Blood
Coagulation
By RATKO BUZINA, M.D.,
After a
AND
Fat Meal
ANCEL KEYS, PH.D.
The influence of meals on the coagulability of blood was studied in experiments on 20 men with
parallel nonfed controls. The whole blood clotting time in siliconized tubes was not altered by a
low-fat meal or by continued absence of a meal, but it was significantly shortened after a meal
containing 100 Gm. of butterfat. The degree of lipemia did not determine the degree of shortening.
There was no important effect on prothrombin time of the fatty meal as contrasted with the nonfatty meal or with no meal. Heparin tolerance results were difficult to interpret.
curred, at which time tilting of the second tube was
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AVARIETY of in vitro experiments have
indicated that lipids influence blood
coagulation,1-3 from which coagulability might be expected to increase in the
lipemic period after a fatty meal. This was,
indeed, reported by Waldron, Beidelman and
Duncan,4 but it was denied by Tulloch,
Overman, and Wright.5 However, the former
group maintained their position on the basis
of further experiments, with special attention
to the avoidance of artifacts,6' 7 and Fullerton,
Davie, and Anastopulos8 found that a fatty
meal shortens the clotting time of recalcified
plasma in the presence of snake venom. But
Manning and Walford9 sided with Tulloch,
Overman, and Wright on the basis of tests with
cream feeding and with a fatty breakfast.
This was the highly controversial picture when
we began to examine the question with carefully
controlled studies on physically healthy men
aged 35 to 60 in a rigidly controlled metabolic
unit in Hastings State Hospital.
similarly started. When clotting occurred in the
second tube, tilting of the third and fourth tubes
was started simultaneously, and the average clotting
time (computed from the time of blood drawing) in
these was taken as the final clotting time. The
standard error of measurement (S.E.M.) between
duplicates* proved to be ±i1.608 min. with 100
samples.
Prothrombin time was estimated at 37 C. in 12.5
per cent oxalated plasma (diluted with 0.85 per cent
NaCl solution). To 0.1 ml. of this diluted plasma,
was added 0.1 ml. of Difco thromboplastin and then,
after a few seconds, 0.1 ml. of 0.02 AI calcium chloride was added and the stopwatch was started.
S.E.M. was ±1.041 see. in 132 samples.
The heparin tolerance test was done with oxalated
blood at 37 C. by a modification of the method of
Sigg.10 To 0.15 ml. of oxalated whole blood was
added 0.15 ml. of fortieth-molar calcium chloride
containing 0.56 units of heparin.t The test was made
in duplicate tubes that were tilted every 60 sec.
S.E.1M. was ±2.108 min. in 100 samples.
All studies were started in the morning, with the
subjects in the fasting, resting state. After drawing
the control blood samples, a "meal" (1080 calories)
consisting of 300 ml. of 40 per cent butterfat cream
was ingested and then, as the subjects continued to
rest, further blood samples were taken at 1.0, 2.5,
4.0, and 5.5 hours from the time of the meal.
Finally, to distinguish between effects of fatty
meals as contrasted with low-fat meals, the experiments were repeated with the same subjects with a
meal that provided 940 calories but only 0.3 Gm. of
fat. This low-fat meal consisted of rice, sugar, and
"divinity" candy and contained 18 Gm. of protein.
The result was a series of precisely parallel values
for the effect of a fatty meal (F), for the nonfat
control meal (C1), and for a fasting control (C2).
METHODS
Blood was withdrawn from an arm vein with siliconized syringes and no. 18-gage needles coated with
Arquad 2 C. If there was any difficulty in obtaining
blood, a new venepuncture was made with a new
needle and syringe. A stopwatch was started as soon
as blood entered the syringe. One milliliter of blood
was added to each of 4 siliconized tubes in a water
bath at 37 C., and 4.5 ml. of blood was added to 0.5
ml. of 0.1 M sodium oxalate; the latter mixture was
centrifuged to provide oxalated plasma.
For whole blood clotting time, one of the siliconized tubes was tilted every 60 sec. until clotting oc-
RESULTS
The results of experiments on 20 subjects are
summarized in table 1 and the statistical
analysis is given in table 2. The whole blood
From the Laboratory of Physiological Hygiene,
University of Minnesota, Minneapolis, Minn.
The work reported here was made possible by research grants from the Schweppe Foundation,
Chicago, by the National Dairy Council, Chicago,
and by the Minnesota Heart Association.
* Computed as S. E. M. = V 2/2N.
t Testagar and Co., Detroit, Mich.
854
Circulation, Volume XIV, November 19.56
855
BUZINA AND KEYS
TABLE 1.-Effect of a Fat Meal on Blood Coagulation
Time in hours
Group
Item
2.5
1
F
F
Cl
Cl
C2
C2
F
F
Cl
Cl
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C2
C2
F
F
Cl
Cl
C2
C2
F
F
Cl
Cl
C2
C2
Whole blood, mean
S.E.
Whole blood, mean
S.E.
Whole blood, mean
S.E.
Prothrombin, mean
S.E.
Prothrombin, mean
S.E.
Prothrombin, mean
S.E.
Heparin,
mean
S.E.
Heparin,
mean
S.E.
Heparin,
mean
S.E.
mean
Density,
S.E.
Density,
mean
S.E.
mean
Density,
S.E.
0.95
±1. 217
-2.92
±1. 179
-0.94
±0.756
3.60
±0.653
1.42
±0 ;775
2.44
±t0.754
0.10
±0.560
-1.14
40.762
0.94
±0.891
0.0256
±0. 0151
-0.0002
±0.00272
-0.0015
±0.00478
4.25
±t1.237
-1.78
±0. 846
-1.83
±-1.299
3.85
--0.924
1.07
±1. 014
1.89
1.038
0.70
±-0. 729
-3.07
±-0.633
0.89
±0. 718
0.2688
0.0358
-0.0030
±=0.00272
0.0039
±-0.00528
4
3.95
±0. 904
-1.71
±-1.273
-1.22
±-1.074
1.60
±=0.998
1.78
±-1.392
2.22
±1. 144
-0.45
±0. 642
-1.00
± 0.688
1.50
-0.t672
0.3058
±-0.0401
-0.0027
±-0.00278
0.0040
±=0.00597
5.5
5.50
±-1.160
-1.92
±1.616
-1.28
±i0. 921
2.75
±= 1.289
4.00
±-0.961
1.94
±-1.478
-1.70
0.858
-2.28
±4-0.774
0.94
±0. 501
0.2734
±0 .0380
-0.0064
±-0.00320
-0.0032
--0.00632
Twenty men received 300 ml. of 40 per cent cream (F) and 12 control men (CJ) received the same amount of
calories derived from carbohydrates and proteins only. (C2) = 18 control, unfed, men. Whole blood coagulation
time shortening ("Whole Blood") is in minutes, prothrombin time shortening ("Prothrombin") is in seconds,
heparin tolerance time ("Heparin") is in minutes. S.E. = standard error of the mean.
clotting time tended to be shortened in all 4 of
the samples drawn after the fatty meal,
though the average difference was slight in the
sample drawn 1 hour after the meal. In contrast, both the low-fat meal (C1) and the
continued absence of a meal (C2) tended to
produce somewhat longer clotting times than
were observed in the premeal sample. The
net result for whole blood clotting is that there
was a highly significant shortening related to
the fatty meal at 2.5, 4, and 5.5 hours, the
net shortening over this period being
6.37 min. with reference to a nonfat meal and
6.01 min. with reference to no meal. The mean
net shortening at 1 hour after the fat meal is
less impressive but this also seems to be real
when comparison is made with both C1 and C2.
In contrast, there was at no time a significant
difference between the coagulation time after
a nonfat meal as compared with no meal.
The prothrombin time after the fatty meal
showed- an average shortening that varied
mean
between 1.60 and 3.85 sec.; this is statistically
significant with reference to the premeal
value. However, both after a low-fat meal and
after continued fasting there was also a
tendency toward shorter prothrombin time,
so that it is dubious whether the fatty meal
has any specific effect on this variable.
The data from the heparin tolerance test
are difficult to interpret. After the fatty meal
there was some tendency toward a prolongation
of the heparin effect, and this tendency was
much more evident after a low-fat meal. In
contrast, when no meal was fed, there was
some trend toward a reduction in the duration
of the heparin effect. The net result is a highly
significant difference between the heparin
tolerance during continued fasting and that
after any meal, though the time relations may
be affected by the type of meal.
The degree of lipemia, expressed as optical
density, showed a slight increase after 1 hour
following the ingestion of cream and a high
856
BLOOD COAGULATION AFTER A FAT MEAL
TABLE 2.-Statistical Analysis of the Results in Table 1
Time in hours
1
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Whole blood, mean diff., F -C
S.E. of diff.
t
Whole blood, mean diff., F - C2
S.E. of diff.
t
Whole blood, mean diff., C, - C2
S.E. of diff.
t
Prothrombin, mean diff., F -C
S.E. of diff.
t
Prothrombin, mean diff., F -C2
S.E. diff.
t
Prothrombin, mean diff., C- C2
S.E. of diff.
t
Heparin,
mean diff., F -C
S.E. diff.
t
Heparin,
mean diff., F -C2
S.E. of diff.
t
Heparin,
mean diff., C- C2
S.E. of diff.
t
Density,
mean diff., 1F -C
S.E. of diff.
t
Density,
mean diff., F -C2
S.E. of diff.
t
Density,
mean diff., C- C2
S.E. of diff.
t
3.88
±1.707
2.272
1.89
±1. 471
1.285
-1.98
±1.346
1.477
2.17
±1.005
2.161
1.16
40. 980
1.184
-1.012
41.088
0.930
1.24
±E0.941
1.320
-0.84
±1.042
0.806
-2.082
±1. 214
1.715
0.0258
±0.0183
1.409
0.0271
±0.0166
1.633
0.0013
±0. 00595
0.219
2.5
4
5.5
6.04
±1.644
3.672
6.08
±1.794
3.389
0.044
±1.658
0.0265
2.78
±1.392
1.996
1.96
±1.139
1.415
-0.819
±1.478
0.554
3.77
41.022
3.689
-0.19
±1.027
0.185
-3.961
5.66
±E1.516
3.736
5.17
±1.395
3.706
-0.494
±1.656
0.298
-0.18
±-1.666
0.111
-0.62
±1.512
0.410
-0.435
±1.786
0.244
0.55
±-0.960
0.572
-1.95
±0.930
2.096
-2.50
±0. 974
2.566
0.3085
±0.0468
6.592
0.3018
±0. 0427
7.059
-0.0067
±0.0072
0.935
7.42
41.934
3.840
6.78
±1.503
4.511
-0.648
±1.764
0.367
1.25
±1.742
0.718
0.81
±1.952
0.415
2.06
±1.882
1.094
0.58
±1.212
0.498
-2.64
±1.025
2.580
-3.22
±0.886
3.634
0.2798
±0. 0442
6.330
0.2766
±0.0405
6.826
-0.0032
±0.0077
1.244
±0.988
4.009
0.2718
±0.0432
6.291
0.2649
±0.0380
6.959
-0.0069
±0.0065
1.063
The pooled variances for the F + Cl and 02 groups are used to compute the standard errors of the differences
between the means, F - C, and F - C2. Abbreviations as in table 1. The following probabilities are associated
with N1 + N2- 2 = 36 degrees of freedom: p = 0.05, t = 2.03; p = 0.01, t = 2.72; p = 0.001, t = 3.59.
degree of lipemia in the 2.5, 4.0, and 5.5 hour
samples, the peak being at the fourth hour.
The association between degree of lipemia and
shortening of clotting time was low, and the
peak times for the 2 variables did not coincide.
The coefficient of correlation between optical
density and whole blood coagulation time
shortening was computed, combining the observations for the periods 2.5, 4.0, and 5.5
hours. The result was an insignificant value,
r = 0.206. Finally, as would be expected, there
was no significant lipemia following the low-fat
meal.
DIscussIoN
While this work was in progress, several
events more firmly established our conclusion
that a fat meal definitely tends to make the
blood more coagulable in the next few hours.
In the first place, we analyzed statistically the
raw data from which Tulloch, Overman, and
Wright5 had concluded there was no effect. The
BUZINA AND KEYS
TABLE 3.-Summary of the Statistical Analysis of the
Data of Tulloch, Overman, and Wrightl on 10 Subjects
Time after meal,
hours ...........
A = mean shortening, minutes.
A/S.E.A ...........
3.0
0.5
1.0
2.0
2.4
1.81
3.1
4.12
4.6 4.3
3.80 3.15
4.0
2.6
2.31
For 9 degrees of freedom, t values are: po.05 =
2.26, po.ol = 3.25, po.ool = 4.78.
Blood was drawn with previously unused syringes
for each sample before and at 0.5, 1, 2, 3, and 4 hours
after a fat meal and whole blood coagulation time
was measured. "Shortening" here is the difference
between the control and the post-meal value.
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statistical analysis, summarized here in table 3
demonstrates clearly that these "negative"
results actually show a statistically highly
significant shortening of the coagulation time,
especially at 1.0, 2.0, and 3.0 hours after the
meal. The error in the previous conclusion
resulted from arbitrarily assuming a value for
method uncertainty and failing to make any
statistical analysis. However, we have no
explanation for the only other negative finding,
that of Manning and Walford.9
1iore important was the discovery of the
work of Salvini and Sordi," of Vandenbroucke
and co-workers,12 and the publication of the
paper of O'Brien.1" Salvini and Sordi drew
blood from 10 men before and at 2, 4, and 6
hours after eating 100 Gm. of butter and
measured the coagulation time of recalcified
plasma, using the method of Quick,14 the
prothrombin activity as estimated by Scardigli
and Mininni,15 the amount of heparinoid
materials by the method of Gibson and
associates,16 and plasma turbidity. They found
maximal turbidity at 4 hours, at which time
there were a moderate shortening of clotting
time and increase of prothrombin activity
together with a marked fall in heparinoid
substances. Vandenbroucke and associates12
also found a significant shortening of clotting
time after a fatty meal.
O'Brienl" found that a fatty meal containing
50 Gm. of animal fat had no effect on clotting
of whole blood in the presence of platelets and
a wettable surface, but in siliconized tubes
857
there was a significant shortening of clotting
time that was not definitely related to the
degree of lipemia. The chylomicron layer
obtained by high speed centrifugation, however, was found to have some shortening
effect on the clotting time, in conformity with
some of the findings of Poole2 in rats. Very
recently, Robinson and Poole3 offered evidence
suggesting that the active materials in chylomicra do not include triglycerides, cholesterol,
lecithin, or phosphatides of serine or inositol,
but that ethanolamine phosphatides may be
suspect.
SUMMARY
Measurements of whole blood coagulation
time in siliconized tubes, of prothrombin
activity, of heparin tolerance, and of serum
lipemia (turbidity) were made on blood
samples drawn from 20 men at intervals of 1,
2.5, 4, and 5.5 hours after a meal of 300 ml. of
40 per cent butterfat cream and after a meal
similar in calories but very low in fats. Parallel
control measurements were made during
continued fasting during the same period.
The whole blood coagulation time was
significantly shortened after the fatty weal,
especially at 2.5 to 5.5 hours, but was essentially unaltered after a nonfatty meal or during
continued fasting. There was a trend toward
shortening of prothrombin time after the fatty
meal but this was of less significance because
a similar, though less pronounced trend
occurred after a nonfatty meal and during
continued fasting. The heparin tolerance
results indicated some relative prolongation
of the heparin effect after both types of meal,
particularly after the nonfatty meal, as
compared with continued fasting. The degree
of shortening of the clotting time after a fatty
meal was not correlated to an important
extent with the degree of lipemia (turbidity).
ACKNOWLEDGMENT
We are grateful to Dr. W. F. Sheeley, Director of
the Hastings State Hospital, for making subjects
and facilities available, and to Dr. J. T. Anderson,
for help with the experiments.
BLOOD COAGULATION AFTER A FAT MEAL
858
SUMMARIO
IN
INTERLINGUA
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Esseva executate mesurationes del tempore
de coagulation de sanguine integre in tubos
revestite de silicium, del activitate prothrombinic, del toleration de heparina, e del lipemia
seral (turbiditate) in specimens de sanguine
obtenite ab 20 homines a intervallos de 1, 2,5,
4, e 5,5 horas post un repasto de 300 ml de
crema a 40 pro cento de grassia de butyro e
post un repasto de simile valores caloric sed a
bassissime contento de grassia. Parallel mesurationes de controlo esseva facite durante le
mesme periodo in statos de jejuno continue.
Le tempore del coagulation de sanguine
integre esseva significativemente accurtate
post le repasto grasse, specialmente a intervallos de 2 ,5 a 5 ,5 horas, sed illo esseva in principio inalterate post repastos non-grasse e
durante le jejuno. Esseva notate un tendentia
verso le reduction del tempore prothrombinic
post le repasto grasse, sed isto esseva minus
significative proque un simile (ben que minus
pronunciate) tendentia occurreva post le
repastos non-grasse e durante le stato jejun.
Le resultatos del tests del toleration de heparina indicava un certe prolongation relative del
effecto de heparina post ambe typos de repasto,
specialmente post le repasto non-grasse, in
comparation con le jejuno continue. Le grado
del reduction del tempore de coagulation post
repastos grasse non esseva correlationate a
grados importante con le grado del lipemia
(turbiditate).
REFERENCES
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2
POOLE, J. C. F.: The significance of chylomicra in
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ROBINSON, D. S., AND POOLE, J. C. F.: The effects
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4 WALDRON, J. M., BEIDELMAN, B., AND DUNCAN,
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13 O'BRIEN, J. R.: Relation of blood-coagulation to
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3
Blood Coagulation After a Fat Meal
RATKO BUZINA and ANCEL KEYS
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Circulation. 1956;14:854-858
doi: 10.1161/01.CIR.14.5.854
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