From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
Quantitative
and Hematocrit
By
J.
ALDULA
Studies
on
MEYERS,
INCE
THE
VIRGINIA
AND
of
the
rouleaux
shown
the
blood
is not
play among
gation.
In the
plasma
and
in
Nichols,29
doctorate
the
and
19l8
ins 1921’#{176}
cell
measure
proteins,
of the
with
into
one;
Council,
the
plasma
asd
rate
simultaiseous
and
protein
of cell
mass
as affectinsg
fibrinogen
but is the
beets done
A more
the
which
sedimentat
ion
of the rouleaux
amouist
upon
sedimentation
any one factor,
Little
work has
volume,
erythrocyte
character
and
agreed
chiefly
the
and Thygesen.35
thesis
upon
the
size
generally
Wardley,’3
Hesearch
rate,
some
most
dependent.
upon
various
factors.
Child
mentation
and
factors
the
Gordon
Leffkowitz,2’
found
in
of disagreement
regarding
obviously
significant:
the
erythrocytes,
are
by
work
clinsical
applications
of the test;
and those
attempting
to
ansd chemical
factors
may be responsible
for the different
found
in health
and in illness.
will h)e made
to review
the literature.
Excellent
reviews
can h)e
Furthermore,
of
Fahreus’
H.
ALFRED
M.D.
thousands
of articles
ins medical
literature
dealing
rate
of erythrocytes.
In general,
these
can be divided
those
proposing
a isew method
or a revisioms
of an existing
From
the welter
rate, two factors
are
by
PH.D.,
MUGRAGE,
been
found
in articles
by Katz
and
complete
review
may
also be
presenit
article
is based.27
ent.
R.
Proteins
Rate
the
physical
discussing
discover
what
rates
sometimes
No attempt
formed
of Plasma
Sedimentation
TREVORROW,
EDWARD
PUBLICATION
there
have
the sedimentation
three
categories:
those
the Influence
Erythrocyte
PH.D.,
M.D.,
WASHBURN,
S
of
the
size
globuliis.
of a givens
result
along
pres-
the
As
sample
of
of a complex
miterthis line of investi-
studies
were
levels
of some
made
of the
sedi-
ninety-five
chil-
dren.
On forty-three
of these,
plasma
protein
analyses
were
donse
1)0th h)y a
modified
Howe
method
and
by electrophoretic
separation.
rFhtts
a gOO(l
opportunity
existed
t.o make
a study
of the combined
effects
of several
factors
upon
the sedimentation
for cell volume,
hoping
THE
PROBLEM
Correction
charts
methods,
by
corrected
(levise
to
such
Three
1.
alsd
nso.
for
nos.
From
Colorado
Submitted
published
beers
the
which
it
our
would
for
the
5 minutes,
Child
February
21,
nsm.
volunses
mark,
below
levels
was
ansd
accepted
this
rensove(l
the
filled
2 and
nsos.
a
50
from
Department
a “correction
CELL
and
at
cell
were
were
FOR
Wintrobe
rate
tubes
I)enver,
19.53;
the
a standard
Smith,3’
method.
cell
Couiscil
Center,
for
at
Smith
plasmssa
Research
Medical
have
modified
26. Five
ito
CHART”
sedimsiemitation
manipulating
1 was filled
to the 50
4 amid 5 to different
centrifuged
of
of
the
value
a chart
for
technics
A “CoRREcTIoN
OF
have
meamss
Experiments
Tube
rate.
We first attempted
to devise
to eliminate
this one variable.
VOLUME
Westergrens
given
cell
volume.
It
893
publication
sedinsenstations
volunse
was
level
our
cans
be
instemitions to
ensployed:
with
3 to
mns.
tuh)es
well-mixed,
differenst
nsark.
rio.
hieparinsized
levels
2
of Clinical
May
8,
above
blood.
this
mark,
the tubes
had beens
3 ansd added
to no. 4
After
and
Pathology,
Cob.
for
(‘hart”
1953.
University
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
894
PLASMA
and
5 to make the
for 3 minutes
mixed
upper
height
of
by
the
gentle
of the red blood
IN
ERYTHROCYTE
column
in
agitation
each
with
during
a
5 minute
cell column
SEDIMENTATION
tube
50
a capillary
mm.
Then
pipet.
the
samples
Readings
were
period
ESR
(mm/5
being
at 5 minute
were
re-
the
the maximum
taken
of
intervals
for an hour,
the erythrocyte
sedimentation
rate
(E.S.R.).
Then the tubes were centrifuged
to obtain their respective
cell volumes.
2. Experiments
27 to 30. The entire sample
of heparinized
blood was centrifuged
for 10
minutes.
All the plasma
was removed
and
1.0 cc. added to each
of five
vials.
Then 1.0, 0.8,
0.7, 0.6, and 0.5 cc. of red blood cells were
added to the vials,
respectively.
After
the vials
had been shaken
for 3 minutes,
two Smith tubes were filled from each, and the sedimentation rate and cell volume determined
as above.
fall
level
PROTEINS
designated
as
mm)
35
29
20
o..’
-
-
32
36
26
a-a
o’.’5
23
25
Hemotocrit
1.-The
FIG.
effect
on
the
P#{149}qaaCy
,e9,a,c,
Rh#{149}umohco’$tn$,$
Ly’apl’obleato’aO
Ca’c”omo
of cs,,,
Nopfm’ls
5.ioemabsC166
()
sedimentation
rate
of
varying
volume
cell
values
3. Experiments
31 to 40. This group
differs
from
the
first
(experiments
1 to 26) in that.
the manipulations
were
carried
out
in small test tubes and vials. These vials were allowed
to stand for 15 minutes,
then shaken for 5 minutes,
after which the Smith tubes were filled.
In all,
forty
individual
curves were
obtained
showing
the relationship
in a given
case
betweens
the sedimentation
1. They
intersect
figure
cases
the se(limentation
drawn
the
showed
types
of
Despite
such
for
same
failure
for
to
rate
decreases
and
by
levelled
use
with
obtain
and
other
type
obtained
curves
criticisms
charts
our
the
rate
each
use
cell
little
Ten
of
various
correction
chart
in
possible
for
No difference
technics
charts
Three
this
curves
pattern,
increases.
of inconsistency.
the
such
common
as the cell volume
methods.
a satisfactory
volume.
show
degree
at
their
the
and
discussed
general,
other
explanations
method:
are
depicted
except
The
that
thirty
was
in
in
curves
might
have
be
not
in
discernible
above.
workers
all
devised
suggested
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
A.
J.
1. Technsics
used
in the present
2. The
Smith
tube
method
of
methods
using
a taller
column
charts
which
3. Correction
gerated
impression
The
first
those
results
obtained
a series
thse
of
curves
then
the
drew
seems
rates
were
is
faulty.
not
as
as
accurate
were
so
constructed
as
to
give
an
exag-
seem
to
be
there
different
the
answer.
was
no apparent
technics
used.
rates
mm.).
100
of
possible
whsose
Our
Also,
were
technics
in
no
are
difference
elevens
of
determined
Thsese showed
the
original
data
explansation
essential
chart,
their
not
to
the
the
experiments
using
greater
similar
between
the
Winstrobe
consistenscy
than
when
charts
were
used.
to
pattern
they
at regular
fit
published
sedimentation
scrutiny
final
lines
to
three
column,
third
curves
for
been
do
the
a close
shows
cell volume
sedimentation
furthermore,
simultaneous
tube (height
of blood
the Smith tul)e was
However,
for mansipulating
determining
had
workers;
with
above,
based
895
AL.
of blood.
explanations
by other
discussed
study
ET
of orderliness.
two
used
MEYERS,
an
parallel
data.
Wintrobe
original
which
true.
was
calculated
intervals,
upon
be
sinssilar
to
average
median
published
and
figure
line
to this
the
Wintrobe
Landsberg4’
1.
To
obtain
from
the
individual
line.
Such
a group
later4#{176} termed
correction
oi)tained
snsoothed
curves,
amsd
of lines
charts
hardly
“crude
and
artificial.”
Rourke
and Ernstene32
have as their
correction
chart
a series
of curves
which
are convergent
at the higher
cell volume
levels.
This
seems
much
more
ims line with the
type
of data
on which
these smoothed
curves
were based.
However,
their origimsal individual
curves
show
much
of the irregularity
which
led us to abandon
the
attempt
to devise
cor-
rection
for
charts
the
for
less,
protein
to
may
of
the
1, more
similarity
between
either
“the
be
of
success
irs
to
to
the
experiment
This
to
devise
by
Gram’5
Xevert
sedimentation
analyses
will
The
these
curves
rates
in
he-
rate
ansd
corrected
ac-
As
of pregnant
may
not
shown
in
processes.
assuming
be
figure
individuals
pathologic
than
Attempts
their
differemice
levels.
a new
and
by Cutler,
place
during
correction
sedimentation
be discussed
unknown
factor
Park,
and
whso state
on
a relatively
Herr7
may
small
be
that
quantity
life.”
chart
for
rate.
The
volume,
treatment
cell
it obviously
of this factor
later.
BETWEEN
VOLUME,
and
obtained.
volume
assumed.
manipulations
take
the
been
cell
statistically,
vitro,
suggested
of curves
and
with
protein
volumes
not
has
of those
plasma
RELATIONSHIPS
CELL
types
of healthy
a satisfactory
statistical
two
amid
mentioned.
if he used
rate
as
large-scale
in the
and
curves
does
in determining
QUANTITATIVE
curves
the
was
involves
which
failure
the
cell
process.
proteins,
ins the
role
Subjects.
inconsistent
insterpret
differences
a significant
into
the
just
the
than
sedimentation
plasma
plays
Materials
rates
for
and
manipulating
into
our
the
groups
a phenomenon
Despite
Westergren3t
ones
between
uncorrected
the
between
exists
due
dilution
of blood,
by
the
than
correlations
suggested
between
of these
Secondly,
introduced
be
levels
without
to
better
if he used
relationship
independent
shape
obtained
consistency
chart.
the
were made
curves
more
found
fractions
his
possibilities
First,
Individual
show
later37
plasma
Two
method.
method
Westergren
cording
in
our
Westergren
AND
THE
PLASMA
RATE,
SEDIMENTATION
PROTEINS
Methods*
subjects
from
whom
blood
was
drawn
for
these
experiments
can
be
divided
categories:
1. Forty-three
of the children
h)eing
studied
in the “birth
series”
of the Child
Research
Council.
Blood was taken at approximately
six month intervals.
The total of seventy-eight
bloods in this group includes
one determination
on each of fourteen
children,
two on each
of twenty-three
children,
and
three
on each
of the remaining
six children.
These
subjects
range in age from 3 to 16 years.
In all but five instances
the children
were
considered
healthy
by a staff pediatrician.
2. Twelve
ward
patients
at Colorado
General
Hospital,
whose
ages
range
from
10 to 71
years.
*
Original
data
are
available
from
the
authors.
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
896
PLASMA
Obtaining
mum
of
blood
stasis.
vial
was
following
tion
of
was
cc.
stoppered
done
in a small
on this
serum
of
venous
containing
about
blood
were
mg.
of heparin
0.3
15 seconds.
hematocrit
drawn
with,
a mini-
made:
values
This
and
(110
portion
of
the
unsits/nsg.).
was
used
erythsrocyte
de-
for
sedimensta-
for
Methods
and
the
bore
throughout
were
the
most
in mm.
3.
in
5.
Plasma
6. Serum
of
in
electrophoretic
out methsod.
the
After
were
taken
the
percentages
Kjeldahl
Concentration.
the
formula:
sample
analysis
calculated
in
turn
to
obtain
The
and
cells
performed
were
in
performed
had
shown
no
had
been
determimsed,
Centrifuge,
divided
Plasma
was
volume.
SB.,
by the height
volume
was
100.
standard
modified
alkaline
hsematin
Morrison’
These
met
hsod,
the
the
of
is
of
on
multiplied
absolute
the
beta
by
values
the
the
(lone
with
thse
with
a pH
of 8.6
globulins
Perry,
and
so-called
gamma
of plasma
serum
deternsined
of the
total
total
protein
Gm./l00
of
The
protein
per
area
concentration
cc.
of
plasma.
used
by
serum,
cent
the
de-
the
par-
for
the
salting-
plsotographs
of cads
photographed;
as
as
consists
Since
was
was
alpha-2
Arnsstrongt
(a);
actually
gbobuhinss.
constituents
measured.
table-
fibrinogems
methods
Hill
ions
were
fraction,
protein
in
the
and
separate
fraction
basis
(y).
that
by
concemstrat
buffer
alpha-i
Edsall,
electrophoretic
content
each
(A);
out
the
each
fibrinogen
representing
by
described
fibrimiogen
analyses
albumin
globulin
usual
portions
study
proposed
method
and
Barbiturate
pointed
of
constant,
Howe
globulin,
White.28
determined:
and
gamma
separation
area
rate
blood
the
albumin,
analysis).
Moore
with
and
blood
interval
blood)
cc.cc. blood)
method).
protein,
($);
analyses
red
range.
This
Hgb.(Gm./100
C.V.(cc./100
-
were
present
was
were
following
Total
sample.
the
the
by
the
experimssents
cell
by
constanst
coborimeter.
this or any
electrophoretic
and
determined
of
were
International
red
foani.
described
with
one 5 minute
temperature
for
of
sedimentation
Tests
in an
from
Hemoglobin
and
fibrinogen
interest
red
initial
shakens
one
descent
used.
previous
this
rpm
l)acked
by
the
sedimentation
cent
fractions
globulin
by
the
of packed
(electrophoretic
ins plasma
own
at 2500
the
of 50 mm.,
values
within
the
was
of
the
then
the
Determinations
two
Our
After
1 hour
column
C H C
Budka,
a mixture
ticular
27 C.).
this,
was
the
reproducible
l)rOdUctiOn
fall in any
rate.
per
described
Armstrong,
ins mm.
rate
(salting-out
proteins
apparatus
termined
to
This
was used.
on each
used.
The
following
globulins
(a2);
beta
and
(22
from
a height
the maximum
obtaims
by
proteins
Trevorrow2#{176}
determined
model
tube
were
an(l
After
was
the
used
clot,
obtain
it
avoiding
and
of the
to
to
routine.
15 minutes;
tube
mm.
Readings
average
order
pipet;
The
sedimentatioms
to
for
a capillary
2.5
allowed
following
and
a rack.
tube,
In
the
motion
the
photoelectric
obtained
aside
the cell volume
Corpuscular
is
set
and
for
sample
M
and
were
0.25 mg. of heparims.
Tisis
the salting-out
metlsod
test
rate.
to
tube.
the
of the
comstent.
Mean
the
value).
entire
proteins.
into
intervals
centrifuged
in mm.
a plain
of about
(hematocrit
a Brocinser-Mass
4.
of
into
closely
in
sedimentation
subtracting
Wintrobe,39
up
the
was
of the
by
drawn
and
temperatures
Hemoglobin
using
by
serum
was
irregular
as
room
height
obtained
length
variation
tube
vial
diameter
samples
volume
1. The
containing
Sedimentation
very
vertically
designated
on
put
and
at 5 minute
1 hour
ordinary
was
placed
internal
made
significant
No.
hsbood
an
1.
the
vigorous
and
it has
cells
tube
determinations
of
adhere
heparin,
a
filled
Smith
was
analysis
to
Immediately
2. Cell
sample
samples.
using
duplicate
the
necessary
5 minutes,
the
of
electrophoretic
analyzing
of blood
during
protein
of
mixing
Smith34;
test
Plasma
was
it
was
in a small
shaken.
sample.
used
tube
placed
WIts
rensainder
results
in
vial
for
arid
SEDIMENTATION
were
shell
shakeis
hemoglobin
and
3. The
of
8 cc.
apportionments
and
the
ERYTHROCYTE
rate.
2. Another
at
placed
stoppered
terminations
IN
Approximately
sample.
The
1. One cc. was
The
PROTEINS
determined
protein
these
by
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
A.
J.
MEYERS,
ET
897
AL.
Results
a. Correction
of sedimentation
plasma.
through
According
between
the
a liquid
of red
blood
body
and
cells
specific
centration
the
aisd
are not spherical,
plied
directly
to
In the
rates
for
variations
to Stokes’
law’2 the
medium
varies
directly
the
liquid.
In blood,
liquid
medium
but irregular
sedimentation
present
study
in
the
solid
body
plasma.
1 -Coefficients
of
Correlation
for
between
estimating
1arious
Sedimentation
(Plasma
Proteins
-Corrected
Fibrimsogens
Globuhims
Ahbumims
gravity
:
E.S.R
of the
plasma.
Mean
cc. cells) was used as an
metic
difference
between
as the factor
“H”.
Obviously
entiating
the various
blood
as the
Inasmuch
be indepenidenit
have beets with
ematical
derivations
Derivation
Total
relative
Salt imsg-out
Method)
be
values
for
the
Influence
between
prediction
these
formula
E.S.R.
+524
+255
-.465
-.184
-.388
+105
-.083
-.421
+304
concenstration
and
the
sedimenstation
+ .960
influence
of
±
Table
variables:
fib.
except
correlations
.008,
rate
should
to the value
it would
was used in the math-
described.
formulae.
1 lists
corrected
+
1.361
The values
of the constansts
preceding
tance
of that
factor.
Albumin
was
cient
/ 100
meaning
only as a basis for differto relative
specific
gravities.
“H”
glob.
the
simple
coefficients
E.S.R.,
+
(Formula
without
(Cm.
conivenience
the arithhas been
labelled
of corplasma/cell
Combinsaleads
to a
levels.
This
is:
(12.368
=
the
-.569
-.590
cell volume,
ratio,
and plasma
proteins
as determined
by the salting-out
method.
tion
of the data
1)y the multiple
correlation
method
of I)oolittle3’
prediction
of E.S.R.
from
the cell volume
value
and the proteins
relation
proteins
confor specific
CV.
hemoglobin
between
to
ap-
P/C
factors,
each
rate was corrected
factor
of 25. This corrected
figure
now
be
Mb.
of cell specific
gravity.
For
two values
(M.C.H.C.-T.P.)
the values
have
samples
in respect.
of prediction
bodies
cannot
plasma
value
which
+557
+348
relationship
of other
au “H”
by
corpuscular
index
these
relative
Factors
Glob.
+901
law
solid
Rote
Determined
Fib.
and
of aggregates
the
Stokes’
gravity
of cells and of plasma
was adopted.
(Gm./100
cc. plasma)
was used
for the
TABLE
of cells
conssists
However,
therefore,
method
gravity
of fall of a spherical
solid
body
the difference
in specific
grait}y
is the
outline;
rates.
indirect
an
in specific
speed
with
the
as
between
results
each
insto
observed
indicated
an
all).
.179
-
C.V.
+
3.377)
K-i)
omitted
it enters
.090
factor
the
later
the relative
imporformulae
since it was
“H”.
Although
represent
from
the
factor
and
predicted
error
probably
sedimentatio,s
ins the
underlying
the
coeffi-
rates
was
assump-
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
898
PLASMA
tion
with
of a linsear relation
between
a high cell volume
gave
of forty-six
forty-four
cases)
cases).
theorized
that
PROTEINS
IN
influence
with
low
of the
[P/C(7.546
=
cell
significaist
was
plasma
be better
measured
in terms
of the
face or volume.
In the prediction
fractions
was computed
in Gm./100
by the plasma/cell
ratio:
E.S.R.
coefficient
predicted
proteins
+
fib.
by
of correlations
by
this
the
formula
TA
formula
for
.722
2 -Coefficients
BLE
glob.
in the
high
Correlation
Proteins
and
between
Determi
E.S.R
+901
low
cell
l’ario
us
glob.
and
which
those
predicted
groups
Factors
mentionsed
In
uence
the
Ansalysis)
gnob.
a,
Mb.
glob.
+442
+195
+730
+744
+114
+673
+ .146
+711
+ .067
+ .066
+ .079
globulins
might
values
volume
a
+ .390
globulins
rates
of the
+.)l
globulins
a
+ .730
P/C
+524
+255
-
.734
-
.656
-
.355
+.
-
.196
+ .067
-
.543
+ .294
Albunsins
*
Fibrinsogens
elect
above
more
gives
properly
an
value
that
but
globulin
consists
even
was
and
high
better
ins
1.
table
No
values
for
or when
may
and
cells
of correlation
were
was
factors
not
Simple
obtained
by
in the
method
fractions.
have
different
therefore
upons
should
coefficients
of correlation
using
fraction
were
the
the data
have
volume
factor,
significant.
elevated,
as when
normal
range.
if prediction
concentration
the
fibrimso-
It is kniown
is not a single
substance,
It seems
probable
that
the
giveus
in table
of protein
rouleaux
rate.
If so,
formulae
amsalysis
are used.
such
data
was done
are
beeni
even
agreememst
only
influences
uponi
the sedimentation
be obtained
electrophoretic
formulae
using
by
as good
were
by the salting-out
of several
globulin
values
found
of prediction
as above.
results
values
all the
this
ins thse text.
This
may
imidicate
that
method
of including
the cell
predicted
as determined
of a mixture
coefficients
upon
protein
Computation
manner
use(1
explained
distribution.
by this
these
various
components
formation
of red blood
higher
samsse as
the
aS
haisdled
observed
gen
are
difference
betweens
+ .960 and + .971 is not
cases
where
the globulin
fraction
was greatly
though
the
In those
between
values
rophoreticallv,
114
+384
.461)
-.650
obtained
of
We
1.809]
sedimentation
globulins
as
of rouleaux
-
Ehectrophoretic
by
y gnob.
Fibrinogens*
y
(thirty
test.
Rate
ned
Fib.
Corrected
high
square
size
.603)
-
Tabulation
.006.
Sedi,nentation
(Serum
on the
observed
±
of
predicted
the Chi
Those
bloods
(twenty-nine
K-2)
between
is + .971
bloods
volumes
by
rate.
rate
conscenstration
of proteins
per unsit of cell surforminla
which
follows,
each
of the protein
cc. of cells by multiplyimsg
its originsal
value
(Formula
The
SEDIMENTATION
cell volume
and sedimentation
a low predicted
sedimentation
while
those
The
difference
the
ERYTHROCYTE
ins the
based
same
2. As before,
fractions
as
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
A.
J.
MEYERS,
ET
Gm./100
cc. of cells; and, as before,
the albumin
little
influence
and was therefore
omitted
from
E.S.R.
[P/C(6.060
=
fib.
+
.866a
+
.326$
899
AL.
fraction
the final
+
was found
formula,
to have very
which
is:
2.091a
+ .183a
1.958)
-
1.327]
-
2o
(Formula
The
coefficient
value
is not
(formula
fractions
K-2),
with
again
by
tions
and
of correlation
significantly
are
the
constants
globulin
exerting
(E S R):
Predicted
data
the
a great
insfluence
precediisg
alpha-2
these
thani
it does indicate
respect
to their
fibrinogen,
beta
using
higher
T-1)
but
difference
upon
the
each
globulin,
little
is + .982
corresponding
factor.
and
.004.
usinig
between
the
sedimentation
The
gamma
±
one
three
globulin,
Although
total
this
globulin
various
rate,
globulin
as shown
most
influential
with
alpha-i
fracglobulin
influence.
[Pfc(6o6oF
+
86614
ES R (mm /5 mm)
3265
+2D9)’+
)83.,-l958).)
32 7]
P1/2
-
//
10
/
‘
regression
Calculated
-
- -
2
standard
(P
line
desmohons
.042*
.
(SD.6.0
9750)
#{149}35)
/
/
,,/
,/
,.
‘
r#{216}o+9e2±oo4,//.,,
Observed
2.-Comparison
FIG.
of sedimentation
E.SR
rates
(mm/S
mm.)
as observed
and
as predicted
by formula
T-1.
In
mined
figure
2, observed
sedimentation
by prediction
formula
T-1.
Discussion
two
rates
are
plotted
against
the
rates
deter-
of Results
It should
methods
are separated
trical
properties.
first be pointed
used are not
on
the basis
In general,
out that
comparable
of solubility,
however,
the protein
because
and
albumin
fractions
of the fact
in the second
as determined
as determined
that
in the
by
first
the
they
on the basis
of elecby the salting-out
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
900
PLASMA
method
mined
PROTEINS
equals,
in amount,
electrophoretically
IN
the
ERYTHROCYTE
albumin,
; and
SEDIMENTATION
alpha-i
“total
amid alpha-2
globulins”
equals
globulinss
beta
plus
as deter-
gamma
glob-
ulins.
The
are
and
three
fractions
fractions
alpha-2
is the
shown
found
which
globulin
antibody-containing
in table
2, however,
fractions
is not
to be most
influential
are most
likely
to chaisge
have been associated
with
necessarily
fraction
of the
it is obvious
upons
proteins.6
that
accompanied
the
sedimentations
rate
with certain
illnesses.
Fibrinogen
tissue
repair24 .33; gamma
globulin
From
the
ani increase
by an
simple
in
increase
correlations
one
any
in either
of these
or both
of the
other.
An elevated
may
be raised.
ment
no.
sedimemitation
For example,
rate gives
experiment
iso insdication
as to which
no. 83 can be compared
of the fractions
with
experi-
87:
G)obulins
No.
Sc
P/C
Fib.
_______________
83
87
9.1)
1.74
1.72
The
sedimentatioms
9.8
error.
On the
No. 83 shows
2.49
0.65
and
plasma
globulin
volume
elevated
of experiments
no.
_________
1.15
0.88
cell
other
hand,
the plasma
a high gamma
globulin,
87 shows
a normal
gamma
Moreover,
a high
cell
sedimentation
rate
despite
comparison
_________
7
.525
.825
rates
Total
______________________________________________
ratios
are
1.17
1.OJ
0.63
0.74
identical,
within
proteins
fractions
with a somewhat
and
3.79
1.87
limits
of
are extremely
different.
elevated
fibrinsogen.
No.
hut a greatly
elevated
(low plasma/cell
ratio)
plasma
proteins.
This
86
glob.
__________
fibrinsogen.
can produce
cans h)e shown
a slow
by
a
89:
Globulins
Sc
P/C
Fib.
6.8
0.5
1.86
0.74
.424
.427
No.
86
81)
This
confirms
rule
out
The
the
high
degree
of other
patient
under
factors
acacia
(formula
was
ence
K-2)
of acacia
have
Comparisons
found
employed,
analysis.
in that
fibrinogen
cliusical
ansd others
aus infectious
workers,2
in the
who
mm.,
1.02
1.26
observations
of
that
but
Numbers,3#{176}
does
ins this
nephritis,
whose
1.07
0.75
a normal
obtained
not studied
therapy
for
3.9
0.90
1.13
emphasize
process.
of correlation
previously
of the
literature
Total
____________________________________________
not
Bannsick
preclude
and
rate
was
rate
the
work.
An example
si-hose
predicted
observed
2.04
2.2.)
0.73
0.40
sedimentation
mm.
his
co-
does
not
possible
is a blood
sedimentations
20.8
g)ob.
exist-
from
a
rate
Such
effects
been
reported
in the literature.426
simple
correlations
obtained
in this
study
with
those
are extremely
difficult
because
of the differeist
technics
both
for sedimentation
rate determinations
and for plasma
Our results
agree
qualitatively
with
those
of other
workers”
positive
correlations
are found
between
the sedimentation
and globulin
levels,
and negative
correlations
between
the
protein
.1719
rate
rate
.25 .39
and
amid
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
A.
albumin
agreement
and
cell
exists
volume
between
(or
J.
MEYERS,
ET
erythrocyte
our
prediction
901
AL.
count)
values.
The same
formulae
and
those
qualitative
of previous
workers.338
ADDITION
The
statistical
or more
results
of the
sedimentatious
the
laboratory
Eight
10 cc.
this
was
were
E S.R
(mm./5
that
cause
an
increase
ins one
of ais
increase
in the
protein
fractions
obtained
to samples
of heparinsized
placed
the
BLOOD
prove
the
and
and
TO
not
purified
added
drawn
centrifuged
Predicted
do
isecessarily
point,
were
study
of blood
tube
is
this
E. J. Cohn*
To
PROTEINS
above
fractions
of Dr.
to
heparin;
described
proteus
rate.
PURIFIED
OF
in
plasma
a
test
removed
tube
and
from
blood.
containing
divided
insto
mm.)
/
Prot.n
fraction
IC
added
to
/
blood
Fibrinoten
*%
a--c
,/
8 - globulin
,#‘
/
/
,
,
/
,
/
,
/
,
7
,///
/
Prediction
,‘
4
foul.
usgd
s .[P/C57546F.722G-603)-l.eo9]I-e25
,/
/
/
-
,
2
Cul...ioisd
- -
2
line
arondard
deviators
,/
/
/
110
..‘
Observed
Fna.
two
3.-Effect
portions.
tions
was
In one
of these
plasma
of these
and
type
0.6
Results
statistical
creasing
cause
*
these
and
wishs
materials.
mm.)
protein
on the
fraction
was
gamma
in the
to
tisank
into
each
vial.
Dr.
performed
using
sedinsentation
s-as dissolved.
rate
Varying
Sedimenstation
rates
From
each vial,
by the salting-out
three
propor-
of five vials,
with
left ins the original
in duplicate
on each sample.
and the proteins
determined
different
the
test
and
finsal
tube
cell
vol-
portions
of
method.
fractiouss-albumin,
globulin.
with these
proteins
analysis.
Addition
amouusts
of fibrinogen
increases
We
the
cc. pipeted
of experiment
fibrinogen,
(mm./5
fractions
plasmas
were pipeted
into
each
always
1.0 cc. Then
the cell mixture
umes
were determined
plasma
were
removed
This
ESR.
proteins
two
volume
mixed
of added
followed
the same
of albumin
causes
and
of gamma
sedimentation
Cohms
and
rate.
thse
Harvard
trend
as that
some
slowing
globulin,
on
Furthermore,
Laboratory
these
for
so
suggested
by the
of the rate.
Inthe other
hand,
increases
graciously
are
supplying
of
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
902
PLASMA
the
same
PROTEINS
magnitude
IN
as the
ERYTHROCYTE
increases
which
would
formula
K-2. This
is showni
in
from
these
additions
experiments
figure
are
regressions
These
predictions
a causal
line calculated
from
experiments
demonstrate
and the
‘estigators
these
sedimenstations
rate
who performed
studies
and
prediction
those
used
us.
Good
ansd confirm
experimensts.5
and
agreement
exists
sedimentation
similar
rate
to
of fibrinogen
those
caused
of the presenst
study,
vitro
experimensts.
studies
ausd therefore
protein
per
was
our
8 times
which
are
.n4
basis
their
proteins
previous
Only
two
16 ,m7 .22 .23 .42
our
of
results
of the
the
of many
addition
proteins
inof
experiments
fractious
results
in the
found
salting-out
as much
that
the
suggest
in a sense
similar
and
rate,
increase
ours.
failure
to devise
cell
a satisfactory
volume,
of the
to
They
with
of blood
can
be
that
experiments
and
on
by the
the
sedimentations
to formula
the statistical
proteins
of adjusting
a critical
investigators,
casts
serious
doubt
on the
Two formulae
are presented
by means
sample
in the
Kylins23
found
rate
K-2, this
results
those
rouleaux
of in
formations,
concentration
of that
CONCLUSIONS
means
together
study.
and
According
between
is determined
AND
present
technsic
in vivo
influence
the sedimentation
red blood
cell.
to a standard
given
‘
between
than
of
SUMMARY
The
work
used
as did additions
of an equal
amounit
of globulins.
ratio is 10: 1. These
ratios
indicate
good agreemenst
These
between
the
with
Mitchell’6
on the
and predicted
the background
formula
K-2.
relationship
quantitatively
Gray
be predicted
observed
againist
addition
of fibrinogen
caused
6 times
as much
increase
in the
rate as did gamma
globulins.
According
to formula
T-1, this ratio
their
experiments
additiois
of albumins
caused
conssiderably
greater
proteins
addition
compared
3, where
plotted
the
in the
decrease
be
formulae.
by
found
that
sedimentation
is 7:1.
In
used
can
in vitro,
similar
SEDIMENTATION
all sedimeistation
analysis
of the
rates
results
of other
validity
of any such correction
of w’hich
the sedimenstations
predicted
ons the
basis
of the
charts.
rate
of a
plasma/cell
ratio
and plasma
proteins
concentrations.
One formula
utilizes
the results
of saltingout analysis
of plasma
proteins,
the other
those
of electrophoretic
analysis
of
serum
proteins.
The most
influential
factors
in determininsg
sedimeistation
rate
are plasma//cell
results
suggest
adequately
Addition
bumin)
to
and suggest
these
proteins
Determination
any
clue
ratio,
fibrinogen,
that the effective
alpha-2
globulin,
amid gamma
globulin.
concenstrationss
of the plasma
proteins
are
expressed
of purified
as concentrations
protein
fractions
the
in
blood
vitro
per
confirm
unit
of cell volume.
(fibrinogen,
gamma
the
finidings
from
the
a direct
cause
and effect
relationship
between
and the rate of erythrocyte
sedimentation.
of the erythrocyte
sedimenstation
rate
as to the
level
the multiple
factors
clinical
significance
of any
one
of the
involved
is essential
of a sedimentations
responsible
for
rate
an adequate
determination.
globulin,
and
statistical
studies
the
conscentration
caninsot
factors.
The
most
give,
of
per
Ans appreciation
interpretations
an(l
properties
S.
H.,
JR.,
of serum
BUDKA,
and
plasma
M.
J.
proteins.
E.,
AND
J.
Am.
K.
MORRISON,
Chem.
Soc.
C.:
69: 416,
Se,
of
of the
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plasma
der
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Bluteiweisse
the
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f#{252}rdie
85: 574,
Scandinav.
T.,
human
T.,
proteins
sedinsentation
rate.
1942.
SHEDLOvSKY,
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the
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J. W.: Suspension
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M.:
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Sedimentation
V.:
to
des
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1941.
AND
Kinderh.
170,
on the
Einfluss
The relationship
H.:
of the
coat”)
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1953 8: 893-904
Quantitative Studies of the Influence of Plasma Proteins and Hematocrit on
the Erythrocyte Sedimentation Rate
ALDULA J. MEYERS, VIRGINIA TREVORROW, ALFRED H. WASHBURN and EDWARD R. MUGRAGE
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