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Microbiologic
By
Determination
of
Derivatives
in Blood
Fuamnn
GRossowlcz,
NATHAN
OLIC
tors.
faulty
important
absorption
intestinal
of
(F.A. ) deficiency
ACID
The most
of the
flora
the
resulting
vitamin
F.A.
family
about
the
in
of
the
vitamin
from
in low
synthesis
body.
F.A.
activity
the
intestinal
blood
Therefore,
tract,
vitamin
to
in the
faulty
several
order
or serum
as possible.
responsible
derangement
and
contains
in
blood
vitamin
lesions
DAVIDOFF
be due to a variety
of facto be unusually
poor
diets,
of the
Human
in human
as many
forms
of the
tion of the biochemical
Rosy
ARONOVITCH
in man
may
are considered
ones
compounds.”2
Acid
MANDELBAUM-SHAVIT,
J.
AND
F
Folic
metabolism
members
obtain
valid
it seems
desirable
Such approach
for, or connected
may
of
the
information
to measure
facilitate
with,
the
detecdeficien-
cy.
In
the
present
laboratory
paper,
to achieve
in our
laboratory
various
this
for
end
several
are
methods36
modified
described.
Most
Lactobacillus-casei
activity,
as
in
this
increase
The
L. casei
it will
to
presumably
culture
is
referred
The
nation
of
siderably
of
due
as
in
been
our
in use
METHODS
F.A.,”
more
sensitive
(see
termination
assay
vorin
(Lederle)
about
50
folinic
cent
halved
in order
Assay
Medium
All three assays
of assay
From
Submitted
the
other
as
the
activity
at correct
of
and
bimonthly,
isolated
ATCC
No.
grown
at
same
laboratory,
derivatives.
8043
polyglutamates.
stored
acid
usually
F.A.)
certain
4 for composition;
in the
in this
(total
utilizes
the further
increase
pancreas
conjugase;
table
and
) are
below
it
other
( see
stabs
acid”
acid
4
C.
is used
for
This
strain
employed
between
medium
for
and
determiis
con-
determina-
citrovorum)
ATCC
No.
tetrahydro-derivatives
standard.
the
of
As
natural
the
synthetic
citrovorum
8081
folic
is used
acid.7
racemic
factor,8
for
product
the
the
Calcium
assay
de-
leucopossesses
figures
are
values.
are performed
with
medium.)
However,
Department
hepta-
tetrahydrofolic
strain
folic
folinic
1).
and
the
of
( see
(Leuconostoc
employed
of
to arrive
position
Jerusalem,
fig.
acid
is
per
deconjugation
9. A strain
“total
and
is given
despite
blood with chicken
transferred
the
)
yeast-peptone-agar
several
than
to measure
(P.G.A.
designation
of human
organisms
No.
cereviriae
of
in
including
below,
Pediococcus
is used
acid
stock-agar),
two
“free
7469
to
maintained
to
other
F.A.
No.
as well.
This
after incubation
similarly
maintained.
b) Streptococcus
faecalis
c)
AND
pteroylglutamic
of P.G.A.
for L. casei
is
be
transfers.
ATCC
addition
polyglutamates
in the activity
tion
have
Organisms
a)
are
developed
years.
MATERIALS
Assay
or
of them
of Bacteriology,
the medium
of Toennies
for the determination
The
Hebrew
et al.3 (See
table
3 for comof folinic
acid
(with
P. cere-
University-Hadassab
Medical
School,
Israel.
Apr.
20,
1962;
accepted
for publication
July
12,
1962.
609
BLOOD,
VoL.
20,
No.
5
(Novmzt),
1982
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
610
GROSSOW1(;Z
visiae),
(1
thyiiidine
acid.9’”
The
matched
18
is Illa(le
x 150
tip
caps
distilled
and
Folic
( to bring
a;id
(
pH
are
of these
7.0
for
60,
are
run
in
different
triplicate.
duplicates
P.G.A.,
are
acid,
us with
providing
Inoculu
from
supplemented
drop
are
of
Twice
folinic
dispensed
distilled
into
the
volume
with
or singly
covered
water
of
are
to
for
between
final
or
F.A.
per
dilution.
duplicates
and
teropterin
Pearl
River,
up
of
Standard
acid
of
usually
these
for
10,
each
20,
assay
40,
curves
activity
the
at
three
triplicates
or
small.
pteroyl
\Ve
NaOH
water;
made
Results
by
O.05N
distilled
or folinic
( SO(lium
Y.
I)rcl)are(l
of
are
acid.
are
N.
arc
(lrOps
concentrations
folinic
their
tubes
aci(l
make
P.G.A.
assayed
a few
100
added
duplicate
lencovorin
with
ml. with
doubly
Standard
solutions
to
either
calciuni
them
frozen.
and
ptg./ml.
and
) were
triglutamate
thank
I)r.
L.
Ellenbogen
for
compounds.
ni Preparation
A loopful
Cultures
vohinie
Laboratories,
these
P.G.A.
of
if kept
pteroic
with
psi.
presence
be assayed,
cotton
( or
to
plugged
the
aliquots
material
15
dissolving
differences
Lederle
the
in
ml.
is used.
)
solutions
averaged;
from
at
the
employing
folinic
obtained
tubes
2.5
AL.
Solutions
month
Extracts
(lilutions,
and
pg/mi.
filling
one
of
minutes
water
growth
and
addition
compounds,
) and
stable
the
strength
10
Acid
100
it enhances
water
for
Folinic
from the concentrated
80, 120, 160 and 200
assay
distilled
deionized
10 ing.
to
solutions
as
double
After
autoclaved
solutions
out
in
tubes.
with
ml.
) afl(l
Concentrated
weighing
test
subsequently
Standard
is added,
is prepared
mm.
5.0
to
aluminum
)
jtg./ml.
medinni
FT
a
with
24-hour
150
L.-casei
of the diluted
stock-agar-culture
ttg./ml.
and S. faecalis
cultures
is used
of
is
P.G.A.
or
are
transferred
folinic
acid
diluted
1 : 100
to inoculate
each
to
and
ml.
5
of
incubated
and
assay
medium
overnight
at
of P. cerecisiac
those
37
1 : 10.
C.
One
tube.
Incubation
tubes
S. faecalLs
are
Assay
the
after
48
hours.
photometer
wide
incubated
assay
for
read
is
Growth
or
at 660
in
hours
24
hours
is
measured
density
m
or 48
24
after
37
C..
at
a Klett-Summerson
accor(ling
while
650
to
the
m1tt
in
colorimeter
the
other
a
with
organism
two
Coleman
an
uSe(l;
assays
adapter
are
read
junior
spectro-
to
18-mm.
hold
tubes.
Release
of Folic
a ) Whole
towards
1 ml.
of
Acid
blood.
the
assay
blood
( 30
acid
Derivatives
In order
to release
organisms,
citrated
Ascorbic
the
the
is hemolyzed
mg.,
freshly
by
psi)
15
Aliquots
double
to
of
strength
stabilize
the
the
clear
medium
and
0.25
for
labile
assayed
for
added
(without
(10
any
prior
to
activity and
mg/mI.
of
preincubation
heating
was,
serum).
at
the
serum
therefore,
C.)
sample
omitted.
of
0.5
volumes
vitamin11
F.A.
activity
(F.A.A.).
NI phosphate
are
water.
( pH
coagulate
pH
6.0,
for
its
not
treated
and
minutes
by
to the
thus
20
did
obtained
serum
added
Samples
for
ascorbate
were
the
) are
is assayed
of
results
active
follows:
distilled
buffer
buffer
autoclaved
supernatant
presence
Similar
them
as
of
and
centrifugation
0.1
render
modified
NI phosphate
by
the
in
17
is
6.0)
and the volume
made
up to 20 ml.
at 37 C. and then
autoclaved
(20
of
the
samples
and
with
and
al.41’
( obtained
their
The
37
it
N NaOH
2 hours
will be referred to as activated hemolysates.
b) Serum.
Samples
are diluted
1:10 with
acid
derivatives
et
2 ml.
forms
supernatant
and
F.A.
Toennies
diluting
)
minutes
at
of
prepared
pH adjusted
to 6.0 with
samples
are incubated
proteins.
various
procedure
are added,
the
The hemolyzed
tion
at
incubation,
ascorbic
at
F.A.A.
further
15
psi
Incubaincrease
Herbert.6
RESULTS
Comparative
No.
8043
Figure
S. faecalis
Growth
and
Response
S. faecalis
1 shows
as compared
the
No.
markedly
to that
to P.G.A.
and
Folinic
Acid
of
S. faecalis
ATCC
9
higher
of strain
sensitivity
No.
8043.
of
Whereas
the
No.
200-250
9 strain
,g./ml.
of
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MICROBIOLOGIC
DETERMINATION
OF’
Ca -LEUCOVORIN (Folinic
F.A.
611
DERIVATIVES
acid)-,u,g/m1
100200
Fig.
(-n
z
w
1.-Response
folinic
acid
of
S. faecalis
ATCC
0
to
P.G.A.
S. faecalis
No.
and
and
9
8043.
-J
4
U
I-.
0
PTEROVL
are
GLUTAMIC
required
40-50
to
are
since
As
with
Pteroic
P.G.A.
with
the
This
basis,
form
racemic
strains
latter
folinic
the
is
the
standard
of
is about
50 per
of folinic
acid.
also
maintained
strain,
the
No.
cent
The
only
9 strain.
as active
difference
in
respect
acid
and
to
as
in
folinic
in figure
causes
the
stimulation
concentrations
between
in the
absence
Effect
of Ascorbic
assay
both
of pteroic
strains
of P. cerevisiae
2. At low
of thymidine
are
the
9 strain;
on Growth
is shown
thymidine
acid,
No.
show
the
teropterin
same
is
specificity
compounds.’2
of Thyinkline
addition
a weight
two
of
response
Acid
and
sensitive
Effect
the
growth
a similar
1).
and
towards
maximum
to obtain
on
between
Teropterin
more
half
it is a synthetic
sensitivity
(fig.
-,,ug/m1
required
leucovorin,
P.G.A.,
acid
produce
,ttg./ml.
Calcium
ACID
of folinic
Acid
concentrations
a four-fold
is
0.1
and
acid
even
two-fold.
acid
in growth,
The
effective
However,
if added
Activity
of Folinic
of folinic
increase
5 ,g./ml.
on F.A.
in Presence
in
of Whole
( 50 p.ig./ml.),
while
at
higher
concentrations
thymidine
of
is ineffective
concentrations
Blood
Acid
as
and
high
as
Activated
50
Blood
Heniolysates
As some
very
biological
types
of the
unstable
in
activity
of
and
7.5-50
compounds-especially
the effect
of ascorbic
of blood
treatment
Methods)
(from
F.A.
air,
were
added
mg.
per
during
studied:
to aliquots
ml.
of blood).
storage
1)
of ascorbic
The
the
acid
tetrahydro
was examined
at 4 C. and
whole
blood
acid
samples
derivatives-are
on the micro-
30 C.,
respectively.
Two
was
hemolyzed
(see
in increasing
were
incubated
concentrations
for
2 hours.
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612
GROSSOWICZ
ET
AL.
00
Fig. 2.-Effect
of thymidine
of P. cerevisiae
(L. citrovorum)
presence
of folinic
acid.
z
U,
-J
on growth
in
8081
4
U
0.
0
01
FOLINIC
at 37 C.
were
and
then
divided
at
citrated
acid.
mg.
7.5
37
C.;
mediately
most
they
for
no
two
) was
(
added
autoclaved.
into
the other
(b
of undiluted,
tion of ascorbic
loss
per
ml.
was
from
tested
and
autoclaved
F.A.A.
The
results,
of F.A.A.
1.-Effect
supernatants
of blood),
then
of as much
Table
(a)
: one
activated
and
which
of untreated
the
are
blood
with
as 65 per
S. faeca&
cent
of the
or
F.A.A.
of Ascorbic
Acid
Hemolysates
(mtg./ml.)
by
on Folic
Stored
(series
Assayed
Blood
of blood)
sample
No.
casei
Assayed
Blood
sample
No.
with
S. faecalis
Assayed
BlOOd
sample
No.
with
P.
samples
F.A.A.,
while
in
acid.
drop
P. cereviiae.
table
the
particularis noticed
other
hand,
a
hemolysates
of Blood
at 30 C.
and
Incubated
at 30 C. for
Blood hemolysates
(series
b)
a)
al-
on incubation
in activated
Acid Activity
at 4 C. and
im-
1, show
This holds
in activity
On
occur
2 hours
assayed
citrated)
ascorbic
a slight
may
for
obtained
Blood
4 Days
Blood
(series
o)
acid
with
L.
for
summarized
Unincubated
Blood hemolysates
(mg./ml.
respective
incubation
extracts
(undiluted,
in the absence
of added
done
with
L. cisei,
while
Ascorbic
the
immediately
incubated
for 4 days at 30 C. prior
to the assay.
2) Samples
blood
were
incubated
at 30 C. for 4 days with
no addiThe samples
were hemolyzed,
a small amount
of ascorbate
it is determined
loss
The
parts
were
at 30 C. for 4 days
ly true for assays
when
1iug/mL
ACID
cerevisiae
7.5
20
88
50
7.5
63
-
88
1
50
-
65
2
8
34
83
85
42
88
88
20
33
50
7.5
38
25.4
49
31.7
57.2
34.1
55.4
82.9
56.4
81
37.8
40
48.1
42
44
1
6.6
8
9.9
5.1
8.4
-
7.2
2
2
2.1
2.6
8.2
1.4
2.1
1.9
2.4
9.0
2.1
8
6
5.2
6.9
7.2
8.8
4.2
4.5
4.4
5.5
1
5.4
7.4
7.6
6.4
4.2
7.0
7.2
7.8
7.8
2
8
1.7
5.1
1.5
4.8
2.0
6.2
2.4
7.0
-
1.1
3.5
1.5
8.8
1.7
4.6
2.1
6.5
10.8
1.7
Whole
blood
was
hernolyzed
(see
Methods)
and to aliquots,
ascorbic
acidS in increasing
amounts
(as
indicated
in the table)
, was
added.
After
incubation
for 2 hours
at 37 C., the samples
were
autoclaved.
The supernatants
of each
sample
were
divided
into
two
parts:
one was tested
immediately
for F.A.A.
(series
a) while
the other
was incubated
for 4 days
at 30 C. prior
Whole
blood
(citrated,
undiluted)
was
incubated
at 30 C. for
Thereafter,
hemolysates
were
prepared,
activated
with
ascorbate
and
The
from
assayed
assays
different
for F.A.A.
activity
were
carried
out
donors.
(series
with
to the assay
(series
b).
4 days
in absence
of ascorbic
acid.
(7.5
mg./ml.
of blood),
autoclaved,
c).
three
different
organisms.
Samples
No.
1,
2 and
3 represent
blood
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
MICROBIOLOGIC
DETERMINATION
Table
2.-Folic
OF
Acid
adults
L.
43
range
Serum
c asei
S.
of Healthy
Blood
Serum
Blood
fa ecalis
Serum
47-149
3.2-15
3.0-20
0.3-1.7
89
8.3
12
0.75
mean
Normal,
613
DERIVATIVES
Activity
in Blood
and
and Infants
(mtg./ml.)
Subjects
No.
Normal,
F.A.
Adults
P.
Blood
cer
evieiae
Serum
1.5-25
0.25-1.3
6.3
0.3
infants
(age
2-24
months)
58
range
mean
incubated
ml.
at 30
of
blood)
more
per
Folic
Acid
ml.
Activity
Table
in
adults
carried
normal
ranges
149
of
obtained
with
cent
protection
the
procedure
L. casei
F.A.A.
The
35
(mean
with
The
adults.
(7.5 mg.
(30 mg.
ascorbate
ascorbic
acid
of the
per
or
activity.
Sub/ects
out
between
mttg./ml.
of low
as
subjects,
more
than
90 per cent
serum
contains
only 5-10 per cent.
or P. cerevisiae.
as in
presence
concentrations
75 to 100 per
affords
results
assays
S. faccalis
3.9
in the
in Healthy
healthy
while
1.0-22
96.4
high
of
2 presents
found
4 days
of blood)
Methods.
In
in the blood,
same
C. for
; addition
35-160
respective
and
than
in blood
160
in
those
mttg./ml.
under
blood
infants
F.A.A.
(mean
is found
activity
is
employing
of healthy
L. casei
outlined
of all F.A.A.
Much
higher
96.4
for
either
is about
the
infants
and
) and
mttg./ml.
47-
89.0).
DIscussIoN
The
S. faecalis
considerably
by
shown
same
No.
more
the
specificity
response
as
(L.
P. cerevLs’iae
9 strain
used
sensitive
assay
the
to P.G.A.
the
to
than
and
standard
folinic
strain
citrovorum)
assay
folic
strain
also
the
been
F.A.
to be
fere
requirement
assayed
with
in these
of microgram
S. faecalis
Preincubation
at pH
The
by
ments
effect
the
longed
of 30 mg.
the
storage
the
30
C.
). Apparently
protection.
Since
acid
acid
on
1 show
(autoclaving
C. The
ml.
in
the
the
the
thymi-
of
in the
samples
would
inter-
of ascorbic
and
10 mg.
medium
added
also
in
of
contains
the
both
buffer
blood.”
recently
The
been
experi-
the F.A.A.
during
the
to the
of serum)
little,
ascorbate
added
of
membrane
has
et al.5
acid
is
shows
intact
serum
Baker
protects
and
ml.
the
in
samples
per
and
blood
phosphate
of
acid
the
and
(F.A.A. ) of
F.A.A.
ascorbic
absence
assay
assays
acid
activity
) of
presented
citrated
the
The
Thymidine,
as it replaces
or thymidine
findings
that
amount
of blood
Undiluted,
at
L. casei
per
addition
presence
ascorbic
folic
previous
experiments
findings.6’15”6
storage
table
heating
at 30
storage
total
in
short
with
the
confirming
exhibits
assays.
of ascorbic
Herbert,6
it
is
as
8043
examined.
assays.10’13
S. faecalis
of thymine
hemolysates
enhances
summarized
during
of
greatly
protective
shown
L. casei
and
No.
1 ), yet
of the
Therefore,
quantities
of blood
6.0
organisms.14
) derivatives
substances
improved;
dine results
in increased
sensitivity
and specificity
however,
should
not be added
to L. casei
and
the
(fig.
acid
towards
has
(
F.A.
ATCC
acid
standard
samples
is ample
agreement
both
pro-
with
(a
in view
recent
if any, loss of F.A.A.
on
(when
assayed
with
red
cell
phosphate
is responsible
and
for
ascorbic
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
I
614
GROSSOWICZ
Table
5%
casein
3.-Composition
of Double
Strength
Assay
Medium*
( Nutritional
Biochemicals
Corp.
Cleveland,
Sodium
Ohio)
acetate,
125
anhydrous
Glucose
ml.
20.0
Gm.
20.0
Gm.
KOHPO4
KHOPO4
3.5 Gm.
3.5 Gm.
Ascorbic
250
mg.
DL-alanine
acid
200
mg-prepare
a 2%
solution;
add
10
ml.
L-asparagine
100
mg-prepare
a 1%
solution;
add
10
ml.
L-cystine
200
mg-prepare
a 2%
solution
MgSO47H2O
200 mg-weigh
out
2.0
10mg.
FeSO47H9O
MnSO44H2O
10mg.
.
weigh
10 111g. j
out
17 mg-prepare
a O.17C/c
Adenine
sulfate
Guanine
hydrochloride
12.5
Uracil
Xanthine
nlg.-prepare
Thiamine
100
mg.
add
a 0.1%
solution;
add
10
ml.
mg.-weigh
out
20
mg.
0.1
mg-weigh
out
10
mg.
0.5
mg-weigh
out
10
mg.
stock
The
pH
The
volume
solutions
of the
*For
assay
final
concentration;
weigh
out
50
mg.
of
each
of
to
to the
ments
not
described
L. casei
conjugase.
the
activity
with
10
thymidine
growth
dissolve
10 rng.
ml.
in
ml.
dissolve
in
of
add
10
200
acetic
1000
in
ml.
ml.
ml.
water
acid;
1000
water;
with
add
ml.;
10
add
ml.
1 ml.
N NaOH;
1
of
stimulation
g./m1.
final
Co.,
\Vilmington,
this
organism
of the
may
ratio
ability
of
concentration.
)
Del.
0.1
in presence
assay
be
(P.
cerevLsioe,
tivity
are
found
active
one
the
For
per
of
organism
organism
paper
have
not
of those
assay
( V/V)
cent
folic
acid.
as shown
sera
that
to
oc-
in the
utilize
and
higher
is assumed
polyglutamates
bypassing
shown
a two
of
substances.5
with
increase
chicken
responds
polyglutamates;2
F.A.
Experi-
to three-fold
after
incubation
that
L. casei
also
of
healthy
L. casei)
of
in
pancreas
to
as
yet
un-
methyltetrahydro-
serum
individuals
is about
and
blood
cells
contain
of
which
Thus,
advisable
the
Similar
(1:25:25).
concentrations
it seems
with
1:2:15.
individuals18
healthy
the
serum.
to 20-fold
L. casei
compounds.17
blood
both
to
10-
with
of F.A.
necessarily
S. faecalis,
in
is about
is found
blood
in the
substances,
than
individuals
activity
in the
in this
compounds
used
cells
highest
of F.A.A.
suggestive
of healthy
The
blood
hemolysates
it is plausible
of
F.A.
acid
blood
out
Powder
enhances
presence
However,
identified
F.A.
add
blood
serum.
than
of
ml.
toluene.
6.45
( Atlas
growth
due
highly
HC1;
ml.
80
Tween
rather
The
500
Tween
of whole
that
folic
to
10
dissolve
j
under
to
1
iN
“)
mg-weigh
4 C.
ml.
and
L. casei#{176}is eliminated.
F.A.A.
be
at
P. cereviriae:
add
a nonspecific
with
to
up
with
S. faecalis:
kept
is adjusted
is made
with
acid,
are
medium
ml.
in 100
ml.
10
0.6
The
10
ml.
0.2
0.01
of
10
mg.
Biotin
ml.
add
mg.
Riboflavin
water
add
0.5
acid
each
solution;
0.5
hydrochloride
10
of
solution;
hydrochloride
P-Aminobenzoic
add
a 0.1%
0.5
Pyridoxamine
of
solution;
acid
Ca-pantothenate
HC1;
dissolve
a 0.125%
10 mg-prepare
mg.
2N
add
10 mg-prepare
Nicotinic
in
Gm.
NaCl
than
AL.
hydrolysate
enzymatic
cur
ET
It
the
are
to
same
much
determine
three
assays
ratios
of ac-
is,
therefore,
assortment
higher
in
F.A.A.
the
in
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
MICROBIOLOGIC
DETERMINATION
OF
Table
Tryptone
4.-
Yeast
(Difco)
Yeast
Stock
Tryptone
2.5
615
DERIVATIVES
F.A.
Medium
Salts
Gm.
extract
(Nutritional
Biochemicals)
Glucose
Sodium
acetate,
Nutrient
anhydrous
agar
25.0
Gm.
25.0
Gm.
1.0
Gm.
KH9PO4
10.0
Gm.
distilled
10.0
Gm.
water
Salts
11.0 Gm.
(Difco)
A:
KOHPO4
B:
Salts
A
5 ml.
MgSO47H2O
Salts
B
5 ml.
NaCl
Distilled
pH
water
500 ml.
FeSO4
6.8-7.0
rather
nounced
than
in
decrease
suffering
The
serum.
of
of
from megaloblastic
simultaneous
assay
tinguish
between
relative
measured
abundancy.
with
the
blood.20
deficiency
due
to
view
blood
three
forms
of the
Thus,
the
P. cerevisiae
activities
0.5
HC1,
1 ml.
conc.
The
differential
lack
of specific
assay
of
used
derivatives
the
in
allows
one
in the
to
and
of
fetal
aid
in
pro-
patients
body,
in
therefore,
more
serum
of the tetrahydro-forms
are much
higher
should,
F.A.
by
that
Gm.
500 ml.
water
organisms
)
assay
Gm.
MnSO44H2O
than
F.A.
Gm.
0.5
0.5 Gm.
is strengthened
whole
anemia.19
with
the
various
maternal
This
F.A.A.
10.0
7H2O
distilled
blood
500 ml.
distheir
F.A.
than
(as
in
discovering
derivatives.
SUMMARY
A
microbiological
blood,
( F.A.A. ) in
P. cerevisiae),
higher
sensitivity
of the
cedure
release
the
F.A.A.
( L.
spares
of extraction
and stabilization
in blood
of
and
the
the
Es
(i.e.
describite
acido
un
folic
of healthy
methodo
in
Lactobacillus
casei,
Streptococcus
le specificitate
es simile.
Le
augmentate
acido
de
pro
per
folinic).
acido
folic
le
Es
listate
adultos
de
addition
ab
valores
derivatos
esseva
compositos
testate.
pro
e infantes.
le
activatate
whilst
The
by
Finally,
are
add-
the
modified
le determination
uso
de
tres
to
proassure
given.
de
folic
acido
pro
modificate
de
acido
pro
folic
considerabile8043,
durante
in
le studio
citrovorum)
elimina
le extraction
cerevisiae).
un
no.
includite
( Leuconostoc
activitate
organismos
e Pediococcus
monstra
ATCC
(que
del
differente
empleate
standard
thymidina
le sanguine
been
infants
faecaloi,
esseva
le linea
Ic technica
a con-
increased
acid.
pro
le
P. cerevLs*ie
de
Finalmente,
e stabilisation del
normal
le vane
sensibilitate
con
que
que
and
shows
INTERLINGUA
microbiologic
e sero
is also
folinic
blood
has
assayed.
adults
IN
sanguine
Le linea
no. 9 de S. faecalis
mente
plus
alte sensibiitate
que
for
acid
activity
S. faecalis
and
No. 8043 strain,
tested
is similar.
) assay
citrovorum
folic
casci,
employed
9 strain
requirement
the F.A.A.
from
of the compounds
serum
of
( L.
ATCC
derivatives
F.A.
SUMMARIO
de
No.
standard
various
P. cerevisiae
which
the
determination
three
organisms
S. faecalis
than
towards
thymidine,
for
with
The
sensitivity
specificity
ing
method
serum
is described.
siderably
the
and
le
del
vector
assecurar
in
es etiam
requirimento
de
de activitate
le le liberation
sanguine
e sero
ab
From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
-e
616
GROSSOWICZ
ET
AL.
REFERENCES
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From www.bloodjournal.org by guest on June 18, 2017. For personal use only.
1962 20: 609-616
Microbiologic Determination of Folic Acid Derivatives in Blood
NATHAN GROSSOWICZ, FREDERIKA MANDELBAUM-SHAVIT, ROSY DAVIDOFF and J.
ARONOVITCH
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