Ascorbic Washko,

Daniel

ABSTRACT and

The

the effect

were of

which

1.0-1

was

was

ofphysiologic

tivity

transport

transport

activity

inhibited

the

transport

activities

ties

had

The

Am

ac-

mmol/L.

Glucose

WORDS

Ascorbic liquid

acid,

acid

by both

tionation, and were performed

buffer

acid

transport

activi-

Nuir

199 1 ;54:

glucose,

the effect

acid,

(1-3)

is found

but its function

facilitate

the

preserve

is not

oxidative

neutrophil

determine

intracellular that

high-

neutrophils

can

tracellular how

ascorbic

(8-10).

These form

phil

membranes. these

However,

studies.

Ascorbic

but

in normal

1 50 jzmol/L

not

found

used

studies acid

plasma

In addition, may

are several

ascorbic

human

have

vs its metabolites

be varied

so

and

exof

acid

analysis

applicable, as described

content tillation

ofcells was spectrometry

analysis viously

of the (3).

by using

acid the

and

range

the

ofascorbic acid

insensitive

assay

accurate

or other

I99I;54:l22lS-7S.

Printed

than

size

the

to

on the transport

strength

ofthe

for changes

and

incubation

in the glucose

samples

was

detection

analysis

done

(19)

by HPLC

with

for dehydroascorbic

previously

(3, 20).

The

with

modifications acid

was also

[14C]ascorbic

acid

determined on a mass basis by liquid scmin conjunction with HPLC (3). Protein

extracted

bars

cells represent

have

of the

was

performed the mean

been

as described ± SD ofat

omitted

when

the

pre-

least SD

three

was

less

to be

ap-

symbol.

Results Neutrophil

express L) basis

94%

millimolar

0.31

volume

zL/l06

(3) as in previous

studies

distribution

which ofthe

acid

This on

acid

was

present

only

ascorbic

within

in the acid

and

volume

was

a concentration

dehydroascorbic acid neutrophils contained

intracellular

determined

[3H]sucrose

(2, 2 1-23).

of ascorbic

acid and I . Isolated

was

cells by using

intracellular ascorbic in all experiments.

acid/L,

associated

intracellular

proximately

Table

neutro-

acid is probably methods

determination

reducing

in USA.

Error

The

substances

(17, 18). lm J C/in Nutr

defrac-

neutrophils

reduced

used

to

(mmol/ and

content are 1 .3 mmol

was cytosolic.

[‘4C]urea

form.

the

shown in ascorbic At

least

Intracellular

dehydroascorbic

dehydroascorbic prevented

ofall

points

samples.

ascorbic

acid

across

problems

glucose the ionic

electrochemical

performed

investigated

dehydroascorbic

transported

was investigated

( 1 1 - 1 5) and

at all (16).

in these

of ascorbic

Transport

that

was

acid

volunteers, subcellular

in plated neutrophils (3). In experiments

to compensate

(3). Where

the

understanding

has been

accumulation described

ofextracellular

maintained

normal

neutrophil

5),

neutrophils.

indicated there

must

an

chromatography

by acting

to intracellular

into

that

(4,

tissues

in neutrophils,

acid

neutrophils

ofextracellular

50-

acid

requires

is transported

of the vitamin

concentrations acid,

This

experiments

the

host

may

the bactericidal products prometabolic respiratory burst (6,

in relation

into

acid

microorganisms

of ascorbic

transport

neutrophils

Ascorbic

protect

of ascorbic

be studied

acid acid

was with

role

concentrations.

Ascorbic

of

and/or

to neutralize during the the

in human

understood.

destruction

concentration

function

amounts

well

integrity,

as a reducing agent duced by neutrophils 7). To

in large

from

volume,

ofascorbic

was

Experimental acid

radioisotope

concentration.

Introduction Ascorbic

distribution

using

liquid

neutrophils

ascorbic acid as previously

accumulation

Glu-

J C/in

dehydroascorbic

chromatography,

and

by

high-performance

ofintracellular

coulometric

performance

a new

accumulation, neutrophils

(19).

of human

termination

determine

122 lS-7S.

KEY

and

assay

Isolation

and

fashion.

ascorbic

in human

Methods

buffer

the low-affinity

ofascorbic

reversible.

the transport,

acid

presence

transport

whereas

Km of6-7

of both

completely

(HPLC)

of ascorbic

high-affinity

accumulation

techniques

94%

by a high-

in a concentration-dependent

inhibition

was

The

in the extracellular

zmoI/L

investigated

Intracellular

form.

concentrations

an apparent and

cytosol.

We

of ascorbic

neu-

at least

was mediated

activity.

acid

transport

isolated

acid/L,

the

acid

Km of2-5

uptake

cose-induced

in

of millimolar

had an apparent

acid

Freshly

in the reduced

Accumulation

a low-affinity

of ascorbic

ascorbic

ofascorbic

intracellularly.

Levine

on ascorbic

neutrophils.

only

amounts

Mark

distribution

unbound

found

led to the accumulation acid

and

.4 mmol

present

acid

and

glucose

in human

contained

ascorbic

uptake

ofextracellular

investigated

trophils

Rotrosen,

,2

© 1991 American

is

the Laboratory ofCell Biology and Genetics, National Institute Digestive, and Kidney Diseases, and the Laboratory of Host Defenses, National Institute ofAllergy and Infectious Diseases, National Institutes of Health, Bethesda, MD. 2 Address reprint requests to M Levine, Building 8, Room 415, National Institutes of Health, Bethesda, MD 20892. I

From

of Diabetes,

Society

for Clinical

Nutrition

l22lS

Downloaded from https://academic.oup.com/ajcn/article-abstract/54/6/1221S/4715197 by Boston University user on 07 January 2019

Philip

acid in human

12225

WASHKO

TABLE 1 Subcellulan distribution

ofascorbic

acid

in neutnophils*

ET

AL

calculation

for [‘4C]ascorbic

component

ofascorbic

acid

acid

in Figure

uptake

had

acid and was saturable. Ascorbic

acid

+

Subcellulan

Ascorbic acid

fractions

dehydnoasconbic acid

n = 3. Data

1 ± SD;

ascorbic acid was saturable.

1.32 ± 0.18 0.01 1.43 ± 0.09

0.0 1 1.34 ± 0.04

activity

0.Ol

1.24 ± 0.12

1.32

± 0.08

1.27

± 0.06

1.25

± 0.01

1.56 ± 0.08

1.27

± 0.03

from

reference

calculation

of[’4C]ascorbic

in the amount

of intracellular concentration data indicated

extracellular linear and

molecular weights in the particulate

fractions. ascorbic from

acid/L

accumulated

initial

concentration

an

uptake

was

sium

1, neutrophils

dependent

in the

the

vitamin

presence

buffer

with

linearly

50 mol

for 210

mm

1.0 to > 5.0 mmol/L.

of

on the

extracellular

incubated

of calcium

This

and

magne-

(24).

Because uptake ofascorbic acid was linear for 2 10 mm, mm intervals were used for studying concentration-dependent kinetics

with

[14C]ascorbic

titation of radioactivity accumulation were

the [‘4C]ascorbic acid,

which

acid.

Uptake

whereas determined

measured

90-

port

into neutrophils

to the

purity

of the

total

Fig 2,

(inset,

ascorbic

acid

acid and transport

Fig 2, D). The

acid

accumulation

ascorbic acid, there was no

or [‘4C]ascorbic

because increase

acid

at any

not shown). of 50-300

transport and accumulation did not occur (Fig 2, B). To

pendent cumulation

was

ascorbic

with acid

by quan-

occur

was

ascorbic

acid

concentrations

for

mmol/L.

by HPLC. did

not

< 2 mmol/L;

second

trans-

concentration-dc-

incubated

at 0.4-15

determined

The

for applying were

acid

against

8 mmol/L.

suitable

Neutrophils

ascorbic

transport

could

acid

was thus

kinetics.

saturation

apparent

ascorbic

activity

20 mm

Ascorbic

with

acid

ac-

As shown

in Figure

3,

occur

extracellular

at

at higher

concentra-

tions

saturation occurred. For ascorbic acid accumulation, the apparent Km of this second transport activity was 6.7 mmol/L by Lineweaver-Burk by Eadie-Hofstee

total ascorbic acid content and by HPLC. More than 90% of

acid transported

corresponded

was

imol/L

total

of ascorbic acid (data that at concentrations

accumulation

extracellular

in Figure

and

ascorbic saturation

this second

whether

of extracellular

As shown

inset,

activity and detera concentration gradient, we incubated isolated neutrophils with millimolar concentrations ofascorbic acid. Neutrophils incubated with cxtracellular ascorbic acid at 8 mmol/L accumulated > 14 mmol/ L of intracellular ascorbic acid (data not shown). In addition, transport occurred linearly for 30 mm when the concentration mine

3.

ascorbic acid was not bound to proteins with 10 000 and virtually none was detectable

(left

for ascorbic high-affinity

extracellular acid addition

extracellular The above imol/L remained

5.4

was acid

are not the result oftrapped immediately after ascorbic

characterize

reprinted

K,,, ofthis analysis

uptake had a high affinity The apparent Km of the

by this

uptake

0.03 ± 0.003 I .42 ± 0.01

0.Ol

The apparent by Lineweaver-Burk

zmol/L

this

obtained human transport

analysis analysis

for [‘4Cjascorbic neutrophils appear activity

(left

(right

inset,

inset,

acid uptake to have both

for ascorbic

Fig 3) and

Fig 3). Similar

6.6 mmol/L results

were

(data not shown). Thus a high- and a low-affinity

acid.

Neither

Time

(mm)

the

intracellular

ac-

was ascorbic

radiolabeled

ma-

terial (data not shown). In addition, extracellular ascorbic acid concentrations did not change (data not shown). Neutrophils were incubated with 1 1 different concentrations of[’4C]ascorbic acid

for

acid

uptake

90 mm.

As can

occurred

be seen

two components. acid accumulation

Similar (Fig

linear at extracellular 300 mol/L (Fig

results 2, B).

ascorbic

acid

from

these

were

points

subtracted

B, to obtain

acid

whether uptake

each and

by regression

from

component

corresponding

saturation

were

for both

acid accumulation Points on each values

curves

I

25 to was not acid

were

lines

analysis

with

ac-

(Fig 2, B). was saturable.

accumulation

25 imol/L,

and total ascorbic through the origin.

substrate

from

in intracellular ascorbic as high as 5.0 mmol/L

concentrations

acid uptake extrapolated

manner

for total ascorbic of uptake was

ascorbic acid concentrations 2, A and B). This linear component

We next determined ascorbic

2, A [‘4C]ascorbic

were obtained One component

due to diffusion and resulted cumulation to concentrations Because

in Figure

in a concentration-dependent

linear

for

constructed [‘4C]ascorbic and of these

in Figure

(Fig 2, C and

were lines

2, A and D). The

FIG 1 . Ascorbic acid accumulation in neutrophils as a function of time. Plated neutrophils were incubated in bicarbonate-free buffer contaming (per L) 1.5 mmol Ca2, 1.3 mmol Mg2, and 50 imol ascorbic acid (pH 7.4) for the times indicated. Intracellular ascorbic acid was determined by HPLC.

Downloaded from https://academic.oup.com/ajcn/article-abstract/54/6/1221S/4715197 by Boston University user on 07 January 2019

4

1.35 ± 0.04 0.Ol 1.34 ± 0.04

2.1

that

for ascorbic transport activity

affinity

C) and 2.2 imol/L by Eadie-Hofstee analysis (right inset, Fig 2, C). Saturation occurred with -40-50 zmol ascorbic acid/L in the extracellular buffer. The calculation for total ascorbic acid accumulation (Fig 2, D) also showed that one component of

mmo//L Homogenate Nuclei + unbroken cells Postnuclear supernatant Particulate fraction (membranes and granules, I 5 000 X g) Supernatant (1 5 000 x g) Microsomes (134 000 X g pellet) Cytosol (134 000 X g supernatant) Cytosol (filtrate, 10 000 MW nominal retention) Cytosol (concentrate, 10 000 MW nominal retention)

was

2, C showed

a high

ASCORBIC

ACID

IN

NEUTROPHILS

l223S

C

0

4-I 5.-

.-

04-I

0

0

Downloaded from https://academic.oup.com/ajcn/article-abstract/54/6/1221S/4715197 by Boston University user on 07 January 2019

80 ..

0 .. 5-. 4-I

C

0

50

100

200

150

External

[14C]

250

Ascorbic

Concentration

300

350

400

350

400

Acid

(tmol/L)

6.00

5.00

5.-

4.00 :C) 0 c

5.-

0-

0

3.00

Is

2.00

1.00

0.00

0

50

100

150 External

200 [14C]

250 Ascorbic

Concentration

300 Acid

(tmol/L)

FIG 2. Concentration dependence of[’4C]ascorbic acid uptake and total ascorbic acid accumulation. Adherent neutnophils were incubated in bicarbonate-free buffer containing 0-300 jzmol [‘4C]ascorbic acid/L for 90 mm: A, uptake of[’4C]ascorbic acid determined by liquid scintillation spectrometry; B, accumulation of total ascorbic acid determined by HPLC: C. substrate saturation curve of the high-affinity transport activity for [‘4Cjascorbic acid uptake (insets, Lineweaver-Burk analysis, left, and Eadie-Hofstee analysis, right): D, substrate saturation curve of the high affinity transport activity for total ascorbic acid accumulation (inset, Lineweaver-Burk analysis). Reprinted from reference 3.

cumulation

oftotal

acid

low-affinity

by the

for by extracellular We expected that

be temperature

ascorbic

acid transport

nor the uptake activity

could

of[’4Cjascorbic be accounted

trapping of ascorbic acid (data not shown). both ascorbic acid transport activities would

dependent

based

on the

above

findings.

To test

this,

neutrophils

200 mol ofascorbic affinity tivity

were

incubated

[14C]ascorbic acid/L. acid was predicted transport

(200

zmol/L),

activity

at 37 and

4 #{176}C with

10, 50, or

Transport ofthese concentrations to be mediated by either the high-

(10 jzmol/L),

or a combination

low-affinity of the

transport two

(50

zmol/L)

ac-

WASHKO

12245

ET AL

0.40 . C

0 .

0.30

.

h.

0-

Downloaded from https://academic.oup.com/ajcn/article-abstract/54/6/1221S/4715197 by Boston University user on 07 January 2019

.20-J . C

50-

go#{176} 0.20

01

.

..

0.10

5-

#{149}

>O7O

C

-------

x

0 00 0

501

.

1 100

1 150

[14C]

External

__6o

1 200

004

ooo

I 250

Ascorbic

I 300

006

008

1 350

400

Acid

Concentration

(tmol/L)

1.00

0

4

0.80

0 5-

i!#{176}k 0.60

I

0

0.40

5-

C

I

0.20

0 I-

0. 300 External (14C] Ascorbic Acid Concentration (tmol/L) FIG 2. (Continued)

(see

Fig 2). Ascorbic

ities

was

inhibited

activities The ascorbic

were effect acid

4. Accumulation

acid

accumulation

90-95%

temperature

by both

by 4 #{176}C (3).

transport

Thus,

both

of 10 zmol/L

ofextracellular

glucose on the accumulation of activities can be seen in Figure

of extracellular

ascorbic

diated

predominantly

greatest ascorbic

dependent.

of extracellular by both transport

ascorbic

mediated almost entirely by the high-affinity transport was maximal for extracellular glucose at 1 mmol/L. accumulation

activtransport

acid

at 200

acid,

activity, Similarly, zmol/L,

me-

by the

for extracellular acid

at both

10 and

extracellular

glucose

accumulation mmol/L had

of ascorbic no additional

compared with We investigated accumulation

low-affinity

glucose between

200 1 and

acid. effect

transport

at 1 mmol/L. zmol/L,

for both

transport

of

inhibited

the

concentrations acid accumulation

1 mmol glucose/L (Fig 4 and whether the effect ofglucose was reversible

was

extracellular

concentrations

30 mmol/L

Glucose on ascorbic

activity, For




incubated

acid

with

in

10 mmol

glucose/L

at 200

1 mmol

or 10 mmol glucose/L incubation in 1 mmol

ascorbic acid the inhibitory transport

[‘4C]ascorbic

were

glu-

for a 1-h glucose/L had

no

was completely Similar results

high-affinity

cubated before

transport

activity

glucose

and

10 jzmol

[‘4C]ascorbic

reversible for the were obtained for

The

and

accumulation

transport

neutrophils

is mediated and

by

temperature

[gIucose

25

30

35

in-

not

shown).

40

mmol/L

FIG 4. Effect of glucose on total ascorbic acid accumulation. Plated neutrophils were incubated for 90 mm in bicarbonate-free buffer contaming either 10 (#{149}) or 200 () zmol ascorbic acid/L and the concentrations of glucose indicated (pH 7.4). Ascorbic acid accumulation was measured by HPLC.

of ascorbic transport

acid

activities,

These

transport

inhibition of [‘4C]ascorbic tnansporterB

in human which

nmoI

ascorbic

are

activities,

acid

C acid/L

60

1 .04 ± 0.06

0.52

120

1.95±0.14

1.11 ±0.09

2.54 ±0.16

1 80

3.70

1 .74

3.66

*

20

(data

were

for 60 mm liter, I mmol

B

mm

15

two

A

Time

0

E E

10

acid

dependent.

TABLE 2 Reversibility ofglucose-induced uptake in neutrophils low-affinity

5

neutrophils

Discussion

saturable

0

when

in buffer containing 10 mmol glucose/L being incubated in buffer containing per

effect

accumulation at 1 mmol glucose/ effect of extracellular glucose in

1 mmol/L activity.

the

Plated

neutrophils

± 0. 1 1

were

incubated

± 0.02

1 .29

± 0.22

in buffer

containing

± 0.07 ± 0.02

200 Mmol

[‘4C]ascorbic acid/L. Intracellular ascorbic acid was determined by liquidscintillation spectrometry in conjunction with HPLC. Initial [‘4C]ascorbic acid concentration was 0. A: cells were exposed to I mmol glucose/L for the times indicated. B: cells were exposed to 10 mmol glucose/L for the times indicated. C: cells were preincubated in 10 mmol glucose/L for I h before ascorbic acid addition; after 1 h cells were washed and incubated, per L, with 1 mmol glucose and 200 mol [‘4C]ascorbic acid for the times indicated.

Downloaded from https://academic.oup.com/ajcn/article-abstract/54/6/1221S/4715197 by Boston University user on 07 January 2019

5-

C

24

12265

WASHKO

however,

have

to function

different

affinities

at different

for

tamin. The high-affinity transport at extracellular concentrations should always concentration transport

ascorbic

extracellular

acid

and

concentrations

activity, of ascorbic

appear

of the

which acid

may

function

to maintain

was saturated > 40 zmol/L,

a minimum

AL

such vi-

as the

intra-

these

(1 , 25).

lular

The

low-affinity

take

transport

of ascorbic

as 1-2

acid

mmol/L, for

neutrophils.

The

the

tivity suggests in neutrophils

which

displayed

not

majority

high

concentration

become acid.

saturated This

at normal

transport

of ascorbic

capacity

linear

of the

activity

acid

up-

action

portant mined

of the

low-affinity

transport

for how human ascorbic ( 1 5). Human ascorbic acid

on urinary excretion repleted with ascorbic concentration

ofthe acid.

vitamin Ascorbic

of ascorbic

may

acid requirements requirements have

acid

when acid

scorbutic appears

in the

plasma

ascorbic

in

by glucose postulated pounds

However,

neutrophils

plasma

the principles acid

much

that

subjects are in urine when reaches

may

precise

trophils

that

crossed

droascorbic acid to the differences periments.

The

high

concentrations

acid

used

in earlier

( 1 1-16).

In addition,

into

actual

is unknown. was the form

neu-

whose

may have contributed and those of earlier exacid

not physiologic

were

nonspecific

and assay

dehymeth-

at the neutrophil membrane is then reduced after it crosses

ofcalcium these

and

cations

of sodium because sence

with

ascorbic

neutrophils ofcalcium

and

by other

cell

of their function and magnesium

system and

required

in the extracellular

required

mechanism calcium

a cotransport

accumulation

magnesium are

uptake, the In neutrophils

acid

ascorbic

acid,

the buffer.

types

similar

acid

in other

cells

accumulate

some

ascorbic

magnesium,

other

factors

ascorbic

acid

the structures

Inhibition

cell types transporter

this is not likely

transport

in these

The acid

complete

uptake

is cation

reversibility

suggests

type

or noncompetitive.

for

diabetics

not well-controlled. tible to infection,

whose

blood

neutrophil

acid

37). It has been for the two com-

results

acid

for the

in marked

temperature occurs

temperature

independent

inhibition

of ascorbic

ofinhibition,

which

could

may

be im-

findings

glucose

deby fa-

concentrations

are

that diabetics are more suscepresult of impaired white blood

integrity

by the

uptake

the case in neutrophils

These

It is known possibly the

and

acid

cell function (41-44). Ifascorbic acid is necessary function, either to promote the oxidative destruction by reducing

oxidative

uptake

free

respiratory

by glucose

less-than-optimal

for neutrophil of microbes

could

radicals

and

burst,

then

offer

a possible

neutrophil

in-

function

seen

in diabetics.

found cytosol

to occur in neutrophils not previously reported.

( 1We

that most of the intracellular ascorbic acid was in the in the reduced form. In addition, it was not protein

bound, at least was it localized bic acid vitamin idants

to proteins to neutrophil

in the cytosol may function that

enter

generated some

tracellular surface.

to deplete

10 000, nor of ascor-

two

from

ascorbic oftrapped the phagosome

the phagosome.

the ox-

Another

pos-

acid transport activities is to ascorbic acid. Disappearance would maximize the effects

to reduce oxidants secretion ofascorbic

near the neutrophil acid was reported for

(45).

proposed

ascorbic

molecular weights granules. The localization

oxidants in this compartment. It is also possible cytosolic ascorbic acid may be secreted into the cx-

environment Nonexocytic

cells

with

in an unbound state suggests that as a protective antioxidant to reduce

the cytosol

sible purpose of the deplete the phagosome of ascorbic acid from

be investigated

(27, 31). as part of

(36,

cells

of glucose

a physiologic

uptake

ofglucose

ofascorbic

and

The

Although

for ascorbic

is unknown may function

that

for the

acid

because

which

of extracelsuggests

similar.

acid

acid by neutroand low-affinity

glucose

cilitated diffusion, (39, 40).

of

presence

that acid

of extracellular

is cation and in neutrophils

other ascorbic

it is possible

acid

accumulation of the vitamin and pendent (3, 24). Glucose transport

that to the

neutroof intra-

in ascorbic

at a concentration

Although ascorbic acid is known 3), its intracellular localization was

or dehy-

of ascorbic

involved

ascorbic

is likely

ascorbic

explanation

shunt, acid

require-

ofascorbic

with

(36, 38). However,

Earlier of the

by reducing

ion

inhibition

reported for other there is a common

ofascorbic

membrane. Maximal

was that

hibition

in other cell types that transport the possibility cannot be excluded

is converted acid, which

This

species

acid and dehydroascorbic acid obtained (1 7, 18). Finally, an oxmechanism for ascorbic acid up-

take has not been demonstrated the vitamin (27-30). However,

are quite

produced

(8-10) and

acid

oxidants

Dehydroascorbic (8-10)

ascorbic

work

the insensitive

ods used to measure ascorbic may have affected the results idation-reduction-dependent

acid

the

monophosphate

Several factors our findings

compete

or to preserve

membrane.

by extracellular

(9, 26). between

at

ascorbic

transport

intracellularly

droascorbic

that ascorbic dehydroascorbic

acid activity acid

by the hexose

increased

acid

The

may

portant

Therefore,

human

because

plasma

to be reduced

supplied was

ascorbic

maximal

be competitive

to be reconsidered. of ascorbic

the

30

of ascorbic the vitamin.

30 zmol/L.

for determining

by each transport that dehydroascorbic

was believed

activity

than

to be determined

that is recognized work indicated

equivalents

have

mechanism

remains

vitamin

to accumulate higher

are the basis

requirements

The

acid

continue

concentrations

deterbased

was

of 1 mmol/L. concentrations

because

be im-

are been

zmol/L. It has been assumed that the appearance acid in the urine indicates tissue saturation with

by higher transporters.

ac-

(33-35),

are

accumulation of ascorbic uptake by both the high-

activities

glucose

be

concentration concentration

activity

events

plasma may

transport

that the intracellular ascorbic acid is dependent on the extracellular

at the cell surface

glucose

accumulated

low-affinity

membranes

as high

vitamin.

The

the

activity,

of ascorbic

responsible

of < 6 jzmol/L

at an extracellular

should

concentrations

ofthe

range

different

functions in relationship

acid and

concentrations. then

replete

of ascorbic

acid

to extracellular We are currently neutrophils

in neutrophils and

must

intracellular

devising

of intracellular

strategies ascorbic #{163}3

acid.

to the transport

(3 1 ,

References

32). However, acid

may

in the be involved,

ab-

1. Crandon JH, Lund CC, Dill DB. Experimental EnglJ Med l940;223:353-69.

human

scurvy.

N

Downloaded from https://academic.oup.com/ajcn/article-abstract/54/6/1221S/4715197 by Boston University user on 07 January 2019

transport

level

scorbutic

with

cation-dependent

Glucose regulated phils. Ascorbic acid

for cell function and the plasma

to the

of transporters

transport.

cellular concentration of ascorbic acid necessary when the dietary intake of ascorbic acid is low falls

existence

ments. Furthermore, because divalent cations facilitate phil adherence, granule exocytosis, and translocation cellular

be saturated in human plasma where the normal range of ascorbic acid is 50-150 zmol/L. This

activity

ET

ASCORBIC 2.

3.

4. 5.

7.

8.

9.

10.

1 1.

12.

13. 14. 15.

16.

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IN

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6.

ACID