fatty acids on
plasma lipoproteins-the
Jerusalem Nutrition Study:
high MUFAs vs high PUFAs13
Elliot
Nathan
M Berry,
Shlomo
Eisenberg,
A Kaufmann,
and Yechezkiel
Twenty-six
ABSTRACT
assigned
acid
to a 24-wk
(MUFA)
12-wk
Yeshiva
crossover
32%
periods.
fat,
Total
were
fatty
fatty
acid (PUFA)
fed alternately
cholesterol
(TC)
diets
(50%
during
two
decreased
cholesterol
(LDL-C)
decreased
groups
an additional
significant
effect between
periods.
Concentrations
of high-density-lipoprotein
cholesterol
did not change
significantly.
LDL-receptor
status in fresh monocytes,
affinity
ofLDL
towards
the LDL receptor
in cultured
fibroblasts,
zonalprofiles,
significantly
higher
different
tendency
and
lipoprotein
between
toward
composition
the diets.
lipid
There
were
on
the
by more thiobarbituric
acid-reactive-substances
on that diet. Dietary
PUFA
results in somewhat
lower
LDL-C
concentrations
whereas
with MUFA
the sus-
TC and
ceptibility
of LDL to oxidative
Nutr 1991;53:899-907.
KEY WORDS
fatty
acids,
Dietary
unsaturated
stress
is lower.
Am
ischemic
fats,
fatty
lipids
constituent
of other
fatty
acids,
acids,
it still remains
major
obstacles
free-living
lipoproteins,
adherence.
environment
associated
the Har-Etzion
campus
where
during
type
lipid
peroxi-
effects
of high
MUFAs
over
We
after nearly
with
40 y of
been
able
studies.
One
of natural
period
on lipoprotein
and
low
fatty
acids
Subjects
and
methods
ofteaching
they
ate diets
l991;53:899-907.
and
and
polyunsaturated
and
and
PUFAs,
cholesterol
and
is
by changes
in the
which
function
fatty
high
of MUFAs
quantity
adherence
acids
keeping
was to
of a diet
(PUFAs)
total
fat,
constant.
to establish
at least
Through
an
in part,
the
ofdiffering
fatty
Printed
in USA.
food
was,
study
and
them
to attend
from
the outside
therefore,
an ideal
with
free-living
ofthe
is very limited.
setting
from
is served
to buy
The ye-
for performing
subjects.
of the academic
the purpose
the college
evening.
All food
and the opportunity
study,
year,
details
all students
ofits
were
as-
performance,
in
I From
the Lipid Research
Laboratory,
Department
of Medicine
B,
Hadassah
University
Hospital,
and the Departments
ofSocial
Medicine
and Nutrition,
School
of Public Health and Community
Medicine,
Hebrew University-Hadassah
Medical School, Jerusalem.
2 Supported
by grant HL-39302
from the National Institutes of Health.
3 Address
reprint requests to NA Kaufmann,
Department
of Nutrition,
Hebrew
University-Hadassah
Medical
School,
POB
1 172, Jerusalem
91010, Israel.
Received
December
13, 1989.
Accepted
for publication
May 30, 1990.
experi-
the prob-
cooperation
acid
situation
requires
to late in the
ofthe
yeshiva
ensuring
of
on the
diet pecontent.
Each student
acted as his own control
in this crossover
study.
Oleic acid has been considered
neutral with regard to its effect
on blood lipid concentrations
( 1 , 2). However,
recent
work sugAm J C/in Nutr
Nutrition
of monounsaturated
or low amounts
structure
(SFAs),
that
At the beginning
sembled
of the
foods
while
Yeshiva,
we set up a metabolic
kitchen
male students
participated
in two l2-wk
which
Jerusalem
while keeping
the
constant.
Dietary
analysis
metabolic
investigation
in its proof.
time
overcomes,
such
high
of the diet
constituents
saturated
shiva
fat is preferable.
of the effect
a long
have
that
dietary
be related
on lipoprotein
structure
and function
over
protocols
are planned
to determine
the effects
of substituting
or eat additional
alter plasma
lipid
research
and dis-
ofdietary
inherent
is the studying
mental
riods
difficulties
populations
dietary
lems
which
a hypothesis
to the
may
monounsaturated
The hypothesis
that diet can predictably
concentrations
continues
to generate
much
is testimony
The
the effects
by food-composition
compare
system
That
between
(3), which
(4).
in detail
about
effect
to investigate
for one
quality
association
disease
Subjects
Introduction
particularly
inverse
heart
J C/in
dation
cussion
be an
hypolipidemic
plans
on blood
with
diet,
as ascertained
formation
may
and
not
PUFA
there
acid
monitored
was a significantly
peroxidation
that
oleic
with
centrifugation
Norman,
gests
Study
diets,
Low-
in both
Yehudit
to a possible
signif-
10% and
16% on the MUFA
and PUFA
Plasma
triglyceride
response
was variable.
density-lipoprotein
Friedlander,
randomly
of monounsaturated
18% protein)
plasma
Yechiel
1991 American
Society
for Clinical
Nutrition
899
icantly by
respectively.
students
study
vs polyunsaturated
carbohydrate,
Dror Haratz,
Stein
BERRY
900
and requirements
by the
investigators
and
participants
the directors
ofthe
were explained
yeshiva.
Thirty
randomization.
Subjects
high
such
blood
with
as diabetes
lipid
were excluded.
of the dietary
endocrine
mellitus,
concentrations,
or any
Twenty-six
subjects
study and completed
22 participants
remained
or
metabolic
hypothyroidism,
distur-
obesity,
other
chronic
or
disease
were included
as participants
the first dietary period, and
in the study
until
the end
ofthe
second
period.
was undertaken
of Health
Protection
consent
and
the
of Human
after
approval
Hadassah
Subjects,
Medical
and
from
the Israel
Center
Board
all subjects
gave
Mm-
items
prepared
Before
each
diet,
the
volunteers
were
stratified
by blood
trays
and
period
(period
1) followed
prepared
subjects
period
MUFA
period
by 4 wk of the
or delivered
were
given
by us). After
diets
in reverse
regular
this
order
yeshiva
wash-out
for
a second
1 2-wk
order
was
reversed
in the other
halfofthe
of Official
Analytical
day.
Thus,
diet
the same
meat
the MUFA
fat was
whereas
added
diet
vegetables
diets.
of olive
However,
oil,
avocado,
was supplemented
of saffioweroil,
12 daily
PUFA
in form
the PUFA
amounts
or the same
and
soy
oil,
and
with
walnuts.
For
of similar composition
but of varied
items
were prepared
and used in rotation.
After the food was
cooked,
portions
were weighed
and put on individual
trays for
each participant.
Subjects
were asked to eat all food on their
menus
to report
was consumed.
all leftovers.
To allow
choice,
several
subjects
could
Usually
all the
the participants
equivalent
items
offered.
kinds
between
various
kinds
of dairy
products
food
a certain
were
choose
cr5 or various
distributed
degree
Thus,
ofbread
of equal
of free
for breakfast
and
crack-
fat and
caloric
content.
Snacks of composition
equal to the daily menu were
prepared
and subjects
could eat them ad libitum
provided
that
the total portion
was eaten and the number
ofsnacks
consumed
was reported.
In this way the participants
could adjust their total
food intake
to their caloric
needs without
changing
the composition
of the diet.
All planning
diet
period,
Association
and
calculations
and
labeling
Very-low-density
students
were
and
done
by a qualified
cooked
under
(VLDL)
and
dietitian
her supervision
of lipoproteins
lipoprotein
low-density
lipo-
protein
(LDL) were isolated,
by ultracentrifugation,
and 1.019-1 .063 kg/L, respectively,
from human
Zonal ultracentrifugation
of lipoproteins
was performed
according
to the techniques
of Patsch et al (7) as modified
by Eisenberg
et al (8). Intermediate-density
lipoprotein
(IDL) and
LDL were separated
from 20 mL plasma by zonal centrifugation.
The amounts
of LDL protein (4-10 mg) were sufficient
for further characterization
whereas
with IDL it was sometimes
nec-
occasions
they
were
given
strict
at their
homes,
and
they
were
providing
most
of the
or walnuts,
which
blood
for plasma
At the end ofeach
studies
specific
instructions
about
what
required
to consume
fats,
as olive
such
to eat
the items
oil,
almonds,
were provided
in take-away
packages.
Fasting
lipid determination
was drawn
periodically.
period, blood was also taken for more detailed
of lipoprotein
structure
and
functions.
aminations
were performed
in 1 2 subjects
lected at random
from all the participants.
every 2 wk throughout
the study.
from
Subjects
equal
d-96
hydrate
acids,
cholesterol,
protein,
and
There
was
300
mg
latter
cx-
essary
se-
on the same
were weighed
carbohydrates,
cholesterol
tubes,
group
were
1 g Na2EDTA/L.
plastic
each
into
These
Diet
urated
taming
in each
into
kept
LDL
under
preparations
and
HDL3
from
nitrogen
from
diet. High-density
HDL2
was sterile
two
lipoprotein
lO-mL
plasma
filtrated,
portioned
at 4 #{176}C.
or even
three
(HDL)
subjects
was separated
samples.
LDL
from
normolipemic
subjects
not participating
in the study was pmpared by ultracentrifugation
as described
above. After dialysis
and concentration,
a sample
of this normal
LDL was radioiodinated
to a specific activity of 3.4-10.2
&iJg
protein
by using
the iodine
labeling
monochloride
Freshly
from
(9) as modified
for lipoprotein
l09/L)
and culture
prepared
30-40
procedures
x
method
(10).
Cell isolation
diet.
to combine
The
and
at d 1.006
plasma
con-
circulating
mononuclear
cells
were
isolated
mL blood collected
under sterile conditions
by the
of Boyum (1 1) and Bilheimer
et al (12). The cells (6
were suspended
in RPMI
1640 (Gibco,
Paisley, Scot-
cholesterol
concentrations
and divided
randomly
into two
groups. After randomization,
subjects were given all food (main
meals and snacks) by the study staff. No other food was allowed.
During the first 4 wk the participants
were given the regular food
ofthe yeshiva including
in-between
meals and snacks. On several
occasions
they were asked to record their 24-h food intake, which
enabled
us to calculate
the individual
energy requirements
of
each participant
and plan his personal
diet accordingly.
At the end ofthis run-in period, the dietary experiment
started.
Two diets, MUFA
or PUFA,
were administered
for a 12-wk
of the
for both
almonds
written
design
the experiment
and
each
MUFA
equivalent
to participate.
Experimental
(not
food
were
for the
their
methods
Chemists
(AOAC)
(5). Duplicate
samples
of the different
diets
were collected
at random
on three occasions
during each period,
the samples
were blended,
and a 5% portion
of each diet was
kept frozen. At the end of each period these subsamples
were
combined
and analyzed.
Fatty acid composition
was determined
by gas-liquid
chromatography
(GLC). The composition
of the
minor
food items was derived
from standard
food tables (6). On
the basis of these data, special food tables for the study were
compiled.
The two experimental
diets contained
the same basic
in the
The study
istry
AL
to standard
stu-
dents volunteered
went a thorough
screening.
Blood
fast at a 3-10-d
bances,
ET
DIETARY
land,
UK)
and
medium
supplemented
5 g protein/L
ie, RPMI
1640-5%
lymphocytes
by
LPDS).
Monocytes
1-h adherence
volunteers
penicillin,
lipoprotein-deficient
Cell viability
was estimated
Human
skin fibroblasts
normal
with
human
and
by trypan-blue
were cultured
used
between
then
Petri
trypsinization,
described
(Biological
100 mL
LDL
3-5
the
5th
and
15th
previously
Industries,
and subcultured
Kibbutz,
Bet
of
passage.
fetal bovine
serum/L
receptor-specific
after
containing
20 mg/L of 251-LDL
in the RPMI
1640-5%
LPDS
LDL
with
by mono-
from a normal-lipidemic
medium.
Degradation
subject
products
tion
nonspecific
of control
Competitive
and
displacement
by experimental
values.
of251-labeled
control
The affinity
LDL receptor
lated cultured
of MUFAand PUFA-derived
LDL towards
the
was determined
by a competition
assay in upregufibroblasts
grown for 48 h in 5% LPDS. Ten mi-
crograms
milliliter
ofcontrol
25I-LDL
protein
was
internalization,
25ILDL
OC
were
by
and
following
proteolytic
proteolytic
determined
degradation
at the end
established
degradation
were
acid-reactive
Conditioning
was carried
simultaneously.
in 1 mL
ofLee
(16).
medium,
data
at 37
of the
out
by incubation
of 50 mg
(Gibco)
without
serum
cultures
of bovine aortic
control dishes were pro-
Thereafter,
(TBARS)
substances
cedure
The
substances
of LDL
LDL protein/L
in Ham F-10 medium
for 24 h in the presence
of confluent
SMC and/or
2.5 mol Cu2/L.
No-cell
cessed
(14).
control
to construct
competition
without
added unlabeled
used
of the
of 3-h incubation
procedures
were
Briefly,
0.5
mL
thiobarbituric
acid-reactive
determined
(1 5) by the modified
pro-
to 1 mL
of 350
plasma
or 50 ig LDL
g tricarboxylic
plasma
or
were
and
procedure
Dieren,
The
Netherlands).
trations
were
determined
lipoproteins
was obtained
tion
collected
triglycerides
in a batch
after
a 12-h
were
determined
analyzer
(Vitalab,
HDL-cholesterol
after
precipitation
with phosphotungstic
by subtraction,
using
overnight
fast.
by an en-
Vital
Scientific,
(HDL-C)
concen-
ofapo
B-containing
acid. LDL-C
(including
IDL)
the Friedewald
et al equa-
(18).
To determine
the composition
procedures
cholesterol,
and
cholesteryl
esters
were
of VLDL,
used
to measure
phospholipids.
LDL,
Unesterified
were
Erythrocytes
separated
from
HDL3,
triglycerides,
cholesterol
were determined
by specific
kit (Boehringer-Mannheim,
with a commercial
and
protein,
and
enzymatic
Mannheim,
methods
FRG).
EDTA-containing
blood
samples
Total
washed
and hemolyzed
by the method
of Dodge
et al (19,
lipids
were
extracted
with
chloroform-methanol
to remove
proteins.
Fatty
acid methyl
esters were
pared
by
transesterification
with
methanolic
nium
hydroxide
(Meth
Prep II, Applied
(2 1). During
the separation
procedures,
(BHT;
5 g/l00
methyl
esters
L) was
Tracor
565 GLC
were
used
column
trimethylammo-
Science,
Deerfield,
2,6-ditert-butyl-9-cresol
as an antioxidant.
separated
and
(1.85
20).
and
pre-
The
quantified
by
m X 4 mm
IL)
fatty
GLC
acid
using
on
acid
dards.
Statistical analysis
included
in the incubation
medium
without
or with different
concentrations (5, 10, 20, and 40 mg/L) of unlabeled
LDL preparations,
derived
from subjects
consuming
MUFA or PUFA diets. Binding,
MDA/L
LDL
LDL
per
(imol
Statistical
analysis
included
t tests
for the comparison
of
changes in lipid concentrations
in response
to dietary treatment
and time period for this basic two-period,
crossover
design according
to Fleiss (22). For subjects commencing
with the MUFA
diet
in period
measures
2. This value
DMP
1 and
changing
to the
PUFA
diet
in the
the change
in response
from period
includes
a measure
of the difference
period
2,
1 to period
between
the
two treatments
regardless
of the period, plus the time trend regardless
ofthe treatments.
Similarly,
DPM
represents
the change
in response
for subjects
in group 2 who followed
the same procedure
in the reverse
order.
The hypothesis
of equal efficacy
of
the two diets was tested
by means
ofa t test comparing
the two
averages,
DMP
standard
error
The
possible
and
DPM
was
used
interaction
(22).
The
to test
sum
the
between
of
plus
existence
the dietary
t test
DM
and
of a period
treatments
(22).
Values
its
effect.
and
pe-
are expressed
protein
(TCA)/L
Results
The baseline
characteristics
ofthe
students
entering
the study
are shown
in Table
1. During each 12-wk period some students
were unavailable
for blood
tests.
The experimental
crossover
design
required
analysis
sets were
available
of those
participants
on
period.
in each
group
whom
there
Such complete
and
therefore
culture
medium
as described
( 1 3). Nonspedetermined
in the presence of25-fold
excess
No-cell
control
dishes were processed
sicells were washed
in protein-free
medium
in 0.5 mol NaOH/L
for protein
determidegradation
was calculated
after substrac-
cific degradation
samples
cholesterol
zymatic
2 mL medium
in
was
of unlabeled
LDL.
multaneously.
The
and were dissolved
nation.
LDL-specific
were determined
Plasma
containing
monocytes
of 25I-labeled
8 h incubation
content
protein).
methods
standard
in fresh
degradation
was determined
medium
Israel)
(13).
degradation
Receptor-specific
cytes
in Dulbecco-Vogt
LDL
All blood
seeded
Haemeq,
equivalent
MDA/g
Analytical
in 35-mm dishes
with 2 mL medium
containing
100 mL fetal calfserum/L.
Cells
were fed three times weekly with the same medium.
Bovine
aortic smooth
muscle
cells (SMC) were prepared
as
After
901
PUFA
from
dishes.
exclusion.
from skin biopsies
X 10 cells were
zmol
(LPDS;
separated
(35-mm)
VS
malondialdehyde
streptomycin,
serum
were
to plastic
MUFA
BERRY
902
TABLE
Baseline
characteristics
of participants
ET AL
TABLE 2
Composition
in study
dietary
Group
1
(n=l2)
Body mass index (kg/rn2)
Number
of smokers
Number of subjects born in Israel
Plasma cholesterol
(mmol/L)
Plasma triglycerides
(mmol/L)
Plasma HDL cholesterol
(mmol/L)
23.3
Group 2
(n=l2)
2.4
21.6
3.8 1
1.02
1.09
0.77
0.22
0.25
3.8 1
0.92
1. 1 1
and house
vs from
Diet
SFAs
Period
0.78
0.27
0.17
diets during
the two
analysis5
MUFAs
PUFAs
Fat
Protein
CHO
35.1
31.4
15.2
19.2
49.7
49.4
% of kca/
SD.
planned
2.2
of the experimental
periods:
MUFA
Planned
From analysis
PUFA
Planned
From
Period 2
analysis
8.5
8.1
17.7
14.9
6.3
6.7
8.1
7.1
17.6
35.2
14.7
50.1
7.2
5.9
14.7
29.6
19.4
51.0
8.9
16.5
6.3
33.8
16.7
49.5
7.8
16.9
8.3
34.9
18.4
46.7
MUFA
carbohydrate
being
nearly
identical.
During
period
1 the amounts
ofboth
MUFAs and PUFAs as percent total calories were slightly
less than planned;
during period 2 the discrepancy
disappeared.
The house diet of the Yeshiva
is shown for comparison.
Adherence
to the diet was monitored
by comparing
changes
in
erythrocyte
membrane
fatty
acid
composition,
in particular
the
ratio between
18: 1 (oleic acid) and 18:2 (linoleic
acid). In both
periods on the MUFA
diet, the ratio increased
(more in period
2), and
3).
on the PUFA
During
period
diet
the ratio
1 the
average
<
determinations
the
0.5
decreased
weight
kg; during
change
period
of the
was
plasma
beginning
ofthe
experiment
(3-10
apart; baseline)
and two determinations
at the end (weeks
10
and 12). After period
1 for both diets there was a significant
decrease
(1 1%) in total cholesterol;
after period 2 this decrease
was more prominent
on the PUFA diet (20%) and less so on
the MUFA diet (8.5%), but both were significant
when compared
with the baseline
values. The data of the crossover
study were
analyzed
to distinguish
between
effects
on plasma
lipids
due
greater
and
a decrease
of 31%
was
observed,
17.1
33.8
16.8
49.5
6.4
17.2
32.3
16.3
51.4
House,
from analysis
10.5
1 1.5
15.8
40.6
15.7
43.7
S SFAs, saturated
fatty acids; MUFAs,
monounsaturated
PUFAs, polyunsaturated
fatty acids; CHO, carbohydrate.
fatty acids;
ducing
a significant
seasonality
effect (P < 0.01). The overall
effect of PUFAs
on LDL-C
concentrations
was significantly
greater
than that of MUFAs
(P < 0.05).
There were no significant
differences
in lipoprotein
compoanalysis
during
either
period
ofthe
diet
study
or between
was,
therefore,
no evidence
for any
change
in
lipoprotein
structure
or function
in response
to the different
diets.
The effect of dietary
MUFAs
and PUFAs
on the tendency
toward
oxidative
stress was assessed
by TBARS
production
in
to
6.4
7.0
cholesterol,
triglycerides,
HDLC, and LDL-C at the beginning
and end ofthe two study periods
are shown in Table 4. Statistical
analysis
of the overall effects
of diet and season
on plasma
lipids is shown in Table 5. The
plasma lipid values reported
represent
the mean oftwo separate
before
8.4
analysis
From
students
2 the change
From analysis
PUFA
Planned
sition
(Table
significantly
Planned
pro-
TABLE
Ratio
Baseline
Paired
ti
from
End of study
period
subjects
on
P5
MUFA
PUFA
Period 2
MUFA
PUFA
S
acid in erythrocytes
0.023t
0.027
1.069
0.801
0.955 0.029
0.981 0.031
1.066
0.885
1.040
0.980
test.
SEM.
0.027
0.026
0.072
<0.001
0.035
0.019
0.033
0.023
DIETARY
TABLE
MUFA
VS
903
PUFA
4
lipid concentrations
Plasma
by study
period
and diet5
Period
Lipid
Period
End of period
Baselinet
End of period
Baseline
mmol/L
Total
2
P
mmo//L
cholesterol
MUFA
PUFA
3.83
3.89
MUFA
PUFA
HDL-C
MUFA
2.30
2.30
1.06
PUFA
Triglycerides
1.10
1.03
0.92
0.31
0.32
3.42 0.24
3.44 0.27
0.008
0.003
3.96 0.32
4.04 0.32
3.62
3.22
0.26
0.25
0.037
<0.00 1
0.28
0.30
1.98
2.05
0.22
0.26
0.015
0.025
2.51
2.58
0.32
0.30
2.18 0.23
1.78 0.22
0.042
<0.00 1
0.07
0.07
1.06
1.08
0.05
0.06
0.898
0.713
1.09
1.09
0.07
0.09
1.08
1.02
0.08
0.1 1
0.85
0.69
0.09
0.08
0.014
0.007
0.79
0.82
0.07
0.10
0.79
0.90
1 .68
LDL-CII
MUFA
PUFA
native
and
conditioned
8). There
after
was
incubation
subjects
LDL
a significant
with
on the
under
SMC
PUFA
diet
various
conditions
or copper
compared
(Table
in native
ions
with
LDL
alone
Chol
(mg/dL)
from
subjects
diet. Because
a priori the tendency
toward
oxidative
stress was thought
to be greater
for PUFA than for MUFA,
a
one-tail
significance
test was used. TBARS
production
was
greater
also in whole plasma samples from subjects on the PUFA
diet than from those on the MUFA
diet (0. 174 0.008
vs 0.164
0.007 imol/L,
respectively).
nutritional
SFA,
MUFA,
and PUFA
intakes
of fatty
testing
ofthe
acids
to the regression
study
is part
of an
ongoing
experiment
to
Chol
any
tion
but
dietary
taneously
applicable
2.74
SFA
from
cholesterol
ofcholesterol
diet,
nipulation
to
Previously,
suggested
a beneficial
12 wk,
and
MUFAs
on plasma
used
were
effect
entirely
of a Mediterranean-
on the pioneer
y ago
In this
MUFAs.
ofcalories
investigated,
initiated
(1)
dietary
in the percent
calories.
Further
ofseveral
variables
revised
equation:
- I .3 1 X PUFA
Hegsted
(2)
et al (2) studied
from
in fat content
wk (usually
three
for the
men
differing
of which
sets of comparisons
equation
by Ancel
were
change
were
analyzed
in plasma
by Keys (29).
It is ofinterest
to compare
predicted
TABLE
et al 30
were
studied
as was
also
done
simul-
from
fatty
the results
ofour
2. Both
diets
equation
but varied
acid
experiments
contained
with
similar
composition
as percent
oftotal
On
calories
Statistical
analysis
of overall
effects
Diet: MUFA
vs PUFA
thesis
Keys
added,
These
equations
were found
to be
population
in the 1960s as shown
recently
in their
12-27
was
items.
to be neutral
experiment,
food
concentrations
work
classic
natural
considered
cholesterol
was based
following
the change
contribution
by Keys et al (28).
to the Israeli
male
the house
Forty-one
X LPUFA
contribution
amounts
data
diet (rich in olive oil) on heart disease but could not prove
a causal relationship
(3). Several studies have recently
investigated the effects ofMUFAs
on plasma lipid concentrations.
They
employed
either liquid formula
(24) or solid food (25-27)
for
periods
of 6 wk. We have extended
the time of dietary
ma-
were
0.902
0.193
-1.35
oftotal
ofthe
(mg/dL)
miologic
type
of9-44%
represented
analysis
those
diets
0.1 1
0.09
X LSFA
X LMUFA
as percent
significance
on each
0.808
0.203
the response
ofplasma
cholesterol
to diet and arrived at a similar
equation.
They also did not include dietary MUFAs
in the equa-
Discussion
(1).
+ 2.76
+ 0.05
without
influence
0.05
0.07
on the
MUFA
This
(18).
in TBARS
alone
when
10 and 12.
equation
increase
Lipid
of diet
and
season
Season:
period 1 vs
period 2
1
Interaction:
diet vs period
t
over a range
4 wk).
rich
and
Sixteen
in MUFAs.
yielded
cholesterol:
the
Total
cholesterol
2.9 1
<0.02
1.68
NS
1.58
NS
LDL-C
2.26
<0.05
2.98
<0.01
2.13
<0.025
HDL-C
1.08
0.22
NS
NS
1.19
3.69
NS
<0.01
0.32
1 . 13
NS
NS
Triglycerides
51SEM.
t Mean of two blood samples, 3-10 d apart.
t Mean ofblood samples at the end ofweeks
904
BERRY
TABLE
ET AL
Composition
oflipoproteins
at the
end
of the
dietary
periods5
Protein
FC
TGs
CE
PLs
VLDL
MUFA,
MUFA,
PUFA,
PUFA,
LDL
MUFA,
MUFA,
PUFA,
PUFA,
HDL3
MUFA,
MUFA,
PUFA,
PUFA,
6)
7)
5)
4)
12.0
13.2
12.0
13.2
0.6
1.7
0.7
1.4
5 1.2
40.3
54.8
48.4
21.8 2.1
24.6 1.5
22.7 2.8
4.6
4.5
5.1
6)
7)
5)
4)
24.0
4.0
4)
4)
4)
4)
51.0 1.9
54.1 2.7
55.7 1.5
54.8 1.1
=
=
1 (n
(n
1 (n
2 (n
1 (n
2 (n
=
=
1 (n
2 (n
=
=
1 (n
2 (n
L, observed
in periods
MUFA-rich
diet
dicted
the
in MUFA,
change
3.2
3.4
3.0
3.0
1.0
1.1
0.8
1.0
3.7 2.0
3.0 0.3
4.4 1.8
3.1 0.7
1.6
CE, cholesteryl
0.2
0.4
0.3
0.2
0.9
1.1
0.9
1.7
26.9 3.0
26.6 3.7
24.5 1.9
27.7 4.5
8.9 0.5
8.3 0.9
9.4 1.0
8.3 0.5
37.4 2.3
36.2 3.7
38.5 2.2
38.1 1.6
26.4 0.8
26.3 3.0
24.4 1.4
25.6 2.1
1.7 0.2
1.4 0.1
1.5 0.2
1.5 0.2
14.8 1.2
15.4 1.6
14.5 2.6
14.5 1.1
28.8 1.4
26.1 1.1
23.9 2.0
26.2 1.7
SFAs:MUFAs:PUFAs.
The PUFA-rich
by average
changes
of -3.4%
in SFA,
+0.2% in PUFA. The predicted
change
using Keys equation
2 would be -0.25
compared
with -0.45 and -0.83 mmol/
I and
2, respectively
relevant
and
changes
-8.3%
in cholesterol
of0.10
mmol/L
mmol/L,
which
2.4
3.4
2.2
5.3
6.8
7.5
5.8
7.8
Values
are percent
contribution
ofindividual
components
mass.
+4.4%
(Table
are
in PUFA.
would
4). For
in -2.6%
Accordingly,
be an increase
the
question
that
workers
is why
equations
the
pre-
-0.34
The
IDL.+ LDL
arises
were
devised
of these
is such
results
and
a discrepancy
results
between
the
was
from
tissue
risen
Israel
(34).
The
10%
<
in the
comparable
1950s,
over
body
in
the
years
a manner
on
subjects
living
in the
l950s
changes
affect
(32).
By
1980
it had
in 1980(US),
may
that
late
States
figures
46%
studies
pose
in light
there
observed
and predicted
changes
in cholesterol
concentrations
after changing
dietary
MUFAs.
There is no clear answer,
but a
possible
explanation
may be found in the changes
in dietary
habits that have occurred
over the past 30 y since the original
in SFA,
in cholesterol
(3.7 mg/dL)
compared
with -0.41
and
was found in periods
I and 2, respectively.
obvious
by other
and
in dietary
cholesterol
41%
in I 986 (Israel).
defined.
in the
In other
TABLE
7
Competition
of 251-labeled LDL degradation
in cultured
fibroblasts
by LDL isolated from subjects on the MUFA
diets at the end ofthe dietary period5
c%J
4-
Concentration
Diet
5 mg/L
of unlabeled
10 mg/L
human
and PUFA
LDL protein
20 mg/L
40 mg/L
100
Zonal
FIG
1. Zonal
in plasma
(-
- -)
300
Rotor Effluent
ultracentrifugation
HDL (bottom)
of the MUFA
200
obtained
and PUFA
(-)
400
500
(ml)
Control
(n
MUFA
PUFA
(n
(n
=
=
=
5)
12)
13)
67
66
66
I l.1
10.5
8.6
S1
51
51
10.8
9.0
8.4
42
39
37
9.1
8.6
8.5
26
22
25
8.1
5.7
8.1
S
SD. Results are expressed as percent ofdegradation
in the presence oflabeled
LDL only. Eleven subjects were studied twice at the end
of each diet period. One subject was studied only on the MFA and two
were studied only on the PFA diet.
to total
1 (n
2 (n
DIETARY
MUFA
905
VS PUFA
-I
I
400510
Unlabeled
LDL
(pg/mI)
FIG 2. Competition
of 25I-Iabeled
LDL metabolism
in cultured
control subjects (healthy
normal humans)
and with LDL obtained
The 25I-labeled
LDL was prepared
from control subjects.
words,
it may be necessary
for the nutritional
ofthe
major
PUFAs
study
ofthis
ofthe
and redefine
population
shows
or MUFAs
cholesterol
MUFA
l990s.
One
investigations
with
on the biological
similar
amounts
of either
a significant
decrease
of plasma
16% and 10% on the PUFA and
during
the two periods)
without
(-
of HDL-C.
of Mensink
study
diets
respectively,
levels
to those
that
produced
concentrations
diets,
changing
ongoing
effects
ofMUFAs
is to collect data sufficient
for addressing
this problem.
The principal
differences
between
this nutritional
study and
other investigations
are that it was carried
out in a free-living
population
eating normal
food (not formula)
over two 12-wk
periods.
The population
studied
was homogenous,
was nonsmoking,
and consumed
negligible
amounts
of alcohol.
This
crossover
aims
to repeat
state
These
and Katan
in a free-living
results
are
in general
population.
The
fact that
interesting
questions
regarding
MUFAs
on plasma cholesterol
arise,
The amount
of MUFAs
in the diet
light
ofrecent
diets
may
findings
cause
On the other
tains
an average
a study
27%
dose
animals
that
of cholesterol
from
MUFAs,
Crete
failed
to show
in the concentrations
understood.
The baseline
values
4) were considerably
lower than
liver
March.
Con-
and
to the baseline
values for
and thereafter
increased
by
July. The effects
of season
on plasma
choconcentrations
in this previous
study were
the differences
found in the present
experi-
until
time
that
dietary
manipulation
as linoleic
acid)
(38),
not clear
it was
also
by changes
in erythrocyte
it has been recognized
for a
will alter
whether
be detected.
membrane
with
Our
essential
erythrocyte
diet
composition
fatty
membrane
acids
compo-
enrichment
with
(Table
3) suggest
findings
may
be altered
MUFAs
that
by MUFAs
(mainly
oleic acid). In both groups the high-MUFA
diet led to
an increase
in 18: 1-1 8:2 ratio whereas
the high-PUFA
diet produced converse
changes.
Oleic acid is the most abundant
fatty
acid
of stored
from
dietary
estingly,
adipose-tissue
sources
stearic
acid
appear
to elevate
TABLE
Thiobarbituric
is unique
plasma
dietary
among
cholesterol
acid-reactive
triglyceride
(30).
or by desaturation
substances
on MUFA
It is derived
of stearic
SFAs
in that
concentrations
in native
both
acid.
(28,
39).
and conditioned
period5
Smooth-
(35).
favorable
of period 2 (Table
in period
1 This may be due
to confounding
dietary factors related to the 1-mo wash-out
period which included
the Passover
holidays.
We (37) investigated
.
November
between
(such
erythrocyte
at the start
mmol
found
a more
oftriglycerides
mmol/L
we found
serum lipoprotein
pattern
in young boys when compared
with
boys from a more westernized
society.
The response
of plasma triglycerides
was variable with a seasonality effect between the two experimental
periods. The effects
ofseasonality
in Israel
of0.07
concentrations
high-MUFA
in the
(36), where
lipid
men a decrease
in
period
0.24
was
on plasma
In middle-aged
April (corresponding
2 in our study) were lowest
centrations
would
of
study.
triglycerides/L
sition
response
ofseasonality
in a previous
similar
in experimental
accumulation
hand,
the
the effect
out their
a somewhat
larger decrease
of total and LDL-C on the PUFA
than on the MUFA diet whereas
Mensink
and Katan (27) did
not may be due to differences
in the design of the diets and the
duration
of the experiment.
The percent decrease in total cholesterol
we observed compares
with the 13% observed
by Mattson
and Grundy
(4), who used
diets of between
27% and 33% MUFAs,
and with the - 12.8%
found by Mensink
and Katan (27), who used a diet of 15.1%
MUFAs.
The concentration
of MUFAs
in our diets was 17%
and although
the control
diets in these experiments
were not
similar,
Diet
Native
Smoothmuscle cells
muscle
+
nmo/
MDA/mg
LDL
cells
Cu2
Cu2
protein
MUFA
(n
5)
1.05
0.03
15.70
4.00
47.00
2.00
7.20
2.00
6)
1.16
0.04t
24.40
2.00
49.80
2.60
18.60
l.40t
I test):
tP
PUFA
(n
<0.0025.
equivalents
(one-tail
<
0.05,
jP
tion
20
906
BERRY
These
indices
of diet
odological
aim
validity
ofresults
adherence
suggest
that
of the study
was achieved
for a free-living
population.
the major
meth-
and strengthens
the
Whether
the results
are applicable
to other populations
and also to females in light
ofthe findings of differing
responses
between
the sexes (27, 40)
requires
further investigation.
Another
point concerns
the issue
ofwhether
the lipid concentrations
would be maintained
or drift
back to the initial values over a longer time period.
There were no differences
in lipoprotein
structure
as shown
by compositional
analysis
or profiles on zonal ultracentrifugation. LDL-receptor
activity
of monocytes
also remained
unchanged
as was the ability ofdiet
LDL to displace
control
LDL
at the fibroblast
receptor.
However,
when the tendency
toward
oxidative
stress was measured,
LDL from the PUFA diet promoted significantly
more TBARS
formation
than did LDL isolated after the MUFA-rich
diet. Recent work showed that unesterified fatty acids may inhibit iron-dependent
lipid peroxidation
(41).
The
effect
of oleic
acid
was
significantly
greater
than
that
because
free
radicals
to be important
atherosclerosis
to the concept
in optimizing
modification
of LDL
factors
In conclusion,
our observations
that
MUFAs
serve
diet
may
lipid
may
considered
lend
as a substitute
concentrations
reduce
are
in the development
(42).
plasma
MUFA-rich
dative
and
pathoetiological
and,
the susceptibility
for SFAs
in addition,
of LDL
of
support
an
to oxi-
stress.
ofthe
for their
enthusiastic
help, the administrative
staffofthe
Yeshiva, and especially
the personnel
involved
in preparation
ofthe special diets. We thank and
appreciate
the student
volunteers
for their devoted
cooperation.
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oflinoleic
acid. It may also be that oleic acid is a poorer substrate
than linoleic acid for peroxidation.
However,
the possibility
that
the MUFA
diet contained
more antioxidants,
such as vitamin
E, cannot
be excluded.
Thus, there may be a theoretical
advantage to the MUFA diet. The possible
adverse effects ofthe PUFA
diet on the tendency
to peroxide
formation
is of major interest
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