From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
Inhomogeneity
of
Hemoglobin
By
D
F.
BENNETT
M.
CARL
more
HORTON,
B.
ROBERT
EVAN
NECHTMAN,
hemoglobins,
important
of these
The
THOMPSON,
NIcHOLs
AND
composed
of
are Hb-Bart’s,
common
most
Hb-F,’7
and
Hb-H,
appear,
therefore,
is to Hb-A.
Small
bloods
to
VI.
of Cord
Minor
Blood
J.
H.
recently
reported
evidence
has
of four
two types
composed
cf
the
blood
of Hb-H
in the presence
two
been
cases
carriers,24
of a gene
in which
presented
both
that
have
been
contrast
found
to the
of polypeptide
of four of the
chains.
y chains
of Hb-A.’M
in the
observed
It would
same
way
primarily
while
Hb-H
has
cf thalassemia.22
Hb-Bart’s
similar
Dozy,
HUISMAN
in
chains
that Hb-Bart’s
is related
to Hb-F
amounts
of Hb-Bart’s
have
been
and in the
principally
occur
composed
M.
ANDREE
TITUS
URING
the past few years,
some
human
hemoglobins
composed
of one single
type
of polypeptide
chain
The
of
Hemoglobin.
Components
and
been
Ramot
Hb-H
combinations
as Hb-H
in cord
reported
et al.23
were
should
present;
occur
more
often.9
During
blood
routine
electrophoretic
of Negro
babies,
ponents
were
noticed
ports
the results
of
terization
of
studies
varying
300
hemoglobin
minor
Fifty-four
cases,
samples
in
of
weeks,
selected
at
the
of
basis
of
AND
random,
present
study
rewith
the charac-
METHODS
babies
were
The
body
com-
components.
Negro
mothers.
cord
Hb-H-like
were
studied
gestational
weight,
date
used
for
more
age
of
electrophoretic
intensively
of
the
expected
studies.
along
newborns
with
was
blood
estimated
confinement
and
date
investigated
five
months
of
last
period.
Seven
babies,
When
who
showed
possible,
Routine
blood
samples
respective
on
menstrual
birth.
the
blood
of umbilical
and
in many
of these
bicod
samples.
The
a more
extensive
investigation
dealing
these
cord
hemoglobin
of Hb-Bart’s-like
MATERIALS
Over
of the
amounts
both
hematologic
cell
specific
parents
studies,
counts,
abnormalities,
were
which
hematocrit
and
were
included
in
included
osmotic
these
the
special
determinations
fragility,
were
after
investigations.
of
total
performed
hemoglobin,
using
ned
standard
proce-
dures.29
Hemoglobin
equal
solutions
volume
centrifuged
wire
performed
using
the
gel
starch
as th
9.0
sodium
0.03
bridge
From
0.5
speed
to
remove
NI
studies
Research
Submitted
were
Mar.
of
Georgia,
Grant
19,
and
the
1962;
Biochemistry,
gel
pH
as a protein
absolute
per
cent
for
the
an
hemolysates
was
preparation
hydroxide
stain
and
alcohol,
hydrogen
and
Pathology,
with
the
electrophoresis’3’26
8.1
acid-Sodium
ml.
30
hours
an
100
and
pH
0-dianisidine
ml.
peroxide)
Obstetrics
of
buffer
0.25
M
for
the
Gynecology,
Ga.
by
American
accepted
200
erythnocytes
20
Starch
Boric
ml.
0.5
washed
After
buffer
used
in
and
Augusta,
debris.
0.1
was
lOB
4.7
supported
of
a
dissolved
the Departments
These
the
triply
toluene.
acid
Amidoblack
pH
the
of
Tris-EDTA-Boric
0-dianisidine
of
mixing
volume
starch)
buffer
College
by
and
solution.
mg.
phosphate
Medical
tional
a
prepared
water
(“Connaught”
(100
reagent
were
distilled
at high
of
U.S.P.H.S.
Research
Cancer
far
Society
publication
Grant,
No.
May
H-5168,
and
the
Institu-
iN-75.
12,
1962.
302
Bi.ooo,
VOL.
20,
No.
3
(SEPTEMBER),
1962
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
OF
INHOMOGENEITY
HEMOGLOBIN
303
vi
CMC-CHROMATOGRAPHy
0D4
-
15
0.0’ N
PHOSPHATE
DEAE-CELL.
-*4-001
N
pH6.5
PHOSPHATE
01 N
CHROMATOGRAPHY
PHOSPHATE,
pH 8644-001
N PHOSPHATE
pHS4
OH6O
+O3M
Nod
0.8
0.7
0.6
0.5
VI
0.4
0.3
‘I
r
0.2
0. I
-CORD
1%
I.
NORMAL ADULT
(35-40mg
of Nb
1L.__4_’
chromotogrophed)
---
20
10
30
40
50
EFFLUENT
demonstration
of
were
the
different
determined
by
the
of
the
Quantitative
analyses
that
described
with
earlier.’
CM-Cellulose,
Amounts
Prior
to
same
buffer
with
the
pH
chromatography,
as
8.4.
During
of
period
for
the
the
When
at
cent,
of
fast
both
being
hemoglobin
the
V,
and
the
quantitative
The
isolated
alkali,”
in the
calculated
as
V2
fractions
V,
fraction
of
alkali.
Hb-H,
Hh-A
was
as
was
showed
and
was
hybridized
fraction,
isolated
from
Bart’s,”
possessed
an
and
the
UV
by
cases
as
the
with
electrophoretic
$
during
V1
with
an
stanch
Hb-fractions
gel
electrophoresis,
of
was
The
of
a large
remain-
second
elution
electrophoresis,
of
amounts
increase
the
V9
0.38
of fast
in
the
method
in
such
the
when
the
hemoglobin
of
the
V1
chain
abnormality
Hb-S.
The
hemoglobin
of
the
V9
mobility
similar
to
that
fraction
of
rate
present
electrophoretic
hemoglobin
the
a high
towards
hemoglobin
as
for
samples.
denaturation
such
moving
percentages
accurate
present
per
to that
of
and
collecting
similar
quantities
Hb-A
the
performed
hybrid
notable
of
6.5.
buffer
(V1)
that
The
Hb-H,
pH
versus
After
gel
and
experiments.6’25
properties
cm.
fractions.
a reasonably
fast
C.
was
the
notable
Hb-H)
4
artifact.
V2
cent
of
NI phosphate
starch
per
allowing
of
and
by
0.2
hybridization
spectrum
produced
with
1),
characteristic
a column
cases
at
one
(V2)
and
amounts
studied
analyses
obtained;
20
chromatographed.
ml./hour.
were
of
of
buffer
0.01
recovered
about
In
(fig.
the
10
an
with
height
Elution
with
the
performed
a modification
overnight
detectable
be
Hb-Bart’s
noted
of
were
were
to
hemo-
were
to a final
exchanger.
second
of
on CMC-
was
phosphate
approximately
was
hemoglobin.
determination
spectral
the
ML)
resistant
1 ml. ) were
dialyzed
percentages
found
(designated
fractions
UV
mobility
fraction
was
fractions
the
Ni
in
cation
fractions
and
components
fraction
of
Hb-A9)
of
0.01
continued
two
proteins,
Hb
rate
was
Hb-F+
moving
V,
a flow
an
the
alkali
used
filled
were
were
of
period,
a calculation
the
solutions
of
fractions
( dissolved
Hb
VOLUME
Visser.19
technic
cm.
x 0.9
with
of
elution
elution
(Hb-A+
no
amount
mg.
Hb
the
first
25
equilibration
non-hemoglobin
allowing
40
buffer
hemoglobin
ing
The
the
used
percentages
hemoglobin
of
50
V2 fractions
and
\7
and
moving
equilibrated
to
The
Jonxis
) chromatography.
was
35
equilibration
number
by
from
ml. of the effluent,
40-45
of
which
varying
of the
of
fast
40
EFFLUENT
zones.
method
Columns
1.21
30
)ML)
hemoglobin
( CNIC
carboxyniethylcellulose
20
0
VOLUME
Fig. 1.- The chromatographic
behaviour
cellulose
and of Hb-A2 on DEAE-cellulose.
globin
BLOOD
SI
Is
of
denaturation
designated
Hb-Bart’s,
as
while
small
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
304
HORTON
( 5-10
amounts
rate
of
identical
to
component
fast
per
by
that
of
was
hybridized
hemoglobin
of
the
we
Bart’s
Hb-F.
components
collected
composed
detectable.
while
in
small
of
of
hybrid
elution.
amounts
of
determined
by
DEAE-cellulose
curve
or
with
in
and
No
the
bulk
these
proteins
the
Hb-A2.
Considering
Hb-A
was
when
electnophoresis
non-hemoglobin
constant
a moderate
hemoglobin
Hb-A
gel
the
showed
adsorption
with
starch
phase
is composed
but
on
by
fraction
spectral
as
abnormalities
second
V1
UV
produced
detectable
the
that
was
chain
The
its
AL.
and
results,
Hb-H,
varying
and
quantities
of
hemoglobin.
The
ing
percentages
the
of
simplified
were
in
chromatographecl
range
of
below
per
cent,
Large
blood
( fig.
1 )
of
amounts
of pure
Freshly
prepared
larger
quantities
of
plied
buffer
to
a 40
rate
of
phosphate
flow
the faster
from
was
abnormal
pH
6.5.
was
ml./hour.
and
V1
the
columns.”
Hb
of
five
out
Bart’s
and
from
fraction
elution
gel
these
was
buffers
solutions
also
were
used
Since
in
dilute
Hb
of 470
and
analysis
with
presence
of
and
determined
508
the
0.01
slow
revealed
cent
solution
of
and
Hb-F
the
use
by
less
than
5 per
Hb-Bart’s
the
method
of
the
pH
of
6.5
at
a
short
CM-
Hb-A
to be
dialyzed
and
apsame
components,
against
0.1
curves of
equilibrium
Bninkman
0.01
separated
of
cent
were
from 6.5 to 7.8. Oxygen
ranging
with
completely
Hb-A
concentrated
) were
buffer
almost
with
the
ml.
20-30
equilibrated
was
moving
chromatogsamples
against
NI phosphate
which
fractions
following
m1
were
studied,
using
a Beckman
Beckman
than
pure
was
to
Hb-A2.
Dirken,2
which
studies.15’2’
solutions
the
less
values
which
0.15
of
blood
in
the
Hb-A2
from
CM-cellulose
dialyzed
( dissolved
the
Hb,
electrophonesis
pH
earlier
lengths
Hb-A
with
carefully
were
with
varying
cord
Hb
of
with
The
present.
Small quantities (4 ml.) of a 1 per
NI phosphate
Gm.
cases
percentages
by
Hb-A2
Hb
hemoglobin;
in
higher
from
were
CM-cellulose
carried
combined
Starch
isolated
fraction,
to
column
100
also
obtained
total
Hb-A2
of
) of
mg
cent
with
follow-
amounts
of
per
with
were
cent
20
samples
solutions,
Three
Elution
moving
collected
cellulose
the
be
small
( 35-40
per
samples
blood
hemoglobin
as
to
blood
adult
very
quantities
found
cord
the
chromatography
only
expressed
was
for
for
Bart’s
cm.
about
were
hemoglobin
x 2.0
buffer.
results
cent
cent
Since
reasonable
method
per
5 per
raphy.
The
.
before.16
samples,
the
10
cent,
and
NI phosphate
were
described
cord
consistency
0.15
per
Hb-A9
method
present
0.5
$
were
of
also
hemoglobin
with
in stating
is
were
reagents,
Fetal
confident
V2
Hb-A
alkaline
hemoglobin
feel
that
) of
cent
denaturation
ET
DK-2
3
Hb-F,
per
all
cent
DU
automatic
cent
of
isolated
per
by
oxygenation
recording
same
measured
at
Mixtures
procedure,
served
the
wave
Spectrophotometric
spectrophotometer
methemoglobin.
the
was
spectrophotometer.2#{176}
revealed
of
Hb-A
and
as
controls.
the
Hb-F,
pure
RESULTS
Over
The
cord
300
hemoglobin
Hb-Bart’s
or
blood
samples
patterns
fell
Hb-H,
moderate
quantities
Hb-Bart’s
and
with
were
into
small
of Hb-Bart’s
Hb-H
were
studied
amounts
and
by
groups,
three
of
Hb-H.
detectable
in over
starch
gel
namely,
electrophoresis.
with
no
detectable
Hb-Bart’s
and
Examples
are
shown
cent
of all
were
arbitrarily
into
Hb-H;
the three
groups:
2)16
cases
with
30
per
Hb-H,
and
with
in figure
the
2.
samples
studied.
Fifty-four
cord
blood
samples,
on the basis
of the starch
1) 31 cases
with no visible
small
quantities
of
amounts
of Hb-Bart’s
are
Results
shown
crease
Hb-Bart’s
and
and Hb-H.
of the quantitative
in figure
3. All
in the
percentage
selected
at random,
gel electrophoretic
amounts
of Hb-Bart’s
Hb-H;
studies
for
three
groups
of
Hb-A2
with
and
pattern
and
3)
7
cases
with
the isolated
fractions
A2,
show
a similar
tendency
a proportional
decrease
divided
moderate
V1 and
for an
in
per
V2
incent
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
INHOMOGENEITY
OF
SOWER
B2
HEMOGLOBIN
30
VI
]I
..-..
BarVs
H
23456
78910112
Fig. 2.-Starch
gel electrophoretic
pattern
at ph 8.1 of representative
hemoglobins
with no detectable
Hb-Bart’s
or Hb-H
( 1 1 ) , small Hb-Bart’s
and
Hb-H
( 6, 10)
and moderate
Hb-Bart’s
and Hb-H
(2) , Cower
II ( 10) , a heterozvgous
B2 carrier
(8),
a homozvgous
Fib-S
(5),
normal
adults
( 1, 3, 4, 7), and
the mother
of the
baby
with
of fetal
Hb-Gower
hemoglobin.
centage
of
of V, with
relationship
increase
The
When
a decrease
could
not
in V,
1.5 per
(12).
cent,
with
were
per
cent
per
cent
of
Hb
was
eluted
decrease
V2 in
as the
of
group
Hb-F,
while
in group
of V2 in this
group
In group
whereas
in
1, the
group
The
ratio
the
ratio
globin,
ranged
indicating
In
per
in group
decreased
a relatively
samples
from
the
with
as has
per
namely,
0.5
compared
about
been
to
with
0.4
pointed
per
out
apparent.
of
before,
V2 percentages
with
decreasing
was
the
cent
the
V,
and
levels
ccnstant
a decrease
per-
The
is
were
levels
of
amounts
cent.
fractions
decreasing
larger
mothers
of V,,
when
relationship
1 remained
the
group
3 a definite
a suggestion
of an
was
conditions,
1.1 to 5.5
at
in
production
centage
of fetal
hemoglobin
was high.
to be constant
at about
3.9. No definite
Hb levels
of cord
blood
and amounts
Blood
2. In
group
2, however,
cent,
and decreased
definite
from
with
about
0.2 per cent
in the percentage
group.
constant
which,
sum of the isolated
2 a decrease
with
of V2/V,
of V2/V,
compared
amounts
in this
experimental
V2 fraction,
3 no
was
group
there
Larger
present
1 remained
our
considered
to be an artifact.
found
to be higher,
0.5-1.0
noted
for
although
Hb-F.
to be
Under
of V
constant
value
for V, of
a slight
but definite
increase
in Hb-F
was
be established,
found
Hb-F.
percentage
the
Hb-F,
a reasonably
in group
1, while
found
was
II
the
of
V2 remained
of Hb-F
about
amount
Hb-Bart’s
was
two.
of
In
constant,
noted.
group
fetal
when
2
hemothe
per-
The ratio
of V2/V,
in group
3 seemed
relationship
was evident
between
total
of fetal
hemoglobin.
of the
54 babies
involved
were
studied
for
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
306
HORTON
Table
1.-The
Blocd
Percentile
of Babies
Amounts
with
of Different
Moderate
Hemoglobin
Amounts
Hb
Pheno-type
Baby
I
cord
A F Bart’s
5 1110.
Baby
II
A F Bart’s
5#{189}mo.
Baby
III
IV
A F Bart’s
A F Bart’s
5mo.
Baby V
( twin 1 )
cord
Baby VI
( twin 2)
cord
“Normal”
cord
the
the
and
from
Hb-Bart’s
mated
fetal
age.
Because
cases
these
two
of V,
The
and
62
-
66
0.31
1.7
5.45
1.7
0.18
0.22
0.18
1.11
3.84
1.9
0.15
0.16
6.5
80
6.5
0.31
1.30
84
0.25
0.20
16
0.22
0.39
1.80
84
2.4
0.17
0.10
10
0.20
0.38
84
.22
of
the
were
of the
as the
amounts
of
more
intensively.
birth
pertaining
and the
the
no close
and
the
including
of
ex-
group
Hb-A2,
(No.
1).
Hb-H
At
present
against
the
existed
between
and
Hb-Bart’s
increased
esti-
increased.
and
Blood
of
levels
relationship
age
Hb-Bart’s
The
plotted
A2 levels
fetal
presence
range.
Two
cases
per cent,
respectively.
normal
were
Hb-H
were
from 6.5 to 16 per
fell within
the normal
pertaining
below
data,
carriers.7
study.
fractions,
Hb-F,
possible
from
7.5 to 15.6 Gm.
cell trait
carriers,
three
Hb-H
in
samples
group
were
3,
these
obtained
from
and
from
their
respective
parents
when
to the babies
is shown
in table
1. Using
CMC-chromatographic
procedure,
no ap-
after
of Hb-Bart’s
V2 were
that
Hemoglobin
studies
the
Hb-B2
within
age.
in table
2. Absence
electrophoresis
and
hematologic
fell
decreased
large
and
ranged
sickle
available
for
hemoglobin
apparent
studied
Hb-A2
values
of four
of
in V2 and
fetal
percentages
amounts
and
3.24
0.0
H
levels
cases
present
amounts
of fetal hemoglobin,
levels
were
increased
and
Results
0.83
0.14
heterozygous
were
not
the various
the babies
five
months
possible.
Analytical
data
starch
gel electrophoresis
presented
starch
gel
6.5
1.8
varied
within
the normal
of Hb-A2
of 3.9 and 5.0
It became
hemoglobin
Hb-H
cent)
amount
of
and
preciable
70
0.33
0.16
total
Hb
consisted
two
per
fathers
in V,,
seven
%
0.07
A F
carriers
(1.6 to 2.9
an increased
babies
and
F
%
2.9
A
concentration,
time,
the
The quantities
fetal
V2
e/,
-
A F Bart’s
abnormalities.
The
The Hb abnormalities
trait
The
H
Vi
(7,
0.14
A
5 mo.
Hb
Hb-A2
hibited
this
A F Bart’s
5 mo.
A2
0.8
A
in
1)
total
Hb-C
H
Present
values,
group
hemoglobin
per cent.
H
AL.
Hb-H_(Group_3)
2.1
A
1110.
cord
( mean
H
and
0.25
A
cord
5’/2
Baby
H
A
cord
Components
of Hb-Bart’s
E’
to the
of Hb-Bart’s
by column
established
the
values
detectable
at this
time.
Small
cent, were
still present.
Hb-A2
range
of 1.7 to 2.4 per cent.
parents
of these
and Hb-H
chromatography.
upper
obtained
limit
was
of
in the
seven
children
are
demonstrated
The
percentages
the
normal
osmotic
range.
fragility
on
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
INHOMOGENEITY
OF
HEMOGLOBIN
tests, were
within
the
of case 2 who exhibited
Gm.
per
cent
tritional
was
or
was
iron
In
a cord
normal
range
with
the exception
of those
of the
a sickle
cell trait
anomaly.
Her low hemoglobin
probably
due
deficiency
a heterozygous
blood
to be
minor
Hb-fraction
sample
found
to be
present
weeks
after
birth
present
her
from
2
with
was
cther
one
)
a slow
a mobility
isolated
for
hemoglobin
One
abnormality
mother,
by
0.3 per
yielded
material
member
DEAE
cent.
of
the
spectrum,
mother,
we,
further
same
Examples
of the
remaining
mobility
feel
that
stressed
sample.
in the
Hb-F
curve
of
showed
constant
to be
1.08
oxygen
affinity
notable
difference
affinity
died
mother,
Hb-A
and
reduced
for
2.1
studied
ultraviolet
ab-
of five
Hb-Bart’s
be
same
cord blood
a ) an enormous
of the
oxygen
Hb-A
dissociation
interaction.
y
was
,
1 +
c)
mixture;
of
The
=
range
absence
and
4. It should
to that
AF
the
at the age
to be normal.
the
Hb’s
in
with
isolated
a wide
the
Hb-B2
identical
s equation
the
in-
absorption
of
heme-heme
over
indicating
The
frcm
the
apparent:
b)
Hill
was
two
in figure
as compared
was
found
the
eight-fold;
Hb-Bart’s
was
of
shown
of Hb-Bart’s
from
and
collected
ultraviolet
of encephalitis
appeared
are
of
was
This
sample
absence
isolated
to be
n, as calculated
for
3,
twins
fraction
Hb-B2.7
is probably
were
always
were
found
a greatly
of Hb-Bart’s
the
curves
and
the
component
( approximately
Hb-Bart’s
interaction
found
oxygen
mixture
white
result.
of
and
this
dissociation
of Hb-F
that both
components
Three
major
differences
increase
and
oxygen
mixture
case
to that
of fetal
hemoglobin,
Unfortunately,
there
was
Because
Hb-Gower
II.#{176}
The baby
subsequently
weeks.
The other
twin,
as well as the
the
a nu-
of
chromatography
of another
the
study.
electrophoretic
at present,
premature
cellulose
Analysis
approximately
for
and
mother
moving
minor
hemoglobin
comparable
to that
of the
sorption
spectrum
of this component
was similar
while
that
of Hb-B2
is similar
to that of Hb-A.
adequate
the
mother
of 8
carrier.
( fig.
7#{189}months
to
anemia.
Hb-B2
about
found
307
VI
of pH
a Bohr
K,,
when
the
values,
no
effect.
Discussiox
The
data
Hb-Bart’s
tities
of
the
fetus;
cord
globin
increased
(y)
a single
study
basis
for
These
amounts
blood
of the
this
demonstrated
of
of the
was
found
in cases
with
deficiency
The
have
Hb-H
in many
cord
abnormal
components
larger
in the
trait,
and
presented
and
these
findings,
chains
a
to be
lowered
production
blood
samples
phenomenon.
therefore,
born
decreased
Hb-F
in the
feature
lacking
premature
Apart
four
cases
heterozygous
two cases
heterozygous
is a common
the
support
human
hemoglobin
and
the
formation,
of normal
of
the
from
the
small
cases
types
were
idea
that
resulting
present
production
indicating
of Hb-H
the existence
and
(a2y2)
failed
to
with
Bart’s
Hb-A
disclose
“a
in the
hemo($.)
of
(012,82).
a genetic
“classical”
a slight
of
The quanthe age of
of
three
cases
heterozygous
no abnormalities
were
fetus,
amounts
Formation
Hb-F
mothers
two
of
cf Negro
babies.
some
extent
with
babies.
for Hb-S,
for Hb-B2,
in the
presence
blood
samples
varied
to
thalassemia
for Hb-C,
encountered.
chain
temporary
deficiency”
forma-
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
308
HORTON
ci
Iz
ci
ci
ci
It)
‘-C
ci
I v
V V V
ci
ci
ci
V V V V
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-
ci
c#{149}d#{149}cc
-
ci
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cic.i
0
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-
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c
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.
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AL.
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
INHOMOGENEITY
OF
HEMOGLOBIN
NO
STARCH-GEL
BARTS
ELECTROPHORESIS
(54
SMALL
+ Nb-H
BARTS
doses)
Nb
309
Vi
MODERATE
+
BARTS
Nb-H
+ Nb-H
(YCoses)
______________________
Is
qn,%
(4
‘3
(2
(0
9
.-..-
39..
#{149}
‘0
50
-1.,_._
08
-..
06
#{149}
,,.,
#{149}-
:‘‘:
04
40
.
.
.
‘-
2O
#{149}
02
.
(0
.
IS
03
#{149}
02
0(
‘44,
#{149}#{149} .
#{149}
O2O””
#{149}.#{149}
.
___________________________________________
04
‘;.-._s#{149}#{149}
.
03
02
--‘#{149}
#{149}#{149}#{149}
Z6 .
.
0l
.
60
.
#{149}---‘..
#{149}
Fig.
80
tion
of small
of excess
Of
larger
1.7
with
of
.
#{149}
- -#{149}#{149}-
.
quantities
of y and
particular
amounts
(00
60
10
80
90
(00
of y
interest
of Bart’s
mongoloid
origin.
Our
the
Hb
tween
ture
of the
offer
the
of the
of Bart’s
in the
parents
cases
evidence
for
eo
70
-
HEMOGLOBIN
isolated
smaller
PER
fractions
amounts
o
00
CENT-
Hb-A2,
of th
are
Bart’s
babies.
hemoglobin
In some
the
could
the
be
similar
presence
in most
abnormality
with
high
incidence
to 5.5 per cent)
results
and
parents;24
hemoglobin
seven
even
the
authors3’5’’#{176}’27’28 have
the infants
being
the
found
Negro
occurrence
and
of
is the relatively
hemoglobin
(up
rickse
et al.8 who have
cord bloods
of Nigerian
is detectable
percentages
60
units
V,,
\2
as a result
$ chains.
per
cent).
Different
similar
abnormalities,
marily
#{149}
#{149}
90
the
.
8
#{149}_-$,#{149}
10
3.-Relationship
Hb-F.
#{149}
#{149}
FETAL
versus
#{149}
established.
larger
production
existence
of a
described
offspring
definite
disease
however,
newborn
of
obtained
in about
10 per
studies
a relationship
of Hb-H
The
newborn
of parents
to those
instances,
of the
of Negro
babies
with
and of Hb-H
(up to
results
Bart’s
and
the
of the
hemoglobin
hemoglobinopathy
or
no
babies
of priby
Hend-
cent
of all
between
thalassemia
connection
be-
hematologic
continued
also
picstudies
failed
in either
to
the
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
HORTON
310
%02
AL.
ET
Saturation
80
BART’S
60#{176}
4
AF
1
Fig. 4.-Oxygen
of the hemoglobin
blood:
#{176}
LI
dissociation
components
Hb-Bart’s
and
the
curves.
of cord
Hb-A
and
F mixture.
20
I
0.5
log
parents
or
the
expression
I
1.5
P#{176}2
five-month
beyond
old
infancy
the larger
failed
to
production
demonstrate
with
amounts
low
I
1.0
infants.
of an
of a chains
any definite
of Hb-Bart’s
This
and
difference
“a
of an independent
one
regulating
the
failure
a
with
chain
control
the
production
of the
with
a large
production
samples
with
increased
of small
amounts
showing
the
family
members
of a Hb-H-like
abnormal
of
$
one
(Augusta
of
investigations,
which
were
that the occurrence
of Bart’s
of Hb-H.
This
finding,
Bart’s
and a corresponding
these
group
for
production
chains
of adult
hemoglobin.
to detect
any alteration
in the
of babies
cord blood
and
deficiences”
of the
the
existence
has
of
This
pattern
a
chains
hypothesis
of the
component
by
babies.
carried
out
Hb is always
The
and
data
with
more
accompanied
a quantitative
to the
two
and
quan-
hypothesis
chain
a
absence
and c)
the
in
cord
the
obtained
refined
by
genes,
a second,
that
was
based
on:
hemoglobins
of the
in both
fraction
of
for
moderate
of
of Hb-F,
absence
responsible
Our studies
also
between
group
2
led
of Hh-Bart’s;
b) the
amounts
of Hb-Bart’s;
1)
be
3 with
which
chains
a
the
could
hemoglobin
types.
for distinguishing
Hb-H,
Hb fractions.
offered
evidence
suggests
which
lacking
basis
tities
of these
abnormal
One
of us’4 recently
in cases
finding
abnormality,
of
the
Hb-H
in
presence
blood
samples
blood
in
technics,
definite
of
a) the
parents
of
the
some
present
indicate
production
together
with
the noted
decrease
in formation
increase
in the amount
of Hb-H
when
the
of Hbper cent
of Hb-F
was lowered,
supports
the existence
of one single
deficiency
in the
production
of a chains
of Hb-A
and Hb-F
in many
newborn
babies
and makes
the above
mentioned
hypothesis
no longer
tenable.
Abnormalities
other
than
Hb-Bart’s
and Hb-H
were
encountered
in cord
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
INHOMOGENEITY
bloods
OF
in our
studies,
the discovery
of
of interest
because
fetuses
only.’#{176}
of this
abnormal
the
and
$“
HEMOGLOBIN
namely,
component.
babies,
The
Bart’s
results
of the oxygen
bear
some
underlying
for
gen
affinity,
a decreased
in
prevented
in the
an
that
in
cord
to
showed
heme-heme
while
those
study
such
blood
as
samples
of the
aspects.
recently
an
interaction
extensive
of
material.
determinations
and theoretical
Hh-types
cases,
abnormalities
presented
comparable
two
one
( white ) baby was
to be present
in very young
demonstrated
present
chain-lacking
Hb-C
in
noting
equilibrium
physiologic
are
cases,
material
been
not
Hb-Bart’s
a
of
is worth
have
were
four
component
is considered
It
which
$4e,12.14
Both
in
lack
Unfortunately,
Negro
Hb-H.’
Hb-S
a Hb-Gower-II-like
this component
a few
obtained
311
VI
Hbresults
described
encrmous
and
isolated
The
the
for
increase
absence
the
in oxy-
of any
Bohr
effect.
It,
therefore,
becomes
tional
abnormalities
physiologic
oxygen
This
might
explain
Lie-Injo
Luan
Eng
The
absence
chains
chain
a--I’
plays
Bohr
effect
suggests
an
for
the
hemoglobin
types
funcunder
tissues.
in Hh-Bart’s
of
and
a
of the
y
or
protein
likewise
interaction
sigmoid
gross
since
to the
composed
between
a direct
the
such
children
described
by
hemoglobin
fraction.4
conformation
interactions
and
newborn
the major
interaction
than
with
malor
component,
be unavailable
of the
formed
in determining
of Hb-F
the
would
in the
rather
role
curves
an
change
interaction,
important
sociation
a hemoglobin
as
in
that
the Bohr
effect.
absence
of heme-heme
The
an
of the
is essential
for
that
not occur
its oxygen
the cause
of death
in whom
Hb-Bart’s
or $ chains
chains
ble
apparent
could
tensions,
y
and
$
responsi-
indicates
between
shape
only
a
that
the
of the
hemes,
oxygen
dis-
Hb-A.
SuIAnY
In
a
survey
of
at
ponents
to Hb-F
Hh-Bart’s,
is discussed.
Hb-H
random
were
possible.
to
more
findings
any abnormality
blood.
The
mately
The
curve
cord
blocd
presented.
In un
del
minor
casos
babies,
the
some
minor
for
absence
a
with
the
of
expression
chain-lacking
54
no
heme-heme
of a minute
amount
of
de
plus
esseva
componentes
300
seligite
hemoglohina
were
com-
these
components
of Hb-Bart’s
and
respective
parents
beyond
hemoglobin
when
infancy
types
of
of
cord
to be approxian oxygen
dis-
interaction.
a prematurely
que
cases
hemoglobin
of Hb-Bart’s
reveals
its high
oxygen
affinity
that of Hb-A,
its lack of a Bohr
effect
and
sample
studio
54
of
along
the
SUMMARIO
negre,
of Negro
occurrence
intensively,
suggest
showing
discovery
bloods
the
responsible
physiology
8-10
times
sociaticn
cord
study
Hb-H,
and Hb-A2.
The
relationship
of
Seven
cases
with
moderate
amounts
studied
The
300
over
selected
IN
of
a Hb-Gower
born
white
Il-like
baby
of
component
in
months
is also
7.5
a
INTERLINGUA
specimens
aleatorimente
Bart’s,
de sanguine
pro
hemoglobina
de
cordon
determinar
ab neonatos
le
occurrentia
H, e hemoglobina
A2.
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
312
HORTON
Es discutite
le relationes
inter
executate
plus
intense
studios
hemoglobina
donatores
Bart’s
e de
hemoglobina
tanto
que
isto
in
absentia-post
pro le presentia
catena
a.
Studios
affinitate
A, (2)
que
iste componentes
de 7 sanguines
le infantia-de
in le sanguine
del
physiologia
H,
esseva
omne
de
de
non
monstra
Es etiam
Gcwer-II-like
ulle
nascentia
prematur
post
7,5
studios
del
Bart’s
un
hem
de un
sanguine
de
pare
del
del anormalitates
responsabile
typos
de hemoglobina
sin
menses
resultatos
parentes
expression
cordon
de
inter
le discoperta
specimen
de
con
Le
e (3)
interaction
presentate
in un
insimul
F. Esseva
quantitates
de
possibile.
hemoglobina
pro oxygeno-approximativemente
absentia
del effecto
Bohr,
e hemoglobina
moderate
con
AL.
ET
revela
in
indicar
illo
(1)
8 a 10 vices illo de
curva
de dissociation
le
le
un
alte
hemoglobina
de oxygeno
e hem.
minute
quantitate
de cordon
ab
de hemoglobina
infante
blanc
un
a
gestation.
ACKNOWLEDGMENTS
The
lege
authors
of
are
Georgia,
Medical
grateful
for
to
Dr.
assistance
Illustration
for
C.
in
S.
Wright,
reviewing
preparation
of
Department
of
hematologic
the
data,
Medicine,
and
to
Medical
the
Col-
Department
of
figures.
REFERENCES
1.
Bcnesch,
R. E.,
R.,
and
of
Ranney,
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the
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and
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born
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in
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in
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P.:
of
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iiniovular
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lite
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the
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Haemoglobin
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variants
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oc-
conjunction
H. Brit. J. Haemat.,
embryo.
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The
in
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“Bant’s”
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Huisman,
T. H. J., and
haemo-
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-‘
A.:
Abnormal
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of
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erythroblastosis
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arteniellen
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6:385,
1960.
der
des
newborn
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E.
A.,
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N.
\Iicrobiol. 10:228,
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314
HORTON
Bennett
F. Horton,
M.D.,
istry,
Medical
College
low,
Division
of
Medical
M.
Research
Medical
Dozy,
Titus
Nichols,
of
Augusta,
B.S.,
Augusta,
Chief
B.S.,
M.D.,
of Georgia
Chief
and
H. I. Huisnian,
Biochemistry,
Technician
of
Ga.
Resident
OB-Gyn
Eugene
Augusta,
College
Memorial
Talmadge
Ga.
D.Sc.,
of
Hospital,
DepartEugene
V.
Research,
A.
Hemo-
Biochemistry,
Ga.
of Georgia
Medical
Augusta,
Ph.D.,
Medical
inadge
Medicine,
Ga.
Medical
College
Biochemistry,
of
Fel-
Ga.
Laboratory,
Department
of
College
of Georgia,
Augusta,
Georgia,
College
Research
M.D.,
Instructor
of Pathology,
Me(lical
College
of Georgia
and
Hospital;
Attending
Consultant,
Carl M. Nechtman,
School,
Department
Evan
of Biochem-
present,
Department
of Georgia,
Hospital,
globin
At
Hematology,
College
Robert
B. Thompson,
nient
of Pathology,
Taimadge
Memorial
Andree
Department
Instructor,
of Georgia..
ET
Graduate
College
Service,
Memorial
of
Medical
Hospital,
Professor
of Pathology
and
Georgia
and
Tal-
Augusta,
Eugene
Ga.
AL.
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
1962 20: 302-314
Inhomogeneity of Hemoglobin. VI. The Minor Hemoglobin Components of
Cord Blood
BENNETT F. HORTON, ROBERT B. THOMPSON, ANDREE M. DOZY, CARL M. NECHTMAN, EVAN
NICHOLS and TITUS H. J. HUISMAN
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