or pressure with
pointed wads of cotton at different points of the medulla in a normal
skate, or in one in which the respiration had stopped, or the move-
ments had been inhibited, gave interesting results; but they alone
cannot be considered reliable for localization of function, because it is
)
244 Lda fH, Flyde.
not definitely known what cells or fibres are acted on by the currents
or pressures, both of which may spread wide of their mark. More-
over, the method of sectioning with the scalpel, across several regions
of nervous tissue, where there is danger of injuring neighboring areas
that may have an influence upon the experiment, is crude and dan-
gerous, unless the function of the entire field sectioned is thoroughly
understood, and it is known either that the neighboring parts bear no
relation to the parts under investigation, or just how the destruction
of those parts would influence the experiment. Before using the
scalpel, to determine the effect of removing or separating certain areas,
their localities were carefully studied and experimented on, so as to
ascertain all possible effects that may arise from a planned lesion,
and to be able to interpret the results correctly. After the operation,
the animals were observed in an aquarium or on the operating board.
They were kept alive by artificial respiration as long as possible.
Later the brains were hardened in formole, and the postmortem made.
More than sixty skates were experimented upon.
SHock EFFECTS.
Shock effects follow any injury or strong electrical or mechanical
stimulations of the different regions of the central nervous system.
The effect produces an inhibition of all or of parts of the respiratory
movements. The inhibition of the movements of the spiracles is
most readily effected, and its duration is always prolonged, lasting
many hours or days. The parts inhibited are not, as is believed,
always posterior to the place of injury, but may be either anterior or
posterior, or both anterior and posterior to the lesion. The nervous
tissue anterior to the medulla moreover does not seem capable of
preventing the shock, since the inhibition, at least upon the respira-
tory mechanism, occurs whether the parts in front of the medulla
have been removed before the operations on the medulla are per-
formed or not.
Attempts to avoid shock effects were not entirely successful. It was
found that cocaine applied locally reduced the time of inhibition, but
it greatly depressed the respiratory movements. Ether used care-
fully while operating, and during the shock, gave more satisfactory
results, although it did not entirely prevent inhibition of certain parts
of the respiratory apparatus. Lesions made with fine red hot needles
SS
Localization of the Respiratory Centre in the Skate. 245
proved less injurious than those made with the scalpel. Intravenous
injections of normal salt solution, or of salt solution with a very small
percentage of calcium and potassium at least served to shorten the
duration of shock. Electrical or mechanical stimulations by pressure
or lesion to other parts of the central nervous system during inhibi-
tion were of apparent benefit in removing the shock. Much depended
upon the condition of the fish. Fish kept in the aquarium for several
days would prove more sensitive than those fresh from the sea.
Sometimes one gill arch only will begin to move a few minutes after
the operation, then other gill arches, or one or both of the spiracles
may begin, but perhaps hours or days later. In speaking, therefore,
of the duration of shock, I mean the time that elapsed between the
operation and first sign of activity of any of the uninjured parts of
the respiratory mechanism. Two operations, performed apparently in
every respect alike, may cause in one case only slight shock effects of
a few minutes’ duration, in another case severe and prolonged effects
that may last days, or even permanently.
Different levels on the antero-posterior axis of the medulla (Fig. 3)
are lettered from a to z, as an aid in describing the regions operated
upon. Many experiments practically alike were carried out, and the
general results of such experiments are recorded in the table or text.
SECTIONS ANTERIOR AND POSTERIOR TO THE MEDULLA.
a. In order to remove the cerebellum from the medulla, to which
it is attached by means of the choroid membrane, and to prevent loss
of blood and injury to the medulla, it was found necessary to tie a
ligature around the base of the cerebellum. In doing so the choroid
vessels are tied, and the cerebellum is separated from the mid-brain
and the medulla. A shock lasting usually fifteen minutes follows,
during which time all respiratory movements cease. At the end of
that time the normal rhythm of all parts began again.
b. The medulla was.entirely cut off from all the parts of the brain
anterior to it, by a transverse section, so that no part of the medulla
was destroyed. After a shock inhibiting all respiratory activity for
about an hour, the spiracles and, later, the gills began to move again.
e. The medulla was severed from the cord without cutting nerves
that emerge from it. For about fifteen minutes following the section,
there were no breathing movements visible, then the spiracles and gills
slowly began their movements again.
246 [da Fl. Flyde.
These experiments prove conclusively that the co-ordinated rhyth-
mical activity of the respiratory mechanism is independent of parts
of the central nervous system outside the medulla.
EXPERIMENTS ON SPECIAL NUCLEI.
a, Rinden-Knoten or nuclei of secondary vagus tract. — (Described
on page 241.) It was of interest to know if this conspicuous nucleus
exerted any influence over the gill arches or spiracles. The anterior
region of the medulla was gradually destroyed. Beginning at the
cephalic end, the middle portion, and then the lateral parts, were burnt
with hot needles without any loss to the respiratory action. That is,
when the region anterior to (a) Fig. 3 was severed from the rest of
the medulla, there followed an inhibition of respiration for about
twenty minutes, after which time the respiration was again resumed.
b. The nucleus funiculi and the commissura-infima-Halleri. — These
are situated at the other end of the medulla. This region was com-
pletely destroyed by burning. The last two gills ceased their rhythm
for about fifteen minutes, and then began again in time with the other
parts. This region, corresponding to the area so near to Flouren’s
respiratory centre, lies outside of the region governing co-ordinated
rhythmical respiratory movements.
MEDISECTION OF THE MEDULLA.
The next problem was to ascertain the effect of dividing the medulla
along the median suture into two lateral halves (Fig. 3). In order
to see the whole extent of the suture, the cerebellum was first removed,
and ether carefully introduced with the sea-water during the operation.
The ether first acts as a stimulant, increasing the rhythm of respira- _
tion; but if continued for some time, it retards the activity. To be
effective, it must be most carefully used. If with a fine hot needle
a cut is made in the median suture along the whole extent of the
fourth ventricle, the respiratory movements may all cease for ten
minutes or more. As a rule, the gill arches of both sides then re-
sume their activity, though it may happen that the arches of only one
side move, and later those of the opposite side begin, or only the
last ones of both, or of one side begin, and several hours later the
remaining arches join them in their rhythm. Occasionally both
spiracles move in the same rhythm with the gill arches, but more
Localization of the Respiratory Centre in the Skate. 247
often one spiracle begins to move with the arches of its side, which
may be in a rhythm very different from those of the opposite side.
If more ether is now given,
the spiracle of the opposite
side may gradually move.
Then both spiracles keep time
with the gill arches of one
side, or the spiracles and gill
arches of the respective sides
may have their own rhythm.
f
There may therefore occa- ho i
. . . Va) 12| gh Z
sionally be no co-ordination WA esa? -
Bese N I
of rhythm among the parts of ISO A
the respiratory organs. Should
the cut, however, be made to
one side of the suture, the
respiratory mechanism of the
injured side and the spiracle
FIGURE 3.— Dorsal view of the medulla. The
cerebellum removed to expose the fourth
ventricle and the lobes of the medulla.
of the uninjured side will
instantly be interrupted and
remain inactive for hours and
even days. In this case it is
certain that the spiracle of the
uninjured side is inhibited.
The inhibition may last sev-
eral days, after which it may
pass off, and the spiracle then
resumes its movements with
those of the gill arches.
The fore part of the brain was cut off at
the posterior junction with the medulla.
a to 2, the levels employed to indicate the
regions of the medulla, as a guide to the
localization of function; 1 to 5 indicating
where injury abolishes the function of the
corresponding gill arches; mes, posterior
limit of the midbrain ; mef, cut edge of the
cerebellum; ms, median suture; 52 to 10v,
corresponding cranial nerves; Z/, lobus
linez lateralis; Za, tuberculum acusticum ;
Lv, lobus vagi; 77, funiculus longitudinalis
posterior ; Sc, spinal cord.
It became evident from the results of median section of the medulla
that the centres for the nervous respiratory mechanism in the skate
were bilateral, each half controlling the movements of its respective
side.
EXPERIMENTS ON THE LOBES OF THE MEDULLA.
It was of interest to ascertain the effect on the respiratory move-
ments produced by lesion in the different regions of the lobes so char-
acteristic of the fish’s medulla (Fig. 3). Furthermore a means was
thus offered for comparing ascertained morphological results with
physiological ones. Since the motor nuclei lie nearer to the ventro-
248 Ida F1. Flyde.
mesial surface, and the sensory nearer the dorsal, distinction must be
made between deep and superficial injury. A deep puncture penetrates
through the medulla, a superficial one aims to penetrate only through
the sensory area. When the cerebellum is removed, the outlines of
the lobes are distinctly seen, and they can therefore be satisfactorily
experimented on from the dorsal side. Operating from the dorsal
side of the medulla is more satisfactory than operating from the
ventral side, because there are on the ventral surface no definite Jand-
marks to aid in defining the limits of the lobes other than the median
suture, the outline at the posterior limit of the medulla, and the exit
of some of the nerves. Ventral lesions had to be verified by dorsal
ones, and were employed only to ascertain the results of special
experiments.
TABLE I.
LESIONS OF THE TUBERCULUM ACUSTICUM (FIG. 3).
Effect on
Effect on injured side. uninjured Remarks.
side.
Tnjury to
area.
no effect superficial puncture.
lst arch stopped deep «
no effect superficial
2d, or 2d and 3d stopped deep
no effect superficial
3d and 4th arches stopped deep
no effect superficial
5th arch stopped deep
Lesions of the tuberculum acusticum and lobus lineze lateralis. —
(@) Superficial injury to the tuberculum acusticum (Fig. 3) caused
only slight inhibitory effects on the operated side. Deep punctures
or incisions occasioned injury to the underlying motor nuclei, or fibres
of either the ninth or tenth nerve, depending upon the area experi-
mented upon. The effect of a lesion, begun at the level 2 and con-
tinued gradually anteriorly to level ¢ in the area of the tuberculum
Localization of the Respiratory Centre in the Skate. 249
acusticum, was permanent suspension of the function of the fifth,
fourth, third, second, and first arches respectively (Table [).
Occasionally the puncture aroused the spiracles and uninjured
arches to increased activities of short duration.
The table on page 248 gives only the general results obtained from
many experiments.
In all the tables, “no effect” means that temporary suspension
of function of some parts may have followed, but no permanent
suspension.
(6) The effect of a superficial lesion beginning at the posterior
limit of the lobus linez: lateralis and extending anteriorly, is to
introduce as a rule more or less prolonged inhibition of the spiracle,
seldom of both spiracles and the first gill arches. Deep puncture,
especially on a level with ¢, d, or c, which caused injury to the under-
lying seventh motor nerve, produced sudden permanent cessation of
the spiracle activity, and shock effect to the arches of the injured side.
TABLE II.
LESIONS OF THE LoBuS LINE LATERALIS.
Effect on
uninjured side.
Injury at
levels. Remarks.
Effect on injured side.
no effect no effect deep lesion.
ie See ae superficial lesion.
deep
superficial
spiracle stopped deep
no effect superficial
spiracle stopped deep
From the fact that in both the tuberculum acusticum and the lobus
linez lateralis, the nuclei and fibres of the special respiratory nerves
are absent, we should not be surprised to learn that they exerted no
direct influence over the respiratory mechanism. The lobes overlie
the important motor fibres that pass to the spiracles and gill arches,
250 [da Ff. Flyde.
therefore deep lesions in their areas cause permanent interruption of
those parts of the respiratory activity whose fibres have been injured.
Lesions of the fasciculus longitudinalis posterior. — As a rule the
punctures and incisions were deep, for superficial injury caused only
temporary suspension of function, which passed away usually before
twenty-four hours. A puncture, as in other experiments with a hot
needle anywhere in the fasciculus between the levels a to ¢, stopped
TABUE sit,
LESIONS OF THE FASCICULUS LONGITUDINALIS POSTERIOR.
Effect on
Effect on injured side. | uninjured Remarks.
side.
Injury at
levels.
spiracle stops no effect | deep lesion.
spiracle on uninjured side in-
hibited 3 hour.
spiracle on uninjured side in-
hibited 5 hours or until death.
spiracle and Ist arch or
spiracle only stops
lst arch only or 1st and
2d stop
3d arch or 3d and 4th ,_| prolonged inhibition of spiracle
stop and one or more arches on un-
injured side.
5th arch stopped
the movements of the spiracle of the injured side, and often the
' opposite spiracle was also stopped, but the gill arches were not seri-
ously affected by lesions above the level of d¢. Destruction of the
tissue just below the region @ suspended the rhythm of the first gill
arch; at /, that of the second gill arch; and at the level of g usually
the third, and often both third and fourth, while the fifth ceased its
activity when the injury was made at the level of & Deep incisions
at g often stopped all respiratory actions of the injured side for many
hours, and often all respiratory movements ceased for about half an
hour. It is seen from Table III and Fig. 3 that the spindle-shaped
area of the fasciculus longitudinalis posterior is capable of division
into distinct regions, each of which controls definite respiratory
ra
Localization of the Respiratory Centre in the Skate. 251
organs. These results were obtained before I knew of Johnston’s!
article, from which I learned that the above results are due to the
destruction of the motor fibres which originate in the definite areas
of the fasciculus, as above indicated.
Lesions of the lobus vagi.— Superficial destruction of the lobus vagi,
in a line proceeding from the anterior limit posteriorly, gave evidence
of localization of the functions of the 7th, 9th, and 1oth sensory areas,
as illustrated in Fig. 3 and Table IV. For control, the punctures were
made both superficially and deep, and the injury in some instances
carried from the posterior limit of the lobus anteriorly. It was seen
that lesions between the levels 4 and d destroyed the movement of
the spiracle of the injured side, the opposite spiracle often, and the
arches occasionally, ceasing activity for several minutes. Destruc-
tion to the area on a level with ¢ usually caused the first arch to stop,
but occasionally the spiracle also became inactive, and it once hap-
pened that both spiracles and all the arches on the injured side
stopped moving for three days, from a deep incision in this region ;
then the arches again resumed their rhythmic movements. A punc-
ture near /, if superficial, usually caused both first and second arches
to stop moving; but if the puncture was deep, it might strike a motor
fibre or cell of the third gill arch, and then the first three arches
would become inactive. Under such circumstances, only the spiracle
and the last two arches of the injured, and all the parts of the un-
injured side, would keep up the rhythm. A superficial injury at ¢
suspends the action of the third arch; if the injury is deep, both the
third and the fourth gill arch; while at the level % the injury is
followed by the interruption of the movements of the fifth arch.
Punctures made both at f and % stopped the second and fifth arches,
while the spiracle and first, third, and fourth arches of the injured
side continued. A deep puncture at % may suspend all respiratory
function for thirty minutes, and then all but the fifth arch may begin
again. Ifa motor fibre or nerve cell of a spiracle is destroyed in the
fasciculus posterior longitudinalis, so that the spiracle no longer
moves, a pressure in the area of the lobus vagi J to d will reflexly
cause the spiracle of the uninjured side to stop moving for ten
minutes or more, the arches being not thereby affected. This proves
that the sensory centre of the injured spiracle is not destroyed, and
that the sensory impulses emanating from the spiracle centre act
1 JOHNSTON: Zoologische Jahrbiicher, Abtheilung fiir Anatomie und Ontologie,
Foor, XV; p. 50.
252 Lda 1. Flyde.
more strongly reflexly on the spiracle than on any other part. When
the respiratory movements are slow and weak, they can be suddenly
spurred to violent activity by touching a hot needle to, or holding it
near any of the areas in the lobus vagi. The organ controlled by
cells of that area will suddenly begin to move most rapidly, and all
other parts of the respiratory mechanism follow in the same rhythm ;
thus the rate of respiratory movements may be set by a spiracle, or
by any one of the gill arches.
TABLE IV.
LESIONS OF THE LOBUS VAGI.
Injury at
Effect on uninjured
level. i
Effect on injured side. ste.
spiracle ceased no effect deep lesion.
“cc “
superficial lesion.
Ist gill arch ceased ‘ &
inhibition of injured side
for one hour; deep le-
sion.
inhibition of spiracle
or no effect
Ist gill arch and spir-
acle stop
2d and 3d, possibly 1st}
2d arch stops 1
3d or 4th, or
3d and 4th arches stop
5th arch ceases
deep lesion.
superficial lesion.
ce iss
deep
deep lesion; all respira-
tion may cease for 30
minutes.
1 In some experiments puncture at # suspended the function of the first and
second gill arches, the spiracle and last three gill arches occasionally continued
moving in a different rhythm.
EFFECT OF TRANSVERSE SECTIONS OF THE MEDULLA.
Since the areas occupied by the sensory nuclei of the principal
respiratory nerves in the lobus vagi can now be mapped out, and the
result of lesions to the other lobes of the medulla understood, it
remains to test the effects of transverse incisions in the medulla on
the respiratory movements. The foregoing experiments enabled us
- We 4
Localization of the Respiratory Centre in the Skate. 253
moreover to determine somewhat whether the phenomena following
an injury to a region of the medulla are in part or whole the result
of shock.
For these experiments the cerebellum was removed. The cut was
either made (a) from the median line to near the edge, or (b) carried
entirely across the median suture of the medulla. (a) If a hemi-
section is made at the level a or 34, the spiracle of the injured side
instantly stops moving. If made at c, usually both spiracles stop, the
one due to injury, the other to reflex inhibition. At d, the shock
effect stops all respiratory movements for about fifteen minutes, then
all but both spiracles continue their rhythm again, one spiracle centre
injured, the other inhibited. At ¢, both spiracles and first arches
stop entirely for twenty-four hours, but sometimes after twenty-four
hours the spiracle and first arch of the injured side are the only parts
whose functions have been suspended. This seems to be influenced
by the method of operation and the condition of the animal. At /,
the first and second arches, sometimes also the third, of the injured
side cease activity due to injury to the cells or fibres. The spiracle
and last two or three gill arches of the injured side as a rule move at
the same rate as the parts of the uninjured side. The respiratory
centres of the organs of the injured side must have been divided into
two divisions, — an anterior, consisting of cells and fibres governing the
movements of the spiracle, and, in two instances, the spiracle and first
arch; and a posterior division, consisting of the last two or three
gillarches. These groups usually move in unison, but each may have
an independent rhythm of its own for a short time. Hemisection at
the level of g usually destroys the action of the three last gills of the
injured side, the fibres, or motor or sensory nuclei of one or the other
being destroyed by the cut. Moreover, sectioning at this or any other
level may be foilowed by a cessation of movements in all gill arches
on the injured side, and of both spiracles for a short time. Then too,
the spiracle of the uninjured side after such experiments may be
inhibited for days, or until the animal dies. Section at the level %
destroys the movement of the last arch of the injured side. (b) Ifa
deep section is made entirely across the medulla at the level a, inhi-
bition of all respiratory movements for one hour ensues, then all parts
continue as before. At the level 4 or c, both spiracles cease. If the
cut is made at the level of @, both spiracles and all the arches may
stop. After twenty-four hours the three, four, or five last arches
continue to move again. Section at ¢ suspends all respiratory action
254 Lda F. Flyde.
for several days. At the end of the third day, all but the spiracles
and first arches are moving. Section at f results in shock effect to
all of the respiratory mechanism. At the end of the third day both
spiracles and the three last gill arches are active, the spiracles mov-
ing occasionally in a different rhythm from that of the last three
arches. Here the respiratory mechanism is divided into two sections,
each having its own rhythm, more distinctly seen and more complete
than when hemisection is made.
In some experiments, section at the level of f produced shock that
resulted in inhibiting all activity for hours, and even after three days,
though all but the first arches were active, the spiracles had not
resumed their movements. The znhibition was anterior to the cut and
was true inhibition, since we know from other experiments that wgury
at this level does not suspend the function of the spiracles. Sections
just posterior to f showed, at the end of ten days, that all but the 3d
and 4th arches were moving. In such experiments the spiracle and
first and second arches moved in rhythm occasionally different from
that of the fifth arches.
Sections at ¢ may cause the last three or four arches to cease, and
the first arches and spiracle to move violently for several hours;
again the last three arches may cease moving for days or until the
death of the animal. Section at % has the effect of stopping the last
gill arches.
The results noted in Table V, obtained by hemisection and com-
plete transverse section of the medulla, were, it is seen, as might have
been expected, not very different; in the one case, cessation occurred
on one side only, in the other on both sides. The reflex inhibition of
the spiracle, following injury to its fellow, was most marked. Section
at (f) or (g) divided the lobus vagi and fasciculus so that the centres
of the respiratory mechanism would be separated into two divisions,
each having for a longer or a shorter time a rhythm of its own.
MEDISECTION FOLLOWED BY HEMISECTION.
When the hemisection followed a median section, which first sepa-
rated the medulla into a right and a left half, and then one of those
halves into an anterior and a posterior division, results were obtained
which corroborated those obtained above. The arches and spiracles
of the uninjured side continued their activities, and the anterior divi-
sion consisting of spiracle, or spiracle and first gill arch, might pos-
oe hae
Localization of the Respiratory Centre im the Skate. 255
TABLE V.
HEMISECTION OF THE MEDULLA.
Injury
at Effect on injured side. Effect on uninjured side. Remarks.
level.
spiracle stops no effect deep incision.
“cc «<
spiracle stops
no effect or spiracle may
stop
é spiracle and first arch] no effect or spiracle stops| “ if
stops
Ist and 2d stop « “ “ «“
£ 3d and 4th, or 3d, 4th, and | spiracle may stop or no| “ -
5th arches stop effect
5th gill arch stops no effect i Z
EFFECT OF A TRANSVERSE SECTION OF THE MEDULLA.
Right side. Left side. Remarks.
a no effect no effect deep cut.
spiracle stops spiracle stops
se “é
“ce “cc
d spiracle or spiracle and | spiracle or spiracle and 1st | prolonged inhibition.
Ist arch stop arch stop
é spiracle and lst arch stop | spiracle and Ist arch stop | inhibited three days.
5h Ist and 2d, or Ist, 2d, and | Ist and 2d, or Ist, 2d, and se “f g
3d arch stop 3d arch stop
g 3d and 4th, or 3d, 4th, and | 3d and 4th, or 3d, 4th, and ve ten oa
5th arches stop 5th stop
h 5th arch stops 5th arch stops deep incision.
EFFECT OF MEDISECTION FOLLOWED BY HEMISECTION.
Fa 1st and 2d, or Ist, 2d, and | no effect spiracle and 4th and
3d arches stop 5th, or 3d, 4th, and
5th active.
g 3d and 4th, or 3d,4th,and| “ “ spiracle and Ist and
5th arches stop 5th arches active.
256 Lda F1. Flyde.
sess a rhythm for a few minutes somewhat different; while the
posterior division, consisting of the last two or possibly three gill
arches exhibited a rhythm which was unlike either of the other
divisions. Again, all three divisions kept the same rate of action
part of the time. The posterior division, being in some cases sepa-
rated by the section, first, from the centres of the seventh and ninth
nerves, second from the centres of the opposite side, and third, from
the ganglia of at least one of the four ganglia belonging to the tenth,
must have been stimulated to action through impulses reaching the
arches from the three remaining or the one remaining ganglion of the
tenth nerve. The movements of these parts were usually more feeble
and of shorter duration than those of the other divisions, but that
they had an independent centre was evident, and this centre capable
of inaugurating and sustaining rhythmical movements of a part of
the respiratory mechanism. Transverse sections destroy not only
sensory but motor nuclei, and may cut into motor fibres of neighbor-
ing arches. For this reason, the function of more than one arch, or
arch and spiracle, may be destroyed, and the injury cause entire
suspension of function of certain parts. The shock effect was more
severe from hemi or complete cross-section than from puncture with
hot needles. The period of inhibition depended upon the vigor of
the animal, and upon the method employed, and whether the opera-
tion was performed with the aid of an anesthetic or not.
CONCLUSIONS.
1. The respiratory movements in the skate are segmental pro-
cesses. The relationship of the respiratory organs and their seg-
mental centres is not so obvious as it is in lower forms. The
developmental changes of shifting and consolidation have begun
to mask the segmental connections of the different parts of the
brain. Where development has proceeded a step further, this connec-
tion would be demonstrated only with difficulty. The results ob-
tained from the experiments on the respiratory centre in the skate,
tend to support Loeb’s! views as to the segmental character of the
respiratory centre.
2. The respiratory centre in the skate occupies definite sensory
and motor areas in the medulla. The sensory cells, comprising neu-
rons of the seventh, ninth, and tenth cranial nerves, are situated
1 LoEB: Physiology of the brain, 1900, p. 144.
Localization of the Respiratory Centre in the Skate. 257
in the lobus vagi; whereas motor cells and fibres are ventrad to
it, as well as in the fasciculus longitudinalis posterior.
3. Each ganglion, through special fibres and cells, controls the
activity of the respiratory muscles with which it is segmentally
related and is capable of initiating impulses that produce co-ordi-
nated rhythmical respiratory movements.
4. The medulla may be severed both from the cord and the regions
of the brain anterior to it, or divided along its median suture, into
two bilateral halves, without impairing the functions of the respira-
tory centre. Each half is capable of sustaining co-ordinated respira-
tory movements which part of the time may be different in rhythm
on the two sides.
5. The ganglia and consequently the respiratory mechanism can
be divided into two or three divisions, each of which may for a time
have its own peculiar rhythm, or all of the divisions may continue
their activity in the same respiratory phases.
6. Not only may either the spiracle and first gill arch, innervated
by the seventh and ninth nerves, or the last four gill arches,
innervated by the tenth, when isolated from the rest of the respira-
tory mechanism by a median and transverse section continue their
movements, but all other than the special part of the respiratory
centre that controls these divisions may be destroyed, and either
the four gill arches or the spiracle and first gill arch will still pursue
their co-ordinated respiratory activity.
7. There is no one spot, the destruction of which is followed by
permanent cessation of respiratory movements, causing sudden death,
provided artificial respiration is maintained until the shock effect
passes off.
8. Any lesion to the medulla may cause a shock or inhibition to
part or all of the respiratory movements for a shorter or longer inter-
val of time. The part whose function has temporarily been sus-
pended may be innervated by nerve cells lying anterior, posterior, or
lateral to the lesion.
9. The shock effects may be shortened or prevented if the animal
is vigorous; if an anesthetic, ¢. g. ether, is carefully administered
during the operation; if a solution of 20 c.c. 3 2 NaCl containing
qs c.c. 2 CaCl,, and 75 c.c. 1 KCl is injected into the blood imme-
diately after the operation; or if a strong electrical or mechanical
stimulus, such as pressure, is-applied to a region that will either
reflexly or directly stimulate the centre of the inhibited part.
258 Ida H. Hyde.
10. The skate illustrates, in its type of respiratory centre, an
intermediate stage, between the simple segmental arrangement of
the neurons presiding over the co-ordinated respiratory movements
found among invertebrates, and the complex, modified, and special-
ized centres existing in higher vertebrates. :
5
4
aeetnP, LOCAL APPLICATION OF SOLUTIONS OF
SALINE PURGATIVES. TO THE PERITONEAL
SURFACES OF THE INTESTINE.
By JOHN BRUCE MacCALLUM.
[From the R. Spreckels Physiological Laboratory of the University of California.|
I. THE PRODUCTION AND SUPPRESSION OF PERISTALTIC MOvE-—
MENTS OF THE INTESTINE BY THE LOCAL APPLICATION
OF SALINE SOLUTIONS.
N a previous paper! I have described the effects of subcutaneous
and intravenous injections of purgative salts on the intestines of
rabbits, and have shown how the peristalsis produced thereby can be
inhibited by similar injections of calcium chloride solution. It was
further pointed out that these actions are analogous to the produc-
tion and suppression of rhythmical contractions in voluntary muscles
as described by Loeb. This analogy is still more clearly shown by
a series of experiments which I have since made on the local applica-
tion of these salts to the intestine.
I have found that all those salts which produced increased peris-
talsis when administered intravenously or subcutaneously, or when
introduced into the intestine or stomach, have the same action when
applied locally to the peritoneal surface of the intestine. The imme-
diate effect of this is essentially local; only those loops which are
moistened by the saline solution are at once set in motion. After
a short time, the other loops also become active. Peristalsis thus
produced may be entirely inhibited by the local application of a solu-
tion of calcium or magnesium chloride. This may be illustrated by
the following experiment : —
The intestines of a rabbit under the influence of morphine were exposed. On
a small group of coils there were poured about 3 c.c. ¥ sodium citrate solution.
1 MacCA.uuM, J. B.: This journal, 1903, x, p- 101; also Preliminary Report,
University of California Publications, Physiology, 1903, i, p. 5.
? Logs, J.: Archiv fiir die gesammte Physiologie, 1902, xci, p. 248.
. 259
260 John Bruce MacCallum.
Almost immediately (within one minute) the loops became very active. Strong
contractions of the muscle coats took place. After a few minutes, the other
loops were also set in movement, so that the whole small intestine showed
active peristalsis. The citrate solution was then washed off by % NaCl solu-
tion, and about 3 c.c. % CaCl, solution poured on the loops. ‘The peristaltic
movements were promptly suppressed, and the intestine remained quiet. By
the further addition of citrate solution, the coils were set in active movement
once more, and by the subsequent application of calcium chloride solution
again inhibited. This was repeated many times (sixteen), and apparently
might have been continued as long as the intestine remained alive.
The same results were obtained by using instead of the sodium
citrate solution, a solution of barium chloride, sodium sulphate,
fluoride, bromide, iodide, phosphate (Na;PO,), oxalate or tartrate.
Local application of solutions of any of these salts produces increased
peristaltic activity. Solutions of sodium chloride have a very slight
action of the same character. On the other hand, the intestinal
movements are equally inhibited by calcium chloride and magnesium
chloride, while strontium chloride has a similar but less powerful
inhibiting action.
In testing those salts with which it was necessary to use dilutions
greater than %, the dilution was made with a neutral fluid consist-
ing of sodium chloride and magnesium chloride. It was found that
% NaCl solution increased to a slight extent the peristaltic move-
ments. By adding to to c.c. ¥ NaCl, 0.5 c.c. ¢ MgCl,, a fluid was
obtained which had apparently neither stimulating nor inhibiting
effects. In addition to solutions made up by dilution with this neu-
tral fluid, others were used in which the salt solutions were diluted
with distilled water. Practically the same results were obtained
in both cases. It was found that 1 c.c. 345 BaCl, solution applied
locally to the intestine is sufficient to cause strong peristaltic move-
ments ina rabbit. This quantity contains about 0.00076 gm. barium
chloride. In the case of sodium citrate, the concentration must be
considerably greater. No solution of this salt more dilute than 74
is active in a rabbit. Of all the purgative salts, barium chloride is
by far the most powerful. If a drop of % BaCl, be placed on the
serous surface of an intestinal loop, or if a small area be moistened
with this solution by means of a camel’s hair brush, the muscle
beneath the moistened area will almost immediately contract so
that a ring-like constriction of the intestine is formed. This often
is so sharply marked that it suggests the effect produced by tying
Lifect of Saline Purgatives. 261
a ligature around the intestine. This constriction remains for a few
moments, and then gradually moves along the loop in the direction of
the normal peristalsis. If the solution be injected into the muscle
of the intestine at any point with a hypodermic needle, a similar
sharp constriction takes place. If also a few drops be injected
directly into a branch of the superior mesenteric artery, all that
part of the loop supplied by the arterial branch will contract vio-
lently. These statements are true also in the case of sodium citrate,
fluoride, sulphate, etc.; the action of these salts, however, is less
powerful.
It must be added here also that the actual passage of faeces may
be produced within an hour by the application of the purgative salts
to the serous surfaces of the intestine. This takes place most
quickly with barium chloride. It is possible to observe directly
through the semi-transparent walls of the intestine the rapid passage
of faecal masses from one loop to another.
The intestines of the rabbit are apparently much more sensitive
to the action of sodium citrate and sulphate than are those of the
dog or cat. Barium chloride, on the contrary, acts with equal
strength in all these animals. In a rabbit, the intestines are always
set in active peristaltic movement by contact with 4 sodium citrate
solution; and even much more dilute solutions are, as a rule, effective.
In a cat, however, it was found that a % solution of sodium citrate
has practically no effect, while a ®% solution sets the intestine in
p y 8
active motion. In a dog also ¥ sodium citrate solution is usually
ineffective. Similarly a % sodium sulphate solution is inactive in
a dog, while a % solution of the same salt starts up distinct peris-
talsis. In the cat and dog, also, the peristalsis may be inhibited
by calcium or magnesium chloride, as shown in the following experi-
ment: The intestines of a cat were exposed in the usual manner,
and a % solution of sodium citrate was applied to the serous surface
of the loops. There was no increased movement. There were
then poured on the loops a few cubic centimetres of a mixture of
5 c.c. ® sodium citrate and 5 c.c. 274 CaCl,. The loops remained
motionless. After waiting a considerable time (fifteen minutes), a
4” solution of sodium citrate alone was poured on the intestine.
Almost immediately they became very active; and the peristalsis
continued until calcium chloride was again applied. The loops then
came to a standstill. The difference in susceptibility to the action of
citrate which exists between rabbits on the one hand, and dogs and
262 John Bruce MacCallum.
cats on the other, may be in some way connected with their being
herbivorous and carnivorous animals respectively.
The action of sodium citrate, sulphate, fluoride, etc., when applied
locally, may be inhibited by the administration of an approximately
equal quantity of calcium or magnesium chloride of the same con-
centration. The counteraction of the effect of barium chloride, how-
ever, requires a much greater concentration of calcium. Using equal
quantities of the two salts, the action of the barium is usually not
inhibited, a fact which I have previously stated. With greater con-
centrations of the calcium chloride, the antagonistic action, however,
is clear. This is shown in the following experiment: Applied
locally to the intestine of a rabbit 1 c.c. 395 BaCl, solution caused
active peristaltic movements. The application of 1 c.c. 349 CaCl,
solution exercised no inhibiting effect whatever. The same quantity
of 5 CaCl, was then poured on the loops, and a slight but distinct
quieting of the loops took place. The addition of 1 cc. ¥ CaCl,
caused the loops to become entirely motionless. After waiting a
considerable time, 1 c.c. % BaCl, was poured on the intestine. Im-
mediately violent nevietadee movements took place. Several c.c.
@ CaCl, exercised practically no inhibiting influence; while 2 c.c.
sas CaCl, solution suppressed the movements entirely for a short
ae
The question concerning the exact seat of action of the purgative
salts remains still unanswered. Whether, upon being absorbed into
the blood, they act on the central nervous system is not known.
There is no evidence to show that this is the case. It seems cer-
tain, on the other hand, from the experiments here described, that
they undoubtedly have a peripheral action either on the peripheral
nervous mechanism or on the muscle cells themselves. It is impos-
sible to prove that there is no action on the central nervous system,
and at present it seems impossible to prove whether the peripheral
action is directly on the nerves or on the muscles. The existence in
the walls of the intestine of the ganglionic plexuses of Auerbach and
Meissner must be taken into consideration; and with the methods
available there seems to be no way of distinguishing the action on
these plexuses and the direct action on the muscle cells. The ulti-
mate effect is on the muscles and glands; and the fact that an
entirely local ring-shaped constriction can be brought about by the
local application of a drop of one of the salt solutions to the surface
of the intestine would seem to indicate that only a small group of the
Effect of Saline Purgatives. 263
circular muscle fibres themselves is affected. The fact that the nerve
plexuses form a continuous network, and are intimately related in
their various parts, would suggest that the occurrence of an action
on these plexuses confined to so small an area is improbable. The
discussion of the exact location of the action is, however, of relatively
little importance, as compared with the main facts shown by these ex-
periments, namely, that z¢ zs possible to produce, by the local application
of a purgative salt to the serous surface of the intestine, a striking in-
crease in peristalsis, and to suppress these movements by a similar appli-
cation of solutions of calcium, magnesium, or strontium chloride.
Il. THe PropucTION oF INCREASED SECRETION OF FLUID INTO
THE INTESTINE BY THE SALINE PURGATIVES.
I have already stated! that, following the subcutaneous or intra-
venous administration of a saline purgative, a distinct increase in the
secretion of fluid into the intestine takes place. Although this in-
crease can be directly observed, a series of experiments had been
planned by which the relative quantities of fluid secreted into an
isolated loop of the intestine, before and after the administration of
a saline purgative, might be determined by actual measurements.
The experiments were interrupted by the summer vacation, and
could not be included in the previous paper. Since then, however,
I have been able to make these determinations with the following
results.
For these experiments rabbits and dogs were used. A loop of
considerable length was tied off, and a cannula inserted in its lower
end. In the rabbit, the upper part of the small intestine was used,
since in this region of the alimentary canal the secretion of fluid
seems to be most active. The upper ligature was placed in each
case below the entrance of the common bile duct, while the lower
ligature and the cannula were placed from twenty-five to thirty centi-
metres below. By gently lifting the successive loops, the fluid could
be easily drained through the cannula. This is made more easy by
placing the animal board at a considerable angle with the table.
After the loop was emptied, the cannula was closed by a clamp, and
the normal secretion allowed to collect in the loop for a period of ten
minutes. The contents were then removed and measured. This was
1 MAcCCALLuM, J. B.: This journal, 1903, x, p. IOI.
264 John Bruce MacCallum.
repeated for successive periods of ten minutes, and an estimate of the
normal rate of secretion was thus obtained in each case. The purga-
tive salt was then administered either subcutaneously or locally. In
the latter case, the solution was allowed to drop from a pipette upon
the loop. Care was taken to have the saline solutions at the body
temperature, and approximately isosmotic with the blood of the
animal. Ten minutes after the administration of the purgative, the
loop was again emptied and the contents measured. This was
repeated several times. In each case special care was taken to have
no interval between the emptying of the loop and the beginning of
the succeeding period of ten minutes. In other words, the loop was
always entirely empty at the beginning of each period. The opera-
tions were performed under morphine; the rabbits were given 5 c.c.
I per cent solution of the hydrochlorate subcutaneously, and the dogs
received the same dose of morphine, and, if necessary, also ether.
The results of the experiments may be seen from the following
reports:
1. Rabbit. Loop 30 cm. long, upper part of small intestine.
Loop contained in beginning’ ~~ . -. . = <= % |g) ))stmeueenencnemes
Fluid removed after rst 10 minutes . . . +. «= 95) 9. on
= = eles ne Me
2c.c. % BaCl, injected subcutaneously.
Fluid removed after 1st 10 minutes following injection . . 40 “
“e “ ce od ce ce ee ce g i 3-4 “ce
ee “ee és 3d “ “c ee “ce . F 3.0 “cc
In this rabbit the increased secretion of fluid was accompanied by
extremely active peristaltic movements. The feces could be seen
passing along the loops of the lower part of the intestine with great
rapidity. Within thirty minutes after the administration of the salt,
the passage of faces to the outside began. This continued for some
time, the faeces becoming constantly softer, until finally they were
almost entirely unformed. 3
As shown by this experiment, and also by the following ones, the
action of barium chloride persists for a considerable length of time.
The action of sodium citrate is more transitory.
1 In all these experiments there was no interval between the emptying of the
loop and the beginning of the ten minute period which followed. The injections
were made as rapidly as possible, and in no case occupied more than a minute.
baie,
——
Lifect of Saline Purgatives. 265
2. Rabbit. Loop 25 cm. long.
Loop contained in beginning 3.0 c.c. fluid deeply bile-stained.
Pek TSE TO MIMUCS: «3, 2 EO YS .
crete | (eh se: or iver —* * SEO
queso 10d
“(eau
tod
/SUIPID
*yuUd0 19g
OLC'S -
queso sad
ues 10d
BENE
SIDYIVID
1aqyng
ysoq
* (Aap
Ayyeried)
posodwoy
TN
272
“WOTJSNquIod Jo yeaTy UdSO1JIN
“SISATVNY GOO
al GLI OAIE
Results of a Smatl Increased Proterd Ingestion. 273
a time, at three’periods during the day, viz., at 9.30 A. M., 3.30 P.M.,
and 9.30 P.M. The crackers, beef (see below), and butter were pre-
pared in sufficient quantity at the beginning of the experiments, and
samples taken for analysis. The whole milk used was obtained each
day from the same cows, thus assuring uniform composition. The
milk was daily tested for fat by means of the Babcock test, and an
aliquot portion of each day’s supply was taken and mixed with similar
portions of the milk of the other days, thus making a composite
sample. The fat was determined in the composite sample, after
partial drying, by the ordinary extraction method. The nitrogen was
determined, by the Kjeldahl method, in each day’s fresh milk and also
TABLE II.
ANALYSIS OF FACES.
| Heat of combus-|_
tion per gram.
| Amount in
Le ate
Period. Subject. grams,
Nitrogen.
per cent small calories
July16-20 80.3 3.30 | 65 5487
lst milk
period 128.5 3.16 | : | 6018
lean
Meat period
26.3 595
July 20-24
2d milk
period
129 3.34
|
(
a
July 20 : 28.9 3:25 5 | 5678
82.6 3.53
in the composite sample. In Table I (page 272) will be found the
results of the analyses of the foods. From this table it will be seen
that the diet furnished 4.764 gms. of nitrogen per meal, or 14.292 gms.
per day, and yielded 915 large calories of energy per meal, or 2745
large calories per day.
Experimental periods and plan of proteid addition.— The uniform
diet was continued for four days, until the elimination of nitrogen was
fairly regular, as seen in Table III and Fig. 1 (pages 274 and 275
On the fifth day, July 20, a relatively small amount of extra nitrogen
was introduced into the morning meal. One hundred grams of milk
and 50 gms. of crackers were replaced by a nearly isodynamic amount
of proteid food in the form of very lean beef. The diet for that
one meal was: 50 gms. crackers, 20 gms. butter, 450 gms. milk, and
100 gms. lean beef. The beef furnished 5.20 gms. of nitrogen, and
274 P. B. Hawk and Joseph S. Chamberlain.
213 large calories of energy. The total nitrogen of this meal was
8.547 gms., and the energy furnished was 857 large calories. The
regular meal furnished 4.764 gms. nitrogen, and 915 large calories of
energy. The nitrogen was thus increased 3.783 gms., while the energy
supplied was decreased by only 58 large calories. At the following
meal, the regular diet was again used, and maintained throughout
the investigation.
FiGuRE 1.— The curves in Figure 1 represent the actual rate of excretion of nitrogen,
and sulphates for each subject. The ordinates represent the rate of excretion in
grams per hour, the abscissz, periods of time. The upper pair of curves represent
nitrogen, and the lower pair, sulphates. In the curves representing sulphates, the
ordinates are drawn to a scale five times greater than in the other curves. C.’s curve
is the full line. H.’s curve is the dotted line. * Sample lost.
In this way the experiments were divided into three periods:
I. First milk period: 6.30 A.M., July 16—6.30 a.m., July 20
(4 days).
Il. Meat period: 6.30 A. M., July 20— 6.30 A.M., July 21 (1 day).
III. Second milk period: 6.30 A.m., July 21 — 6.30 A. M., July 25
(4 days).
Feces and urine. — At the commencement of the investigation, and
also at the end of each separate period, charcoal, in gelatin capsules,
was taken, in order to facilitate the separation of the feces of one
period from those of the period following. The faeces were dried in
an air bath at 100° C., carefully weighed, and the nitrogen and heat
of combustion determined in the sample of each period.
The urine was analyzed for nitrogen, sulphates, and phosphates.
The samples of urine were collected in three hour periods during the
day, and a nine hour period at night, excepting on two days, July 20
and 24. During the preliminary days, three hour periods were con-
sidered satisfactory, the samples being taken at 9.30 A. M., 12.30 P. M.,
Results of a Small Increased Proterd Ingestion. 275
3.30 P. M., 6.30 P. M., 9.30 P.M., and 6.30 A.M. the next morning. On
July 20, the day on which the extra proteid was ingested, half periods
of one and one-half hours duration were used during the day, between
8.00 A.M. and 9.30 P.M., in order to secure more exact information in
TABLE III.
ANALYSIS OF COMPOSITE URINE.
Amount
of om Nitrogen. P.O. SOs.
grams.
| per cent grams | percent | grams percent |
July 16 | 641.5 1.52 9.76 0.241 1.540 0.271
17 739.0 | 1. 11.58 0.299 2.210 0.258
136 || 885.0 12.10 | 0.276 | 2445 | 0.209
19 BS 11.69 | 0.253 2.461 0.189
20° | | |) 15:28 0.288 3.349 | 0.199
21 | lee lke | 14332) 9|) 0!286 2.890 0.204
| | 130252 O57 2.509 0.205
13.79 | 0.248 | 2756 | 0.176
11.84 | 0.248 | 2.406 | 0.203
10.61 | 0.200 1.720 0.209
166, 1" 0/279" ||) 3905 | 0 271
W222 0.325 2.248 | 0.270
12.76 0.287 2.205 0.251
15.45 0.310 2.700 | 0.280
13.85 0.319 2.340 | 0.279
11.82 0.298 2.120 0.279
11.66 0.347 2219 0.272
11.96 | 0.316 | 2.330 | 0.243
| | |
regard to the time interval between the ingestion of proteid food and
the maximum rate of excretion following it. Then in order to secure
a normal day of half periods to compare with the day of increased
proteid ingestion, the short periods were again used on July 24, after
the nitrogen excretion had again reached a normal uniform rate.
276 P. B. Hawk and Joseph S. Chamberlain.
The urine was placed in a refrigerator for a definite time, to cool to a
uniform temperature, after which it was weighed and aliquoted for the
composite sample. Analyses were then made of the original sample
as soon as possible, and at the end of the day the composite sample
TABOR ive
URINE VOLUMES.
Period.
was analyzed. The heat of combustion was determined in the
composite sample.
Analytical methods. — The methods of analysis were those in com-
mon use in this laboratory.!. The nitrogen and phosphate determina-
tions were all duplicated, and the totals checked with the analysis of
the composite. The sulphate analyses were not made in duplicate,
but were checked with the composite sample as in the other cases.
1 See SHERMAN and HAWKE: Loc. cit.; Hawk: Loc. cit.
Results of a Small Increased Proterd Ingestion. 277
IV. RESULTS.
In Table III, page 275, will be found the data for the analysis of
the composite urine samples of each subject, and in Table IV, page
276, are given the urine volumes for each period of the several days.
Tables V-IX, inclusive, on pages 279, 281, 283, 284, and 285, re-
spectively, give tabular data regarding the nitrogen, sulphate, and
FicureE 2.— In Fig. 2 the upper pair of curves represent the actual rate of excretion of
phosphates for each subject. The ordinates represent the rate of excretion in grams
per hour, the abscissz, periods of time. The one-day curves on the 20th and 24th
represent the rate of excretion of, nitrogen, sulphates, and phosphates as measured by
three hour periods, the same as used on all the other days. The regular curves rep-
resent, o7 these days only, periods of one and one-half hours.
The ordinates of the curves for sulphates and phosphates are drawn to a scale five
times as great as those of nitrogen.
phosphate excretions, and Table X, page 286, shows the relation be-
tween the nitrogen content and heat of combustion of the urine.
In Figs. 1 and 2, pages 274 and 277, the excretion in grams is
represented graphically. In these figures the curves are in pairs, in
order that the similarity in the course of excretion of the two subjects
may be observed.
In Fig. 3, page 280, and Fig. 4, page 282, the graphic representa-
tion of the course of excretion for each subject is shown by itself,
in order to bring out more clearly the relations between the three
substances. .
In Fig. 5, page 287, the average rate of excretion of the two sub-
jects is represented as in Figs. 3 and 4.
278 P. B. Hawk and Joseph S. Chamberlain.
Elimination of nitrogen. — From Table V and Fig. 1, it will be seen
that the excretion of nitrogen during the days July 17, 18, and 19,
was fairly uniform in both cases, and that this same regularity is again
attained to a certain degree on July 22, two days after the proteid
ingestion. On the day when the extra proteid food was ingested at
the morning meal, the rate of excretion of nitrogen rose rapidly, even
during the first short period after the meal. The maximum rate of
excretion was reached in the case of both subjects during the third
short period, that is, from three to four and one-half hours after
ingestion.
Sherman and Hawk (/oc. c7t.) found that the maximum excretion
of nitrogen occurred in from six to nine hours after ingestion, whereas
Hawk (/oc. cit.) found the maximum rate of excretion in the case
of one subject to occur in from six to nine hours, and in the case of
the other subject in from nine to twelve hours. The occurrence
of the maximum rate of excretion at an earlier hour was no doubt
due in part to the relatively small increase in the ingested proteid,
as well as to the use of shorter periods for the collection of the
urine.
By an examination of Table V (page 279) or Fig. 1 (page 274), it
will be seen that in the case of H., if the collections of urine had been
made in three hour periods on the 20th as on the other days, and as
in the investigations previously mentioned, the maximum rate of excre-
tion would have fallen in the period from 12.30 P.M. to 3.30 P. M., Six
to nine hours after the ingestion, instead of between 9.30 and I1.00 a. M.
In each case after the maximum was reached, there were two other
rises during the day, and at night the rate of excretion was still high.
The maxima for each subject occurred in the third, sixth, and tenth
periods, or from three to four and one-half hours, seven and one-half
to nine hours, and thirteen and one-half to fifteen hours after the in-
gestion. This seems to agree more nearly with the maxima found by
Falck, and also with those of Voit, except that in the latter case the
first two maxima are advanced about three hours in each case.
The total amount of nitrogen eliminated in the urine, by C. and H.,
was almost the same for the two days, the 20th and 21st. During
the three following days, the rate of excretion, in the case of C., was
much lower than on the 2oth, although considerably higher than
before the extra proteid ingestion, and it did not regain the normal
until the fourth day following the increased excretion. In the case
of H., however, the first day after the ingestion was the only one in
279
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280 P. B. Hawk and Joseph S. Chamberlain.
which excretion remained above normal, and the uniform rate was
regained on the 22d. Thus in the case of C., the increase in the
excretion of nitrogen after the ingestion of extra proteid food took
place, in general, as the previous work from this laboratory has
shown; about 80 per cent of the extra nitrogen ingested being ex-
creted during that day, and the maximum rate of excretion occurred
shortly after the ingestion, while a marked increase in the rate of
excretion was observed for several days. In the case of H., about
FicurE 3.— In Figs. 3, 4, and 5, the curves represent variations in the rate of excretion
of nitrogen, sulphates, and phosphates expressed in per cents of an assumed average
rate of excretion, viz.: 0.5 gram per hour for nitrogen and 0.1 gram per hour for
sulphates and phosphates.
The ordinates represent per cent, and the abscissz, periods of time. The curves
representing nitrogen and sulphates are drawn together for comparison, nitrogen
being the full line, and sulphates the dotted. The curve for phosphates is the full
2
line at the bottom. Fig. 3 shows the curve for C. Fig. 4 shows the curve for H.
Fig. 5 shows the curve representing the average rate of excretion of C. and H.
the same per cent of the extra nitrogen ingested was excreted during
that day, but on the day following, viz.: the 21st, and practically dur-
ing the first half of it, the greater part of the extra nitrogen was elim-
inated and the rate of excretion fell suddenly to the normal.
By consulting Table IX (page 285), in which the nitrogen balance
is given, it will be seen that, during the second and third days after
the extra ingestion, C. was metabolizing a larger per cent of the nitro-
gen ingested than was H., if we may measure the relative amounts of
nitrogen metabolized by the amounts excreted in the urine. This fact
may account for the difference in time between H. and C. in regain-
281
Results of a Small Increased Proterd L[ngestion.
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282 P.B. Hawk and Joseph S. Chamberlain.
ing the normal rate of nitrogen excretion, H. regaining it quickly, near
the close of the 21st, and C. not until two days later.
Elimination of sulphates. — Comparing the excretion of sulphates
with that of nitrogen for each subject, as shown in Figs. 3, 4, 5 (pages
280, 282, and 287), it will be seen that the rate of excretion of sulphates
followed more closely that of nitrogen than did the rate of excretion
of phosphates. There is, however, not an exact parallelism. The
daily maximum of the nitrogen excretion was usually in the second
or third period, while the maximum rate of excretion of sulphates
FIGURE 4.— See explanation of Fig. 3. + Sample lost.
usually appeared in the fourth or fifth. Also, while the rate of nitro-
gen excretion rose immediately after the morning meal, that of the
sulphate excretion fell. Thus the maximum rate of sulphate excre-
tion for the day on which the extra proteid food was ingested oc-
curred with C. during the fifth long (three hour) period, and with
H. during the third. In the case of H., the rate of excretion of sul-
phates followed, in general, that of nitrogen on the day following the
extra ingestion, and fell to the normal one day later. With C., the
rate of excretion of sulphates regained its normal after four days, as
was also true of the nitrogen. In this point the rate of excretion of
sulphates was contrary to that observed by Garrett in regard to re-
gaining its normal quicker than nitrogen and reaching its maximum
earlier.
In Table VII (page 283) the ratio of nitrogen to sulphates is
1 GARRETT: Journal of physiology, 1898, xxiii, p. 150.
Results of a Small Increased Proterd Ingestion. 283
shown. It will be noticed that in the case of C. the ratio is notice-
ably lower on the day of the increased proteid ingestion than on any
of the other days. With H., however, the ratio on that day is an
approximate average of the entire period, and the lowest ratio is on
the following day.
Elimination of phosphates.—In the investigations already cited,
previously carried out in this laboratory, only one rise and fall per
TABLE VII.
RATIO OF NITROGEN TO SULPHATES.
Date, 1900. oe : i Average.
July 16 | TOO IGS 100 : 15.4
17 100 : 15. | 100 :
18 : 15. 100 : 14. | 100 :
19 : 15. 100 : 14. | 100 :
20 ga 100 : 15. | 100 :
100 : 14. 100 :
LOO Me. cc 100: .
100 : 15. 100 :
100 : 15. | 100 :
1 Sample lost.
day in the rate of excretion of phosphates was observed in one case,
while in the other, in nearly every instance, two distinct rises were
recorded.
In the present investigation, however, with both subjects, two such
daily maxima were observed, and on the days when short periods were
used there were three. These variations in the rate of excretion were
more marked in the case of phosphates than with nitrogen or sulphates,
and were also more regular. Especially prominent, as shown by the
graphic representations, Figs. 3, 4, 5 (pages 280, 282, and 287), was
the fall in the rate of excretion during the first morning period imme-
diately following the morning meal. On comparison with the days of
short periods, it will be seen that the greater part of this fall in the
rate of excretion occurred during the first hour and a half. In most
P. BL. Hawk and Joseph S. Chamberlain.
284
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Results of a Small Increased Proterd Ingestion. . 285
cases, the daily maximum occurred during the third period, but some-
times during the fifth. In the excretion of C., the rate usually fell
regularly from the fifth period, and continued during the night, the
lowest point occurring during the first period of the morning. With
. TABLE IX
NITROGEN BALANCE.
N
Date, 1900. | in food.
N
in feces.
N
in urine.
Gain or loss
(Con):
July 16
17
18
19
20
21
22
23
24
otal.
Avg. per day
14.29
14:29
14.29
14.29
18.07
14.29
eee)
14.29
14:29
0.663
0.663
0.663
0.663
1.306
0.518
10.240
11.955
12.520
11.920
15.480
14.400
‘13.130
13.760
11.872
1S 277
July 16
Sealy
18
19
20
21
2,
23
24
+3.38
+1.67
sell
+1.71
+1.284
—0.628
+0 642
+0.012
+1.900
+11.083
+1.231
+2.520
+1.450
+0.845
+0.485
+1.004
—0.534
+1.436
+1.856
+1.637
Total .
Avg. per day
+10.699
and Joseph S. Chamberlain.
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Results of a Small Increased Proterd Ingestion. 287
H., however, the fall began somewhat later, but reached its point of
minimum excretion at the same hour, viz.; during the first period of
the day.
The maximum rate of excretion on the day of the increased proteid
ingestion was reached during the fifth short period or third long one,
one period later than the nitrogen maximum, and earlier than that of
the sulphates. The normal was regained with each subject at almost
exactly the same time as in the case of the nitrogen.
Nitrogen balance. — Table IX (page 285) gives the nitrogen balance
of the experiments. From an examination of the income and outgo
FIGURE 5.— See explanation of Fig. 3. * Sample lost.
of nitrogen, we see that just preceding the ingestion of extra proteid
food, both C. and H. were storing nitrogen in about equal amounts.
On the three days following the ingestion, C. was approximately in a
condition of nitrogen equilibrium, while H. was storing a small quan-
tity. During the whole experiment, however, each subject averaged
about the same gain of nitrogen. If now we recall the fact that, in
the course of the experiment, H. lost 300 gms. in weight, while C.
gained 100 gms., we see that the subjects were not similarly affected
by the diet maintained.
Heat of combustion and nitrogen content of urine. — The ratio be-
tween the nitrogen content of the urine and the heat of combustion
of its unoxidized material was found to vary between I : 10.2 on normal
days, and 1: 8.7 on the day of the increased proteid ingestion. These
ratios are somewhat higher than others determined in this laboratory
288 P. B. Hawk and Joseph S. Chamberlain.
under similar conditions of diet, but in every instance the charac-
teristic minimum ratio, on the day when the extra proteid was eaten,
was observed. (Table X, page 286.)
V. SUMMARY.
1. The rate of excretion of nitrogen, measured by three hour
periods, showed two maxima daily. The rate rose from the morn-
ing meal until midday or a little later, then fell, and rose again about
the time of the evening meal. The real maximum rate was usually
that of the midday rise, and’ the minimum rate of excretion took
place during the night. Measured by shorter periods, however, three
maxima were observed, the real maximum rate coming at a slightly
earlier period, and the third rise between this and the evening rise.
2. The rate of excretion of sulphates followed in general a parallel
course to that of nitrogen, the main difference being that the mini-
mum rate of excretion was reached after the morning meal, and the
maximum late in the. afternoon. Frequently three maxima were
observed on normal days with three hour periods.
3. The phosphates differed decidedly in their rate of excretion
from either the nitrogen or the sulphates. Two very distinct rises
were shown each day, and in one instance, that of H. on July 20, an
indication of a third rise was seen. The maximum rate of excretion
was reached after the midday meal, and was usually the first of the
two rises. With one subject, C., the rate of excretion fell from about
the fifteenth hour through the night, reaching the minimum during
the first period of the morning. With H., the fall in the rate of ex-
cretion was later in beginning, but resembled the excretion of C., in
reaching the minimum point in the first period of the day.
4. After the ingestion of a small extra amount of proteid food at
the morning meal, the rate of excretion of nitrogen reached its maxi-
mum within three to four and one-half hours, after which it fell to its
normal rate; in one case slowly, after four days, and in the other
rapidly, after two days. It would seem that the time required to
reach the maximum excretion of nitrogen, after increasing the pro-
teid of a diet, was more or less directly proportional to the amount
of proteid ingested, the length of time being greatest when the quan-
tity was large.
5. With each subject the maximum rate of sulphate excretion dif-
Results of a Small Lucreased Protecd Ingestion. 289
fered from that of nitrogen only in reaching its highest point about
six hours later.
6. In one subject the ratio of nitrogen to sulphates was lowest on
the day of increased proteid ingestion; in the other, on the day after
this ingestion.
7, The maximum rate of phosphate excretion due to the increased
proteid ingestion fell in a period between those in which the maxima
of nitrogen and sulphate occurred.
8. In the case of H., the normal rate of nitrogen and phosphate
excretion was regained on the second day following the increased
proteid ingestion; the normal rate of sulphate excretion was regained
one day later, z.¢, three days after the ingestion. With C., the
normal rate of excretion was not regained in any case until the
fourth day following the increase in proteid food.
g.. The ratio between the heat of combustion of the urine and its
nitrogen content was lower on the day of increased proteid ingestion
than on normal days.
THE RELATION BETWEEN SOLUTION. f232e,
ATOMIC VOLUME, AND THE PHYSIOLOGiIG#s=
ACTION: OF “THE -ELEMENa:
By ALBERT P. MATHEWS.
[Hrom the Hull Physiological Laboratories of the University of Chicago, and the Marine
Biological Laboratory, Woods Holl.)
THE RELATION BETWEEN SOLUTION TENSION AND PHYSIOLOGICAL
ACTION.
IY Gin attempts have been made to correlate the physiological
action of the elements with their physical or chemical proper-
ties, but with only partial success. Thus several writers! have
established with certain elements and compounds a relationship
between physiological action and atomic or molecular weight. Many
higher alcohols, for example, are more poisonous than ethyl or methyl
alcohol; the sodium salts of iodine are more poisonous, molecule for
molecule, than the corresponding but lighter chlorides. Unfortunately,
however, this rule of increase of action with an increase of molecular
or atomic weight is found to have so many exceptions as to indicate
that the relationship is not a simple one. To cite two examples, it is
well known that strontium or barium are for most forms of protoplasm
less poisonous than the corresponding but lighter metal calcium.
Upon the nerve, calcium acts many times as powerfully in inhibiting
the action of sodium chloride, as either magnesium or strontium. It
is well known besides that the same element will, under different con-
ditions, exhibit very different physiological actions, while its atomic
weight does not vary. Ferric salts are more active than ferrous salts.
Copper is poisonous for many plants in minute amounts, and, as a
rule, far exceeds in action the heavier metal lead.
The attempt to find a connection between physical properties and
physiological action was given a great impetus by the ionic hypothe-
* BLAKE: Chemical news, 1887, lv; Proceedings of the Royal Society, London,
1841; Journal of physiology, 1884, v, p. 35; GruTzNeR: Archiv fiir die gesammte
Physiologie, 1893, lii, p. 83.
290
Solution Tension, etc., of the Elements. 291
sis. It has been shown by the work of Kahlenberg and True, Kronig
and Paul,! and other botanists, and by Loeb and other animal physi-
ologists, that the metals are most active when in an ionic form.
Thus, silver as an ion is more poisonous than silver in a complex ion,
such as a silver-cyanogen ion, or in combination with albumin. These
and many other facts which are well known led to the hypothesis
which has been particularly developed by Loeb and the writer in this
country, and by Pauli and others abroad, that the elements acted
principally by means of their free electrical charges, although some
action has been ascribed to the atoms themselves. Loeb and the
writer have particularly emphasized the importance of the number of
the charges, or valence of the ion in determining its physiological
action. The chief difficulty in the way of ascribing action to the
number of charges, is the fact that some monovalent ions, like silver,
are enormously active, while some trivalent ions, like aluminium, are
relatively inert. These facts led me to propose the alternative hypoth-
esis that the movement of the charges was the main cause of the
physiological action of ions, and that the different action of two mon-
ovalent ions was due primarily to the fact that the movement of the
charge varied either as regards its path or speed of rotation about
the atom, and I attempted to bring this conception into line with
the spectra of the elements.
As this relationship is a difficult one to follow, I have examined a
suggestion made two years ago by Professor J. Stieglitz of this univer-
sity, that the difference between the action of bivalent and trivalent
cations and anions described by Loeb and myself might be due not
to the difference in number of the charges, so much as to the ease with
which the charge is given up from the atom, a trivalent ion like the
ferric ion being more efficient physiologically, for the reason that one
or more of its charges are given up more easily than are the charges
in the bivalent condition. The importance of this factor for other
properties of salts had been indicated by Bodlander. That this idea
might be fruitful, was indicated by the table of solution tensions of
the elements, as given by Nernst? and Wilsmore.?
1 KAHLENBERG and TRUE: Botanical gazette, 1896, xxii, p. 91; KAHLENBERG
and Austin: Journal of physical chemistry, 1900, iv, p. 553; KrONnIG und PAUL:
Zeitschrift fiir Hygiene, 1897, xxv, p. 1; PaAuLi: Hoffmeister’s Beitrage zur
chemischen Physiologie, 1902, iii, p. 225; LoeB: This journal, 1902, vi, p. 411;
MATHEWS: Science, 1902, xv, pp. 492-498; /bzd., 1903, xvii, p. 436.
2 NERNST: Theoretische Chemie, 3d ed., 1901, p. 370.
3 WILSMORE: Zeitschrift fiir physikalische Chemie, 1901, xxxvi, p. 92.
292 Albert P. Mathews.
In a preliminary paper,! it has already been indicated that such a
relationship between the solution tension of the metal and its poi-
sonous action exists for the motor nerve. In support of such a rela-
tionship there was the following reasoning: it is conceivable that the
elements act on protoplasm, either by giving up their electric charges
or valencies to compounds in the protoplasm, or by taking charges
away from these compounds. For example, ferric iron, as is well
known, has a strong affinity for,a negative charge; it gives up its
positive charge and goes very readily into the ferrous state. Now if
a ferric ion comes in contact with a molecule which has a negative
charge held less firmly than the ion can hold it, the ferric ion will
seize that electric charge for itself, and thereby bring about the same
kind of a physical or chemical change in the protoplasm as would be
produced by an electrolysis.2 The complex protoplasmic molecule,
thus deprived of its charge, would necessarily undergo a rearrange-
ment of equilibrium, in the course of which new compounds would
appear, energy be liberated, and protoplasmic movements produced.
It is easy to see that the power of any element to produce such
a change in protoplasm might depend on its affinity for its positive
or negative charge, leaving for the present undetermined the cause of
that variation in affinity. Potassium or sodium, for example, having
a great affinity for their positive charges, give these up with great
difficulty ; they would leave the negative charges on the protoplasm
unaffected, because sodium and potassium ions, unless they are
present in great numbers, have a lower affinity for such charges than
has the protoplasm.
There was, therefore, good reason for thinking that the physi-
ological action could thus be measured by the affinity of the element
for its charge, those elements, as already said, being relatively indif-
ferent which hold their charges most firmly. This affinity may be
measured electrolytically, or it may be computed approximately from
the heat of formation of salts. For example, the tension which must
exist on the electrodes to separate the elements from aqueous solu-
tions of their salts, may be measured. Such measurements show,
in equivalent solutions of completely ionized salts, that it requires a
much higher tension to deposit sodium or potassium than to deposit
1 MATHEWS: Science, 1903, xvii, p. 436.
2 This action of a ferric ion may be seen if a little ferric chloride is added to
a solution of potassium iodide. The ferric ion takes the negative charge from
the iodine.
ivy
Solution Tension, etc., of the Elements. 293
mercury or silver. The greater the affinity of the element for its
ionic charge, obviously the greater voltage will it take to separate
that charge from it. This measure is also a measure of the solution
tension; that is, of the tendency of the elements to go into solution,
and thus to acquire a charge. The following table, taken from
Wilsmore,! shows the relative solution tensions of different elements
for normal ionic solutions. The figures in parentheses have been
computed by me from the heat of formation of the salt by the formula
arr = volts, where g is the heat of formation of the salt.
TABLE, I:
SOLUIION TENSIONS IN VOLTS FOR NORMAL SOLUTIONS.2
kK "+ 2.92 Co — 0.045 r= 0797
Nal -- 2.04 Ni — 0.049 Br — 1.270
Rb + (2.54)(?) Sn <— 0.085 Cl — 1694
Ba 4 2204 Pb — 0.129 O — 1.396
Sr + 2.49 H — 0.277 Fl (—2.24)
Ca -— 2:28 Cu — 0.606
Li + (2.369) As <--0.570
ee 2.26 Bi <— 0.668
1.214(?) Sb <— 0.743
Al + 0.999 Hg — ‘1.027
Mn + 0.798 Ag — 1.048
r Zn + 0.493 Pd <— 1.066
Cd + 0.143 Pt <—1.140
Be + 0:063 Au <—1.356
We. se (OAS
An inspection of the foregoing’ table is sufficient to show that the
elements when arranged according to their solution tensions are also
arranged approximately according to their poisonous action. Thus
we have at one end of the list the extremely poisonous metals, copper,
silver, gold, mercury, and platinum; and at the other end, sodium,
potassium, strontium, and barium, which are relatively inert. Arsenic,
1 WILSMORE: Zeitschrift fiir physikalische Chemie, 1901, xxxvi, p. 92.
2 While these figures do not measure the absolute affinity of the charge for the
atom, except under certain theoretical conditions, they may be used for compara-
tive purposes. The solution tension of oxygen is that of oxygen in acid solutions.
294 Albert P. Mathews.
antimony, and bismuth occupy their proper positions as poisonous
elements. Beyond gold, we have the negative elements, iodine,
bromine, and chlorine, which become less poisonous as their solution
tension or affinity for the negative charge increases. The position of
hydrogen, which by every other classification is out of place, is here
where it should be, near lead and copper. I have already compared
this table with the inhibitory and stimulatory effects of salts on the
nerve, and showed that the inhibitory effect of the cations and the
stimulating action of the anions increased as their solution tensions
diminished. A more complete discussion of this relationship will soon
be published. At Woods Holl during the summer I was able to get
much more readily the data of the physiological efficiency of the salts
from a study of the action of salts on the eggs of the fish Fundulus
heteroclitus. The eggs of this fish are peculiarly adapted to this
work, for the reason that they appear to be easily penetrated by nearly
all sorts of ions, and they are quite insensitive to variations in osmotic
pressure of the solutions. I determined the least concentration of
the various salts which would just prevent the formation of the
embryo as a visible line on the egg, that is, a concentration which
would kill in about thirty-six hours. The end point, however, is not
very sharp. I have confirmed these observations by following the
embryos, and determining the concentration which will just prevent
pigment formation, or which will kill in about sixty-four hours. The
results of the two series were on the whole harmonious. The results
here given represent the mean of a large number of experiments,
which agreed very well with one another. The eggs were fertilized
in sea-water, and then allowed to remain there for from two to four
hours, until they were in a 2-8 cell stage. They were then repeatedly
washed in fresh and distilled water, and allowed to remain for one
hour in a large amount of distilled water, so that the salts already in
the egg! could dissolve out. About twenty eggs were then transferred
with a sieve, so as to carry over little water, to 100 c.c. of the various
solutions in finger-bowls. It is important to have the eggs well
scattered, not bunched, and to have not more than fifty eggs in this
quantity of solution. The room temperature varied from 19° to
26° C. No corrections have been made for temperature, beyond
repeating the experiment at different times and taking the means.
Observation showed, however, that a small increase in temperature
1 A determination of chlorine in eggs so washed, showed, however, that a con-
siderable quantity of chlorine remained in the egg.
Solution Tension, etc., of the Elements.
TABLE II.
Salt i aria | la coal ere
1. AIC], mn 52 11 ||PbC,(H3O.)e) 1355
(continued)
|| aotnect m+ 18 0
"2 100 | 95% |\ fs m 3 31 1
7 | © Seo | 50 ake | ce mz 26 5
1 The ferrous chloride solution was used immediately after making it up. On
standing two days or more, its activity increases. The salt was a very good
Kahlbaum preparation.
Solution Tension, etc. of the Elements. 297
TABLE II — (concluded).
| i]
| No. of | No. of i No. of | No. of
i eggs. jembryos.
eggs. jembryos.
| | |
|
MgCl, (cont.) 40
30
20
25
65
34
36
increases the poisonous action markedly. I hope next summer to
secure the data necessary to calculate the temperature coefficient.
Tables II and III give the concentration in fractions of molecular
and in some instances normal solutions, which will just prevent the
formation of embryos at a temperature of about 20° C. Most of
these salts have been tested several times. The experiments quoted
will indicate fairly the character of the results.
It will be seen from the foregoing summary that the chlorides of
the metals arrange themselves in nearly the same order, so far as
their poisonous action goes, as they are arranged by their solution
tension, and we may, therefore, express this fact in the following
statement:
The physiological (potsonous) action of any cation varies inversely
with its solution tension. —It will be noticed, however, that there is
298 Albert P. Mathews.
an enormously greater difference between the poisonous action of
mercury and potassium than might have been expected from their
difference. in solution tension. Furthermore, a difference of one-
tenth of a volt near the end of the table determines a greater differ-
ence in physiological action than a corresponding difference near the
top of the table. This difference is perhaps to be expected, since a
difference of one-tenth of a volt when the tension is already low, is
greater proportionally than when the tension is high.
Certain exceptions to the general law will be perceived. For
example, cadmium is nearly as poisonous as copper, whereas its
TABLE III.
SUMMARY. UNCORRECTED FOR DISSOCIATION.
Least Least Least
poisonous poisonous poisonous
dose. dose. dose.
AIC fy ak HCl
MnCl, | | Cue,
ZnCl, st | AuCls
CdCl, [eee | HeCl,
FeCl, | tt AgNO,
CoCl, 3 | FeCl,
NiCl,
Pb(C3H30.)o |
solution tension brings it above iron. Ferrous salts are less poison-
ous than one would expect from the table. Zinc is somewhat out
of place, being more poisonous than it should be, and lead, instead
of just preceding hydrogen, comes just after it. Gold, again, from
the few observations made upon it, was found to be not quite so
poisonous as mercury, although it ought to be more poisonous.
These exceptions are to be explained in part by the fact that the
different charges on a polyvalent ion are not equivalent, that is, they
have different solution tensions, and in part, no doubt, they are to
be explained by variations in dissociation. The table just quoted
probably gives at the best a mean value for the different charges.
Consider, for example, the difference between ferric and ferrous
,
Solution Tension, etc. of the Elements. 299
ions. The ferric ion is more poisonous, equivalent for equivalent,
than is hydrogen. This is to be explained by the fact that the
third positive charge of the ferric ion leaves the atom with ex-
treme ease. The solution tension for the charge is not, there-
fore, represented by the figure 0.063, but by a figure more nearly
that of copper; in fact, by the figure —o.314 volts, as calculated
from the heat of combination of ferric chloride. The ferrous ion is
less poisonous, for the reason that the solution tension of the two
remaining charges is higher than that given in the table, being
approximately 0.076. A ferric ion has, therefore, a much greater
affinity for a negative charge than has a ferrous ion. This explana-
tion is confirmed by observing the action of aluminium. The three
charges on this element are more nearly of equal value, as aluminium
does not form two series of salts, like the ferrous and the ferric salts,
and it occupies its proper place in the table of poisonous action.!
As regards zinc, this element is probably rendered somewhat more
poisonous than its position indicates, by the fact that it forms so
many free hydrogen ions in its solution. The hydrogen, having
a low solution tension, increases the poisonous action of the zinc
chloride and sulphate. Cadmium is a marked exception. It is
nearly as poisonous as copper, and this is the result obtained also by
Kahlenberg and True? for seedlings, and by other authors.® It is not
impossible that the cadmium chloride examined had a trace of silver
or mercury in it, but as the difference is so marked, and as it corre-
sponds to the figures of Kahlenberg and True, I am inclined to
believe that it is the cadmium itself which is an exception. The
explanation of this action is, I believe, the same as in the case of
iron. Cadmium has a marked tendency to form double and complex
ions, in this respect resembling mercury. The tendency to form such
ions has been shown by Abegg and Bodlander* to be correlated in
the case of any element with a low solution tension. The fact that
cadmium forms these double compounds indicates, therefore, that
one of its charges has a lower solution tension than the Table shows.
The same explanation will hold for lead, but the lead charges are
more nearly equal than are those of cadmium or iron. That this
1 As is shown farther on, aluminium is prevented from exercising its full poison-
ous action by the protection afforded by the egg envelopes.
2 KAHLENBERG and TRUE: Loe. cit.
8 KrONIG and PAUL: Zeitschrift fiir Hygiene, 1897, xxv, p. I.
* ABEGG and BoLANDER: Zeitschrift fiir anorganische Chemie, 1899, xx, p- 453.
300 Albert P. Mathews.
is at least an explanation of some of these exceptions, is indicated
also by a study of the anions.!
TeABEB sive
SOLUTION TENSION OF ANIONS.
Cy [—0.014] ; [—0.109] ClO, [—1.138]
Oxalic ‘ :
OH —1.157 [—4.105 ] CnS_ [—0.83] (?)
I —0.797 ‘ —1.694(?) NO; —2.229
Acetic
Br —].270 ' [—4.682] BrOg [—0.727]
Cl —1.694 Sulphuric —2.177(?) IO3 [—2.536](?)
O —0.557 Fl —2.24(?)
The figures given for the anions show the relative affinity they
possess for the positive charge; oxygen, for example, having in an
alkaline solution a much lower solution tension as regards the nega-
tive charge than iodine or chlorine. The following table gives the
least poisonous dose of some salts of sodium and potassium:
TABLE V.
Least fatal }| Anions arranged in the
dose. order of poisonous action.
Least fatal
NaOH !
KOH
} Ba(OH),
NaCl
Nabr
KI
Na B rOs
KBrO,
NaF
Na,SO,
Na acetate
NaNOsz
KNO,
KCyS
NagC.0,4
K,C,0,4
KC1O3
NaHCOg
CaClOs
Naz citrate rio 2
> a tj
K3FeCy,