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The metabolism of the hexitols

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THE METABOLISM OF THE HEXITOLS
A Thesis
Presented to
the Faculty of the Department of Biochemistry
University of Southern California
School of Medicine
In Partial Fulfillment
of the Requirements for the Degree
Master of Science
by
Cornelia Hendrick Johnston
May 1941
UMI Number: EP41275
All rights reserved
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UMI
" OSsssrtatton Pjbfeh'ng
UMI EP41275
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T h i s thesis, w r i t t e n by
.........................C m N ELIA ..H EIfflSIC K ..JQ H IST.Q N ..............
u n d e r the d i r e c t i o n o f h
F a c u l t y C o m m it t e e ,
a n d a p p r o v e d by a l l it s m e m b e r s , has been
presented to a n d accepted by the C o u n c i l on
G r a d u a t e S t u d y a n d R esearch in p a r t i a l f u l f i l l ­
m e n t o f the r e q u ire m e n ts f o r the degree o f
-MASTEK~GE~SC.imC.E~
D ean
Secretary
1241
F a c u lty Com m ittee
/J
Chairman
The writer gratefully'acknowledges her indebtedness
to Dr. H.J. Deuel, Jr., for his guidance, and to him and
Miss-Lois Hallman for their generous assistance in carrying
out the laboratory work on which this thesis is based.
TABLE OP CONTENTS
CHAPTER
I.
II.
III.-
PAGE
I N T R O D U C T I O N ............ ......................
1
..........
LITERATURE
EXPERIMENTAL
4
............................
12
Experiments on the ketolytic action of
the hexitols
...........................
12
Experimental results of the ketolytic
action of thehexitols,
...............
Statistical treatment of results
Experiments on glycogen formation
.
...........
15
16
24
Experimental results on the glycogenforming ability of sorbitol, mannitol,
and dulcitol
■ IV.
¥.
DISCUSSION
SUMMARY
..................
...............................
27
. .
................................ . . .
B I B L I O G R A P H Y ...................................
42
4849-
LIST OF TABLES
TABLE
I.
PAG-E
Ketone Bodies in Urine of
Rats Receiving Sorbitol
II.
Ketone Bodies in Urine of
Fasting Female
. .
..........
Fasting Female
Rats Receiving M a n n i t o l ..................
III.
Ketone Bodies in Urine of
Ketone Bodies in Urine of
22
Summary of the Effects of the Hexitols on
K e t o n u r i a .................................
VII.
21
Ketone Bodies in Urine of Fasting Female
Rats Receiving Sodium Chloride ............
VI.
20
Fasting Female
Rats Receiving Glucose . . . . . . . . . . .
V.
19
Fasting Female
Rats Receiving D u l c i t o l ...........
IV.
18
23
The Liver Glycogen in Pasting Male Rats
after the Intraperitoneal Injection of
Different Concentrations of Sorbitol at
Different Time Intervals . . . . . . . . .
VIII.
125
The Liver and Muscle Glycogen in Fasting
Male Rats 6 Hours after, the Intra­
peritoneal Injection of a 25 per cent
Solution of Sorbitol, 1 cc. per 100
sq. cm.
.................................
29
iv
TABLE
ix.
PAGE
The Liver,, and Muscle Glycogen in Blasting
Male Rats 6 Hours after the Intraperi toneal Injection of a 25 per cent
Solution of M'annitol, 1 cc. per 1QQ
sq.
x.
cm,
......................
30
The Liver and Muscle Glycogen in Fasting
Male Rats 6 Hours after the Intraperitoneal Injection of a 25 per cent
Solution of Glucose, 1 cc. per 100
sq.
XI,
c m ..........................................
31
The Liver and Muscle Glycogen in Fasting
Male Rats 6 Hours after the Intraperitoneal Injection of Sodium Chloride,
1 cc. per 100 sq. cm.
XII.
......................
32
The Liver and Muscle Glycogen in Fasting
Female Rats 6 Hours after the Intraperi toneal injection of a 25 per cent
Solution of. Sorbitol, 0.5 cc. per 100
sq.
cm.
.....................
33
TABLE
XIII.
The Liver and Muscle Glycogen in Pasting
Female Rats 6 Hours after the Intraperi toneal Injection of a 25 per cent
Solution of Mannitol, 0.5 cc. per 100
sq. cm.
XIV.
. . . . . . .
....................
The Liver and Muscle Glycogen in Fasting
Female Rats 6 Hours after the Intraperi toneal Injection of a 25 per cent
Solution of Glucose, 0.5 cc. per 100
sq. cm.......................................
XV.
The Liver and Muscle Glycogen in Fasting
Female Rats 6 Hours after the Intraperitoneal Injection of a Sodium Chloride
Solution, 0.5 cc. per 100 sq. cm.
XVI.
. . . .
The Liver and-Muscle Glycogen in Fasting
Male Rats 6 Hours after the Intraperitoneal injection of a 9 per cent
Solution of Dulcitol, 1 cc. per !GG;
■vi
TABLE
XVII.
PAGE
The Liver and Muscle Glycogen in Fasting
Male Rats 6 Hours after the Intraperi toneal Injection Of a 9 per cent
Solution of Glucose, 1 cc. per -100
-
sq. cm.
XVIII.
«-
. . . . . . . . . . . . . . . .
-38
The Liver and Muscle Glycogen in Fasting
Male Rats 6 Hours after the Intraperitoneal Injection of a Sodium
Chloride Solution, 1 cc. per 100
sq. cm.
XIX.
....................
39
Summary Table Giving a Comparison of
Liver Glycogen of Fasting Rats 6
Hours after the Intraperitoneal
Injections of
XX.
the H e x i t o l s ..................
40
Summary Table Giving a Comparison of
Muscle Glycogen of Pasting Rats 6
Hours after the Intraperitoneal
Injections of
the H e x i t o l s ..................
41
CHAPTER I
INTRODUCTION
The hexitols are hexahydric alcohols which may be
obtained by reduction of the aldehyde group of a hexose
sugar.
Thus sorbitol is the hexitol of glucose, mannitol
of mannose and dulcitol of galactose.
These polyhydric
alcohols are widely distributed in plants from which they
can be crystallized.
They may also be obtained commercially
by reduction of the corresponding hexose by various reducing
agents such as sodium amalgam.
The reaction proceeds very
slowly, two hydrogen atoms being added to the hexose.
At
the present time the hexitols are' used in the manufacture
of explosives and are obtained by the electrolytic reduction
of hexose sugars.
By this latter process it is possible to
obtain them in large quantities and at a greatly reduced
price.
In recent years the hexitols have become increasingly
prominent, claiming the attention of investigators and filling
various needs of the layman.
For example, In Europe, many
clinicians have advocated the use of sorbitol as a carbohy­
drate substitute in the treatment of diabetes.
of the opinion that It is to be avoided.
Others are
Within the past
few years the use of sorbitol in food preparations has been
extended as the price has been greatly reduced.
It is now
2
used in the manufacture of candies,
and as a moistening
agent in the preparation of bakery products.
A study of the metabolism of the hexitols was there­
fore thought to be of interest.
There are several methods that can be used in studying
the metabolism of carbohydrates and glucose producing sub­
stances.
Probably the most accurate method is to feed the
substance under question,
stop the metabolism at a certain
point by means of pancreatectomy or phlorhizin,
gate the elimination products.
and investi­
By this means it is possible
to determine what products are changed to glucose.
animal is unable to utilize glucose,
Since the
any substance that is
changed to glucose will be excreted quantitatively as such
in the urine.
Another method used in metabolism studies is
to follow the blood sugar curve.
substances are changed to glucose.
This also indicates what
A procedure followed by
some workers is to determine the effectiveness of the material
under study in reviving an animal after insulin shock.
A
widely used method is'to fast a n ’animal until,the liver
glycogen is used-up.
The substance under question is then
given and the liver glycogen is determined.
Thus any com­
pound that is converted to glucose will cause an increase
in the liver glycogen.
Another manner of investigating the
ability of a substance to be converted to glucose in the
3
animal body is the study of its effect on ketonuria.
If
carbohydrate oxidation is cut to a minimum, the breakdown of
fatty acids, in the animal is disturbed.
Their complete
combustion to carbon dioxide and oxygen does not take place
with the result that so-called ketone bodies or acetone
bodies a r e ,eliminated in the urine.
Any substance capable
of going to glucose will, however, produce a ketolytic effect
causing a decrease in ketonuria.
namely,
The last two procedures,
the determination of the ability of the hexitols,
sorbitol, mannitol ‘
a nd dulcitol, to produce glycogen and of
their effect on ketonuria were carried out.
CHAPTER II
LITERATURE
In 1883 Jaffe found that the urine of dogs contains
mannitol as a normal constituent.
After mannitol was fed to
dogs, large quantities were recovered unchanged in the urine.
Embden and Griesbach (1914) demonstrated that
d-sorbitol is converted into d-lactic acid on perfusion
through the liver of a fasting dog.
In the phlorhizinized
dog sorbitol is changed Into a mixture of fructose and
glucose.
Mannitol, dulcitol and inositol form neither sugar
nor lactic acid under similar conditions.
Upon plotting blood sugar curves after feeding normal
fasting colored men one hundred grams of various carbohy­
drates, Field (1919) found an increase of 10 mg. per cent
in blood sugar when mannitol was fed.
There was a 40 mg.
per cent increase when the equal amounts of glucose were fed.
Roth gave the same time curve, however.
Many investigators are of the opinion that sorbitol
can safely be given diabetics as a sweetening agent.
Kaufmann
(1929) recommended sorbitol as a carbohydrate substitute in
the treatment of this disease. . He claimed that It was sweet,
easily absorbed,
spared protein as It could be oxidized, led
to glycogen formation and did not elevate blood sugar.
Reinwein (1929) found no rise in blood sugar and no decreased.
5
carbohydrate tolerance.
He claimed that only traces of sorbi­
tol were excreted unchanged.
A rise in respiratory quotient
and the disappearance of hypoglucemic symptoms indicated that
the hexitol was utilized.
The effect of sorbitol on blood sugar of two juvenile
diabetics was, studied by Payne, Lawrence and McCance (1933).
The rise in blood sugar after sorbitol administration was
found to be slight compared with that of glucose.
The
acetone excreted in the urine was determined by Rotheras'
test before and after feeding sorbitol in order to see if it
produced any effect on ketosis.
The differences were not
marked but slightly less acetone was found when sorbitol was.
administered.
An attempt was made, to determine the effect
of the sorbitol in relieving insulin hypoglycemia but no
change was noted.
Sorbitol also failed to increase liver
glycogen in starved rats.
For this reason it is thought
that sorbitol has a place in the diabetic diet.
Roche and Rayboud (1933)
are of the opinion that;
sorbitol is not metabolized by normal or diabetic individuals.
Their experimental results on fasting rabbits indicate that
sorbitol is not transformed into glycogen,
on insulin hypoglucemia in rabbits.
and has no effect
It did appear, however,
to be utilized to some extent by phlorhizinized animals.
On the other hand, Donhofer (1930) gave a normal
fasting individual and a fasting diabetic 50 gm. sorbitol,
and determined the blood glucose and blood sorbitol at
varying- intervals.'
In the normal individual approximately
equal gains were observed in both glucose and sorbitol.
the blood •of' the'diabetic, however,
In
the amount of glucose
increased enormously, but the amount of sorbitol was no
greater than in the normal subject.
The conclusions drawn
are that sorbitol is easily synthesized into glycogen and the
sorbitol hyperglucemia seems to result from the same causes
as hyperglucemia following the ingestion of glucose or other
carbohydrates.
(1932)
This is in agreement with the work of Labbe"
who claims there are no clinical data available to
justify its use in the diabetic diet.
In a later paper,
Bertrand and Labbe" (1934) report that sorbitol Is poorly
absorbed and is no better tolerated than glucose.
It was
noticed that the Ingestion of sorbitol caused gastric
disturbances.
Because of its lower dietetic value Raybaud
and Roche (1934) are likewise of the opinion that sorbitol
can not be used as a substitute for glucose.
Lecoq (1934) fed rations rich In lipids and containing
mannitol and sorbitol to the extent of 35 per cent, of t h e •
total diet to pigeons.
completely utilized.
Both sorbitol and mannitol were
When the mannitol diet was given, less
7
than the usual amount of vitamin B was required.
When
given a ration containing 66 per cent mannitol, the pigeons
died in 8 to 15 days with symptoms of polyneuritis, even
though extra amounts of yeast -were added.
A 66 per cent
sorbitol ration produced the same results except that the
survival period was 17 to .50 days.
Lafon (1937) reports that mannitol is utilized by
mice to a very slight extent.
When fed in large quantities
the mannitol proved to be toxic.
diet,
Up to 30 per cent of the
sorbitol, however, was not toxic and seemed to .be
almost completely metabolized.
In this country as in Europe, there has been great
divergence of opinion concerning the fate of the hexitols.
Carr, Musser,
Schmidt' and Krantz (1933) found additional
glycogen storage when fasted rats were given a weighed
supply of cacao-butter and mannitol to the extent of 33
per cent of the diet.
The animals were maintained on this
diet for 80 hours and then sacrificed.
The controls were
given only the cacao-butter and sacrificed after the same
period.
It was found, however, that mannitol has no effect
on the respiratory quotient of rats.
Silberman and Lewis
(1933-34), on the contrary,
fasted rats for 24 hours and fed two or four cc. of a
15 per cent solution of manni.tol by stomach tube-.
The rats
8
were killed and the liver, glycogen was determined after
absorption periods of 2, 3, 4 and 6 hours.
No significant
increases in the glycogen content of the liver after oral
administration of mannitol were noted as compared with the
values obtained for the control series.
Following this work of Silberman and Lev/is, Carr and
Krantz (1938)
mannitol.
again endeavored to' study the metabolism of
The animals were treated as before— being fed-the
mannitol in cacao-butter.
Again they were able to demonstrate
the production of liver glycogen by mannitol.
The work of
Silberman and Lewis was repeated and confirmed.
Todd, Myers and West (1939) were able to demonstrate
that in dogs intravenous injections of mannitol cause a drop
in the blood sugar curve.
It was suggested' that this drop
might be the result of blood dilution.
When mannitol was
given either by stomach tube or by intraperitoneal Injection,
no Increase in liver glycogen was noted.
The time interval
between the administration of mannitol and the sacrificing
of the animals was 5 hours.
They did find, however, that if
mannitol was fed along with cacao-butter- there was a definite
glycogen increase.
In studying the metabolism of dulcitol, Carr and
Krantz found that when white rats were fed mixtures containing
33 per cent dulcitol with a basal diet of cacao-butter there
y
was an increase in liver glycogen over the controls.
tissue glycogen was reduced, however,
The
and there was no
noticeable effect on the. respiratory quotient.
’Dulcitol
likewise failed to increase significantly the-blood sugar of
rabbits when administered orally.
They were able, however,
to demonstrate an increase in liver glycogen following
administration of dulcitol by stomach tube.
Contradictory to the report of Payne, Lawrence and
McCance (1933) that sorbitol fails to increase liver glycogen
in the rat, Waters (1938) reported an increased liver glyco­
gen in rats following intraperitoneal injections of sorbitol.
He was unable to obtain any increase in glycogen following'
administration by stomach tube.
Contrary to the results of
Roche and Raybaud (1933) considerable glycogen deposition
in the liver was found in guinea pigs after intraperitoneal
injections of sorbitol,
Waters found only a mild transitory
hyperglycemia in normal fasting dogs following t h e ’intra­
venous administration of sorbitol.
He demonstrated that
intravenous injections markedly depress the glucose tolerance
curve of the normal dog and also of the depancreatized dog
receiving a steady supply of insulin.
According to Waters,*
only one-other substance, namely, fructose, has been found so
far to have this effect.
It is suggested that perhaps sorbitol
is oxidized to fructose in the liver.
This, of course, is
10
compatible with the early perfusion v?ork of Ernbden and
G-riesbach where a mixture of glucose and fructose was shown
to exist after perfusion of sorbitol through a phlorhizin-ized
dog liver.
Waters 'seems to feel that the effect of sorbitol
on the glucose tolerance curve is caused by the stimulation
of the glycogen synthesis mechanism.
Todd, Myers and West (1939) demonstrated that follow­
ing the. administration of sorbitol to fasted rats by stomach
tube or by intraperitoneal injection,
glycogen occurs within 8 hours.
a deposition-of liver
When sorbitol was fed with
cacao-butter there was, likewise,
an increase In glycogen,
though not so great as deposited after intraperitoneal
injection.
An increase in blood sugar was also shown follow­
ing the Intravenous injection of sorbitol.
Comparing their
studies on sorbitol with those on mannitol, Todd and his
coworkers conclude that sorbitol is m u c h more readily converted
into glucose and glycogen in the animal body than Is mannitol.
Carr and Forman (1939) found that fasting rats fed
one third sorbitol mixed with two thirds cacao-butter have
higher liver glycogen than do the controls' fed on the
cacao-butter alone.
Blatherwick, Bradshaw, Ewing, Larson and Sawyer (1940)
fasted rats for different periods.
Various amounts of
sorbitol were given by stomach tube and the effects of
11
3 and. 6 hour absorption periods studied.
When the animals
were fasted 48 hours and were then given large doses of sor­
bitol male rats appeared to store liver glycogen but the
females did not.
According to the.above workers,
the' inabil­
ity to show glycogen formation after oral administration of
sorbitol is probably due to its low absorption coefficient.
Cacao-butter lessens peristalsis•and prevents the diarrhea
which almost always accompanies the ingestion of large
quantities of aqueous solutions of sorbitol.
When fed with
cacao-butter the sorbitol remains1 in the gut for longer
periods and probably for this reason more is absorbed.
CHAPTER III
EXPERIMENTAL
Albino rats from the stock colony weighing between
100 and 215 grams were- used throughout.
The animals had
previously been on the stock diet snd were all in good
nutritional condition at the start of the fast.
Wherever
possible litter mates were used for comparative tests.
EXPERIMENTS ON KETOLYTIC ACTION OF THE HEXITOLS
The rat differs from man-in that it does not normally
develop ketosis during fasting.
An exogenous ketonuria can
be produced, however, by the method of Deuel, Hallman,
and
Murray (193b) by feeding the sodium salt of butyric acid.
Deuel, Gulick and Butts (1932) and Butts and Deuel
(1933)
demonstrated a marked sex difference in the degree of
ketosis.
Because female rats develop ketonuria much quicker
and to a greater degree than males,
they were used through­
o u t ’the experiments.
.The metabolism cages.used consisted of round wire
m e s h containers which fitted over large 10-inch funnels.
A
wire cone fitted into the funnel prevented any fecal or
extraneous material from passing through.
A small container,
provided with a stop-cock was attached to the stem of the
funnel.
This served to collect and deliver the urine.
13
Approximately one cc. of mineral oil was placed In the
bottom of the container to prevent the loss of acetone bodies
by evaporation.
The animals were fasted for 48 hours, weighed,
placed i n .individual metabolism cages.
and
They were allowed
free access to water throughout the experimental period.
An exogenous ketonuria was produced by the method of Deuel,
Hallman and Murray (1938).
The rats were given the sodium
salt of butyric acid in a total amount equivalent to 150 mg.
as acetone per 100 sq. cm. of body surface per day, or
461 mg. of butyric acid per 100 gm. of rat per day.
The
desired concentration of the salt was made by neutralizing
S2.5 gm. butyric acid with NaOH and making up to a volume
of 100 cc. with water.
Since general metabolism is thought to be proportional
to the superficial area of an animal, it would seem likely
that the production of ketone bodies is proportional to the
surface area.
Butts and Deuel (1933) obtained more uniform
action in animals of different weight where this method was
employed.
For this reason, the doses administered were
calculated on the basis of surface area in mg. per square
centimeter.
The surface area was determined by the formula-
of Lee, S _ k¥/°*^, where S — surface area in square meters,
W — weight of animal in kilograms,
and k = constant which
14
varies In the different .species.
For the rat k = 0.125.
Solutions of sorbitol, mannitol,
and dulcitol were
made up so as to be the equivalent of 25. mg, of glucose per
0.5 cc.
Thus, 5.056 grams of each hexitol was made up to a "
volume of 100 cc. separately.
The rats were divided into five groups corresponding
to the materials■being administered.
All received by stomach
tube 0.5 cc. per 100 sq. cm. of the sodium butyrate twice
daily.
In addition and Immediately following the butyrate,
part of the rats were given 0.5 cc. of the different hexitol
solutions per 100 sq. cm.
Two sets of controls were run: To
one group, 25 mg. of glucose per 100 sq. cm.
given in addition to the butyrate,
(0.5 cc.) were
and to the other.group
only the butyrate was administered.
The urine was collected the following morning In a
100 cc. volumetric flask and made up to volume by repeated
washings of the funnels.
The determination for acetone
bodies was made by the usual Van Slyke technique on an aliquot
of the urine.
The remainders of the urine samples were saved each
day and stored in the ice box.
On the last day of the experi­
mental period, urinary nitrogens were determined by the
Kjeldahl procedure.
In case the nitrogen was abnormally
high, the result on ketone bodies for that particular animal
15
for that day was discarded.
(If the urinary nitrogen is
greatly increased the decrease in ketone bodies can not be
attributed solely to the ketolytic effect of the materials
being studied, for in such cases allowance must be made for
the ketolytic effect of the carbohydrate produced by the
breakdown of tissue protein.)
If one day's urine were
discarded, Kjeldahls were run on the previous day's urine
sample for that particular animal to determine whether or
not that r e s u l t ■should be discarded as well.
EXPERIMENTAL RESULTS OF THE KETOLYTIC ACTION OF THE HEXITOLS
Tables: I, II, and III give the results of feeding
sorbitol, mannitol,
and dulcitol in the doses mentioned above.
Table IV gives the values obtained when an equivalent amount
of glucose was administered,
and Table V the degree of
ketosis developed by animals receiving only sodium butyrate.
A summary consisting of a comparison of the average acetone
body excretion In the urine during the administration of the
hexitols with that for.the glucose and fasting controls is
given in Table VI.
16
STATISTICAL TREATMENT OF RESULTS
The reliability of - the results/obtained for the
comparative ketolytic ability of the hexitols
determined by statistical treatment.
studied was
The mean difference .
of any two groups is significant if the observed difference
is a real one.
When the ratio of the difference In the means
obtained In two comparative groups
to the standard error of
the mean difference Is 3 or over, the results are considered
significant.
Absolute reliability Is obtained if the ratio
is 4 or above.
The method of calculating the ratio is as follows.;
After the mean is calculated,
the deviation (d) from the
mean is determined for each experiment.
obtained and the sum of the d
p
O
Next, d
[Z.& ) is found.
2
Is
The standard
error of the mean (S.E.M.) is calculated by the formula;
S.E.M. =
V
a2 :
n
\I n
in which n is the number of experiments.
The standard error of the mean difference
(S.E.M.D.) is
equal to the square root of the sum of the squares of the
standard errors of the means: |!(S.E.M. )2_2 + (S.E.M.)g2 ' .
..
If the ratio
M,D.(Mean difference)
---- g - .. -------
the results are significant.
.
.
is greater than 3
17
Another statistical treatment that may be used is the
Fisher t.
■
—
x 'l
_ 1(x-| )
n-^
s2 =
_
_ l(xP )
x2 “ n g
*
<* 2 ■
i da! 2 *!■■%
(n-^-1) + (ng -l)
t=
*3 I P_i__*_n.£
S
V n! + n2
x = mean
x^ and x g = results of individual experiments on the
comparative group.
n = number of experiments.
d
2
•
= square of the deviation from the mean.
The significance of the t value found is then
interpreted by reading from the Table' of t of Fisher when
the jo value is 0.01, or when there is one chance in 100.
of the difference being due to experimental error.
Each investigator may choose the value for jo which
he considers significant.
In many cases a value of 0.05
(one chance in 20) is accepted as satisfactory.
However,
by choosing the value of 0.01, there can be no question
about the significance of the results.
The use of the Fisher t method for statistical
evaluation in cases where the number of observations is
17 A
small (under 30) is generally considered to tie more
reliable than the first procedure described (determination
of ratio of mean difference to standard error of mean
difference), while the latter procedure is equally
acceptable if the number of cases exceeds 30.
TABLE I
Ketone Bodies in Urine of Fasting Female Bats Receiving Sorbitol
Expt. Uo.
Body Wt.
Surface Urinary Ef per
Area
100 sq. cm.
Acetonuria per 100 s^. cm. as acetone
4th day
1st day .
2nd day
3rd day
gms.
sq.. cm.
mg.
mg.
mg.
«mg.
m8Sh
1
125
228
94.6
78.7
---
----
2
105
205
35.0
109.0
79.0
9
156
260
35.5
73.8
68.2
60.7
10
141
244
37.3
77.0
60.2
74.0
19
144
247
28.3
131.5
88.6
88.3
—— **
20
154
257
33.8
78.6
----
29
157
261
36.3
94.7
96.4
76.2
59.6
30
143
246
40.2
56.7
58.5
51.2
51.8
38
196
298
30.9
73.8
67.2
75.2
---
39
186
288
26.7
80.4
63.3
83.7
----
151
253
30.4
87.0
±6.4
73.3
±3.8
72.8
±4.0
55.7
±2.8
Average
------
TABLE IX
Ketone Bodies in Urine of Fasting Female Hats Receiving Mannitol
V ♦
4
Body, Wt. Surface Urinary, IT per _______ Acetonuria per 100 sq. cm. as acetone
_________ Area
100 sq. cm.______ 1st day,_____2nd day,
3rd day,
4th day.
gms.
sq. cm.
mg.
mg.
”g» ■
Mi
Mi
‘
-----
3
135
238
32.0
184.0
131.0
11
155
259
33.5
111.0
114.0
131.0
12
138
241
32.0
112.4
102.2
101.0
13
136
239
29.8
90.5
94.2
108.0
21
152
256
29.9
105.2
95.7
.95.6
22
214
314
29.1
79.6
68.4
71.0
-----
31
116
217
40.3
77.2
82.3
■84.5
73.7
40
172
275
29.5
95.5
68.4
79.3
41
184
287
•25.4
83.9
90.5
57.0
076
154
258
99.0
67.6
077
165
268
----
76.5
93.7
088
145
348
----
84.6
92.5
157
258
31.2
99.9
+8.1
91.7
+4.9
-
-----
'
90.9
+7.7
73.7
TABLE III
Ketone Bodies in Urine of Fasting Female Eats Eeceiving Dulcitol
Expt. Ho.
Body, Wt. Surface
Area
Urinary, N per
100 sq. cm.
Acetonuria per 100 sq. cm. as acetone
1st day.
3rd day
4th day,
2nd day,
gms.
sq. cm.
mg.
mg.
mg.
•mg.
mg.
23
174
377
31.0
91.5
73.7
89.0
----
34
165
369
27.6
115.0
144.0
141.5
33
133
335
38.0
97.7
105.2
33
134
336
40.8
75.4
84.2
85.6
74.0
34
118
330
34.9
57.2
64.3
----
----
43
134
337
36.7
109.8
89.5
104.7
43
116
317
38.7
82,0
84.7
82.3
44
136
328
38.9
80.8
69.8
71.4
135
237
35.9
88.7
♦7.4
89.4
£8.4
96.1
♦7.9
Average
98.0 ■
' . 90.7
----
82.4
♦5.9
TABLE IV
Ketone Bodies in Urine of Easting Female Bats Receiving Glucose
Expt. Ko.
Body Wt,
Surface Urinary, N per
Area
100 sq. cm.
Acetonuria per 100 sq. cm. as acetone
1st day,
2nd day
3rd day,
4th day.
SSL
mg.
mg.
mg.
84.8
83.7
----
----
31.5
80.4
61.7
284
29.4
39.5
47.7
50.4
156
260
27.0
15.0
28.3
12.1
25
166
269
26.6
66.4
47,4
63.0
26
168
271
32.4
54.3
46.2
47.2
35
123
225
36.4
45.6
32.5
18.5
----
45
118
220
39.5
60.0
34.9
33.8
—
148
251
31.8
55.8
±7.5
47.8
±5.9
37.5
±7.3
•••■ w e e
gms.
sq. cm.
5
108
208
6
130
233
14
181
15
Average
mg.
-
—
-
----
----
-
TABLE V
Ketone Bodies in Urine of Pasting Female Bats Receiving Sodium Chloride
Expt. No.
Body Wt.
gms.
Surface Urinary N per
100 sq. cm. .
Area
sq. cm.
mg.
7
112
213
16
174
277
17
168
18
Acetonuria per 100 sq. cm. as acetone
1st day,
2nd day
. 4th day,
3rd day.
mg.
mg.
mg.
117.0
117.8
37.5
108.0
115.5
124.0
271
35.6
116.2
143.2
139.4
134
237
36.7
64.2
78.1
104.3
171
274
31.6
101.7
105.0
101.7
28
172
275
31.6
135.5
114.0
103.7
36
107
207
37.2
109.8
101.2
112.0
37
125
227
34.2
80.1
85.3
83.4
46
134
226
40.5
123.5
109.3
113.3
135
238
42.4
104.2
88.3
98.4
-----
130
232
98.8
108.9
141
243
105.4
*5.7
106.1
±5.4
108.9
±4.9
98.2
27
47
083
Average
.
‘
36.4
98.2
' TABLE VI
Summary, of the Effects of the Hexitols on Ketonuria
Substance
Fed
Acetonuria per 100 sq. cm,, as acetone
1st day
2nd day
3rd day
4th day,
Ho.
Expts.
mg.
Ho.
Expts.
mg.
Ho.
Expts.
Controls
11
105.4
±5.7
11
106.1
±5.4
9
Sorbitol
10
87.0
*6.4
9
73.3
±3.8
Maanitol
12
99.9
±8.1
12
Dulcitol
8
88.7
±7.4
Glucose
8
55.8
±7.5
»
♦With £»0.01
Average
Acetonuria
over 4-day,
period
t
M.D.
Cal­
Differ­
S.Jj.M.D, cu­
ence
lated
t
Theo­
ret­
ical"
Ho.
Expts.
mg.
mg.
108.9
±4.9
1
98.2
106.5
±2.9
7
72.8
±4.0
2
55.7
±2.8
76.9
±3.3
29.6
6.53
6.53
2.58
91.7
±4.9
8
90.9
±7.7
1
73.7
93.9
±3.9
12.6
2.57
2.51
2.58
8
89.4
±8.4
7
96.1
±7.9
2
82.4
±5.9
90.5
±4.1
16.0
3.19
3.14
2.58
8
47.8
±5.9
6
37.5
±7.3
0
---
47.9
± 4.3
58.6
11.32
11.20
2.58
mg.
— —
.
24
EXPERIMENTS ON GLYCOGEN FORMATION
Determinations were made of the amount of glycogen
deposited in the liver and the gastrocnemius muscle of rats
fasted 48 hours and-receiving intraperitoneal injections of
the hexitols.
Groups of rats used in these experiments were
either all male or all female,
not to use mixed groups,
since it was thought best
as Deuel,
Samuels,
and Gulick
(1932-33) reported that there is a sex difference in the
glycogen content of the livers of fasting animals.
The concentration of the materials
determined by preliminary experiments.
administered was
Different concentra­
tions were injected and different time intervals allowed to
elapse before determination of the liver glycogen.
The
results of these experiments are listed in Table VII.
From
these results, the optimum dose was found to be a 25 per cent
solution,
administered in two doses three hours apart with a
6 hour time interval between the first dose and the removal
of the liver.
Dosages were based on surface area here as
in the ketosis work for the reasons previously given.
TABLE VII
The Liver Glycogen in Pasting Male Rats
after the Tntraperitoneal Injection of
Different Concentrations of Sorbitol
at Different Time Intervals
Time
Liver
Expt. Rat Body Surface Cone, of Doses
Glyco­
Solution Given interval Wt.
Wt.
No.
No.
Area
gen
Adm.
after
1st dose
gms.
sq.cm. per cent
No.
hours
gms. per cent
3
600.
130
233
25
1
3
5.21
1.80
4
648
190
292
25
1
3'
5.88
1.34
5 .
610
165
268
50
1
3
6.58
2.12
6
601
165
268
50
1
3
4.91
1.88
7
669
175
278
25
1
4
5.79
2.21
8
620
180
283
25
2
6
5.31
3.70
9
695
170
273
50
1
■6
5.80
2.72
26
After the preliminary experiments, one cc. per 100
sq. cm. of rat of a 25 per cent solution of sorbitol and
mannitol was injected intraperitoneally into two different
groups of animals.
As a comparison with' the effect produced
by glucose, a similar concentration of this material was
injected into another group of fas'.ting rats.
The control
animals received one cc. of physiological saline per 100
sq. cm.
The injections were started in the morning and
spaced 10 minutes apart for the different animals.
Six
hours later, one cc. of a one per cent arnytal solution per
100, grams rat w a s 1 injected into the peritoneal cavity.
After several minutes,
tized.
the animals were completely anesthe­
The muscle was exposed without stimulation, frozen
in situ in solid carbon dioxide-ether mixture; the liver
was then removed followed by the frozen muscle.
The
removed parts were quickly placed in the freezing mixture,
to prevent glycogenesis.
After quickly weighing the dried
frozen tissue, potassium hydroxide was added to it and the
mixture digested in a water bath.
After digestion, the
glycogen was precipitated by alcohol, centrifuged, reprecipi­
tated and recentrifuged.
For the determination of glycogen
the method of Good, Kramer and Somogyi (1933) was followed.
After the glycogen was hydrolyzed with 0.6 N.HC1 for 3 hours,
the glucose was determined by the Shaffer-Hartman method.
27
EXPERIMENTAL RESULTS ON THE GLYCOGEN-FORMING
'ABILITY OP SORBITOL, MANNITOL, AND DULOITOL
Tables VIII and IX give the results of the intraperi tone al injections of one cc. per 10Q sq. cm. of surface
area of a 25 per cent solution of sorbitol and mannitol on
the liver and muscle glycogen of male rats after a fasting
period of 48 hours.
Table X gives the values obtained when
the same amount of glucose is administered by the same route
while Table XI shows the amount of glycogen that has been
retained by the fasting,
control animals.
Another set of experiments was carried out on female
rats receiving 0.5 cc. per 100 sq. cm. surface area instead
of one cc. as previously given.
The concentrations of the
solutions were the same, however.
The results of this work
are given in Tables XII through XV.
As it was impossible to obtain more than a 9 per cent
aqueous solution of dulcitol because of its relative insolu­
bility, two groups of animals were given one cc. per 100
■sq. cm. surface area of 9 per cent solutions of dulcitol
and glucose, respectively.
which received NaCl.
A control group was also ruin
The results are found in Tables XVI
through XVIII.
Summaries of the comparison of liver and muscle
28
glycogen of rats receiving intraperltoneal Injections of the
above mentioned dosages and'concentrations of the different
hexitols with those receiving like amounts of glucose are
given in Tables XIX and XX,
The only statistical treatment used was the Fisher jb
as the number of experiments was too small to apply the
ratio of the difference in the means to the standard error
of the mean difference.
TABLE VIII
The Liver and Muscle Glycogen in Pasting Male Rats
6 Hours after the Intrap'eritoneal Injection of
25 per cent Solution of Sorbitol,
1 cc. per 100 sq. cm.
Expt.
No.
Rat
No.
Body
Wt.
‘Surface
Area
gms.
sq. cm.
Wt.
Liver
Glyco­
gen
gms. per cent
Wt.
Muscle
Glyco­
gen
gms. per cent
8
1120
180
283
5-. 31
3.70
----
----
15
1051
225
322
6.50
- 4.37
----
----------------
16
1046
120
221
4.27
3.51
----
----
17
1067
110
210
4.56
5.47
----
----
18
1057
110
210
4.57
3.86
----
----------------
19
1157
160
264
4.96
3.38
---------------
_ _ _ _
14170
8678
140
243
• 6.13
3.14
---------------
----------------
171
8687
141
244
5.51
3.86
0.92
0.37
172
8696
165
268
7.21
1.87
1.04
0.29
173
8700
134
237
5.28
3.40
---------------
—
174
8709
185
288
6.43
3.79
----------------
----------------
175
8718
137
240
5.69
4.07
----------------
----------------
176
8727
153
256
6.47
4.21
0.89
0.39
177
8740
137
'240
5.78
4.51
---------------
----------------
178
8736
141
244
4.89
3.05
0.69
0.32
149
250
5.57
3.75
0.20
0.88
Average
±
±
- —
0.34
0.02
TABLE IX
The Liver and Muscle Glycogen in Pasting Male Rats
6 Hours after the Intraperitoneal Injection of
25 per cent Solution of Mannitol,
1 cc. per 100 sq. cm.
Bxpt.
Rat
. Ho. ' •No...
Muscle
Glyco­
. Wt.
gen
Body
Wt.
Surface
Area
Liver
Glyco­
Wt.
gen
gms.
sq. cm.
gms. per cent
gms. per cent
20
1050
235
332
5.30
1.39
----
---- _
21
897
210
310
4.82
0.42
----
----
22
769
115
216
3.98
0.31
----
----
23
1060
113
214
4.66
0.67
----
24
1148
120
220
4.12
0.40
----
----
25
1061
170
273
3.99
0.08
----
----
14179
8679
140
243
4.56
0.44
----
----
180
8688
137
240
4.69
0.23
0.82
0.20
181
8697
160
264
6.82
0.71
0.78
0.26
182
8706
128
230.
4.33
0.33
0.76
0.30
183
8710
133
236
5.08
0.38
1.03
0.26
184
8719
168
271
5.51
0.15
1.01
0.18
185
8728
142
245
5.14
0.26
0.82
0.28
186
8737
141
244
5.44
0.85
151
252
4.88
,0.47
tO.09
0.87
0.25
lO.02
Average
TABLE X
The Liver and Muscle Glycogen in Fasting Male Rats
6 Hours after the Intraperitoneal Injection of
a 25 per cent Solution of Glucose,
,1 cc . per 100. sq. cm.
Expt.
No.
Rat
'No.
Body
Wt.
Surface
Area
gms.
sq. cm.
Wt.
Liver
Glyco­
gen
. gms. per cent
Wt.
Muscle
Glyco­
gen
gms. per cent
30
1151
225
323
6.45
1.64
----
----
31
1176
100
199
3.58
3.07
----
----
32
1149
235
332
6.52
0.98
----
----
33
749
120
220
4.41
3.45
----
----
.34
768
120
220
4.80
3.69
----
----
35
1164
190
292
6.07
3.32
----
----
L4161
8677
157
260
6.51
2.83
0.69
0.63
162
8686
150
254
6.17
4.03
----
----
163
8690
145
248
6.88
3.67
----
----
164
8699
130
233
5 .60
3.06
0.68
0.68
165
8708
153
256
5.72
3.09
0.94
0.59
166
8717
137
240
6.20^
4. 37 .
1.00
0.44
167
8726
138
241
5.57
3.87
----
----
168
8730
187
289
7.85
3. 50
1.50
0.60
169
8739
160
264
5.83
3.11
----
----
154
255
5.90
3.18
±0.20
0.96
0.59
+0 *04
Average
TABLE XI
The Liver and Muscle Glycogen In Fasting Male Rats
6 Hours after the Intraperitoneal Injection of
a Sodium Chloride Solution,
1 cc. per 100 sq. cm.
Expt:.
NO.
Rat
No.
Body
Wt.
Liver
Glyco­
gen
Surface
Area
Wt.
gms.
sq. cm.
gms. per cent
Wt.
Muscle
Glyco­
gen
gms. per cent
----
1
1087
168
271
4.50
0.14
2
1091
150
254
4.52
0.05
11
1096
190
292
4.34.
0.00
12
1170
100
199
3.74
0.53
----
----
13
1158
110
210
'4.03
0.12
----
----
14
1150
120
222
3.90
0.39
L4152
8676
135
238
5.10
0.26
0.50
153
8680
167
270
7.25
0.23
---
154
8689
150
254
6.01
0.12
----
.----
155
8698
151
254
5.48
0.12
1.26
0.13
156
8707
162
266
4.83
0.17
-----
—
157
8716
193
295
7.30
0.27
0.61
0.36
158
8720
135
238
5.15
0.16
----
----
159
8729
129
232
5.27
0.20
0.81
0.24
160
8738
135
238
5.35
0.36
0.92
0.34
146
-249
5.12
0.21
±0.03
0.82
0.26
±0.04
Average
----
-------
----
0.25
----
—
TABLE XII
The Liver and Muscle Glycogen in Pasting Female Rats
6 Hours after the Intraperitoneal Injection of
a 25 per cent Solution of Sorbitol
0.5 cc. per 100 sq.' cm.
Expt.
No.
Rat
No.
Body
Wt.
Surface
Area.
gms.
sq. cm.
Liver
Glyco­
Wt.
gen
Muscle
Glyco­
Wt.
gen
gms. per cent gms. per cent:
.4122
10365
84
179
3.60
2.56
----
----
123
10264
113
214
4.76
2.55
0.80
0.24
124
10272
118
220
5.07
2.53
0.91
0.36
125 .
■10311
132
235
5.70
2.27
0.80
0.28
126 "
10315
126
.228
4.40
2.19
0.59
0.44
127
10323
97
195
4.80
2.07
0.58
0.32
128
10333
114
215
4.58
1.61
0.72
0.27
129
10342
107
207
4.56
2.96
0.60
0.32
130
10351
111
.212
5.02
3.02
0.78
0.25
131
10355
117
218
4.33
1.78 •
0.80
0.26
112
212
4.68
0.76
0.30
±0.01
Average
2.35
±0.14
TABLE XIII
The Liver and Muscle Glycogen in Pasting Female Rats
6 Hours after the Intraperitoneal Injection of
a 25 per cent Solution of Mannitol
9
0.5 cc. per 100 sq. cmi
.
Expt.
No.
Rat
No.
Body
Wt.
.
Sms*.
Surf ace
Area
Liver
Wt.
Glyco­
gen
Muscle
Glyco­
Wt.
gen
sq. cm.
gms. p■er cent
gms. per cent
L4132
10364
93
.190
3.74
0.00
0.51
0.26
133
10263
102
201
4.11
0.46
0.78
0.54
134
10271
135
238
4.67
0.09
0.77
0 .10
135
10275
132
235 ■
4.82
.0.11
--
----
136
10314
•90..
186
4.12
0.17
0.60
0.20
137
10321
103
202
3.93
0.23
---
-
----
138
10332
94
192
3.55
0.00
0.54
0.29
139
10341
105
205
3.88
0.00
140
10345
101
200
3.70
0.09
---
---
141
10354
100
199
4.06
0.16
----
----
106
205
4.06
0.13
±0.05
0.64
0.28
±0.07
Average
TABLE XIV
The Liver and Muscle Glycogen in Fasting Female Rats
6 Hours after .the Intraperitoneal Injection of
a 25 per cent Solution of Glucose,
0.5 cc. per 100 sq. cm.
Expt.
No.
-Rat
No.
Body
Wt.
gms.
Surface
Area
Liver
Glyco­
Wt.
gen
Muscle
Wt.
Glyco­
gen.
sq. cm.
gms. per cent
gms. per cent
.4112
10261
109
209
4. 65
2.53
0.87
0.28
113
10265
119
221
4.78
----
0.93
0.33
114
10273
136
239
5.48
2.25
----
----
115
10312
118
• 220
5.16
2.03
0.72
0.36
116
10321
107
207
•4.90
1.36
0.62
0.40
117
10325
96
194
4.00
1.22 '
0.42
0.46
118
10334
107
207
4.21
1.47
0.70
0.40
119
10343
116
217
4.41
2.39
0.66
0.28
120
10352
107
207
4.30
2.24
0.66
0.34
121
10361
97
195
3.77
2.23
0.58
0.47
111
212
4.57
1.97
±0.15
0.71
0.57
± 0.02
Average
TABLE XV
The Liver and Muscle Glycogen in Fasting Female Rats
6 Hours after the Intraperitoneal Injection of
a Sodium Chloride Solution,
0.5 cc. per 100 sq. cm.
Surface
Area
Liver
Glyco­
Wt.
gen
Mus cle
Wt.
Glyco­
gen
gms.
sq. cm.
g m s . per cent
gms. per cent
10563
■89
185
3.47
0.53
----
----
145
10262
107
207
4.67
0.00
0.65
0.21
144
10274
130
233
4.90
0.00
0.73
0.24
145
10313
130
233
4.77
0.05
0.83
0.20
146 '
10325
108
208
----
----
_
----
147
10332
96
194
3.53 ' 0.00
0.53
0.22
148
10335
111
212
4.50
0.00
----
----
149
10544
108
208
4.38
0.12
0.76
0.09
150
10353
114
215
4.31
.0.00
0.76
0.21
151
10363
89
185
3.20,
0.00
0.70
0.33
108
208
4.19' '' 0.08• ■ ■ 0.71
'±0.06
0.21
±0.03
Expt.
Ho.
.4142.
Average
Rat
No.
Body
Wt.
■
_
_
_
TABLE XVI
The Liver and Muscle Glycogen in Pasting Male Rats.-.
6 Hours after the Intraperitoneal Injection of
a 9 per cent Solution of Dulcitol,
1 cc. per 100 sq. cm.
Expt.
No.
Rat
No.
Body
Wt.
m s-
S urface.
Area
Wt.
Liver
Glyco­
gen
sq. cm.
gms. per cent
gms. p er c ent
Wt.
Muscle
Glyco­
gen
----
L4217
8680
223
322
7.68
0.43
-----
218
8337
281
370 •
8.67
0.68
. 1.65
0.26
219
8349
223
322
7.27
0.26
----
----
220
1084
243
339
6.38
0.09
1.26
0.23
221
1119
144
247
5.00
0.39
0.90
0.29,
222
8356
291
377
7.84
0.46
----
----
223
8340
254
348
7.26
0.30 .
1.50
0.28
' 224
1093
.210
310
5.75
0.10
1.22
0.22
225
1138
150
254
4.88
0.30
1.01
0.14
226
8360
216
316
6.25
0.00
1.35
0.16
224
321
6.70
0.30
±0.06
1.27
0.23
±0.02
Average
TABLE XVII
The Liver and Muscle Glycogen in Pasting Male Rats
6 Hours after the intraperitoneal Injection of
a 9 per cent Solution of Glucose
•-
Exp t.
No.
Rat
No.
1 cc. per.100 sq. cm. .
Surface
Area
Liver
Glyco­
Wt.
gen
Muscle
Wt.
Glyco­
gen
gms.
s q . 'cm.
g m s . per cent
g m s . per cent
Body
Wt.
14207
8333
244
339
7.46
1.66
1.55
0.43
208
8336
273
263
9.00
1.29
1.66
0.33
209
8346
207
308
6.57
1.73
1.00
0.33
210
8047
237
334
7.21
1.74
1.37
0.38
211
1115
148
251
5.82
2.41
----
----
212
8356
233
330
6.60
1.28
1.31
0.36
213
8338
225
313
6.32
1.28
1.35
0.31 -
214'
8348
231
329
7.65
.1.59
1.53
0.26
215
1168
202
303
7.64
0.94
.1.33
216
8357
368
434
10.57
1.38
227
320
7.48
1.53
±0.12
Average
1.38
0.35;.
0.34
±0.02
TABLE XVIII
The Liver and Muscle Glycogen in Pasting Male Rats
6 Hours alter the Intraperitoneal Injection of
a Sodium Chloride-Solution,
1 cc. per 100 sq. cm.
Expt.
No.
Rat
No.
Body
Wt.
Surface
Area
gms.
sq. cm.
Liver
Wt.
Glyco­
gen
£
2 S *_ per cent
Muscle
Glyco­
Wt.
gen
g m s . per cent
.4227
----
227
325
6.93
0.49
1.63
0.18
228
8339
263
355
8.04
0.20
1.11
0.29
229
8350
222
321
6. 61
0.00
1.35
0.22
230
1057
264
356
8.56
0.14
231
1135
134
237
4.85
0.40 ■
222
319
6.99
Average
0.22
±0.08
---- .
1.36
0.23
±0.03
TABLE XIX
Summary, Table Giving a Comparison of Liver Glycogen of Pasting Hats
6 Hours after the Intraperitoneal Injection of the Hexitols
Material
Injected
t
t
Calculated Theoretical*
Concen­
tration
Amt. per
100 sq.cm.
per cent
cc.
0.9
1.0
15
Male
0.21 * 0.03
----
Sorbitol
25
1.0
15
Male
3.75 ± 0.20
3.54
0.57**
Mannitol
25
1.0
14
Male
0.47 ± 0.09
0.26
2.70
2.77
Glucose
25
1.0
15
Male
3.18 ± 0.20
2.97
12.98
2.76
0.9
0.5
9
Female
0.08 4 0.06
----
Sorbitol
25
0.5
10
Female
2.35 k 0.01
2.27
0.38**
Mannitol
25
0.5
10
Female
0.13 k 0.05
0.05
0.61
2.90
Glucose
25
0.5
9
Female
1.97 * 0.15
1.89
11.15
2.92
0.9
1.0
5
Male
0.22 ± 0.08
----
Dulcitol
9
1.0
10
Male
0.30 k 0.06
0.08
•0.73
3.01
Glucose
*9
1.0
10
Male
1.53 * 0.12
1.31
6.75
3.01
Sodium chloride
Sodium chloride
Sodium chloride
♦With p * 0.01
♦♦Compared with glucose.
Ho.
Expts.
Sex
Liver
Glycogen
Difference
per cent
per cent
16.40
1.96**
2.76
2.76**
— —
13.881.77**
Other comparisons are with sodium chloride controls.
2.90
2.90**
TA3LE XX
Summary Table Giving a Comparison of Muscle Glycogen of Fasting Hats
6 Hours after the Intraperitoneal Injection of the Hexitols
Material
Injected
Sodium chloride
Concen­
tration
Amt. per
100 sq. cm.
Ho.
Expts . Sex
Muscle
Glycogen
Difference
per cent
per cent
per cent
cc.
0.9
1.0
5
Male
0.26 i 0.04
Sorbitol
25
1.0
4
Male
0.34 ± 0.02
0.08
Mannitol
25
1.0
6
Male
0.25 i 0.02
_
Glucose
25
1.0
5
Male
0.59 ± 0.04
0.33
0.25***
0.5
7
Female
0.21 * 0.03
---
Sodium chloride
0.9
t
1
Calculated Theoretical*
1.54
3.50
---
5.73
5.05***
3.36
3.50***
-- -
Sorbitol
25
0.5
9
Female
0.30, i 0.01
0.09
3.53
2.98
Mannitol
25
0.5
5
Female
0.28 i 0.07
0.07
1.02
3.17
Glucose
25
.0.5
9
Female
0.37 i 0.02
0.16
0.07***
6.12
2.90***
2.80
2.92***
Sodium chloride
0.9
1.0
3
Male
0.23 £ 0.03
— —
---
Dulcitol
9
1.0
7
Male
0.23 ± 0.02
___
---
Glucose
9
'1.0
8
Male
0.34 ± 0.02
0.11
3.17
•With £ = 0,01
••Values too low to compare with control.
•••Compared with sorbitol. Other comparisons are with sodium chloride controls.
3.25
CHAPTER IV
DISCUSSION
If a substance can be demonstrated to relieve ketosis
in an animal and to cause an increase in live? arid muscle
glycogen, then the particular substance must have been
absorbed from the intestine, gone into the blood stream,
entered into the metabolism of the animal.
and
Since it has
been demonstrated by many workers that only carbohydrates
or carbohydrate-producing substances are capable of cutting
down the ketone-body excretion of an animal, It seems fairly
certain that any substance which acts in a ketolytic capacity
is convertible to glucose in the animal organism.
If the
material under study Is shown to cause an increase in liver
and muscle glycogen in a fasting animal, It is a confirmation
that the substance in question has been converted to glucose,
which in turn has been changed to glycogen.
In the experiments, that were carried out on fasting
female rats receiving sodium butyrate alone, a high degree
of ketosis was obtained. - This was maintained throughout
the experimental period with' an average daily excretion of
106.5 + 2.9 mg. total acetone bodies per 100 sq. cm.
Acetonuria of the animals receiving the butyrate and glucose
was greatly reduced as was expected owing to the ketolytic
43
effect of the glucose.
The average dally excretion was
47.9 ± 4.3 mg. per 100 sq. cm.
While the Retone-body output of the animals receiving
sorbitol was greater than that of those receiving equimolecular amounts of glucose, it was, none the less, much lower
than that of the controls, viz., 76.9 * 3.29 mg. per 100
sq. cm.
Mannitol, on the contrary, was not found to produce
the ketolytic effect that sorbitol did,
as the average daily
excretion of ketone bodies was 93.9 i 3.94 v/hen mannitol
was fed.
While this value is lower than that of the controls
by 12.6 mg. per 100 sq. cm., it is not statistically
'significant,
as the ratio of the difference in the means
of the controls and the animals receiving mannitol,
standard error of the mean difference is 8.57.
to the
Neither
is it significant if the Fisher _t is applied when the jo
value is 0.01 (one chance in 100 of the difference being
due to experimental error),
as the t value found is 2.51
while the theoretical t value is 2.58.
If, however, a j> ■
value of 0.02 is used (one chance in 50 of the difference
being due to experimental error), mannitol is significantly
ketolytic as the t- value Is 2.35.
While dulcitol was like­
wise found to be inferior to sorbitol in relieving acetonuria,
it appears to be a slightly better ketolytic substance than
mannitol.
The average acetonuria over the 4-day period is
44
90.5 £ 4.1 mg. per 100 sq. cm. which is statistically
significant.
The ratio of the mean difference to the
standard error of the mean difference is 3.19 and the _t
value calculated is 3.14 while the theoretical t value of
Fisher for a jc value of 0.01 is 2.58. ■
The experimental results of the effect of the
hexitols on the formation of liver and muscle glycogen
after intraperitoneal administration are much more striking
with respect to sorbitol.
The liver glycogen in the fasting,
control male animals was found to be 0.21 ± 0.03 per cent.
The value obtained for glucose is 3.18 t 0.20 per cent 6
hours after the injection of one cc. per 100 sq. cm. of a
25 per cent solution.
Glucose, of course, is highly
significant in causing glycogenesis.
The calculated t
value is 12.98, while the theoretical _t value for a p
value of 0.01 is 2.76.
After the administration of a like
amount of sorbitol, the liver glycogen values obtained are
even higher than those for glucose, being 3.75 t 0.20.
In
this case the calculated t value is 16.40 while the p value
is the same as for that of glucose.
When 0.5 cc. per 1QQ
sq. cm.’ of a 25 per cent solution of glucose Is given, the
liver glycogen Is 1.97 +■ 0.15 per cent with a calculated
t value of 11.15 while the theoretical t_ value is 2.92.
When the same amount of sorbitol is.given, the liver
45
glycogen Is again greater than that of glucose, viz.,
2.35 ± 0.01.
Here the calculated _t value is 13.88 while
the theoretical t value is 2.90.
Although the values found
following sorhitol are higher than those obtained when
glucose was given the difference is not quite significant
if the Fisher _t is applied and a jo value of 0.G1 used.
But
the slight superiority of sorbitol over glucose as a glyco­
genic substance does not hold true for the percentage of
muscle glycogen found after administration of the above
amounts of the two substances.
In this case it was found
that the muscle glycogen formation in the fasting animal
receiving sorbitol Is not statistically different from that
of the controls, while glucose is highly glycogenic
In
the muscle when one cc. per 100 sq. cm. of a 25 per cent
solution is administered.
If, however, doses of 0.5 cc.
per 100 sq. cm. are given fasting females,
the muscle glyco­
gen following sorbitol is statistically significant when
compared with the controls.
In this case, the calculated
j; value Is 3.53 with a theoretical _t value of 2.98.
For
the same amounts of' glucose, the calculated t. value is
6.12 with a theoretical t value of 2.80.
The muscle glycogen
for glucose administered animals Is statistically higher than
for those receiving sorbitol when the t formula of Fisher is
applied.
46
Thus it would appear that in the doses given follow­
ing a 6-hour time interval between the first dose and the
glycogen determinations, more liver glycogen is present
when sorbitol is given than when glucose is given.
There
is, however, more m u s c l e .glycogen following glucose than
following sorbitol.
Prom this it would appear that the liver
glycogen is more rapidly formed in the case of glucose with
the result that after a 6-hour period the liver glycogen
has undergone a certain amount of glycogenolysis resulting
in an increased amount in the muscle.
Although there was some increase in the liver glycogen
following mannitol and dulcitol,
significant statistically.
the increases were not
An increase in muscle glycogen
is found in fasting female rats given 0.5 cc. per 100 sq. cm.
of a 25 per cent solution of mannitol.
It is difficult to compare the results on glycogen
formation of dulcitol with those of the other two hexitols,
as the concentration used was so much less, owing to its
insolubility.
It did appear to have a positive effect on
relieving ketonuria, however.
As dulcitol is the hexitol
that is converted to galactose on oxidation, it is interesting
to compare the metabolism of these two, substances.
In
contrast to the limited ability of dulcitol to relieve
ketosis, Deuel, Gulick,
and Butts (1932) found that on
human subjects galactose exhibited a superior ketolytic
'action.to that of glucose.
Clark and Mur1 in (1936) demon­
strated that galactose has a greater ketolytic ability than
glucose on ketosis produced by prolonged high-fat .diet and
injection of anterior pituitary extract.
Butts (1934)
found that galactose exhibits a superiority to glucose in
ketolytic action, in rats exhibiting ketonuria produced by
the administration'of sodium acetoacetate.
The glycogenic
superiority of galactose over dulcitol is likewise an
argument against the ready convertibility of dulcitol to
galactose.
Deuel, MacKay, Jewel, Gulick, and Grunewald'(1933)
showed that the glycogen retention in the livers of rats
fasted for intervals up, to 72 hours was actually greater if
the previous diet had been high in galactose than if glucose
had been the predominating dietary sugar.
Higher values in
liver glycogen also were found in dogs when galactose wa.s
given than when glucose was fed.
Thus the metabolism of
these two chemically similar compounds does not appear to be
related in the animal body.
The relative inability of mannitol to relieve ketosis
would not seem caused by its failure to be absorbed from the
intestine alone, as it was likewise unable to increase
48
glycogenesis to any appreciable extent following intra■peritoneal.injection.
In some respects mannitol appears to
behave much like mannose.
Deuel, Hallman, Murray,
In studies on this monosaccharide,
and Hilliard found that mannose was
absorbed at a rate of'12.3 compared w i t h glucose as 100,
was also able to form small amounts of glycogen.
'It
Even when
glucose and mannose were absorbed at the same rate, glucose
■was definitely a better glycogen former.
When mannose was
fed to fasting rats, having an endogenous ketonuria produced
by a previous
high-fat diet, the ketone body excretion was
only-slightly lowered, while glucose depressed it to about
one-third its original level.
From this it would appear
that mannitol and,mannose behave similarly when taken into
the animal body.
In general a parallelism between the ability to reduce,
ketonuria and the ability to be deposited as liver glycogen
has been observed in earlier experiments from this laboratory
when various metabolites were fed.
In the present tests,
greater amounts of glycogen in the liver follow the administra­
tion of sorbitol while the ketolytic effect is decidedly less
than after glucose.
this discrepancy.
There are two possible explanations for
In the first place the rise in muscle
glycogen is much slower with sorbitol than with glucose.
If
the disappearance in butyric acid is primarily related to the
49
level of glucose precursors In the muscle,
answer the discrepancy.
then this would
One must then conclude that a
ketolytic effect is related to the glycogenic action of an
intermediate in the muscle rather than in .the liver.
In
general the increases in muscle and liver glycogen are parallel
but this apparently does not always occur,- as in. the case of
sorbitol.
A second possible explanation is that the rate ofabsorption of glucose Is much greater than that of sorbitol.
In the exogenous type of ketonuria,
the butyric acid may be
completely metabolized before sorbitol is absorbed; on the
other hand glucose will already have been completely utilized.
The best method to obviate the difficulties in the discrepancies
in absorption rate is to compare the ketolytic activity of these
substances in an endogenous, ketonuria when a constant production
of ketone bodies -Is taking place.
Such experiments are now in
progress.
Prom all of these results,
of the three hexltols
it might be concluded that
studied, sorbitol is more completely
metabolized than are mannitol and dulcitol.
If sorbitol is
so readily convertible to glycogen following an intraperitoneal
injection, but is not as ketolytic as glucose following oral
administration,
It.would seem t h a t 'sorbitol Is not as readily
absorbed from the intestine as is glucose.
These results may
50
be explained by postulating an enzymatic change ’taking place
in the liver.
The. liver may contain'an oxidase which is
capable of o x i d i z i n g ’sorbitol to glucose.
It is also
possible that sorbitol is oxidized in the liver to fructose.
The work of Waters
(1938) demonstrating the similar effects
of sorbitol and fructose on the glucose tolerance curve, .
and the early perfusion work of Embden and Greisbach (1913)
where a mixture of glucose and fructose is shown to exist
after perfusion of sorbitol through a phlorhizinized dog
liver would make It seem highly probable 'that sorbitol Is
converted Into fructose in the liver.
CHAPTER V
SUMMARY
Exogenous ketonuria In fasting male rats Is appreciably
decreased following 'the oral administration of sorbitol.
Dulcitol was found to relieve ketosis but to a lesser degree
•than sorbitol.
Mannitol, while causing a somewhat lower
level of ketosis, was not statistically significant as a
ketolytic agent.
Six hours following the intraperitoneal injection of
a 25 per cent sorbitol solution, the values for liver
glycogen were greater following the administration of
sorbitol than for like amounts of glucose in both fasting
male and female rats.
The muscle glycogen, on the contrary,
was higher following glucose.
Although-mannitol caused a
slight increase in liver glycogen, and, in some cases, of
muscle glycogen, the difference was not great enough to be
significant.
In the concentrations
failed to show glycogenic activity.
administered, dulcitol
BIBLIOGRAPHY
1
.
2
.
Bertrand, R . , and M. Labbe", "Sur l ’emploi de la sorbite
dans 1 1alimentation des diabetiques,11 Bull. Acad,
m e d . , 112: 8, 1934.
;
Blatherwick, N.R.., P. J.- Bradshaw, M.E. Ewing, H.W. Larson,
and S.D. Sawyer, "The Metabolism of d-Sorbitol," J.
B i o l . Ghem., 134: 549, 1940.
3.
Butts, j.S.., "The Comparative Ketolytic Effect of
Galactose, Glucose and Lactose in Rats,” J. Biol.
C h e m . , 105: 87, 1934.
4.
Butts, J.S., and H.J. Deuel, Jr., "The Metabolism of
Diacetic Acid in Pasting Rats and Guinea Pigs," J.
B i o l . Ch e m ., 100-: 415, 1933.
5.
Carr, C . J . , and S.E. Forman, "The Pate of d-Sorbitol,
Styracitol, and 1-Sorbose in the Animal Body," J. Biol.
C h e m . , 128; 425, ,1939.
.
6
7.
8
.
9
.
Carr, C.J., and J.C. Krantz, Jr., "The Pate of Dulcitol
and Dulcitan in the Animal Body," J. Biol. Chem.,
107: 371, 1934.
Carr, C.J., and J.C. Krantz, Jr., "The Pate of Polygalitol
and Mannitol in the Animal Body," J. Biol. Chem.,
124: 221, 1938.
Carr, C . J . , R. Musser, J.E. Schmidt, and J.C. Krantz, Jr.,
"The Pate of Mannitol and Mannitan in the Animal
Body," J. B i o l . Chem., 102: 721, 1933.
Clark, D . E . , and J.R. Mur1 in, "The Effects of Glucose,
Fructose and Galactose on Ketosis," J. Nutrition,
12; 469, 193S.
.
Deuel, H.J., Jr., M. Gulick, and J.S. -Butts, "The
Comparative Ketolytic Action of Glucose, Galactose,
Fructose, and Sucrose," J. B i o l . Chem., 98; 333, 1932.
11
.
Deuel, H.J., Jr., L.F. Hallman, and S. Murray, "Ketolysis
versus Antiketogenesis: as an Explanation for the
Action of Carbohydrate on Ketonuria," J. Biol. Chem.,
124: 385, 1938.
12.
Deuel, H.J., Jr., L.P. Hallman, S. Murray, and J. Hilliard,
"The Comparative Metabolism of d-Mannose and d-Glucose,"
J. Biol. C hem., 125: 79, 1938.
10
53
13.
Deuel, H.J., Jr., E.M. MacKay, P . W . ‘jewel, M. Gulick,
and C.F. Grunewald, “The Comparative Glycogen
Formation and Retention after the Administration
of Glucose, Galactose, and Lactose," J. Biol.
Ch e m ., 101: 301, 1933.
14.
Deuel, H.J., Jr., L. Samuels, and M. Gulick, "Sexual
Variation in the Carbohydrate Metabolism of Rats,"
P r o c . Soc. E x p e r . B i o l ., and M e d ., 50: 27, 1932-33.
15.
Donhoffer, S., "Uber den Mechanismus der diabetischen
glykamischen Reaktion," Deut. Arch. klin. Med.,
169: 242, 1930.
16.
Donhoffer, S., and M. Donhoffer, "Uber die klinische
Untersuchung des Kohlehydratstoffweehsels mittels
d-Sorbit," Deut. A r c h , k l i n . M e d ., 167: 257, 1930.
17.
Embden, G . , and W. Greisbach, "Uber den Abbau der
d-Sorbose," Z. Physiol. Chem., 91: 251, 1913.
18.
Field, C. W . , "Blood Sugar Curves with Glucose, Lactose,
Maltose, Mannite, and Cane Sugar," P r o c . Soc.
E x p e r . B i o l , and M e d ., 17: 29, 1919.
19.
Fisher, R . A . , Statistical Methods for Research W orkers,
6th ed.
London: Oliver and Boyd, 1936..
20.
Good, C. A . , H. Kramer, and M. Somogyi, "The Determina­
tion 'of Glycogen," J. B i o l . Chem. , 100: 485, 1933.
21
.
22
.
Jaffe, M . , "ijber das Vorkommen von Mannit im normalen
Hundelharn," Zeits.chr. f. physiol, chem., 7: 297,
1883.
Kaufmann, E . , "Ein neuer Kohl ehydr at ersatz zur
Diabetesbehandlung," K l i n . W o c h .., 8: 66, ,1929.
23.
Labbe, K . , "Une ann£e d ’observation medicale chez
diabetiques," B u l l . A c a d . M e d . , 107: 426, 1932.
24.
Lafon, M . , " S u r ,1’utilisation alimen Faire des hexitols
par la souris," Compt. rend. Soc. biol., 126; 1147,
1937.
25.
Lecoq, R., "La valeur alimentaire de la mannite et de
la sorbite en rapport avec l ’equilibre de la ration,"
Compt. rend, acad. Sci., 199: 894, 1934.
54
26.
Lee, M.O.,' ’’The Determination of the Surface Area of
the White Rat with its Application to the
Expression of Metabolic Results," Am. J. Physiol.,
89: 24, 1929.
27.
Payne, W . W . , R.D. Lawrence, end R.H. McCance, "Sorbitol
(Siomon) for Diabetics," Lan c e t , 225: 1257, 1933.
28.
Raybaud, A., and A. Roche, "Valeur dietetique.de la
sorbite dans la cure diabete sucre'," Presse m e d . ,
42: 172, 1934.
29.
Reinwein, H . , "Uber die Verwertbarket des d-Sorbitol in
der Behandlung des Diabetes mellitus," Deut. Arch,
klin. M e d . , 164: 61, 1929.
30. Roche, A., and A. Raybaud, "Sur 1 1utilisation de la
sorbite par 1 ’organisme," Compt. rend. Soc. b i o l . ,
113: 320, 1933.
31.
Silberman, A.K., and H.B. Lewis, "Glycogen Formation
after Oral Administration of Mannitol to White
Rats," Proc. Soc. Exper. Biol, and Med., 31: 253,
1933-34.
32.
Todd, W . R . , J. Myers, and E.S. West, "On the Metabolism
of Sorbitol and Mannitol," J, Biol. Chem., 127: 275,
1939.
33.
Waters, E . T . , "Metabolism of Sorbitol," P r o c . XVI
Internet. P h ysiol. C o n g ., Zurich, 122, 1938.
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