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ORIGINAL ARTICLE
Effects of lupin-enriched foods on body composition
and cardiovascular disease risk factors: a 12-month
randomized controlled weight loss trial
R Belski
1,2
, TA Mori
1,2
, IB Puddey
1,2
, S Sipsas
3
, RJ Woodman
4
, TR Ackland
5
, LJ Beilin
1,2
, ER Dove
1,2
,
NB Carlyon
1,2
, V Jayaseena
2,6
and JM Hodgson
1,2
1
School of Medicine and Pharmacology, University of Western Australia, Perth, Western Australia, Australia;
2
The WAIMR
Centre for Food and Genomic Medicine, Perth, Western Australia, Australia;
3
Department of Agriculture and Food, Western
Australia, Australia;
4
School of Medicine, Flinders University, Adelaide, South Australia, Australia;
5
School of Sport Science
Exercise and Health, University of Western Australia, Perth, Western Australia, Australia and
6
School of Public Health,
Curtin Health Innovation Research Centre, Curtin University, Perth, Western Australia, Australia
Background: Regular consumption of diets with increased protein or fibre intakes may benefit body weight and composition
and cardiovascular disease risk factors. Lupin flour is a novel food ingredient high in protein and fibre.
Objective: To investigate the effects of a lupin-enriched diet, during and following energy restriction, on body weight and
composition and cardiovascular disease risk factors in overweight individuals.
Design: Participants (n¼131) were recruited to a 12-month parallel-design trial. They were randomly assigned to consume
lupin-enriched foods or matching high-carbohydrate control foods. All participants underwent 3 months of weight loss,
1 month of weight stabilization and 8 months of weight maintenance. Body weight and composition and cardiovascular disease
risk factors were assessed at baseline, 4 and 12 months.
Results: Lupin, relative to control, did not significantly influence (mean difference (95% CI)) weight loss at 4 months (0.1kg
(1.2, 1.4)) and 12 months (0.6 kg (2.0, 0.8)), maintenance of weight loss from 4 to 12 months (0.7 kg (1.83, 0.48)) or
measures of body fat and fat-free mass. Relative to control, 24-h ambulatory systolic (1.3 mm Hg (2.4, 0.3), P¼0.016) and
diastolic (1.0 mm Hg (1.9, 0.2), P¼0.021) blood pressures were lower at 12 months but not at 4 months; fasting
insulin concentrations and homeostasis model assessment (HOMA) scores were significantly lower at 4 months (1.2 mU l
–1
(1.3, 1.1), P¼0.004 and 0.6 units (1.0, 0.19), P¼0.004) and 12 months (1.3 mU l
–1
(1.4, 1.1), Po0.001 and
0.7 units (1.1, 0.24), P¼0.002).
Conclusions: A diet higher in protein and fibre derived from lupin-enriched foods does not enhance weight loss or improve the
maintenance of weight loss. However, such a diet may provide cardiovascular health benefits in terms of insulin sensitivity and
blood pressure.
International Journal of Obesity (2011) 35, 810–819; doi:10.1038/ijo.2010.213; published online 12 October 2010
Keywords: lupin; weight loss; blood pressure; insulin; lipids
Introduction
Data suggest that a higher protein or fibre diet can enhance
satiety and reduce energy intake acutely, and contribute to
body weight loss in the longer term.
1–4
Increasing both
protein and fibre in the diet can be difficult because popular
low-carbohydrate, high-protein diets tend to have quite low
fibre intakes.
5
Thus, there are limited data on the effects of
regular consumption of diets higher in both protein and
fibre at the expense of starch. Protein and fibre can
independently increase satiety,
1–5
and these effects are likely
to result via different mechanisms. Fibre appears to have
a role in satiety and satiation through energy dilution,
increased mastication, gastric distension, delayed gastric
emptying and changes in orexigenic hormones.
3
The
mechanisms for the effects of protein are less clear but
protein may influence satiety via orexigenic hormones.
6
Therefore, it could be anticipated that their combined
effects on body weight and composition would be additive.
Received 15 June 2010; revised 12 August 2010; accepted 23 August 2010;
published online 12 October 2010
Correspondence: R Belski, School of Medicine and Pharmacology, Royal Perth
Hospital Unit, GPO Box X2213, Perth, Western Australia 6847, Australia.
E-mail: regina.belski@uwa.edu.au
International Journal of Obesity (2011) 35,810– 819
&
2011 Macmillan Publishers Limited All rights reserved 0307-0565/11
www.nature.com/ijo
A practical approach to increasing both protein and fibre
content of processed foods is to incorporate high protein
and fibre ingredients into commonly consumed high-
carbohydrate foods. There is evidence that this approach
could also improve cardiovascular disease risk factors such
as blood pressure
7
and blood lipids.
8,9
Lupin flour is a novel food ingredient derived from the
endosperm of lupin. It contains 40–45% protein, 25–30%
fibre and negligible sugar and starch.
10
It is commonly used
as a minor food ingredient in baked foods, but can be used to
partially replace wheat flour in foods such as bread and
pasta, resulting in significant increases in protein and fibre
levels.
11,12
These foods remain palatable and are acceptable
to consumers.
12
We have previously shown that high protein and fibre
lupin flour-enriched bread, with 40% wheat flour replace-
ment, significantly reduced appetite and energy intake
acutely.
8
We also showed that the lupin-enriched meal
acutely suppressed plasma ghrelin,
8
an orexigenic hormone
thought to stimulate appetite.
13
If consumption of lupin
flour-enriched foods has similar effects on energy intake in
the longer term, such effects could translate into weight loss.
Diets higher in plant protein and fibre may also benefit
blood pressure,
14–17
serum lipids
18,19
and glucose and insulin
metabolism.
20,21
In a previous 16-week trial, we showed that
consumption of lupin flour-enriched bread reduced 24-h
systolic blood pressure by 3.0 mm Hg.
22
Lupin-enriched diets
have also been shown to reduce blood cholesterol concen-
trations in animals
23–25
and humans.
26
Furthermore, acute
reductions in postprandial glucose and insulin have been
demonstrated with lupin flour-enriched bread consump-
tion,
12,27
but longer-term effects are not clear.
11
The effects
of a lupin-enriched diet on body weight, body composition
and cardiovascular disease risk factors in the setting of
weight loss and longer-term maintenance of weight loss
have not been investigated.
Therefore, the objective of this study was to investigate the
effects of a lupin-enriched diet, during and following energy
restriction, on body weight, body composition and cardio-
vascular disease risk factors in overweight men and women.
Subjects and methods
Participants
Overweight and obese (body mass index 27–35kg m
–2
),
otherwise healthy volunteers aged 20–71 years were
recruited from the Western Australian (WA) population
through newspaper advertisements from August 2007
to May 2008, into a single-site study. Exclusion criteria
included history of cardiovascular or peripheral vascular
disease, diabetes, history of asthma, renal disease, liver
disease or gout, a psychiatric illness, history of major gastro-
intestinal problems, other major illnesses such as cancer,
hypertension (systolic blood pressure 4150 mm Hg or
diastolic blood pressure 495 mm Hg), use of antihyperten-
sive agents, total cholesterol 46.2 mmol l
–1
or triglycerides
42.0 mmol l
–1
, use of lipid-lowering medications, women
who were pregnant or intended to become pregnant, history
of food allergies, current/recent weight loss/gain (change of
46% body weight over last 6 months) and alcohol intake
4140 g alcohol per week for women and 4280 g alcohol per
week for men. In addition, individuals with no history of
diabetes, but with fasting plasma glucose concentrations
X6.0 mmol l
–1
were excluded. Of the B800 telephone
respondents screened by questionnaire, 231 were eligible
and willing to attend the Research Unit for a screening visit.
Of those 231 participants, 134 met the inclusion criteria and
131 were randomized and commenced the study (68 men
and 63 women). All procedures followed were in accordance
with institutional guidelines. The study was approved by the
University of Western Australia Human Research Ethics
Committee, and all participants provided written informed
consent. The study was registered with the Australian
New Zealand Clinical Trials Registry ACTRN12607000434493.
Study design
A randomized, controlled, double-blind parallel design trial
was performed. Eligible participants were randomized into
either a control group (consuming control foods) or a lupin
group (consuming foods enriched with lupin flour). Eligible
individuals were matched for body mass index, age
and gender and randomly assigned (1:1) using computer-
generated random numbers to either the control or lupin
group. The randomization was performed by an independent
person and group allocation was sealed in opaque envelopes.
Participants and researchers responsible for dietetic inter-
vention and assessment of outcomes measures were blinded
as to group assignment. The researcher responsible for food
distribution was not blinded to group assignment.
After randomization, participants commenced on a
3-month weight loss program (B35% energy restriction)
designed by a dietitian to achieve an average weight loss
in all participants of between 7 and 8% of body weight.
This incorporated the consumption of the assigned foods,
monthly dietetic visits and fortnightly dietary phone
consultations. This was followed by a 1-month weight
stabilization period where participants ceased weight loss
and maintained body weight within 1.5 kg. During the
following 8-month weight maintenance stage, participants
followed an ad libitum diet incorporating the assigned foods.
The aim during this period was to maintain the weight loss
achieved. The primary outcome variable was weight, with
the secondary outcomes being body composition, analysed
by dual-energy X-ray absorptiometry, including body fat
mass and fat-free mass, and cardiovascular disease risk factors
including blood pressure, fasting blood lipids, glucose and
insulin. All assessments of outcome variables were under-
taken at baseline, 4 months and 12 months. The study was
run with two cohorts: the first cohort completed the study
Effects of lupin on body weight and CVD risk
R Belski et al
811
International Journal of Obesity
from October 2007 to November 2008, and the second
cohort completed the study from April 2008 to May 2009.
Foods
Lupin or control foods were provided to participants and
were consumed in place of other cereal-based food products
normally used in the diet. The three foods provided were
bread, biscuits and pasta. The bread and biscuits were baked
at Bodhi’s Bakery (Fremantle, Western Australia, Australia).
The bread was sliced and supplied fresh or frozen to the
participants every month, commencing from baseline.
Participants were required to freeze the bread. The biscuits
were provided fresh every month starting at the 4-month
appointment. The pasta was produced at Belmar Foods
(Balcatta, Western Australia, Australia) and was provided
fresh every month starting at the 6-month appointment. The
food introduction was staggered to keep the initial weight
loss period of the study simple, and to aid retention in the
later phases of the study by increasing variety.
The lupin flour in the lupin products was substituted for
wheat flour, primarily wholemeal, in the control products.
The incorporation rate of lupin flour into lupin products
was 25–40% by weight. The two sets of food products were
matched as closely as possible in colour, taste and texture,
and sensory acceptability and energy, fat and sodium
content in order to assist in blinding of participants to their
treatment group allocation (Table 1).
Dietary assessment
Dietary intake was assessed using a 3-day food diary designed
by a qualified dietitian, based on a previously validated 3-day
estimated food record.
28
Participants were given both verbal
and written instructions on its completion using house-
hold measures to quantify intake. The food diaries were
completed on two weekdays and one day of the weekend.
A weekly log of alcohol intake was included in the food
diary, where participants were asked to record their alcohol
consumption over 7 days. Food intake data were analysed
using FoodWorks Professional 2007 Software (Xyris, Brisbane,
Australia) based on the Australian Food Composition
Database to determine average daily energy, protein, total
fat, cholesterol, carbohydrate, dietary fibre, mineral and
alcohol intake.
Physical activity assessment
Participants were instructed to maintain their current
physical activity levels for the first 4 months of the study
(during the weight loss period and stabilization). During
the 8-month weight maintenance stage, participants were
free to alter their usual physical activity if they wished.
Physical activity was assessed using the Stanford 7-day
Recall Interview
29
and the International Physical Activity
Questionnaire, which has previously been shown to have
an acceptable test-retest reliability and criterion validity.
30
Body weight and composition
Body weight was determined using Wedderburn digital scales
(20–200 kg) (Wedderburn, Perth, Western Australia, Austra-
lia). Participants were weighed with minimal clothing,
without shoes, and weight was recorded to the nearest
0.1 kg. Body composition was measured with dual-energy
X-ray absorptiometry (GE Lunar Prodigy, GE Lunar
Corporation, Madison, WI, USA) by a trained researcher.
11
The exclusion criteria for the dual-energy X-ray absorptio-
metry scan included a chance of pregnancy, a recent barium
test or nuclear medicine scan. At the start of each session,
quality assurance and quality control tests were performed.
Participants removed jewellery/metal objects, and wore
minimal clothing or a hospital gown. Participants were
supine on an X-ray bed, centred within the scan line with
their head 3 cm from the top border. The dual-energy X-ray
absorptiometry scanner reliability is high with coefficients of
variation of 0.6% for fat tissue and 4.5% for lean tissue.
31
Blood pressure
The 24-h ambulatory blood pressure measurements
were performed using Spacelab monitors (Model 90217;
SpaceLabs Medical Inc., Issaquah, WA, USA). The monitor
was programmed to take an oscillometric reading every
20 min during waking hours and every 30 min during
sleeping hours. The monitor cuff was fitted to the non-
dominant arm approximately 2.5 cm above the antecubital
fossa. The Spacelab machine was calibrated by taking a
reading with Spacelab monitor connected to a mercury
sphygmomanometer and at least three readings recorded by
the monitor were within ±7 mm Hg of the readings observed
on the sphygmomanometer after correcting for bleep stop
value. A valid 24 h recording was defined as a minimum
of 80% successful readings, with hourly mean blood pressure
recordings missing for o4 h of the 24 h period. The 24-h
ambulatory blood pressure provides greater statistical power
than casual (clinical) measurement. Small effects on blood
pressure may be missed using single clinical measurements.
This is partly a result of increased power with multiple
measurements over 24 h, but may also be the result of
Table 1 The composition of the lupin flour-enriched foods and control foods
per 100 g
Nutrient (per 100 g) Food provided
Lupin flour enriched Control
Bread Biscuits Pasta Bread Biscuits Pasta
Energy (MJ) 0.9 1.5 1.3 1.0 1.8 1.4
Carbohydrates (g) 27.1 35.0 48.0 42.0 64.8 66.2
Fat (g) 2.8 16.4 2.8 2.9 16.1 1.2
Protein (g) 16.0 19.0 20.1 7.6 4.2 10.7
Fibre (g) 11.1 11.2 11.0 5.4 3.6 3.2
Effects of lupin on body weight and CVD risk
R Belski et al
812
International Journal of Obesity
measuring a different parameter not influenced by the ‘white
coat’ effect.
32
Biochemistry
Venous blood samples were collected following a 12-h fast
from the antecubital vein of the forearm. Blood was collected
into BD Vacutainer (Franklin Lakes, NJ, USA) serum separator
tubes and EDTA tubes. All biochemical and haematology
analyses were performed in the PathWest Laboratory at Royal
Perth Hospital (WA, Australia). Analyses of lipids, insulin and
high-sensitivity C-reactive protein were performed on sera
stored at 80 1C in a single batch to reduce variability.
Full blood picture was performed on venous blood
collected into an EDTA tube on an Abbott Cell-Dyn 4000
(CD4K) instrument (Abbott Laboratories, Abbott Park, IL,
USA). Serum total cholesterol, high-density lipoprotein
(HDL) cholesterol and triglyceride concentrations were
analysed with a fully automated analyser (Architect
c16000; Abbott Laboratories). The assay coefficients of
variation were 0.9% for total cholesterol, 1.7% for triglyce-
rides and 2.6% for HDL cholesterol. Serum low-density
lipoprotein cholesterol was calculated using the Friedewald
equation.
33
Serum glucose was measured using a hexokinase/
G-6-PDH method (Abbott Laboratories) using a fully auto-
mated analyser (Architect c16000). The assay coefficient of
variation was 2.7%. Serum insulin was analysed by immuno-
assay (Abbott Laboratories) using a fully automated analyser
(Architect c16000), with an assay coefficient of variation
of 1.5%. The homeostasis model assessment (HOMA) score
was calculated with the following formula,
34
(serum glucose
(mmol l
–1
)serum insulin (mUml
–1
)/22.5), to estimate
changes in insulin sensitivity. High-sensitivity C-reactive
protein was analysed using BN Systems (Dade Behring,
Germany) with a BNII analyser, assay coefficient of variation
of 4.4%.
Statistics
Statistical analyses were performed using SPSS 15.0 software
(SPSS Chicago, IL, USA) or Stata 11.0 software (StataCorp,
College Station, TX, USA). The sample for this study was
calculated on the primary outcome of body weight. With
a¼0.05, 50 participants per group provided 480% power to
detect a 2-kg difference between groups. This would also
provide 480% power to detect a 10% difference in fasting
glucose and a 10% difference in low-density lipoprotein
cholesterol. Secondary outcome measures included body fat
mass, fat-free mass and fasting serum concentrations of
cholesterol, triglycerides, glucose and insulin. To allow for
dropouts, we planned to recruit at least 130 participants to
this study. The primary analysis included participants who
completed the intervention. Intention-to-treat analysis was
also performed. For descriptive data, results are presented as
mean±s.d., except for insulin and triglycerides that were
log-transformed and are reported as geometric mean and
95% confidence interval (CI). The baseline-adjusted 4- and
12-month values and between-group differences are
presented as mean (95% CI) with Po0.05 being the level of
significance in two-tailed testing. At baseline, characteristics
of participants in the two groups were compared using the
independent-samples t-test and the w
2
test for categorical
variables. The Stata ‘xtmixed’ and ‘margins’ commands were
used to assess baseline-adjusted between-group differences
at 4 and 12 months. Fixed effects in each model (except for
24-h blood pressure) were baseline value of the variable,
an indicator variable for month 4, month 12, and for
treatment group, a treatment group month 4 interaction
term and a treatment group month 12 interaction term.
A subject-specific random intercept term was also included.
For 24-h blood pressure, fixed effects were treatment group,
month (as a categorical variableFthat is, with values 0, 4 or 12),
month treatment group and hour (as a categorical vari-
able). A subject-specific random intercept and a subject-
specific random slope for hour (treated as a factor using the
R.hour notation in Stata) were included as random effects.
Results
Participants
A total of 131 participants (63 control, 68 lupin) aged 22–71
years were randomized and commenced the study. In all, 110
(55 control, 55 lupin) participants completed the trial to
4 months and 93 participants (47 control, 46 lupin; 71%)
completed the trial to 12 months (Figure 1). The reasons for
withdrawal from the lupin group included: not able to
commit to time requirements of study (n¼10), relocation to
other city (n¼5), commencement or change in medication
or health status (n¼4), inability to consume foods provided
in required amount (n¼2), and one participant failed to
disclose a pre-existing condition prior to randomization and
was withdrawn immediately. The reasons for withdrawal
from the control group included: not able to commit to time
requirements of study (n¼11), relocation to other city
(n¼1), commencement or change in medication or health
status (n¼3) and inability to consume foods provided in the
required amount (n¼1).
The two groups (control and lupin) were well matched at
baseline (Table 2). There were no significant differences in
baseline characteristics between the participants who completed
the trial to 4 months or 12 months and those who withdrew.
There were no reported adverse effects from eating either the
control or the lupin foods during the 12-month study.
Energy and nutrient intake and physical activity
Energy and nutrient intakes and levels of physical activity
were well matched between groups at baseline (Table 3).
During the 3-month weight loss period, both groups were
placed on B35% energy-restricted diet. During the 8-month
ad libitum weight maintenance period, the energy intakes of
Effects of lupin on body weight and CVD risk
R Belski et al
813
International Journal of Obesity
both groups increased but remained below baseline energy
intake. Estimated energy intakes during the weight loss and
weight maintenance periods were not significantly different
between groups (Table 3). At both 4 and 12 months, there
were higher protein and fibre intakes and lower carbohydrate
intakes in the lupin group relative to control. At 4 and 12
months, respectively, mean protein intake was higher by 19 g
per day (95% CI: 8, 29; Po0.001) and 15 g per day (95% CI:
3, 28; Po0.001); fibre intake was higher by 9 g per day (95%
CI: 6, 12; Po0.001) and 14 g per day (95% CI: 10, 18;
Po0.001); and carbohydrate intake was lower by 26 g
per day (95% CI: 47, 5; P¼0.016) and 27 g per day
(95% CI: 54, 1; P¼0.041). The differences in protein, fibre
and carbohydrate intake are close to those estimated to be
observed based solely on intake of the lupin flour-enriched
food provided. Differences in energy, fat and alcohol intakes,
sodium and potassium excretion and physical activity at both
4 and 12 months were not significant (Table 3).
Body weight and composition
Body weight and body composition measurements were not
different between groups at baseline (Table 2). The baseline-
adjusted body weight and body composition measurements
in the control and lupin groups at 4 and 12 months and the
between-group differences are presented in Table 4. There
were no significant differences between treatment groups in
body weight, fat mass, fat-free mass, android and gynoid fat
percentages at 4 or 12 months. Furthermore, there were no
significant differences between treatment groups in the
maintenance of body weight loss (0.7 kg; 95% CI: 1.8,
0.5) and fat mass loss (0.4 kg; 95% CI: 1.4, 0.6) during the
weight maintenance period (from 4 to 12 months). Subgroup
analysis according to gender and baseline age (above and
below 50 years) and an intention-to-treat analysis did not
alter the interpretation of these results (data not shown).
Cardiovascular disease risk factors
Blood pressures, and fasting blood lipids, glucose and insulin
concentrations were not significantly different between
groups at baseline (Table 2). Baseline-adjusted blood pres-
sures, and fasting blood lipids, glucose and insulin concen-
trations at 4 and 12 months are presented in Table 5. For
lupin relative to control, mean 24-h systolic and diastolic
blood pressures were significantly lower at 12 months, but
not at 4 months. There were no differences between groups
in total cholesterol, low-density lipoprotein cholesterol,
Recruitment respondents:
screened by telephone
n ~ 800
Screened in-person
n = 231
Randomized
n = 134
Commenced trial
n = 131
(68 men, 63 women)
Control Group
Baseline
n = 63
(34 men, 29 women)
Lupin Group
Baseline
n = 68
(34 men, 34 women)
4 month assessment
n = 55
(32 men, 23 women)
4 month assessment
n = 55
(29 men, 26 women)
Completed trial
n = 47
(27 men, 20 women)
Completed trial
n = 46
(25 men, 21 women)
Withdrew
n = 8
Withdrew
n = 8
Withdrew
n = 13
Withdrew
n = 9
Withdrew prior to
baseline assessments
n = 3
Figure 1 Flowchart of participants at each stage of the trial.
Effects of lupin on body weight and CVD risk
R Belski et al
814
International Journal of Obesity
triglyceride and glucose concentrations at 4 and 12 months.
HDL cholesterol was significantly lower for lupin relative to
control. This difference was almost entirely because of an
increase from baseline to 12 months in HDL cholesterol
within the control group (0.07 (95% CI: 0.01, 0.13) mmol l
–1
,
P¼0.03), with no change within the lupin group (0.00
(95% CI: 0.06, 0.06) mmol l
–1
,P¼0.94). At both 4 and
12 months, the lupin group had significantly lower fasting
insulin concentrations and HOMA scores relative to
control (Figure 2). Subgroup analyses according to gender,
baseline body mass index (above and below 30 kg m
–2
) and
baseline total cholesterol (total cholesterol above and below
the median of 5.3 mmol l
–1
) and an intention-to-treat
analysis did not alter interpretation of these results (data
not shown).
Discussion
We have investigated the effects of 12 months of regular
consumption of a lupin-enriched diet, during and following
energy restriction, on body weight, body composition, and
cardiovascular disease risk factors in overweight and obese
men and women. The higher protein and fibre lupin-
enriched diet did not significantly influence body weight
and body fat and fasting total cholesterol and glucose
concentrations. However, the lupin-enriched diet resulted
in significantly lower fasting insulin concentrations by
16 and 21%, and HOMA scores by 30 and 33% at 4 and
12 months, respectively. We also found that the lupin-
enriched diet resulted in lower blood pressure at 12 months.
There is now strong evidence that increasing the protein
and fibre content of the diet can reduce appetite acutely.
2,3
In line with this, we have previously shown that increasing
the protein and fibre content of bread with lupin flour
significantly reduced appetite and energy intake acutely.
8
In
addition, lupin fibre-enriched foods have been shown to
increase satiety.
35
Data from studies using ad libitum diets
with increased protein
36–38
or fibre
3
intake are limited. We
recently reported that an ad libitum lupin flour-enriched diet
higher in dietary protein and fibre consumed over 4 months
did not significantly influence body weight or composition
in overweight individuals.
11
There is some evidence that
longer-term regular consumption of increased protein
36,37
Table 3 Mean energy and nutrient intakes, physical activity levels and urinary analytes of participants in the lupin and control groups at baseline, 4 and 12 months
a
Control Lupin
Baseline
(n¼63)
4 months
(n¼55)
12 months
(n¼47)
Baseline
(n¼68)
4 months
(n¼55)
12 months
(n¼46)
Energy intake (MJ per day) 9.8±2.5 7.9±1.7 8.3±2.4 9.9±2.5 8.0±1.9 8.2±2.2
Total fat intake (g per day) 92±30 63±21 66±29 96±32 69±27 69±27
Protein intake (g per day) 105±29 86±26 95±29 106±29 105±26 110±27
Carbohydrate intake (g per day) 236±83 208±53 218±64 230±68 182±49 191±55
Fibre intake (g per day) 24±826
±825
±723
±835
±839
±12
Alcohol (g per day) 15±17 11±16 11±17 20±21 10±13 9±13
24 h creatinine excretion (mmol per day) 14.0±5.0 14.4±5.4 16.8±11.9 13.5±3.4 13.9±3.7 14.7±4.5
Sodium excretion (mmol per mmol creatinine) 11.4±4.6 11.3±4.5 11.5±4.4 10.7±4.0 11.3±6.1 11.9±4.7
Potassium excretion (mmol per mmol creatinine) 5.8±1.8 5.7±1.9 5.7±2.3 5.7±1.7 6.2±2.6 5.7±1.6
Activity (kcal per kg per day) 40±744
±945
±941
±644
±11 45±7
a
Values are presented as mean±s.d.
Table 2 Characteristics of participants in the control and lupin groups at
baseline
a
Control (n¼63) Lupin (n¼68)
Men/women (n) 34/29 34/34
Age (years) 46.7±9.4
b
46.5±10.1
Height (m) 1.72±0.11 1.71±0.10
Activity (kcal per kg per day) 32.4±7.3 33.2±6.4
Body weight and composition
Body weight (kg) 93.7±15.2 91.8±13.5
BMI (kg m
–2
) 31.4±2.8 31.3±2.7
Body fat mass (kg) 34.7±7.3 34.1±7.2
Body fat (%) 39.3±7.7 39.1±7.3
Fat-free mass (kg) 57.5±13.3 56.6±11.6
Android fat (%) 48.3±6.3 47.7±6.7
Gynoid fat (%) 40.7±9.7 40.8±9.0
Blood pressure
24 h Systolic pressure (mm Hg) 121±10 122±9
24 h Diastolic pressure (mm Hg) 75±875
±7
Fasting biochemical assessments
Total cholesterol (mmol l
–1
) 5.33±0.84 5.18±0.84
LDL cholesterol (mmol l
–1
) 3.33±0.76 3.29±0.77
HDL cholesterol (mmol l
–1
) 1.34±0.27 1.33±0.28
Triglycerides (mmol l
–1
)
c
1.30 (1.18, 1.44)
c
1.12 (1.02, 1.23)
Glucose (mmol l
–1
) 5.03±0.51 5.11±0.45
Insulin (mU l
–1
)
c
7.83 (6.94, 8.83) 8.29 (7.28, 9.44)
HOMA-IR (units) 2.0±1.0 2.3±2.4
High-sensitivity C-reactive
protein (mg l
–1
)
2.93±4.37 2.93±3.27
Abbreviations: BMI, body mass index; HDL, high-density lipoprotein; HOMA-IR,
homeostasis model assessment of insulin resistance; LDL, low-density lipo-
protein.
a
Between-group differences analysed using the independent samples
t-test and the w
2
test for categorical variables. There were no significant
differences between groups for any of the variables reported.
b
Values are
presented as mean±s.d.
c
Geometric mean (95% confidence interval).
Effects of lupin on body weight and CVD risk
R Belski et al
815
International Journal of Obesity
or fibre
39
diets can assist weight loss with energy restriction
in overweight individuals. However, several intervention
studies have failed to demonstrate benefit.
We have shown that a lupin-enriched diet did not
significantly influence body weight, body fat mass or weight
regain. The lack of a significant between-group difference in
Table 4 Mean baseline-adjusted body weight and composition measurements and between-group differences of participants in the control and lupin groups
at 4 and 12 months
a
At 4 months At 12 months
Control (n¼55) Lupin (n¼55) Difference Control (n¼47) Lupin (n¼46) Difference
Mean (95% CI) Mean (95% CI) Mean (95% CI) Mean (95% CI) Mean (95% CI) Mean (95% CI)
Body weight (kg) 85.4 (84.5, 86.3) 85.5 (84.6, 86.4) 0.1 (1.2, 1.4) 85.8 (84.8, 86.8) 85.2 (84.2, 86.2) 0.6 (2.0, 0.8)
Body fat mass (kg) 29.5 (28.6, 30.3) 29.0 (28.1, 29.9) 0.5 (1.7, 0.7) 29.9 (28.9, 30.8) 28.9 (28.0, 29.9) 0.9 (2.2, 0.4)
Fat-free mass (kg) 55.7 (55.3, 56.2) 56.2 (55.8, 56.6) 0.5 (0.1, 1.1) 55.9 (55.5, 56.4) 56.1 (55.6, 56.6) 0.2 (0.5, 0.8)
Android fat (%) 43.7 (42.8, 44.7) 44.1 (43.1, 45.1) 0.4 (1.0, 1.7) 44.3 (43.3, 45.4) 44.0 (43.0, 45.1) 0.3 (1.8, 1.2)
Gynoid fat (%) 38.7 (38.0, 39.4) 38.4 (37.7, 39.1) 0.3 (1.3, 0.7) 38.7 (37.9, 39.4) 38.2 (37.5, 39.0) 0.5 (1.5, 0.6)
a
Values are baseline-adjusted mean or mean between-group difference (95% confidence intervals).
Table 5 Mean baseline-adjusted blood pressure, fasting biochemical measurements and between-group differences of participants in the control and lupin groups
at 4 and 12 months
a
At 4 months At 12 months
Control (n ¼55) Lupin (n¼55) Difference Control (n ¼47) Lupin (n¼46) Difference
Mean (95% CI) Mean (95% CI) Mean (95% CI) Mean (95% CI) Mean (95% CI) Mean (95% CI)
24 h Systolic ABPM (mm Hg) 118.5 (117.8, 119.2) 119.0 (118.4, 119.7) 0.51 (0.5, 1.5) 120.9 (120.1, 121.7) 119.6 (118.8, 120.3) 1.33 (2.4, 0.3)*
24 h Diastolic ABPM (mm Hg) 73.4 (72.9, 74.0) 73.6 (73.1, 74.1) 0.19 (0.6, 1.0) 74.3 (73.7, 74.9) 73.3 (72.7, 73.9) 1.0 (1.9, 0.2)*
Total cholesterol (mmoll
–1
) 5.03 (4.90, 5.16) 5.08 (4.95, 5.21) 0.05 (0.13, 0.24) 5.04 (4.90, 5.18) 5.08 (4.94, 5.22) 0.04 (0.16, 0.24)
LDL cholesterol (mmoll
–1
) 3.20 (3.09, 3.31) 3.23 (3.12, 3.34) 0.03 (0.12, 0.18) 3.18 (3.06, 3.30) 3.20 (3.09, 3.32) 0.03 (0.14, 0.19)
HDL cholesterol (mmoll
–1
) 1.39 (1.35, 1.42) 1.32 (1.28, 1.36) 0.06 (0.12, 0.01)* 1.42 (1.38, 1.46) 1.34 (1.29, 1.38) 0.08 (0.14, 0.02)*
Triglycerides (mmol l
–1
)
b
0.95 (0.89, 1.02) 1.03 (0.96, 1.11) 0.08 (0.02, 0.18) 0.96 (0.89, 1.04) 1.05 (0.97, 1.13) 0.08 (0.02, 0.19)
Glucose (mmol l
–1
) 5.12 (5.03, 5.21) 5.06 (4.97, 5.15) 0.05 (0.18, 0.07) 5.13 (5.03, 5.22) 5.08 (4.98, 5.18) 0.05 (0.18, 0.09)
Insulin (mU l
–1
)
b
7.18 (6.61, 7.81) 6.03 (5.54, 5.55) 1.19 (1.34, 1.06)* 7.56 (6.90, 8.28) 5.96 (5.44, 6.53) 1.27 (1.44, 1.11)*
HOMA-IR (units) 1.96 (1.68, 2.24) 1.37 (1.09, 1.66) 0.59 (0.99, 0.19)* 2.06 (1.75, 2.37) 1.39 (1.08, 1.70) 0.67 (1.11, 0.24)*
High-sensitivity CRP (mgl
–1
) 3.68 (2.80, 4.56) 3.12 (2.22, 4.01) 0.56 (1.82, 0.69) 2.71 (1.74, 3.68) 2.56 (1.60, 3.51) 0.15 (1.52, 1.21)
Abbreviations: ABPM, ambulatory blood pressure monitoring; CRP, C-reactive protein; HDL, high-density lipoprotein; HOMA-IR, homeostasis model assessment of
insulin resistance; LDL, low-density lipoprotein.
a
Values are baseline-adjusted mean or mean between-group difference (95% confidence intervals).
b
Geometric
mean (95% confidence interval). *Statistically significant (Po0.05).
04
12
5.4
5.2
5
4.8
4.6
4.4
0
HOMA- IR (units)
Glucose Concentration (mmol/L)
Months
0412
Months
Control
Lupin
0412
Months
2.3
2
1.9
1.8
1.7
1.6
0
2.2
2.1
2.4
Insulin Concentration (mU/L)
10.5
10
9.5
9
8
7
6.5
0
7.5
8.5
Figure 2 Glucose and insulin concentrations and HOMA scores at baseline, 4 months and 12 months for participants in the control (X) and lupin (O) groups.
Values presented are mean and s.e.m.
Effects of lupin on body weight and CVD risk
R Belski et al
816
International Journal of Obesity
body weight suggests that effects of lupin-enriched foods on
satiety may be, in part, counterbalanced by other dietary,
lifestyle and/or environmental factors that influence energy
balance in the longer term.
40
The observed differences in
body weight in our study were small (o1 kg). Although
effects of this magnitude may be important for long-term
maintenance of healthy body weight, our study was only
powered to detect a 2-kg body weight difference between the
groups. A recent meta-analysis suggests a small benefit of
higher protein diets on fat-free mass retention (B1.2 kg), but
not on body weight or fat loss.
4
The observed differences in
fat-free mass in the current study were not significant. The
difference in protein intake of 15–19 g per day in our trial
was modest compared with most previous trials in which
differences have usually been 430 g per day.
2
However, the
higher protein intake was concurrent with higher dietary
fibre intake (additional 9–14 g per day), which would be
regarded as considerable.
3
The total fibre intakes of
B35 g per day during weight loss and B39 g per day during
weight maintenance in the lupin group would also be
regarded as high and substantial enough to impact satiation
and satiety.
3
The type of fibre present in lupin may also be
important. Lupinus angustifolius, the species of lupin used in
these studies, contains B70% insoluble fibre.
41
Insoluble
fibre may be less effective than soluble fibre in reducing
appetite.
3
However, the results of this trial when taken
together with results of our previous trials
11,12
bring into
question the importance of acute effects of dietary nutrients
on appetite and energy intake for longer-term effects on
body weight and composition.
In the current study the lupin treatment resulted in a
significant reduction in fasting insulin concentrations and
HOMA scores following weight loss at 4 months. These
differences were maintained during weight maintenance to
12 months. In a recently published study, involving a similar
population of overweight men and women, we found that
4 months of regular consumption of an ad libitum lupin-
enriched diet did not alter fasting glucose or insulin
concentrations.
11
This suggests that weight loss in over-
weight individuals may be important to attain improve-
ments in insulin sensitivity with a lupin-enriched diet. If a
lupin-enriched diet can improve insulin sensitivity, the
components responsible remain uncertain. It is possible that
both the fibre and protein in lupin flour could contribute to
improved insulin sensitivity. We and others have previously
shown that lupin flour-enriched foods can acutely reduce
postprandial glycaemia and insulinaemia.
12,27
Such effects, if
maintained in the longer term, may contribute to improved
insulin sensitivity. Data from the Framingham Offspring
Cohort study suggest that an extra 14 g of dietary fibre per
day would improve HOMA scores by 0.6 units.
42
In the
present study the difference in fibre intake at 12 months was
14 g per day and the difference in HOMA score was 0.7 units.
Acutely, dietary protein enhances insulin secretion,
43,44
leading to increased glucose disposal. Although data from
epidemiological studies indicate that this may not result in
improved insulin sensitivity,
45,46
positive effects of higher
protein intakes have been observed in some intervention
studies.
21,47,48
The lack of effect on glucose concentrations
in the current study may be because of population selection.
All participants were non-diabetic and had otherwise
normal fasting glucose concentrations at screening of
o6.0 mmol l
–1
.
Regular consumption of lupin flour-enriched foods
resulted in lower systolic and diastolic blood pressure at
12 months, but not at 4 months. Available data suggest that
higher plant protein
15
and dietary fibre
17
intakes, and
substitution of carbohydrate in the diet with protein and
fibre can benefit blood pressure.
22
We previously showed
that 4 months of regular consumption of an ad libitum
lupin flour-enriched diet resulted in lower systolic blood
pressure by 3 mm Hg in overweight individuals.
22
The lack
of difference in systolic blood pressure at 4 months and
the estimated systolic blood pressure difference at 12 months
of 1.3 mm Hg indicates that weight loss resulting in lower
blood pressure may have obscured some of the effects of the
lupin flour-enriched diet. Another possible explanation of
the smaller observed effect size is that the lupin flour was
substituted primarily for wholemeal wheat flour. Most
previous trials have used a refined carbohydrate as the
control.
14–16,22
Limited data from human and animal studies suggest that
a lupin-enriched diet may benefit serum lipids and lipo-
protein.
24,49
However, we previously showed that an
ad libitum lupin flour-enriched diet over 4 months also had
no benefit on the serum lipid profile.
11
The lack of effect on
blood lipids in our studies may be because of a number of
factors. Most of our participants had normal lipid profiles at
baseline. Nevertheless, subgroup analyses in participants
with baseline total cholesterol above or below 5.3 mmol l
–1
(median) failed to show any between-group differences,
which is a result similar to our previous trial.
11
The lack of
changes in serum lipids may also be because of the physical
and structural characteristics of the lupin fibre, which is
primarily insoluble.
41
It is primarily soluble/viscous fibres
that have been found to significantly reduce total cholesterol
concentrations.
41
The findings of a significant increase in
HDL cholesterol from baseline to the end of intervention in
the control group, with no change in the lupin group, are
also consistent with our previous study.
11
The mechanism
for this is uncertain.
Our study was designed to investigate the functional
effects of 12 months of regular consumption of a lupin-
enriched diet. The study design did not allow demonstration
of the effect of lupin per se, independent from any generic
effects of increasing protein and fibre. However, the use of
lupin flour is a novel approach to increase the consumption
of both protein and fibre in significant amounts with a single
food ingredient. This may be more difficult using more than
one ingredient, or using selected high-protein and high-fibre
foods. In addition, most of the available evidence for
effects of protein and fibre on weight management and
Effects of lupin on body weight and CVD risk
R Belski et al
817
International Journal of Obesity
cardiovascular disease risk factors derives from studies
investigating these dietary components independently from
one another. The current study is therefore unique in
studying the functional effects of a higher protein and fibre
lupin-enriched diet, following both an energy-restricted
weight loss diet and an ad libitum weight maintenance diet.
In conclusion, regular consumption of lupin flour-
enriched foods during and following energy restriction did
not significantly influence body weight. The changes in
insulin concentrations and insulin sensitivity at 4 and 12
months indicate that even among non-diabetic individuals
with normal blood glucose levels, lupin flour-enriched foods
may improve insulin sensitivity in overweight individuals
when combined with weight loss. The observed effects
on blood pressure are consistent with a growing body of
evidence suggesting that diets higher in plant proteins and
dietary fibre lower blood pressure.
Conflict of interest
The authors declare no conflict of interest.
Acknowledgements
This study was funded by The Western Australian
Government, Department of Industry and Resources. We
thank Dr Kay Cox for her assistance with recommending
appropriate physical activity assessment tools. We also thank
Bodhi’s Bakery, Fremantle, WA, for baking the bread and
biscuits and Belmar Foods, Balcatta, WA, for manufacturing
and providing the pasta.
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