From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
Morphological
Cells from
with
atid
a Patient
Special
Histochemical
Studies
of the Leukemic
with Atypical
Myeloblastic
Leukemia
Reference
to Intracytoplasinic
Mucopolysaccharide
Fibrillar
By G.
B
AI0LIH
ECAUSE
a
OF
the
tiicl
with
I)ttieIlt
highly
cytoplasniic
vacuoles
data
further
of the
tocheniical
of
mnieleoprotein,’
which
vacuoles,
leukemic
cells
a
variation
the
cells.’
nature
l)een
Detaile(l
.
froiii
leukemic
In
and
from
comprehen-
mi tiate(I
‘as
have
AND
nitra-
addition,
development
observed
in
of
k’uke-
certain
green
) and
method
md succinic
Electron
iinstuned
and
activities
enzyme
1)1 11c( polysa(
) ,‘
peroxidase
dehydrogenase.’
vital
ilistocheniieal
p(Iltoseiluel(oprot’i1L’
hhme
METhODS
iiueroseop.s.
acid-Schiff
reaction)
,‘
lipids,’
ammo
groups,#{176} protein-l)011n(1
1)01111(1
the
leukemia,
the
concerning
formations
( neutral
red
and
JanUS
contrast
and bright
field
the following
sul)staneeS
phase
(periodic
cytoplasniic
of
in
KNou1’F
#{149}s11,1,l(,19
Stipravital
stration
certain
(listiflct
descril)ed
obtained
fibrillar
a
A.
IIALPII
investigation
revealed
MATERIALS
the
of
many
myeloblastic
heretofore
l)een
so-called
cells.
inic
has
his
vacuoles
and
AND
in
aty)ical
aII(l
the
of
Gii.sso,
nature
Present
fulminating
eXalliiflatitl)fl
A.
unusual
orgaiielles
nlorplu)logical
S1V(.
Josipli
ACKELIMAN,
inclusions
Vacuoles
Formation
( Graham’s
were
the
glveogen
histidine,’
indoles,la
proteinhemoglobin
( Icuco-patent
groops.4’
(stcrase,_!.I
leucin(
aininopeptidase,’
1
1
ulieroscolne
fixation
(1’ inon-
(leOxvpefltOse-
omit:
sal va-labile
examination
was
performed
on routinely
prepared
whole
blood for 20 minutes
with 2 per cent osniiuui
tetroxmdc
to p1 1 7.4 with
veronal
acetate.
The plasma
proved,
in this instance,
adequate
oroper
tonicity.
1lo
fixe(l
svliole
1)100(1
sp(’ciilleIi
\V1S
SlIi)se(kIently
centrifllge(l
huffy
coat
removed.
Specimens
were
embedded
in ii 1)Imtyl-nletharcrylate
and
with the RCA E \ I U-2b
electron
microscope.
following
with
exaIIlii)e(l
for
carried
and
1
).
method
were
chtri(leS
plasmologen,’
siilfhiydryl
fihims
procedures
specimuells
of
buffered
miaintaiil
to
and
the
examined
OBSERVATIONS
The
atyl)ical
show
feren
( figs.
cell
1-4)
and
room
lbs
tutes
\%e
of
a thin
health,
wish
Suh,,mitted
(14
round,
was
to
thank
1)ec.
ii
22
,.)
ks
are
and
degree
rarely
to
one
Ohio
a research
Time
State
round
to
(11-4061)
dif-
pr1111iti’
regular
delicate
cyto-
chromatin
irregular
Columbus,
from;
the
[mnilersit,,.
fr(!?mt
a
and
or
most
a smooth
with
three
maturation
‘I’he
with
nucleus
variation
of
observed.
rounded
indented
by
morphological
some
membrane;
Ohio.
National
shaped
insti-
.\Jd.
the
?1i(’m?ibCTs
L Tnicersity
21,
to
of Anatoini,
supported
Bethesda,
Laboratory,
grati
slightl’
nuclear
considerable
m(hcating
s1)ecific
is large
time 1)epartiicnt
incestigation
Research
changes
although
contour,
pattern
exhibit
structural
tiation,
plasmic
I
myeloblasts
certain
19.59;
lbs
accepted
the
of
pita!,
fey
i1c,n,toboy
for
!)irisiolg
their
assistance
J)Uhlic(ltU)fl
Marc/i
1253
(111(1
111
18,
this
1960.
th(
ilerioao
iii icStig(ltioli.
1!ostcr
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1254
nucleoli
show
marked
cytocentrum
is less
exceedingly
are
large
scattered
in
portion
myeloblasts
( figs.
some
in size and optical
than
the remainder
leukemic
the
of
and
1-6)
the
evident.
many
nucleoli
and
more
the number
primitive
cells
processes
Electron
the
though
leukemic
marked
cells.
loss
32)
between
moderately
dense
finely
well-developed
been
observed.
cells
sociated
small
are
scattered
cyte
granules
dense
with
are
less
Fibrillar
Golgi
dense
throughout
seen
thin
few
cytoplasm
cortical
32,
33);
of
cristae
many
al-
of the
mitochondrion
(figs.
31, 32),
(fig.
dense
(3)
and
and
in
between
structures
inconspicuous
20,
26);
19,
cells
(fig.
the
so-called
rare
containing
Transition
abundant
(figs.
leukemic
in
24),
of moderately
and
19,
the
membranes
24).
is
(figs.
the
in very
dense
is small
granules
23,
mitochondrial-like
reticulum
RNA
the
of cristae
23,
dense
system
endoplasmic
of
appearance
(figs.
these
than
20,
mitochondria
(figs.
numerous
present
19,
of
have
dense
usually
many
free
20,
31)
in
and
27).
and
the
asRNA
Promyelo-
appear
very
rims.
Formations
Large
cytoplasmic
fibrillar
structures
formations’2
vital
films
to
cytoplasm
and
nuclear
fail
limiting
rounding
tend
to
some
located
in the
into
or
instances
central
with
the
of
these
strands
portion
by
in viscosity
Faintly
laminated
several
mitochondria
the
fibrillar
structures
of faintly
and
of
and/or
(fig.
suggesting
with
11).
PAS-positive
5).
Large
azurophilic
In
one
material
the
sur-
organization
granules
(fig.
small
near
cytoplasmic
structures,
fibrous-like
few
structures
surrounding
cytoplasm
as compared
formations
a
the
in
stained
ovoid
the
membrane,
these
studied
Wright’s
than
limiting
occupied
basophilic
in
poorly-defined
proximal
developed”
cells
and
basophilic
apparent
region
and
portion
within
the
a difference
faintly
delicate
but
“fully
leukemic
1, 5)
and
is no
matrix.
appear
localized
in
some
(figs.
opaque
there
to move
in
prominent
less
localize
Although
of
observed
microscope
These
are
cytoplasmic
in
be
11).
barrier
evident
suggestive
been
phase
length)
hof.
organelles
formations
the
(fig.
8 is in
some
have
with
preparations
structure
the
(figs.
evidence
material
and
The
while
with
granules
may
granular
no
are
mitochondria
curling
the
and
indicative
6)
cells
mitochondrial
cristae
or
4,
these
swelling
the
and
little
mitochondria
leukemic
the
with
the
and/or
of
poorly-defined
in
of
(1)
32)
cristae
(figs.
structure
in
invagination
of the
the
mitochondria
(fig.
and
of density
instances,
have
observed
include:
borders
is serrated
Mitochondria
majority
fine
more
round
round,
slightly
smaller,
membrane
and more
is greater
the
classical
fragmentation
(fig.
material
that
the
alterations
are
These
alterations
31, 33, 34),
(2)
exhibit
the
the
cells
“blebs”
myeloblasts.
vacuoles
reveals
cells
dissolution
34);
immature
microscopy
leukemic
(figs.
the
of cytoplasmic
myeloblasts.
in
cytoplasmic
of these
and
of the
and
is
mitochondria
between
irregular
of these
cells are
a thicker
nuclear
The region
cytoplasm
concentrate
forms
borders
AND KNOUFF
spherical
to
possessing
cytoplasmic
protoplasmic
than
fine
tend
Transitional
of active
pinocytosis.
The nuclei
a more
coarse
chromatin
network,
round
density.
of the
Many
but
cytoplasm.
The
thin
cells.
cytoplasm
myeloblast-like
are
possesses
variation
basophilic
throughout
proximal
(4
GRASSO
ACKERMAN,
instance,
was
is
may
be
fibrillar
granules
a large
found
and
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
MORPHOLOGICAL
AND
considered
(fig.
to represent
16)
is apparently
structures
nor
their
fibrillar
formation.
amino
groups
and
similar
to that
of the
negative
histochemical
images
could
The
fibrillar
methods.
devoid
of peroxidase,
Electron
microscopy
esterase,
indicates
in size
and
and
complexity
(figs.
20-25).
“fully
developed”
swollen
and/or
structures
of
show
internal
reticulum
may
(figs.
21-25).
endoplasmic
uncommon
(figs.
RNA
granules
branes
regions
the
The
reticulum
in
(figs.
fibrillar
may
show
formations.
reticulum
stage
in the
small
Parallel
is evident
formation
of
26 and
These
vacuoles
cells
presence
irregular
in contour
represent
and
the
their
of twenty
14),
(1 to 4 is)
are
faint
the
material
vacuoles
the
of lipid
common
optically
lar
internal
internal
are
some
of
endo-
extremely
minimal
vacuoles
basophilic
(fig.
17),
rich
than
(fig.
(fig.
early
the
in
phase
15).
18)
fibrous
of
strands
the
leucine
content
cytoplasm
is
less;
and
of
may
appear
dense
vacuoles
appears
granular
(fig.
to be
only
reveals
microscopy
masses
which
and
Their
histidine
Electron
many
5-8)
and
of internal
surrounding
and
irregular
to
microscope
mucopolysaccharides
activity.
the
atypical
individual
round
29).
derived
The
from
29).
moderately
circular
found
evident
refractile
vacuoles
with
distinct
limitmembrane-like
and/or
distinct
granuin a few
of the leukemic
cells
(fig. 7). The
in supravital
films
and
stain
stain
light
pink
in Wright’s
films
areas.
They
exhibit
a moderate
and
in
(fig.
16)
delicate
Large
the
esterase
greater
of very
(figs.
are
is most
seen
within
an
They
appear
in supravital
(figs.
to have
a suggestion
peroxidase
is
27-30).
(2 to 4 is)
and prominent
organization
sulfhydryls
memin
sacs
an
under
are
strong
sulfhydryls
composed
(fig.
structure
neutral
red. The
have
moderately
for
their
comprising
flattened
vacuoles
vacuoles
demonstrable
reticulum.
spherical
membranes
sacs
and
strands
represent
dense
is present
in variable
amounts
in
are enclosed
by a double
membrane
ing
fibrous
to
possess
groups
for
are
granular
endoplasmic
Large
reactivity
vacuoles
somewhat
and
amino
their
traces
that
moderate
activity
protein-bound
although
of
fibrillar
structure.
most
the
exhibit
aminopeptidase
of
along
density
number
varies
considerably
to thirty
is not uncommon.
Histochemically,
13,
strands
the
of flattened
“blebs”
long
may
and
stain
orange-red
and
pink
respectively
Wright’s
(fig. 10) films. Some vacuoles
appear
(figs.
and
fibrillar
formation
is not
disassociated
from
their
the
alignment
in figure
or
appear
of
alignment
in electron
resemble
circular
infrequently
region
of
vary
membranes
the
granules
central
and
vary
of
not
and
Vacuoles
leukemic
cell; the
structure.
center
RNA
the
by
activity.
markedly
laminated
and
protrusions
may
of a fibrillar
Mucopolysaccharide
in
association
(figs. 23, 24);
the membranes
exhibit
minimal
density
and
plasmic
21-24)
Free
found
close
the
these
stained
sudanophobic
aminopeptidase
formations
be
fibrillar
neither
in films
are
and
fibrillar
sulfhydryls
the
since
formations
found
of
in
distinguished
within
and
around
the
These
sacs loose
or become
23-25).
and
be
modification.
be
hyaloplasm
of delicate
formations
content
present
of a series
may
fibrillar
The
histidine
dehydrogenase
that
the
consist
Mitochondria
endoplasmic
1255
STUDIES
a large
protein-bound
structures
these
HISTOCHEMICAL
weak
to
moderate
PAS
reactivity
orange-red
(fig. 10)
to strong
and
with
and may
reaction
peroxidase
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1256
GBASSO
ACKEBMAX,
AND
KNOUFF
31
;
.
-.‘
‘.‘
r.
Figs.
Acute
1-9-Supravital
Myeloblastic
films
Leukemia
examined
X 1650.
with
the
phase
contrast
microscope.
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
MORPHOLOGICAL
AND
activity.
Histidine,
minimal
amounts
Under
the
HISTOCLEMICAL
prctein-l)ound
and
electron
reflection
of
electron
density
and
microscope
than
vacuoles
chemically
with
these
and
general
cytoplasm
1)
phase
are
appear
( fig.
descril)ed
above
and
electron
only
not
observed.
be
formed
material
)
28
vacimoles
contrast
I)resemt
is
to
cytoplasmic
(nuicopolysaccharide
the
lipids
activity
vacuoles
about
vacuoles
and
groups
dehvdrogenase
reticulum
the
between
these
(niucopolvsaccharide
Fat
amino
esterase
endoplasmic
1257
STUDIES
have
by
of
and
been
the
greater
Transitional
.
II)
in
stages
the vacuoles
ol)Serve(l
histo-
microscopes.
Droplets
Small
moderately
cytoplasm
stain
refractile
of many
deep
red
in ethanol
with
Sudan
chemical
(acidic)
technics
and
Ingested
Erythrocytes
Large
Fic.
in
except
1 -Primitive
films.
their
round
deeply
throughout
and,
in some
droplets
may
be
non-reactive
are
stained
with
non-specific
Electron
lipoid
are
color
with
These
occasional
structures
distributed
1, 3, 9)
vapor
fixation
droplets
are
a greenish-yellow
m’eloblasts
are
(figs.
acid
The
methods.
nature
of these
microns)
appear
droplets
cells
supravital
for
dehydrogenase
homogenous
(6-8
and
fat
leukemic
but
after
osmic
black
(fig.
15).
esterase
osmiophilic
cells
of the
in
present
1)0th
basophilic
cytoplasm,
with
the
established
32, 33).
in a few
unstained
highly
soluble
successfully
other
histo-
staining
microscopy
droplets
(figs.
the
instances,
of the
vital
the
leukemic
and
supra-
vesicular
nuclei
amid prominent
nucleoli.
A large
oval
poorly-defined
region
(if)
is considered
represent
a large
fibrillar
formation.
Fat droplets
(d) also indicated.
Fic. 2.-Primitive
mveloblast
(A)
and
a more
differentiated
myeloblastic
(B)
with
to
cell
serrated
(p)
are
borders.
Mitoehomidria
(m)
and
small
mucopolvsaccharide
vacuoles
evident.
Fmc.
3.-M’eloblast
showing
clear
c’toplasmic
vesicles
(c)
and
fat
droplets.
A prominent
cvtocentruin
(i)
is
indicated;
small
mucopolvsaccharide
vacuoles
also max’ l)e seen.
Fmc. 4.-Large
mveloblast
with
fine
mitochondria,
scattered
dark
fat droplets
and clear cvtoplasmic
vesicles
(c),
ms’eloblastic
cell containing
many
fine mitochondria
and small
mucopolvsaccliaride
vacimoles
(p)
and
has
man
thin protoplasmic
processes.
Fic. 5.-Mveloblast
containing
an oval somewhat
fibrous
structure
(if)
which
represents
the fibrillar
formation.
Several
dark
fat droplet
and
tin
mitochondria
may
be seen within
this structure.
Fmc. 6.-Mveloblastic
cells
containing
large
mucopolvsaccharicle
vacuoles
(p).
Some evidence
of internal
structure
within
the vacuoles
is
indicated
(p).
Micromveloblast
indicated
at
A.
Fmc. 7.-Leukemic
cell with many large mucopolvsaccharide
with internal
organization
clearly
evident
within
the vacuoles
Fic. 8.-Leukemic
cell containing
an erythrocvte
(r),
as
mucopolysaccharide
vacuoles.
Fic. 9.-Myeloblast
containing
many
dark fat droplets
(d)
vacuoles
(v)
which
are considered
to represent
ervthrocvtes
disintegration.
vacuoles
(s).
well as
(Type
a few
II)
dark
and two amorphous
in late
stages
of
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1258
ACKERMAN,
p
3’
ph
3
p
I
-‘a
15
14
F
0
Figs.
10-18-Acute
Myeloblastic
Leukemia
0.
X 1650.
GRASSO
AND
KNOUFF
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
MORPHOLOGICAL
vital
films.
cyte
and
AND
( fig.
sulfhydryls,
13
)
The
histochemical
and
fail
responds
very
evidence
phous
brane
These
closely
vacuoles
show
bo’md
are
faint
)
12
with
similar
and
and
and
They
and
reactivity,
exhibited
by
of
an
erythrohistidine,
are
PAS-negative
dehvdrogenase
as
films
that
hemoglobin,
lipids.
supravital
erythrocytic
to
for
peroxidase
those
neroxidase
faintly
ve1l
)
of
as
their
these
mature
activity.
tinctorial
structures
cor-
erythrocytes.
Such
nature.
groups
and
scribed
above
and
and
and
structures
Structures
traces
vacuoles
negative
for
between
have
may
represent
pale
pink
histidine,
and
protein-
mucopolysaccharides
but
morpho-
ervthrocytes
It seems
erythrocytes
a moderate
apparent
ingested
observed.
(fig.
exhibit
Some
the
been
appear
for
activity
of hemoglobin.
transitions
vacuoles
and
slightly
de-
probable
undergoing
that
chemical
dissolution.
considered
degradation
10.-Located
are
stain
react
dehydrogenase
faint
these
or
and
by Wright’s
vacuoles
lipids,
and
histochemical
degradation
of
The
esterase
reaction
logical
colored
basophilia.
amino
FIG.
groups
esterase,
pattern
their
sulfhydryls,
stages
density
reactions
amino
( fig.
indicates
optical
vacuoles
( 4 to 7 is ) are seen occasionally
which
appear
to be amoror
have
minimal
internal
structure,
possess
no distinct
limiting
memand are optically
less dense
than
the surrounding
cytoplasm
( fig. 9).
Large
these
an
histochemical
staining
1259
STUDIES
have
to exhibit
( Wright’s
properties
14)
They
.
positive
protein-bound
( fig.
may
)
8
exhibit
HISTOCHEMICAL
to
correspond
have
been
with
observed
ingested
with
the
ervthrocytes
electron
in
microscope
various
(fig.
in the lower
portion
of the cell a prominent
mucopolvsaccharide
dark central
regiomi probably
represent
the Type
II vacimole.
Smaller
vacuoles
(Type
I) (p) also are evident.
Wright’s
stain.
X 1650.
prominent
lightly
basophilic
oval structure
(if) which
appears
some-
vacuole
(p) with
and more irregular
Fic.
11.-A
what
fibrous
and
a well-developed
contains
fibrillar
several
azurophilic
formation.
Wright’s
gramnmles.
stain.
This
structtmre
represents
X 1650.
Fic.
12.-Leukemic
cell
containing
a phagocvtized
ervthrocvte
(r)
amid clear
cytoplasmic
vesicles
and
fat droplets
(c).
Wright’s
stain.
X 1650.
FIG.
13.-Dark
glycogen
(g)
granules
are
evident
in the
myelohlastic
cells.
Mucopolysaccharide
vacuoles
(p)
and a large
clear
vacuole
(v)
(phagocvtized
erythrocyte)
are indicated,
PAS reaction.
X 1650.
14.-Meloblastic
with
prominent
mucopolsaccharide
vacuoles
(p)
and
(g) granules.
An erythrocvtic
vacuole
(v) is evident.
PAS reaction.
X 1650.
FIG.
15.-Leukemic
cells
containimig
sudanophilic
mitochondria
(rn),
sudanophilic
mucopolsaccharide
vacuoles
(p) and dark staining
fat droplets
(d).
Sudan
black
B. X 1650.
FIG.
16.-Mucopolvsaccharide
vacuoles
(p)
are numerous
and
are less
SH
reactive
than the hvaloplasrn.
The ctoplasmic
structure
(if) may represent
a fibrillar
formation
or a large cytoplasmic
vacuole.
DDD
reaction.
X 1650.
FIG.
glcogen
Fic.
17.-Mveloblasts
exhibiting
a
moderate
cvtoplasmic
reaction
for
protein-
boumid sulfhvdryl
groups.
The damaged
cell has only a minimal
sulfhvdrl
reaction
and contains
several
large sulfhvdryl
positive
vacuoles
(s) which
correspond
with
the mucopolsaccharide
vacuoles
(Type
II). DDD
reaction.
X 1650.
FIG.
18.-Peroxidase
negative
mveloblast
(A)
and myeloblast
(B)
containing
many
vacuoles.
round
peroxidase
Graharns
positive
Peroxidase
vacuoles
Reaction.
corresponding
X 1650.
to
the
mucopolysaccharide
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1260
ACKERMAN,
GRASSO
(#.
AND
KNOUFF
‘--“‘
iV
.
‘a’
‘,
..
AI
4’
Cij.
p
.
Figs.
19-27-Myeloblastic
minutes
in 2 per cent
Bars indicate
1 micron.
osmium
Leukemia.
tetroxide
Electroi
buffered
micrography.
to pH 7.4
with
Fixation
veronal
time
20
acetate.
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
MORPHOLOGICAL
35,
)
36
These
.
variation
in
mnay
reveal
double
mnay
brief
charicle
extremely
structure.
They
other
and
frequently
be
morphological
1 and
leukemic
derived
tppear
compared
from
with
of
marked
and
single
amid
reticulum.
dense
These
material
36).
ingested
so-called
of
electron
comparison
and
the
endoplasmnic
( fig.
exhibit
fragmentation
foldings
masses
granular
and
undergo
marked
the
histochemical
formations
dense
to
of
to l)lrtition
highly
and
fibrillar
electromi
appear
composed
probably
surround
vactmoles,
table
in
are
fine
structure
1261
STUDIES
structures
their
membranes
which
HISTOCHEMICAL
internal
mnemTfll)ranes
A
AND
of
erythrocytes
granule-vacuole
the
mnucopolysacis
bodies
presented!
seen
in
cells.’
DIswSsIoN
Detailed
scopy
)
charide
swollen,
primitive
Fic.
vacuole
( phase
morphological
and
histcchemical
vacuoles,
modified,
leukemic
19.-Group
located
studies
fat droplets,
and altered
cells
from
contrast,
reveal
bright
the
field
presence
of
and
unusual
electron
micro-
mucopolysac-
fibrillar
formations,
ingested
erythrocytes,
and
mitochondria
in the cytoplasm
of many
of the
a patient
with
atypical
myeloblastic
leukemia.
of mveloblasts.
in the nuclear
hof
Arrow
indicates
of a mveloblast.
delineated
by
arrows.
a large
mimcopolvsaccharide
X 3,300.
The laminated
double
memreticulum
(e)
are
indicated.
Fic.
20.-Fibrillar
formation
bramies
are
evident.
Prominent
sacs
of endoplasniie
X 15,000.
Fic.
21.-Fibrillar
formation
delineated
b
arrows.
A large
flattened
sac of
endoplasmic
reticulum
(e)
shows
loss of its membrane
resolution
and
appears
to
be condemisimig
with
the
fibrillar
formation.
A large
altered
miiitochondrion
with
cristae
is located
in the central
portion
of the fibrillar
formation.
X 19,200.
Fic.
22.-Fully-developed
circular
fibrillar
formation
containing
a large
altered
mitochondrion.
X 13,000.
Fic. 23.-The
region
of the fibrillar
formation
is indicated
1w arrows.
Mitochondna
and
modified
dense
mitochondria
(iii)
are located
within
the fibrillar
structure.
Prominent
sacs
of endoplasmnic
reticulum
(e)
appear
to be coming
condensed
imito
the
fibrillar
formatiomi.
A small
protrusion
of the
membrane
of the
endoplasinic
reticulum
is eviclemit
at e. X 10,800.
Fic.
24.Same
as figure
23. Endoplasmic
reticular
sacs,
mitchondria
and
the
modified
dense
miiitochonclrion
are shown
iii the central
portion
of the fibnillar
formation.
Au osmophilic
particle
is present
within
the
modified
mriitochondrion.
Note
the delicate
lamimia
of the fibnillar
formation
and
the small
protrusions
or
blebs
(arrow)
on the
endoplasmic
retieimlar
sac.
X 21,000.
FIG.
25.-Portion
of a fibrillar
formation
showing
the delicate
strands
of poorlydefined
double
mnemimbranes
and
dark
strands
of endoplasmnic
reticulum
associated
ivth
dark
RNA
granules.
Some
RNA
granules
are n(it directly
associated
with
the
endoplasmic
reticulum.
Arrow
indicates
omie of several
points
where
the strands
of
endoplasmic
reticulum
and
those
of the fibrillar
structure
appear
to be continuous.
X
22,000.
Fic.
(indicated
the
Fic.
material
in
26.-Strands
flattened
sacs
of
may
possibly
formation
of a fibrillar
structure.
X
27.---Mucopolvsaccharide
vacuole
and
surrounded
by a thin limiting
between
of
arrows)
endoplasmic
represent
15,000.
(arrow)
membrane.
an
reticulum
aligned
early
developmental
containing
X 12,400.
finely
parallel
stage
granular
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1262
ACKERMAN,
GRASSO
AND
KNOUFP
rib.
r
‘..‘i.’
..j
4
p1
-.
k.
_aL’_
..
‘I,
r,
..i
Figs.
28-36-Myeloblastic
minutes
in 2 per cent
Bars indicate
1 micron.
osmium
Leukemia.
tetroxide
Electron
buffered
Micrography.
to pH 7.4
with
Fixation
veronal
time
20
acetate,
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
MORPHOLOGICAL
AND
Such
cytoplasmic
changes
possible
relationships
activity
of
striking
structural
reticulum.
as
small
appears
sacs
to
vacuoles
be
and
directly
erythrocytes
and
The
abundant
fibrillar
actively
is associated
mucopolysaccharide
have
light
not
been
microscopic
in the
the
the
However,
of
in
presand
the
endoplasmic
of
ingested
pinccytosis.
in
either
the
granules
destruction
of
the
involve
mucopolysacchariie
addition,
the
Perhaps
is usually
RNA-rich
In
in
process
cells
reticulum
formation
and
cells.
This
evidence
erythrophagocytosis,
leukemic
without
observed
previoumsly
technics.
or
evidence
metabolism
mitochondria.
the
formations.
vacuoles
described
in
participate
with
morphological
with
normal
lipophanerosis,
with
with
the range
of cellular
insight
into the infergrowth,
endoplasmic
associated
the
to
addition,
cellular
alterations
concerned
perhaps
appears
reticulum
In
alterations
The
either
indicate
provided
in
as compared
of mitosis,
intra-cytoplasmic
endcplasmic
cells
high
and
occur
cells
rate
primitive
pinocytosis
variations.
alterations
a relatively
active
1263
STUDIES
and alterations
cells
and have
of these
marked
these
includes:
ent
modifications
in leukemic
of some
that
suggests
most
HISTOCHEMICAL
of
the
normal
many
or
leukemic
et
al.11
DiMayorca,
myeloblastic
has
cells
by
depicted
a
FIG. 28.-A
vacuole
forming
(delimieated
by arrows)
from
comidensed
cytoplasm
and becoming
enclosed
by concentric
strand
of emidoplasmic
reticulum.
This
vacuole
represents
the mucopolsaccharide
vacuole
tYpe 11 in contrast
to the t’pe
I vacuole
(p)
and
those
seen
in figures
19, 27, 29, 30. X 20,000.
Fic.
29.-Mucopolysaccharide
vacuole
surrounded
1w
a limiting
membrane.
Arrow
indicates
a region
where
this membrane
is continuous
with
the endoplasmic
reticulum.
The internal
structure
of the vacuole
appears
granular,
as well
as somewhat
fibrous.
X 10,000.
Fic.
30.-Two
mucopolsaccharide
vacuoles
showing
slightly
different
stage
of development.
Circular
double
membrane
is indicated
by arrow
in upper
vacuole
and small sacs are evident
(arrow)
in the lower vacuole.
X 12,000.
Fic.
31 .-Mveloblast
exhibits
niarked
alterations
in its mitochondria.
Cristae
have
disappeared
and
only the fragmented
terminal
portion
of the cristae
are
evident
(m). Some cristate
have become
coiled.
Prornvelocvte
with dense neutrophil
granules
(g) and many
sacs of endoplasmic
reticulum.
X 4,300.
Fic. 32.-A
mitochondrion
(o) showing
some inter-crista
electron
dense
material
and a mitochondrion
(m)
showing
a loss of resolution
and electron
demisity.
A
mitochondrion
by the arrow.
Fic.
decreased
showing
an
Fat droplet
invagination
of the
(d).
X 20,500.
33.-A
swollen
mitochondrion
electron
density
and area
mitochondrial
(arrow)
of inter-crista
with
electron
membrane
cristae
density
is indicated
showing
between
areas
of
them.
X 20,000.
Fmc. 34.-Mitochondrion
showing
invagination
of their
inner
and outer
membranes.
Mitochondria
showing
fragmentation
as well
as curling
of their
cristate
also
are
evdent.
X 20,300.
Fic.
35.-A
large
electron
dense
structure
showing
areas
of rarefaction.
This
structure
is considered
to represent
an ingested
ervthrocvte.
X 5,800.
Fmc. 36.-Au
electromi
demise
structure
showing
sacs
of endoplasmic
reticumlum
surroumndimig
amid apparently
material.
The outer
membrane
siderable
internal
folding
and
sidered
cell. X
to represent
12,000.
a late
partitioning
areas
of the electron
demise granular
of the vacuole-like
stnmcture
(arrow)
shows
conis discontinuous.
This vacuole-like
structure
is constage
of erythrocyte
degradation
within
the leukemic
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1264
ACKERMAN,
CRASSO
+
I
+
.
1
4Z:f+
.3
+
a
ii
+
-‘-
E
ri
+1
+1+
-1
+Z
1
-
I-
I
I
.E
I
+
++
+
I
+
++±+
+±++;‘-
It
1
.
.
__
.
.
b
4
+
.“
+
+
-
E
.‘E
±
tL
+
I
I
+
--
4-
+
-f
I,,
.
.LJ
±
-4++
I
I
.
..
+
+
-
-r+
±
+-+
.;.
i
i_++Ii-itiii
±!
+
ii
-
+
.
ti
I
+
1
+
I ‘1-’
+
t+tIiiI
a
.
.2
0
0.
0
.s.E
.
AND
KNOUFF
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
MORPHOLOGICAL
large
finely
graph
of
those
seen
The
AND
IIISTO(I1EM
granular
vacumoles
of
double
the
membrane.
and
the
high
activities
the
the
The
of
the
vacumolar
general
mucopolysaccharide
ture,
are
not
chari(le
or
vacuoles
vacuoles,
have
these
least
similar
occur
cells
occasional
1v smaller
cytes
in miormal
tions
in all stages
fibrous
bomie
fibrillar
scope
and
scopic
of the
evidence
endoplasmic
In
condensation
or
or
zomies
the
in
livimig
to
possible,
however,
in the
sanie
or
‘hich
at
iiiay
flattened
either
the
described
have
examinedi
been
mass.
There
any
to
(listiliguish
from
no
the
phase
micromicro-
modifications
arise
by
to
RNA
indication
granules
of
that
or
the
alignimig
and
a poorly-
the
employed!
vacuoles
the
the
electron
formiiation
techniques
mucopolysaccharicle
of
reticulum
their
leumkoforma-
in mamiy
represent
appear
resultant
the
certain
in
fibrillar
that
emidloplasmic
is
of
seemi
the
with
feel
We
of
in
and
observed
formations
structures
sacs
with
vacimoles
been
microscope”
films.
the fibrillar
The
fibrillar
fibroims-like
related
lipoid
mmicopolysac-
alterations
have
cells
stained
laminated
not
struc-
of a malformation
is
organization
and complexity
it has proven
possible
defined
are
it
have
to one another,
becoming
disassociated
a loss of electron
density
with
the
resemble
tile
result
the
dm
internal
a surrOun(ling
with
electron
exhibiting
formations
possess
metabolic
structures
\Vright’s
strandis
have
variatiomis
\Ve
suggests
that
reticumlum.
of
as
cvtoplasmic
1)0th
routine
arise
marrow.121
addition,
formations
in
contrast
represemit
with
of development,
miiveloblasts.
not
in
cOm1)OsitiOmi
tile-vacuole
cells.
studied
human
large
miiav
or “crescentic
leukemia
do
metabolic
chemiiical
may
granules.1
modification
formations
in acute
leukemic
actually
leumkemiuic
fibrillar
to
cytoplasmic
cvtoplasniic
in certain
The
conSi(lere(1
certaimi
structures
tWo
and
bodies,
high
formatiomi.
gran
they
stages
a poorly-defined
the
its
and
that
latter
mucopolysaccharides
with
so-calledi
in
granules
beemi
of
or
the
differ
granule-vacuoles
diegemieratiomi
that
with
associated!
The
Iiiem’nl)ramie.
of
imi m’norphology
and
vacuoles
by
of
cytoplasmic
the
suggests
material
similarities
rnimcopolysaccharide
mOdlifie(1
amounts
vacuoles
membranes
durmg
is enclosed
large
these
with
vacuoles.
by
of
an(l
vacuole
of
of
micro-
corresponds
formed
niasses
the
activity
with
the
of
be
amorphous
presemice
enzymatic
associated!
Imi s1)ite
of
substance
electron
an
vacuole
mucopolysaccharide
to
surroundi
becomes
in
This
.
the
appear
which
subsequently
development,
of
vacuoles
reticulum
which
material
membrane
)
leukemia
micrographs
mucopolysaccharide
emicloplasmiiic
a limiting
with
oumr electron
1263
STUDIES
( myelocytic
a myelocytes
in
ICAL
fibrillar
and
they
granule-vacuole
bodies.
The
cells
extremiiely
been
has
piltiemlt
have
with
been
glol)in
to
uncomiimiion
to occur
atypical
proven
to
myeloblastic
erythrocytic
reaction
those
and
exhibit
have
been
changes
described.
have
Electron
been
noted
erythrocyte
of the
origin
Large
since
histochemical
stages
and
of
with
the
yield
and
electron
of
leukemic
from
reactions
diegradation
histochemically
similar
microscope.
size
this
strumctures
a positive
tinctorial
erythrocytic
structures
by
cells
cvtoplasmic
they
morphologically
dense
ingestion
myeloblast-like
leukemia.
in
Various
observed
of
in some
other
of erythrocytes.
solution
erythrocytes
process
observed
and
These
hemosimilar
and!
dIiS-
and
the
contour
structures
to
From www.bloodjournal.org by guest on June 15, 2017. For personal use only.
1266
ACKERMAN.
are
presumed
lion.
to
Electron
is associated
the
with
process
gations
erythrocytes
these
Braunsteiner
represent
cells
ingested
The
indicated
that
more
m:tochondria
their
and
Pakesch,9
their
of
that
appears
In
degradation
the
and
Tanaka19
some
KNOUFF
and
dissolu-
endoplasmic
to be
recent
inclusions,
actively
electron
have
reticulum
concerned
with
microscopic
investi-
characterized
of which
in
of
problems
tools
evident.
comprehensive
leukemic
the
metabolic
morphological
and
of techniques
elucidation
and
destruction.
and
stages
indicates
structures
of approaching
variety
various
AND
some
were
of
considered
to
erythrocytes.
necessity
a wide
in
evidence
of erythrocyte
phagocytic
the
be
microscopic
GRASSO
studies
cells
various
is clearly
of the
are
warranted
alterations
that
in
hematology
This
endoplasmic
and
may
must
exist
with
investigation
has
reticulum
prove
in
and
fruitful
leukemic
in
the
cells
and
processes.
SUMMARY
A
detailed
study
(phase
the primitive
leukemic
has been
presented.
been
observed
In addition,
structure
and
this
and
the
cells.
contrast,
cells from
An unusual
and characterized
study
has yielded
chemical
alterations
in
in these
of
erythrocytes
abnormal
studio
detaliate-per
e electronic-del
charido,
etiam
massas
es
br
de
information
fibrillar,
ingestion
per
esseva
observate
tiones
in
un
vitiate
cellulas
presentate.
Un
esseva
observate
histochimicamente.
mationes
relative
al
de
histochemically.
concerning
formations,
their
ingestion
fat
by
have
a contrasto
leucemic
ab
vacuolo
inusual,
metabolismo
un
been
the
the
droplets
leukemic
observed
and
in the mitochonaltered
metabolism
iste
con
structural
clar,
leucemia
mucopolysaccomo
considerabile
chimic
del
del
erythrocytos
forpost
intramitochondrial
myeloblastos
endoplasmic
campo
atypic
morphologicamente
producite
e al composition
anormab
a
continente
ha
Alterationes
in
phase,
patiente
e al alteration
in numerose
e be reticubo
de
tanto
studio
structura
grassia,
leucemic.
e characterisate
be mitochondrios
o alterate
fibrillar
of
leukemia
vacuole
has
IN INTERLINGUA
e characterisate
In
plus,
le
al guttettas
be cellulas
microscopy)
myeloblastic
myeloblasts.
Changes
reflect
impaired
or
microscopia
primitive
myeloblastic
electron
cells.
SUMMARIO
Un
and
alterations
of the primitive
reticulum
may
leukemic
the
fcllcwing
structural
in many
endoplasmic
field
both
morphologically
and
considerable
information
composition
Intramitochondrial
characterized
dna
and
bright
a patient
with atypical
mucopolysaccharide-containing
primitive.
reflecte
cellulas
Altera-
possibilemente
leucemic.
REFERENCES
1. Ackerman,
G.
A.:
Morphology
and
cytochemistry
of the granule
vacuole
body
of leukemic
cells. Blood
12:433447,
1957.
2. -:
Demonstration
of non specific
esterase
and
dehydrogenase
activity
in blood
cells by means
of a “supra-
3.
-:
4.
-,
vital”
procedure.
1959. Abstract.
Histochemical
indoxyl
acetate
normal
human
row.
Lab.Invest.
and Sneeringer,
Anat.Rec.
133:355,
demonstration
of
esterase
activity
in
blood
and bone
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1960 16: 1253-1267
Morphological and Histochemical Studies of the Leukemic Cells from a
Patient with Atypical Myeloblastic Leukemia with Special Reference to
Intracytoplasmic Mucopolysaccharide Vacuoles and Fibrillar Formation
G. ADOLPH ACKERMAN, JOSEPH A. GRASSO and RALPH A. KNOUFF
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