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(pvright
Levels
of Platelet
KIRK
KINDSFATER.
M.D.t.
vestigation
Calmodulin
ABSTRACT:
tor
protein
tems
regulates
the
of skeletal
muscle
and
calmodulin
olescents
ship
were
THOMAS
severity
levels
of idiopathic
enteen
patients
severity
who
had
The
the
was
study
idiopathic
two normal
volunteers.
Platelets
were
isolated
all
adolescents.
centrifuged,
tant was
and
isolated.
measured
ploys
with
use
of
(more
than
twelve
months)
‘9-labeled
showed
patients
10 degrees
(3.83
venous
blood
native,
in the
of
gram
*No
benefits
in any
have
been
received
or will
unlabeled
calcurve
previous
of pro-
levels
nano-
to be an
be received
80262.
:I:Woodridge
Orthopaedic
Clinic,
3550 Lutheran
Parkway
West,
201 . Wheatridge.
Colorado
80033.
§Kempe
Research
Center,
The Children’s
Hospital,
1056 East
19th Avenue.
Denver.
Colorado
80218.
9jSouthern
Colorado
Orthopaedic
Clinic.
4233 Academy
Place.
Suite
101. Colorado
Springs.
Colorado
80909.
#Orthopaedic
Center
of The Rockies,
2500 East Prospect
Road.
Fort
Collins.
I 186
Colorado
80525.
The
Chi/dren
WEINSTEIN.
progressive
was
curve
had
curve,
the
0.69
a Risser
average
=
sign
level
per
who
did
spine
predictor
Risser
of pro-
sign
of
alone.
0 or
per
In
1 and
of platelet
microgram
nanogram
patients
let calmodulin
vature
of the
acute
the
nanograms
with
in the
more
a
calmod-
of protein,
microgram
had a Risser
0.65 nanogram
and a stable
curve
and
of protein
in those
who had
2 and 5. A single
determination
r
MDI.
ter
than
who
4.42
compared
Den
possibly
of the
patients
tein
‘s Hospita/,
and
gression
the
DAVID
of pro-
sign
of 0 or 1
per microgram
a Risser
sign of between
of the level
of plate-
not predict
the degree
of cur(Pearson
correlation
coefficient,
0.30).
CLINICAL
RELEVANCE:
appears
of the
curve
to be a useful
The
level
predictor
of platelet
calof progression
scoliosis.
If future longitudinal
studies corroborate
our
results,
these levels could be used clinically
to avoid
unnecessary
irradiation
in patients
who have a low risk
em-
from
a commercial
party
related
directly
or indirectly
to the subject
of this article.
Funds
were
received
in total
or partial
support
of
the research
or clinical
study
presented
in this article.
The funding
source
was the Kempe
Research
Center.
The Children’s
Hospital.
Denver.
Colorado.
tDepartment
of Orthopaedics.
University
of Colorado
Health
Sciences
Center.
42(X) East
9th Avenue.
B-202.
Denver.
Colorado
Suite
Center.
PH.D.11.
COLORADO
that
of protein)
(p < 0.01);
the
and the control
group
(0.69
form
LAWELLIN.
DENVER.
Scoliosis*
modulin
per microgram
per microgram
of protein)
were similar.
The level of platelet
calmodulin
appeared
of Progression
supernawas then
tein) was significantly
higher than the level in the patients who had a stable curve (less than 5 degrees
of
progression
in the previous
twelve months)
(0.60 nanogram
per microgram
in the stable
group
DAVID
and
calmodulin.
that the level
of platelet
who had a progressive
of progression
of
patients
who
problems
homogenized
between
nanograms
5ev-
of varying
a radioimmunoassay
binding
and
results
in the
or the
consisting
the calmodulin-containing
The level
of calmodulin
competitive
calmodulin
The
modulin
the
were
lizeorporated
Idiopathic
ulin
a relation-
scoliosis
group
from
platelets
of
ad-
included
ten age and sex-matched
subjects:
eight
were being managed
for non-scoliosis-related
and
sys-
levels
progression
The
Surgery.
independent
recep-
in twenty-seven
and a control
The
Research
protein
there
and
,,,d Joi,,i
M.D.#.
at Kempe
platelets.
scoliosis.
and patterns
M.D4.
AKMAKJIAN.
contractile
whether
these
LOWE.
JACK
performed
measured
to determine
between
of Bo,u’
for the Prediction
is a calcium-binding
that
platelet
Jourizal
i/U’
of Adolescent
AND
Ill
by
Calmodulin
and Severity
BY
I 994
in patients
of progression
rately
the need
have
of the curve
and
for management
ative
intervention,
risk
of progression.
Idiopathic
who
or both,
scoliosis
deformity
in children
ology
and pathogenesis
and
adolescent
idiopathic
to predict
more
accuwith a brace
or oper-
in patients
who
have
a high
is a common
cause
of spinal
adolescents:
however.
the etiare still unknown.
Idiopathic
scoliosis
is probably
a multifactonial
abnormality
in
which
genetic,
biochemical,
and neuromuscular
factors
are
involved7’9”4’9.
Abnormalities
in skeletal
in patients
who have idiopathic
postulated
that
this
muscle
have
been
found
scoliosis,
and it has been
is a possible
Many
investigators
have also found
structure
and function
of platelets
idiopathic
scoliosis8’3’5”9’23.
Because
etiological
factor2.
abnormalities
in the
in patients
who have
the contractile
pro-
tein systems
(actin
and myosin)
of platelets
muscle
resemble
each other,
it makes
sense
and
that
these
elements
would
be
derlying
systemic
disorder
is some
uncontractile
systems.
increase
Cohen
in the
affected
if there
involving
these
skeletal
both of
et al., in 1985, found
a 2.5 to threefold
level of calmodulin,
an essential
media-
ton of calcium-induced
patients
who had
THE
contractility,
idiopathic
scoliosis.
JOURNAL
OF
BONE
in the
They
AND
platelets
of
also found
JOINT
SURGERY
LEVELS
OF
PLATELET
TABLE
DvI’A
IOR
I’IIE
SIvENIIIN
WEb
PA’rIINi’s
HAD
I
SCOLIOSIS
IDIOI’ATIIIC
1 187
CALMODULIN
AND
THE
TEN
SUBJE-IS
Wuo
WERE
IN
‘tIlL
CoN’IIo1.
GRouv
Progression
Date
of
Birth
Case
Sex. Age
at Time
of Study
(Yrs.)
Risser
Sign
Diagnosis
of Curve
in 12 Mos.
before
Study
(Degrees)
Primary
Curve
(Degrees)
Type
of
Curve
Level
of
Platelet
Calmoduhin
(ng/.tg
Protein)
Brace
Idiopathic
scohiosis
group
I
12/78
F, 13
0
Double
35
>10
Yes
12/77
F. 14
0
Double
35
>10
No
15
3
48
28
>10
No
1.47
<5
Yes
0.45
0.87
6/76
M.
4
10/76
F, 15
4
Thoracic
Double
10.67
1.56
‘5
6/77
F. 14
2
Thoracolumbar
30
<5
Yes
6
5/76
F. 1 5
3
Thoracic
26
<5
Yes
0.11
7
7/78
F. 13
1
30
<5
Yes
Yes
0.57
0.57
S
10/76
9
5/77
It)
2/78
4
Double
Lumbar
16
<5
14
0
Thoracic
15
<5
No
0.09
F. I 3
5
Thoracic
16
<5
No
0.31
14
1
Double
58
>10
Yes
3.25
0.67
F. 15
M.
6/77
M.
12
I 0/71
F. 20
5
Double
50
<5
No
13
I 1/80
F, 1 1
0
Thoracic
90
>10
No
2.20
14
6/77
F. 14
0
Double
<5
Yes
0.53
15
7/76
5
Double
35
22
<5
No
0.72
16
6/78
M.
13
0
Thoracolumbar
24
<5
Yes
1.10
17
6/79
M, 12
0
Double
20
<5
Yes
1.16
F. 15
thoracic
Control
group
18
12/80
F. I 1
19
12/79
F, 12
20
12/80
Congen.
plasia
hip
M.
I I
F. 19
21
5/72
22
6/75
23
2/75
F. 16
24
1/77
F.l3
25
1/81
M.16
M.Il
dysof
(hilat.)
1.23
Overlapping
5th toe
0.78
Osteochondroma
Plica
0.32
Tear,ant.
cruciate
Meniscal
0.57
1.13
hg.
tear
0.29
None
(normal
volunteer)
None
(normal
volunteer)
0.63
0.72
26
12/73
F. 18
Cerebral
palsy
0.57
27
5/78
F. 13
Cerebral
palsy
0.65
that
the level
of calmodulin
was associated
with
the
severity
of the spinal
curve.
These
findings
have clinically
relevant
implications.
modulin
Not
factors3”.
only
research.
can
platelets
but
theoretically,
be
used
as a model
if the
levels
for
of
muscle
calmodulin
and
predictor
of the
severity
be made
available.
This
would
young
patients
to avoid
multiple
of the
allow
exposures
disease
predict
more
accurately
the need
for management
with
a brace
or operative
intervention,
on both,
in patients
who have a high risk of progression
of the curve.
At the time of the study of Cohen
et al., only indirect
were
VOL.
NO.
76-A.
available
S. AUCJLJST
to
1994
measure
the
levels
tive than
developed”.
can
many
of these
to radiation,
as fewer
nadiographs
would
be needed
for patients
who
have a low risk of progression
of the curve
(a low level
of calmodulin)’2.
Surgeons
also could
use these
levels
to
methods
activity.
their
assays
of cal-
new
These
Methods
of
that
are
previously
The purpose
techniques
Cohen
methods
et al.
enteen
dence
direct
used
questionable,
as
by multiple
measurement
more
and
to
better
prediction
scoliotic
Materials
The
time-consuming,
accurate
by
and
techniques
have
radio-
more
sensi-
recently
been
of the current
study
was to use these
to reproduce
and extend
the findings
of
calmodulin
in the
sion of adolescent
Selection
were
accuracy
and sensitivity
were
are susceptible
to interference
immunoassay
in the platelets
of patients
who have
idiopathic
scoliosis can be measured
easily,
a conveniently
obtainable
marker
and
the
define
the
role
of the severity
curves.
and
of
and
platelet
progres-
Methods
of the Subjects
study
comprised
patients
who
of idiopathic
twenty-seven
adolescents:
had radiographic
and clinical
soliosis
of varying
severity
sevcviand
KIRK
1188
patterns
and who had
one year, and a control
sex-matched
in the
subjects
been
followed
for a minimum
group
consisting
of ten age
(eight
Orthopaedic
patients
Clinic
non-scoliosis-related
teens)
(Table
I).
at The
problems
control
All
in
each
twenty
scoliosis
seven
female
group.
The
group
was
and eleven
and control
The
primary
who
had
Children’s
of
and
been
spine,
and
patients
had
fourteen
years
curves
had
a
34
to
degrees
(range,
curves
except
one.
five, a single thoracic
Eight
patients
had a double
curve;
two, a thoracolumbar
curve;
curve;
and one, a double
thoracic
classified
as being
stable
had progressed
to more
curve.
or prothan
30
with more
than 5 degrees
of progression
year, it was classified
as progressive.
degrees,
preceding
tion,
if a curve
had
progressed
more
than
right
30 degrees
in the
Ten
to -70
assay.
When
all
hydrochloric
in
12 and 14) that
to more
than
patients
were
being
managed
with
was determined
Patients
in whom
a brace
ysis was visible,
whom
as much
visible
having
as having
as 50, 75,
the
for
sample
was
hours
then
and
force
of gravity
for
had been
disrupted
from
the
All
blood
and
a Risser
sign of 1; those
in
on 100 per cent of the iliac
appeared
sign
drawn
(Becton
coated
tube
was
adequate
The tubes
of gravity
rich
plasma.
from
fused
to
warmed
One
adolescent,
mixing
were
for
The
sulfate,
and
acid.
The
of a Bronson
California)
at
one-minute
pulses
at 50 degrees
at
Celsius
15,000
hundred
times
the
all cells
debris
microliters
and
the
samples
were
coded
of su-
so that
tual
assay was done
in a blinded
fashion.
The levels of calmodulin
were measured
the
calmodulin-I’
ben NEK
RlAgent
-018;
Dupont,
System
New
the
with
use
kit (catalogue
England
acof
num-
Nuclear,
Boston,
C
the
0
0
C
0
of 2, 3, and
apophysis
iliac
wing,
0.3
as
0.5
5
1
Unlabeled
Samples
adolescents
had
an antecubital
vein
Dickinson,
Rutherford,
with
ethylendiarninetetna-acetic
invented
one-millimolan
five minutes.
to ensure
that
and to separate
the cellular
supernatant.
prorinsed
Tnis
pernatant
was pipetted
off and used for protein
analysis2. The remaining
supennatant
was used to measure
the
levels of calmodulin.
Two samples
were assayed
for each
of 5.
of the Blood
twenty-seven
tube
was
ready
and
Francisco,
centrifuged
Calmodulin
gently
two
five
milliliters
into
a Vacutainer
New
or three
times
Jersey)
acid.
of
that
The
to ensure
of the anticoagulant
with the blood.
then centrifuged
at 200 times
the force
fifteen
minutes
to obtain
a plateletsupernatant,
free
of
red
and
white
Graph
noassay.
modulin
this
whereby
assay
sites.
of tracer
THE
a
>Icalmoduhin
= -14.5
ln
utilizes
When
(labeled
JOURNAL
OF
radioimmu(unlabeled
cal-
competitive
a radioactive
calmodulin)
and a non-radioactive
labeled
calmodulin)
compete
antibody-binding
fixed amount
20
(nanograms/milhihiter)
showing
a standard
curve
for
Per cent
bound
‘25I-calmoduhin
concentration)
+ 58.6.
Massachusetts);
binding,
10
1
FIG.
Preparation
collected
containing
San
in
were
were
thawed,
of tcn-millimolar
power,
employing
three
sample
was kept on ice.
twelve
they
three-millimolar
magnesium
ethylenediaminetetra-acetic
then homogenized
with use
Scientific,
minutes
E
a Risser
sign of 0;
of the iliac apoph-
visible,
as having
a Risser
sign
and those
in whom
the entire
a Rissen
until
been
7.8,
centnifuged
five
from which
the superpellets
were then fno-
platelet
pellets
in a solution
pH
off and
Scien-
at the
with
use of standard
radionone
of the iliac apophysis
was visible
were
classified
as having
those
in whom
as much as 25 pen cent
was
had
were
for
Celsius
samples
acid,
The
0
apophysis
was
4, respectively;
tubes
pellet,
platelet
pipetted
tubes
(Fisher
past.
time of their entry
into the study, and no patient
had had
any operative
intervention
for the curve.
The Rissen
sign of each patient
who had idiopathic
scoliosis
graphs’7.
These
of gravity
degrees
(VWR
while
in the
In addi-
10 degrees
then
100 pen cent
the preceding
year, it was considered
to be rapidly
progressive.
All other
curves
were considered
to be stable.
With use of these
criteria,
five curves
were found
to be
rapidly
progressive
at the time
that the blood
sample
was obtained.
The remaining
twelve
curves
were
found
to be stable,
although
two curves
(Cases
were
identified
as stable
had progressed
at -40
for
sonicaton
with
a lumbar
curve:
The curves
were
gressive.
If a curve
being
force
was
Eppinger
to obtain
a platelet
was decanted.
The
dithiothreitol,
one-rnillimolan
platelets
were
idiopathic
curves
the
cessed,
the frozen
once, and suspended
15 to 90 degrees),
one,
thonacic
eleven
the
Colorado).
times
zen
idiopathic
(range,
averaged
all
Denver,
1000
order
natant
volunnormal
and three
male subjects
were
average
age of the adolescents
to nineteen
years
for
groups,
respectively).
AL.
blood-cell
contamination,
placed
into 1.5-milliliter
at
for
none
ET
tific,
seen
Hospital
and two normal
subjects
had had
findings
on examination
of the
family
history
of scoliosis.
Twelve
female
and five male
scoliosis,
and
in the control
KINDSFATER
antigen
for
antigen
a fixed
protein(I’25-labeled
(native,
number
unof
unlabeled
antigen)
antigen
and a
are allowed
to
BONE
JOINT
AND
SURGERY
LEVELS
OF
PLATELET
E
had
a stable
o,3
more
than
0
let
0
E
Cu
cn2
0
gram
C
Cl)
z
curve
and
sixfold
was
0
of protein).
to the
levels
The
.c,
divided
tients
0
E
Cu
00
Control
Scohiosis
Fw;.
Bar
graph
comparing
the
control
group
with
scohiosis.
The I bars show
react
with
a constant
and
decreasing
amounts
as the amount
of
use
centnifugation,
radioactivity
curve
noted
This
from
with
and
which
may
of the
paired
be
values
from
the
total
counts
of calmodulin
less
to
are
concen(Fig.
be
than
a 15 per
reported
in
with use
correlation
was
had
had
calmodulin
had
groups,
immature
highly
a stable
similar
to
and the
relative
4).
scoliosis
in order
osseous
a
significant
curve
results
(Fig.
idiopathic
had
had a stable
pen micro-
were
compare
indices
then
the pa(a Risser
a threefold
with
0.65
increase
in the
compared
with
sign of between
nanogram
5); however,
this
per
average
microgram
difference
level
of
the eight
patients
2 and 5 (2.3 com-
was
not
of protein)
significant.
100
r=0.30
80
Cl)
60
0
0
0
0)
a)
0
40
20
0
as nanograms
performed
the Pearson
who
group
who
those who
nanogram
the
0
of calmodulin
per microgram
of protein.
This normalizes all samples,
as any differences
in the levels
of calmodulin
secondary
to different
platelet
counts
in the
platelet-rich
protein
are accounted
for.
Statistical
analyses
were
Student
unpaired
t test and
efficient.
patients
with
0.60
A
of plate-
0
1). All
included
the
group.
level
.
of cala standard
unknown
to have
than
After
and
the
a gamma
concentrations
to construct
for
(Fig.
in
control
of 0 or 1) with those
who had more
mature
ossedevelopment
(a Risser
sign of between
2 and 5).
nine patients
who had a Rissen
sign of 0 on 1 had
pared
free
system.
by interpolation
had
in
analysis.
The levels
known
are used
obtained
samples
difference
separated
antibody
into two
who
had
more
of antibody,
supennatant
is discarded
pellet
is measured
with
Standards
are assayed
in
who
the
average
difference
of platelet
patients
platelet
calmodulin
who had a Risser
amount
are
a second
the
in the
trations
cent
of
calmoduhin
had
idiopathic
are bound
to the antibody
antigen
is increased.
The
complexes
with
counter.
modulin
limiting
of tracer
unlabeled
antigen-antibody
antigen
2
the average
level
of platelet
that
in the
patients
who
the standard
errors.
sign
ous
The
for
in the
curve
compared
compared
with
(p < 0.01). The patients
subjects
in the control
C
0
those
increase
calmodulin
progressive
curve
(3.83
Cl)
1189
CALMODULIN
of the
co-
2
4
Calmoduhin
6
(Nanograms/Microgram
FIG.
Graph
showing
the
modulin
and the degree
had idiopathic
scoliosis
correlation
of curvature
(r = 0.30).
8
10
12
Protein)
3
between
of the
the level
of platelet
spine
in the patients
calwho
E
Cl)
0)
0
Results
Analysis
scoliosis
of
and
the
the
variables
control
C.,
of
groups
age
and
showed
sex
no
for
the
‘a
E
significant
Cl)
0)
difference.
The average
0
C
levels
than twice
as high
in the control
group
of platelet
calmodulin
in the patients
(1.55 compared
were
Cl)
more
who had scoliosis
with 0.69 nanogram
z
levels
of calmodulin
with
lation
coefficient.
This analysis
the data points,
with minimum
variables
(n = 0.30) (Fig. 3).
The
data
for the patients
curve
VOL.
were
76-A.
compared
NO.
8. AUGUST
with
1994
use
of the
Pearson
come-
showed
a wide scatter
correlation
between
who
those
had
for
the
in
the
a progressive
patients
a)
0
0
per microgram
of protein)
(Fig. 2). However,
this result
was not significant
(p > 0.05).
The degree
of curvature
was evaluated
in relation
to
the
C
as
who
-
E
Cl)
0
Controls
Stable
Curves
Progressive
Curves
FIG. 4
Bar graph
comparing
the control
group,
that
and that in the patients
standard
errors.
the average
level of platelet
calmoduhin
in
in the patients
who had a progressive
curve,
who had a stable
curve.
The I bars show
the
1190
KIRK
KINDSFATER
ET
group
that
had
divided
again,
( four patients)
The
into
and
those
those
E
C)
patients).
-a
E
Cl)
AL.
3
This
difference
between
C
Cl)
z
analysis
in the
the
a Rissen
sign
0 or
1 was
who
who
a progressive
a stable
curve
curve
(five
revealed
average
two
had
had
a more
levels
groups
a)
0
cult
(4.42
to
interpret
the
results
are involved.
There
was
average
levels
of platelet
tients
who
had a Risser
.
E
than
of platelet
when
sixfold
calmodulin
compared
nanogram
pen microgram
of protein)
mal statistical
analysis
was performed,
0
of
with
0.69
(Fig. 6). No forsince
it is diffisuch
small
numbers
virtually
no difference
calmodulin
between
sign of 0 or 1 and
in the
the paa stable
curve,
RisserOorl
Risser2-5
FIG.
those
who had a Risser
sign of betwen
2 and 5,
the control
group
(Fig. 6).
We also compared
the difference
in the levels
of
platelet
calmodulin
between
the ten patients
who were
and
5
Bar graph
comparing
the average
level
of platelet
calmodulin
in
the patients
who had a Risser
sign of 0 or 1 with that in the patients
who
had
a Risser
sign
of between
2 and
5. The
I bars
show
the
standard
errors.
managed
not wear
with a brace
a brace.
The
an increased
pared
with
E
and the
patients
average
level
those
who did
pared
with
1.0 nanogram
This difference
was not
seven
who
of platelet
not wear
patients
who
wore
a brace
calmodulin
a brace
(1.93
per microgram
significant
(Fig.
did
had
comcom-
of protein).
7).
Discussion
spinal
per
Idiopathic
deformity
cent
of
scoliosis
is the
in adolescents,
all
types
Despite
numerous
sis of this disease
It has
Controls
Risser
0 or 1
Risser
2
-
5
FIG.
Risser
0 or 1 &
Progressive
& Stable
6
Bar graph
comparing
the average
level
of platelet
calmoduhin
in
the control
group,
the patients
who had a Risser
sign of 0 or 1 and a
stable
curve,
the patients
who had a Risser
sign of between
2 and 5,
and the patients
who
had a Risser
sign of 0 or 1 and a progressive
curve. The I bars show the standard
errors.
scoliosis
studies,
remain
been
suggested
osis21’2.
identified
Multiple
in the
that
disorder
anatomical
be
and
surprising
in patients
action
of calcium
tor protein
lam calcium
enzymatic
scoliosis,
ofmuscles
FIG.
7
level
of platelet
a brace
with that
calmodulin
in
in the patients
may
be
that
that
abnormalities
the
of platerecep-
of eukaryotic
celluof many
important
who
have
the contractile
its interaction
its regulation
of
reticulum.
Cohen
idiopathic
properties
with actin
calcium
fluxes
from
et al. found
a 2.5 to
in the activity
of calmodulin
who had idiopathic
scoliosis.
was measured
with
a kinetic
JOURNAL
OF
BONE
5cc-
intracellular
a calcium-binding
systems”4’#{176}. In patients
can
with
properties
controls
be
muscle
homeostasis
of the contractile
factor
it is
would
in which
these
calcium
calmodulin
regulates
and platelets
through
THE
been
idio-
therefore,
a disorder
is calmodulin,
threefold
increase
platelets
of patients
ever,
this activity
also
have
in platelets
that is a critical
mediator
function
and a regulator
and myosin
and
the sarcoplasmic
have
who
similarities;
have
to altered
underlying
changes
patients
abnormalities
who
dysfunction
lets. The
the average
managed
with
with a brace.
series.
pathogene-
Platelets
and muscle
both contain
actin and myosin
and have many
functional
that
attributed
ondary
Bar graph
comparing
the patients
who were
who were
not managed
a muscle
of
function
is also abnormal.
It is important
to note
No Brace
large
and
pathological
platelets
pathic
scoliosis8”3”5’8.
the contractile
proteins
found
Braced
in most
the etiology
unknown7”4”6.
of primary
importance
in idiopathic
scoliosis2’.
Elevated
levels
of intracellular
calcium
in the panaspinal
and axial muscles,
along
with
altered
properties
of myosin,
have been
found
in patients
who have idiopathic
scoli-
not
E
of
most
common
cause
of
accounting
for 75 to 85
AND
JOINT
in the
Howassay,
SURGERY
LEVELS
OF
PLATELET
1191
CALMODULIN
employing
acceleration
of bovine
heart,
calmodulindeficient,
3’S’ cyclic-phosphodiesterase
activity.
This
assay involves
multiple
steps, which are susceptible
to
interference
at several
levels”. Because
the ability of the
phosphodiesterase
assay to detect
calmodulin
has been
questioned3”,
we believed
that additional
studies
would
help to validate
on refute these important
findings.
Only recently
has it become
possible
to obtain
specific calmodulin
antibodies.
This has allowed
the development
of sensitive
and specific
nadioimmunoassays
believe
that
crogram
of protein).
On the basis of the results
for the
patients
who had a progressive
curve,
it may be sunmised
that, at some
time during
the course
of prognes-
can
Such
directly
an assay
Our
measure
was
results
the
used
levels
of
in the current
demonstrated
not
well
This
but
ceding
study.
that
had
sion
who
tients
who had adolescent
with the control
group.
earlier
findings
of Cohen
had
a progressive
curve
who had
maturity.
and
a Rissen
sign
a stable
curve
and
This result indicates
of 0 on
were of the
that platelet
two
been
these
stable
and
0.53
patients
more
idiopathic
This result
et al. More
of
pen
mi-
than
levels
a twofold
in the
pa-
scoliosis
compared
corresponds
to the
importantly,
when
be a better
acute predictor
than the Rissen sign alone.
demonstrates
that the levels
of platelet
calmodulin
appear to reflect
active
progression
of a curve
and that the
outcome
of any statistical
comparison
of a control
group
current
study
was
static
had a stable
control
group,
of the curve
in nature,
because
it
involved
assessment
at only one point in time. Prospective studies
are required
to define
the dynamics
between the level of platelet
calmodulin
and progression
of
the
curve
as
a function
whether
the levels
the natural
course
of
of platelet
of idiopathic
time.
It
is
calmodulin
scoliosis;
calmodulin
pre-
level
the
The
platelet
year
calmodulin
is a factor
independent
of the Rissen
sign in
the prediction
of progression
of the curve.
On the basis
of our results,
we believe
that platelet
calmodulin
may
of progression
of
of platelet
have
than
elevated
calmodulin
levels
to
more
a low
had
our
patients
nanogram
had
the
with
in the
had
did
of
of our
increase
levels
level
supported
of platelet
calmodulin.
Our
data
also demonstrated
in the
the
severity
patients
(0.67
levels of platelet
calmodulin
in the patients
who had
adolescent
idiopathic
scoliosis
that was actively
progressive. They also demonstrated
substantial
differences
in
the levels of platelet
calmodulin
between
the patients
1 and those
same skeletal
absolute
However,
These
curve,
this probably
of
had a curve
that appeared
in the past (that
is, it was
calmodulin
of the
and
be rigidly
of data.
14)
30 degrees)
platelet
the
cannot
amount
the
are dynamic,
determination
with
theory
(Cases
12 and
been
progressive
in the
levels
a single
correlate
limited
calmodulin”.
increase
these
why
curve.
study.
a marked
that
explains
and a group
of patients
who
point
in time,
may
depend
curves
that
in
curve
were
compared
the difference
was
are
actively
the
patients
with
minimum.
have
solely
who
those
of the
This again
scoliosis,
on the
at a single
number
of
progressive.
unknown
vary during
however,
we
The authors thank Cheryl
Alongi.
MA..
the graphics
included
in this paper.
NOTE
for providing
for her help
in the statistical
analyses
and
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JOURNAL
OF
BONE
AND
JOINT
SURGERY
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