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Guanidinosuccinic
Effects
of
Acid
Exogenous
on
Urea,
Aggregation
By James
W. Davis,
increase
LEEDING
times
in optical
TENDENCY
are
may
frequently
occur
prolonged
usually
associated
with
creased
(PRP)4’5
retention
in glass
bead
or whole
blood.3’5”
in
when
adequate
platelet
aggregation
was
decreased,
platelet
aggregation
was
normal
O’Shea’#{176} also
groups
uremic
patients.
Morris
in vitro
From
City,
have
inhibits
the
Research
Mo.,
and
Submitted
June
Presented
Boston,
in
Mass.,
James
the
part
April
W.
Davis,
Administration
University
of
Technician,
pital,
Kansas
Veterans
City,
388
and
Medical
of
the
4,
City,
Hematology
at
may
July
49th
Veterans
School
Annual
29,
1977;
Session
Kansas
accepted
September
the
five
or creatinine
added
aggregation,
and
Hospital,
Medicine,
of
but
of some
glass
patients.
urea
Administration
of
Hutton
in the PRP
irreversible
or ADP-induced
Services,
Kansas
(ADP)-
patients.
aggregation,
of uremic
de-
plasma
thrombin-
diphosphate
of uremic
platelet
blood
a level
have
American
City,
8,
College
to
Kansas
Kans.
1971.
of
Physicians,
1968.1
M.D.:
Clinical
School
investigator
Kansas
of
Phyllis
City,
Medicine,
Hematology
Mo.
bleeding
are
patients
adenosine
PRP
as to whether
to glass
revised
1971;
at
Kansas
Administration
Technician,
adhesion
Hospital,
Formerly,
reports
University
17,
but
in the
in the
conflicting
platelet
whose
counts
Such
ADP-induced
aggregation
been
patients
platelet
found
it to be decreased
Heslop’#{176} reported
decreased
and
platelet
There
normal
of investigators2’2’5
bead-induced
ans
reported
E. Phillips,
filters
of platelets
in either
platelet-rich
Salzman
and
Neri7
reported
that
induced
other
uremic
their
hemostasis.2’#{176}
induced
and
Phyllis
lagen-,
and
norepi
neph rine-i nduced
platelet
aggregation
were
shown
to be
inhibited
1 hr after
urea was added
to
platelet-rich
plasma
to produce
urea
nitrogen
increments
of 100-300
mg/
100 ml.
Increasing
concentrations
of
creatinine
by 10 or 20 mg/100
ml did
not inhibit
platelet
aggregation.
Guanidinosuccinic
acid,
in a concentration
in the range
found
in uremic
plasma,
also had no effect
on platelet
aggregation.
Through
inhibition
of platelet
aggregation,
elevated
blood
concentrations
of urea may be one of the causes
of the
bleeding
tendency
of uremic
platients.
density
of platelet-rich
plasma.
This
precluded
the
use
of
turbidimetric
techniques
for
the
measurement
of
platelet
aggregation
after
addition
of
urea,
until
the optical
density
bocame
stable.
Adenosine
diphosphate-,
col-
B
and
A. Graham
three
compounds
that are found
in increased
concentration
in the blood
of
patients
with
renal
failure.
The
addition of urea was followed
by an imme-
but transient,
Creatinine,
R. McField,
Barbara
Patients
with severe
uremia
may have
a bleeding tendency
associated
with
a
prolonged
bleeding
time and an adequate
platelet
count.
We have tested
the effects on platelet aggregation
of
diate,
Platelet
In Vitro
James
and
Human
and
Mo.,
Kansas
Research
E.
Phillips:
Hospital,
Kansas
Research
Laboratory,
Chief
and
of
City,
Technician,
Mo.
Veterans
James
Veterans
of
R.
VeterMedicine,
McField,
B.S.:
Administration
Hematology
Barbara
Section,
Professor
Kansas.
Laboratory,
City,
Hematology
Associate
Research
A.
Graham,
Administration
HosLaboratory,
B.S.:
Formerly,
Hospital,
Kansas
Mo.
Blood,
Vol.
39, No. 3 (March)
1972
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PLATELET
AGGREGATION
our knowledge
aggregation
ported
no
with
PR?.
and
the
then
of
other
enhances
ADP-induced
but
retention
seemed
platelet
these
previously
addition
in glass
a direct
These
sera
of uremic
let factor
3 activation20
or connective
tissue.21
was
it
platelet
al.4
that
PR?
decreased
the
and
Castaldi’3
on ADP-induced
their
preliminary
of exogenous
for
incubated
inhibits
found
Stewart
effect
that
a need
creatinine
because
creatinine
further
on
investigation
on platelet
aggregation.
Guanidiits concentration
is elevated
reported
to inhibit
aggregation
MATERIALS
Blood
effect
suggested
and
and
filters.
deleterious
reported
inhibitory
reports
patients,19
et
to normal
bead
platelet
Neri7
re-
urea
was
urea initially
Hellem
in vitro had no
and Cortellaro’8
to suggest
adhesiveness.
when
that
of creatinine,
of platelets
added
Praga
collagen-induced
Salzman
and
aggregation
et al.’7 stated
aggregation.
the
of the possible
effects
of urea and
nosuccinic
acid was also investigated
the
compounds
on
investigated.
ADP-induced
hand,
Somer
not
that creatinine
aggregation.
results
of
ADP-induced
of urea,
stated
platelet
effects
not been
have
inhibition
On the
addition
389
ADP-induced
induced
AND
by
ADP,
in
plate-
epinephrine,
METHODS
Samples
Blood
and
was
obtained
creatinine
Instances
we
mining
After
with
no
used
urea
nitrogen
sodium
achieve
no
and
any
mixture
Test
male
and
0.1
pH
the
mixtures
of
reagent
mg/100
of
ml,
under
35
urea
nitrogen
respectively.
yr
of
age
In
few
a
without
deter-
concentrations.
solution
(saline)
concentration
than
concentrations
1.2
workers
creatinine
highest
of
plasma
ml
hospital
or
more
with
mg/100
chloride
the
was
men
20
from
0.9%
there
trol
blood
mixing
to
fasting
than
plasma
PRP
paper,
from
more
and
of
unit
each
difference
containing
each
test
test
between
a
test
compound
compound
the
in
reported
pH
of
saline
in
the
this
saline
con-
substance.
Compounds
Test
compounds
Calbiochem,
were
Los
oratories,
Angeles,
New
York.
mesh,
of Bio-Rad
did not reduce
aggregation,
or
The
urea
the
creatinine
were
acid
with
Richmond,
of ADP-,
that
and
a
from
mixed
bed
resin
Research
was
not
due
to
Lab-
(AG5O1-X8,
Calif.)
just before
use in the PRP
collagen-,
or norepinephrine-induced
inhibition
from
purchased
Mann
contamination
20-50
of two men
platelet
of
urea
with
ions.
used
treated
Urea
guanidinosuccinic
of
Laboratories,
the inhibition
other
urea
was
and
Purification
indicating
cyanate
grade.
Calif.,
in
with
all
the
the
experiments
resin
on
just
collagen-induced
before
use.
platelet
Untreated
urea
aggregation
was
at
used
in
37#{176}C
all
other
experiments.
Platelet
The
Aggregation
decrease
aggregating
fied
optical
that
of
Born
in
With
(where
ml
0.8
of
and
(OD)
as
an
Cross22
at
index
and
600
of
ms
differs
25#{176}C) and
of
platelet
was
from
not
citrated
PRP
aggregation.
theirs
in
after
agitation
Our
that
method
aggregation
recorded.
continuously
It
with
was
an
modi-
occurred
at
been
de-
has
elsewhere.23
exception
ml
used
(about
detail
the
Temperature
density
was
temperature
scribed
0.1
in
agent
from
room
at Room
of
a solution
PRP
of
of
the
and
0.2
the
test
experiments
ml
of
compound
reported
additive
were
in
saline
in
used),
or
the
0.9
with
last
ml
two
of
the same
rows
PRP
was
volume
of
Table
mixed
of saline
1
with
as
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
390
DAVIS
ET AL.
U-)
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oc’JOON-QOL()
dodciddcocid
V V V V V V VVV
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E
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C
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From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
PLATELET
AGGREGATION
Fig. 1. Changes
of PRP following
in OD
at 600
addition
of saline
of urea in saline.
ings
were
made
Multiple
linked
tometer.
Value
of
zero
assigned
to OD
tions
were
made.
a
control.
After
0.1
tion
60
ml
10
essentially
the
as
“one-half
of
were
by
method
was
incubated
was
in
for
saline.
55
at
1800
at
rpm).
agents
37’C.
described
Statistical
Each
A
in
to
agitation
agent
or
a
agents
were
the
latter
kept
in
be
of
with
referred
frozen
OD
by
(Levoprepared
preparation
will
the
solu-
trihydrate
suspension
al.24
decrease
produced
a
bitartrate
collagen
of
et
of
was
diphosphate
l-norepinephrine
1 :1 dilution
OD
of Born
and
volume
of
ml)
of
temperature
a
PRP
to
as
until
use.
The
after
agitation
centrifugation.
and
stirrer
Cross.22
PRP
test
5 mm
at
was
set
1
mm,
at
volume
37’C.
We
to
record
ml
with
used
one
saline
or
PRP
a
were
Chrono-Log
light
transmission
speed
of
platelet
of
saline
maximum
0.04
in which
0.1-0.2
or
recorder
approximately
Mixtures
and
compounds
potentiometric
magnetic
was
of
Packham
maximum
(0.4
to
for
above
A
prior
aggregating
adenosine-5’-
saline,
by
that
room
stirring
al.24
0.8-0.9
linked
After
of
/Lg/ml
8
6
MINUTES
C
37
Mixtures
mm
The
in
decrease
contained
Aggregometer
continuously
of
immediately
aggregating
essentially
tested
substance
Platelet
et
of
cent
20
described
The
at
used
aggregation
test
per
Aggregation
The
Packham
and
added.
salt
or
York)
determinations
as
was
disodium
Mo.),
collagen.”
expressed
Platelet
the
New
solution
duplicate
temperature
Louis,
described
4
agent
of
St.
Tyrode’s
strength
means
room
Laboratories,
modified
addi-
aggregating
Co.,
Winthrop
arbitarily
before
itg/ml
Chemical
phed,
was
at
an
trac2000
Recorder
Spectropho-
PRP
mm
of
containing
(Sigma
of
IU,
m
and
Superimposed
by a Gilford
Sample
Absorbance
a Beckman
DU
to
mixture,
391
(approximately
of
the
aggregating
added.
Methods
test
mediately
substance
after
aggregating
was
its
saline
agents.
used
an
equal
to
compensate
control
The
t test
for
number
paired
of
for
times
any
samples25
immediately
effects
was
of
used
in
before
aging
analysis
and
im-
of
the
PR!’
and
of
the
data.
RESULTS
Preliminary
Studies
Figure
1 shows
transient
ing
the
urea
concentration
in saline
was
result
of adding
of the
two
as
expected
peared
that
when
urea
The
of urea
upper
in OD.
increase
one
mixtures
because
unaggregated
to detect
a difference
added.
The transient
to nine
part
of saline
of
after
in their
increase
addition
the
PRP
200
mg/100
of
PRP.
The
parts
same
of the
urea
size or shape
from
in OD observed
effect
ml
was
was
tracing
The
but
lower
than
and
shows
final
microscopy
solution,
a rapid
OD of increaswhen
one part
of
on
lower
of PRP.
and
phase-contrast
there
the
by
to nine
By
to
shows
parts
essentially
dilution.
added
tracing
nitrogen
added
was
was
OD
the
base
platelets
we
were
those
to which
saline
after
addition
of the
the
of each
had
urea
line
ap-
unable
been
solu-
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392
DAVIS ET AL.
Fig.
2. Superimposed
tracings
of
platelet aggregation
at 37#{176}C
show effects of incubation
of PRP with increment of urea nitrogen
of 300 mg/100
ml for 1 hr. ADP was added
at arrow
to obtain
final concentration
of 1.7 M.
Tracing
of control
PRP begins
below
and reaches
peak above
that of PRP
to which
urea
was added.
There
is
more rapid platelet disaggregation
in
PRP with added
urea.
Control
50
1
MINUTES
tion
to
PRP
addition
required
of the
When
an
was
chloride
in saline,
magnitude
was
much
PRP
changes.
stabilized.
PRP
from
urea
of
nitrogen
200
By phase-contrast
controls
on
ADP-Induced
20
the
aqueous
greater
was
effect.
effects
on
not
due
inhibitory
effect
guanidinosuccinic
(200
acid
mg/100
of
osmolarity
urea.
(4.5
ml),
were
of
mixed
with
PRP
from
these
and
by
not
detect
an
PRP.
by
rows
of
adding
200
mg/100
increasing
4.5
mg/100
figures
urea
urea
by
or
100
that
acid
in
creatinine
ml
in Table
saline
had
1
and
no
com-
urea
in
in saline
did not have
urea
at this concentration
a
1),
PRP
from
guanidinosuccinic
Increments
mg/100
agent
increment
was
could
of
of urea
of PRP.
in Table
or
an
in stirred
of PRP
horizontal
shown
creatinine
of
nitrogen
aggregation
the
in OD
aggregating
we
of platelets
added.
sensitive
effects
distinguish
Thus,
aggregation
two
(not
whether
not
with
saline.
Since
of platelet
aggregation
to increasing
experiments
to determine
nitrogen
platelet
solution
isomotic
effect,
inhibition
In other
urea
last
solution
saline.
or guanidinosuccinic
The
urea
of
Temperature
platelet
ml
an
caused
was a
water
been
was
changes
had
the
could
with
at Room
increasing
mg/100
demonstrable
we
there
by
had
osmotic
without
in saline
aggregation
ADP-induced
or
pare
that
mm
1
use
sus-
to differentiate
of PRP
the
in
was
it
volume
unable
to
after
urea
approximating
of PRP
produced
OD
the
potassium
in that
saline
the
attempt
hr after
1
agitated
spontaneous
shows
1
10
been
Aggregation
Table
inhibited
ml and
were
to which
that
not
urea
microscopy
had
PRP
for
of
after
by
or
an equivalent
we
until
stirred
mg/100
that
of urea
from
aggregation
was
OD
differing
substituting
us to conclude
addition
in
but
with nine parts
decrease
in OD
by
did
observed
chloride
increase
microscopy
we
not
produced
sodium
solution,
added
led
was
to that
of
immediate
urea
produced
been
men
their
by
that
the
PRP.
ml
the
Therefore,
four
after
effect
an
by
of platelet
immediately
equal
of water
was mixed
decrease
in OD.
The
had
and
plasma.
solutions
was
experiments
as a monitor
platelets
of PRP
adding
By phase-contrast
water
above
to osmotic
PR!’
part
than
water.
to which
The
had
there
greater
for
by
produced
When
one
and sustained
saline
of
to platelet-poor
of osmolarity
obtained
that
presence
solution
increase
solution
tamed.
rapid
the
urea
of
ml),
no more
creatinine
together
effective
ten
acid
(20
with
than
men
could
mg/100
an
increment
urea
alone.
were
tested
enhance
the
ml)
or
of
of
urea
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
PLATELET
AGGREGATION
393
Fig.
3. Superimposed
tracings
of
platelet
aggregation
at 37#{176}Cshow effects
on
collagen-induced
platelet
aggregation
of incubation
of PRP with
increment
of urea nitrogen
of 200 mg/
100 ml and
increment
of guanidinosuccinic
acid
of 4.5 mg/100
ml for
lhr.
ADP-Induced
The
Aggregation
effects
the
was
PRP
100
ml
of
urea
(p
and
crement
of
presence
6.8
acid
did
(Fig.
2).
not
appear
Collagen-Induced
Table
2
platelet
ml.
mg/100
of urea
succinic
of the
20.8
shows
that
acid
mg/100
ml had
In other
experiments
to determine
An increment
nitrogen
mg/100
Collagen-Induced
effects
of urea
platelet
increase
in per
urea
nitrogen
37.9
for
shown
had
ml)
and
induced
in-
in the
of guanidino-
PRP
inhibited
strength
mg/100
more
effective
col-
of 4.5 mg/100
ml
A higher
concentration
effect.
2)
ml
mg/100
a one-half
A creatinine
from
ten
than
urea
did
increment
men
were
tested
enhance
the inhibitory
effect
ml) together
with an increment
of
acid
tested
of urea.
of urea
alone.
acid
PRP
for
37.0
increment
the
of
strength
of eight
saline
(p <0.01,
ml
mg/100
200
on one-half
in the
was
Aggregation
that
urea
guanidinosuccinic
acid nor
control,
collagenThe
mean
25.8
for
inhibition),
30%
mg/100
4.5
men.
ml.
An
at Room
example
Temperature
inhibited
norepinephrine-induced
acid
creatinine
did not potentiate
demonstrably
platelet
this
inhibited
aggregation
Aggregation
acid
at
aggrega-
inhibition.
Neither
norepinephrine-
increments
curves
(Fig.
from
4).
C
37
4.5
to
9.1
mg/100
ml had
a
and
aggregation.
Guanidinosuccinic
platelet
in
acid
occurred
presence
200
or
inhibitory
could
transmission
Norepinephrine-Induced
on
duplicate
transmission
of 300 mg/
3.
shows
tion and that
guanidinosuccinic
of
a standard
effect.
in Table
were
Norepinephrine-Induced
2
in the
controls.
acid increment
either
preparation.
no
in
at 37#{176}C
a guanidinosuccinic
Table
tested
usually
the
guanidinosuccinic
increment
in Fig.
5
4
a guanidinosuccinic
increment
a small
was
aggregation
cent
3
Temperature
either
demonstrable
(not shown
Aggregation
induced
2
MINUTES
in per cent
increment
curves
from
nitrogen
whether
creatinine
of creatinine
(20
(200
The
no
for
24.6
at Room
by
were
disaggregation
different
a urea
20
Collogen
I
acid
rapid
lagen
preparation.
A guanidinosuccinic
not inhibit
aggregation
induced
by
of
0
mean
increases
for a urea nitrogen
Disaggregation
induced
of guanidinosuccinic
0
and
More
Aggregation
aggregation
strength
1/2
guanidinosuccinic
inhibition),
16%
<0.05,
4O
at 37#{176}C
of six men. The mean
for the saline
control,
24.9
-Control
#{176}
50
no effect
is
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
394
DAVIS ET AL.
Fig.
z #{176}
4O
30
Large
4.
second
let aggregation
PRP that had
.
phase
of plate-
occurred
at
been incubated
37#{176}Cin
with in-
crement
of guanidinosuccinic
acid of
9.1 mg/lOU
ml for 1 hr. Norepinephrine
was added
at arrow to obtain
final con-
Norepinephrine
20
centration
of 5.4 M.
Saline
control
aggregation
curve
is not shown,
as it
is almost
identical
to one shown
for
guanidinosuccinic
acid.
10
-
C
MINUTES
DISCUSSION
Our data show
that
ment
of 200
mg/100
norepinephrine,
the
or
reports
of
induced
inhibits
incubation
of PRP for
ml inhibited
platelet
collagen
Salzman
by
and
platelet
aggregation,
and later
enhances
apparent
discrepancy
means
Neri7
aggregation
Figure
is
that
those
immediately
at a time
shows
that
1
altered
indicator
the
of platelet
Lovett26
lets)
“are
hr with
aggregation
that
urea
data
did
not
contrast
with
inhibit
ADP-
who stated
that urea
A likely
explanation
investigators,
who
either
increADP,
initially
of this
simultaneously
PRP’7
or did not state
the period
of incubato use OD changes
of 1’RP as an index
of
when
the
the mere
OD
a urea
nitrogen
induced
by
These
12-30%.
stated
and Somer
et al.’7
platelet
aggregation.
mixed
urea and ADP
with stirred
tion of urea
with
PRI’,7
attempted
platelet
reason.
of
who
1
curve
OD of PRP was unstable
for
addition
of a urea
solution
and
temporarily
invalidated
its
another
to PRP
use
as
an
aggregation.
reported
that when
mixed
with
solutions
bacteria
(which
of urea
they
approximate
undergo
an
the
initial
size of plateosmotic
de-
hydration
which
produces
an increase
in light
scattering
and then,
as urea
enters
the cell,
osmotic
equilibrium
is regained.”
Fantl27
found
a transient
increase
in the OD of PRP after urea was added
in much
higher
concentrations
than were
of platelets
PRP
was
shown
used
by
approximately
in Fig.
osmolarity
25%
in
platelet
in OD
that
due
the
mM
71
that
If this
relationship
ml,
volume,
which
that
we
could
It seems
occurred
to osmotic
after
of 150
of investigators.
g of urea
time and
ing time
decreased
increased
into
Hellem
a subject
in our
by
to the
raising
only
approximately
explain
our
failure
to conclude
was
to
that
added
in
experiment
when
45%
down
produced
in saline
shrinkage
of urea
decreased
expect
reasonable
urea
used
volume
is linear
we
might
Castaldi’3
observed
the concentration
increment
platelet
approxi-
the
plasma
3%
detect
the
deit
by
transient
to PR!’
(Fig.
1)
shrinkage.
That
our demonstration
of inhibition
vitro
may be relevant
to in vivo situations
groups
and
when
found
mg/100
microscopy.
probably
times
in osmolarity
200
phase-contrast
increase
400%.
increase
nitrogen
crease
was
ten
Lundberg28
1.
increased
mately
urea
in our experiments.
Stewart
phase-contrast
microscopy
et al.4
with
of
platelet
is suggested
reported
normal
that
hemostasis
aggregation
by the
the
by
results
intravenous
prolonged
platelet
adhesiveness.
Eknoyan
et al.3 found
that
and platelet
adhesiveness
decreased
in normal
urea
in
of two
infusion
the
bleeding
the bleedvolunteers
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
PLATELET
who
AGGREGATION
ingested
urea.
administration
ness,5
raised
than
100
1
hr
Hellem
24 hr
did
bleeding
not
time
maintain
platelet
an
Eknoyan
increase
effect
Cronberg,5
platelet
inhibitory
function2
of normal
higher
concentrations
found
that
as was adafter
only
the
bleeding
hr of experimental
8-10
failures
of Cronberg5
and
Castaldi
may
been
due
concentration
of
urea
more
have
long
than
enough
Several
guanidino
aggregation
ADP-induced
platelet
dinosuccinic
compounds
have
in
PRP.293#{176} Their
citrated
factor
acid.
This
uremic
patients,19
tion was completely
but
occur
in vivo,
to occur.
Rabiner
Horowitz
succinic
been
addition
shown
of
to
inhibit
large
recently
blocked
as
9.1
2.5
effect
In their
platelet
and
found
as large
inhibit
either
(Fig.
4).
We
found
of guanidinosuccinic
by increments
of
dinosuccinic
6.8
acid
no
by
acid
reported
in
the
sera
4.5
2).
ml)
guanidino-
induced
either
first
after
by
by
or
incubation
ml.
mg/100
2.5
a consecond
with
Increments
as
of norepinephrine-induced
mg/100
The
highest
15
uremic
of
celite,
induced
of collagen-induced
of
had
utilizing
mg/100
AD!’
6.8
of
patients
aggregation
phase
inhibition
ml (Table
mg/100
as
sera
3 activawould
not
activation.
(1.75
M
1.7
of
coagulation
uremic
systems
3
be
guani-
in the
to allow
inhibited
aggregation
no inhibition
of
acid
aggregation
assay
factor
of platelet
induced
not
ml)
found
of their
mM
0.1
phase
aggregation
ml did
range
75%
of guanidinosuccinic
mg/100
may
compounds
inhibition
mg/100
(3.5
insufficient
that
that
second
the
was
found
reported
the
mM
0.2
is within
Hrodek3’
by increments
aggregation
ml and
slight
concentration
patients
inhibition
of guani-
was
5.4
mg/
mi.’9
The
et al.2’
reason
is not
aggregation
tion
of
aggregating
for the apparent
conflict
of our results
clear.
It may be related
to their
inclusion
in their
methods20’2’
agents
test
the
second
imidazole.32
phase
system
as a buffer.
that
they
dissolved
in imidazole-buffered
tion of imidazole
to 0.5 mM.
inhibition
of collagen-induced
mM
testing
ADP-induced
inhibitory
of ADP-induced
platelet
factor
mM calcium2#{176} and presumably
factor
3 activation.
acid did not inhibit
platelet
increments
by
of calcium
,M AD!’ or 1.4 j.M epinephrine
nective
tissue
suspension.
We
phase
activation
concentration
and
et al.2’
acid
3
the inhibition
reversed
by
as this
decreased
platelet
guanidinosuccinic
of
et al.2
to failure
before
antagonized
by calcium
that appears
to compete
with the guanidino
to form
a complex
with
AD!’.3#{176}Horowitz
et al.2#{176}
have
reported
let
but
after
50%
function.
platelet
100
times
azotemia,
increased
administration
the
or adhesivesubjects
to less
volunteers
The
of urea
reported
urea intravenously
platelet
adhesiveness
et al.3
after
who
achieved
twice
as much
who measured
azotemia.
an
and
effect
on
concentration
of six other
of experimental
to demonstrate
al.2
et al.4 presumably
azotemia.
volunteers
mean
et
more
than
by Cronberg,5
of experimental
the
Castaldi
to be without
urea
nitrogen
ml.
administered
orally
of four
to
Both
of urea
serum
the
mg/100
as they
ministered
395
of
norepinephrine-
It appears
both
saline,
Recent
experiments
platelet
aggregation
and
with
those
of
of an inhibitor
from
guanidinosuccinic
bringing
the
in our
by
ADP-induced
their
final
laboratory
0.5
mM
Horowitz
of platedescripacid
and
concentra-
have
imidazole
aggregation
shown
and
by
1
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
396
DAVIS
Hemostasis
hal
depends
substances
exposed
by
about
been
as
vascular
platelet
suspected
ADP-,
upon
such
injury.
and
in
acid,
vitro
aggregation
it appears
and
Since
uremic
by
released
from
the
platelet
AD!’,
and
discovered
to subendotheand
microfibrils33
adherent
platelets
et al.4
not
after
to its
coupled
that
the
with
the
prolonged
patients
with
administration
uremia
by urea,
which
and
of urea,
of
urea
are
at
inhibits
to
people,
in
guanidino-
inhibit
were
acid.
least
the
play
urea (in conaggregation
that
not
platelet
part
in
to
vivo
urea,
data3’4
make
it seem
likely
frequently
seen
in
to
oxidative
due
Their
the present
paper
also
and mucosal
bleeding
due
also
did
may
sera of uremic
patients,’9
found
by Eknoyan
et al.3
guanidinosuccinic
in vitro
data
of
bleeding
times
in vitro,
norepinephrine
in the
times,
brings
plug.
It has
potentiate
have
shown
that
human
platelet
found
bleeding
metabolite,
severe
aggregation
aggregation
metabolite
in vitro
in concentrations
likely
that
the increased
and
platelets
membranes,
our experiments
patients)
inhibited
collagen,
a recently
Hellem
itself,
AD!’
thrombin-induced
a role in hemostasis.34’35
centrations
found
in
succinic
of circulating
basement
aggregation
and formation
of a hemostatic
platelet
that
catecholamines,
which
in low concentrations
collagen-,
induced
adherence
collagen,
ET AL.
inhibition
of
metabolism
platelet
of platelets.36
ACKNOWLEDGMENT
We
Dr.
gratefully
acknowledge
Kenneth
T.
N.
the
many
helpful
suggestions
of
Dr.
David
V.
Cohn
and
Yue.
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From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
1972 39: 388-397
Guanidinosuccinic Acid on Human Platelet Effects of Exogenous Urea,
Creatinine, and Aggregation In Vitro
James W. Davis, James R. McField, Phyllis E. Phillips and Barbara A. Graham
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