Nutrition 22 (2006) 367375

www.elsevier.com/locate/nut
Applied nutritional investigation

Effects of liquid carbohydrate/essential amino acid ingestion on acute

hormonal response during a single bout of resistance exercise in
untrained men
Stephen P. Bird, B.H.M.S. (Hons.)*, Kyle M. Tarpenning, Ph.D., and Frank E. Marino, Ph.D.
School of Human Movement Studies, Charles Sturt University, Bathurst, New South Wales, Australia

Manuscript received March 31, 2005; accepted November 8, 2005.

Abstract Objective: The primary objective of this study was to investigate the influence of nutritive
interventions on acute hormonal responses to a single bout of resistance exercise in untrained young
men. Specifically, the aim was to determine whether the acute hormonal milieu conducive to
enhancing skeletal muscle hypertrophic adaptation to resistance training could be created. The
potential role of cortisol in inhibiting training-induced muscle growth is of particular interest, as is
whether exercise-induced cortisol release can be attenuated by nutritive interventions.
Methods: After a 4-h fast, 32 subjects performed a single bout of resistance exercise (60 min),
during which they consumed a 6% carbohydrate (CHO) solution, a 6-g essential amino acid (EAA)
mixture, a combined CHO EAA supplement, or a placebo beverage. Blood samples were
collected every 15 min throughout the exercise bout, immediately after exercise, and 15 and 30 min
after exercise for analysis of total testosterone, cortisol, growth hormone, insulin, and glucose.
Results: No significant change in glucose or insulin was observed for placebo. CHO and CHO EAA
ingestion resulted in significantly (P 0.001) increased glucose and insulin concentrations above
baseline, whereas EAA resulted in significant postexercise increases (P 0.05) in insulin only. Placebo
exhibited a significant increase in cortisol within 30 min (P 0.01), with a peak increase of 105% (P
0.001) immediately after exercise, and cortisol remained 54% above baseline at 30 min after exercise
(P 0.05). Conversely, the treatment groups displayed no significant change in cortisol during the
exercise bout, with CHO and CHO EAA finishing 27% (P 0.01) and 23% (P 0.05), respectively,
below baseline at 30 min after exercise. No between-group differences in exercise-induced growth
hormone or testosterone concentrations after nutritive intervention were present.
Conclusion: These data indicate that CHO and/or EAA ingestion during a single bout of resistance
exercise suppresses the exercise-induced cortisol response, in addition to stimulating insulin release.
We conclude that the exercise-induced hormonal profile can be influenced by nutritive interventions
toward a profile more favorable for anabolism. 2006 Elsevier Inc. All rights reserved.

Keywords: Resistance exercise; Carbohydrate/essential amino acid supplementation; Acute hormonal response

Introduction proteins and are intimately involved with protein synthesis

Biochemical (hormonal) responses play a central role in
signaling the cellular remodeling process of myofibrillar
This research was supported in part by Musashi and the Gatorade
Sports Science Institute.
In memory of the late Kyle Tarpenning, Ph.D.
* Corresponding author. Tel.: 612-6338-4155; fax: 612-6338-
4065.
E-mail address: sbird@csu.edu.au (S.P. Bird).
and degradation [1,2]. A multitude of hormones that exert
anabolic effects such as testosterone, growth hormone, in-
sulin, and insulin-like growth factor-I regulates protein syn-
thesis [35], whereas catabolic hormones, principally cor-
tisol, increase protein degradation [6] in an attempt to
support glucose synthesis. This dynamic balance between
anabolic and catabolic milieu will ultimately influence pro-
tein turnover [1] and result in a positive or negative muscle
protein balance. One way to influence muscle protein bal-
ance after resistance exercise is by nutritive interventions

0899-9007/06/$ see front matter 2006 Elsevier Inc. All rights reserved.
doi:10.1016/j.nut.2005.11.005
368 S.P. Bird et al. / Nutrition 22 (2006) 367375
[79]. Research indicates that carbohydrate (CHO) and/or
essential amino acid (EAA) ingestion around the time of
exercise modifies the acute biochemical events associated
with resistance training, shifting the exercise-induced hor-
monal profile toward a profile more favorable for positive
protein balance. Specifically, it is the response of insulin
and cortisol that has received much attention because they
are intimately involved in protein turnover.
An acute bout of resistance exercise performed in con-
junction with liquid CHO ingestion during the exercise bout
[10], 10 min before and after exercise [11], and 60 min after
exercise [12,13] results in significantly increased insulin
levels. Roy et al. [12] suggested that consumption of a CHO
supplement after an acute bout of resistance exercise in-
creases insulin concentrations, thereby enhancing muscle
protein balance. This is most likely due to the inhibitory
effect of insulin on muscle protein breakdown [14], al-
though this effect may be entirely restricted to globular
proteins because myofibrillar proteins are unaffected by
systemic hyperinsulinemia [15]. Therefore, insulins mech-
anism of action on skeletal muscle is controversial [16,17].
In the absence of an increase in amino acid concentra-
tion, an increase in insulin has only a modest effect on
muscle protein synthesis [18]. Therefore, supplying exoge-
nous EAA during the exercise bout should allow expression
of the stimulatory effect of insulin. Accordingly, Tipton et
al. [19] showed that rates of protein synthesis and net
protein balance are higher when amino acid availability is
increased after an acute bout of resistance exercise than
when subjects are fasted. As a result, amino acid supply to
the muscle intracellular compartment through enhanced
amino acid transport into muscle is suggested to be an
important regulatory factor in determining the rate of mus-
cle protein synthesis [20]. However, recent data presented
by Bohe et al. [21] have indicated that protein synthesis is
modulated by extracellular, not intracellular, amino acid
availability.
Research has shown that different resistance exercise
protocols result in an acute increase in cortisol [10,22,23].
However, Conley and Stone [24] suggest that excessive
cortisol levels exhibited after a single bout of resistance
exercise may elicit physiologic responses detrimental to the
positive adaptations attributed to resistance training. Liquid
CHO ingestion during the exercise bout is suggested [10] to
increase the exogenous glucose load, resulting in inhibition
of the glucose-regulatory response of cortisol. Further, ad-
dition of EAA (CHO EAA) may augment this response
by providing a potent stimulus, increasing extracellular
amino acid availability and insulin release. Such an envi-
ronment may provide greater potential for protein accretion
and skeletal muscle growth, thus counteracting the stimula-
tory effect of cortisol on skeletal protein degradation.
The present investigation examined the influence of nu-
tritive interventions (a 6% CHO solution, a 6-g EAA mix-
ture, or a combined CHO EAA beverage) on acute
biochemical responses to a single bout of resistance exercise
in untrained young men. Specifically, the objective of this
investigation was to determine whether the acute hormonal
milieu conducive to enhancing skeletal muscle hypertrophic
adaptation to resistance training could be created by ingest-
ing one of these interventions during the exercise bout. The
potential role of cortisol in inhibiting training-induced mus-
cle growth [10] is of particular interest, as is whether exer-
cise-induced cortisol release can be attenuated by such nu-
tritive interventions.
Materials and methods
Subjects
Thirty-two untrained young men (18 to 29 y old) volun-
teered to participate in this investigation. After a full expla-
nation of all procedures and possible risks of the investiga-
tion, written informed consent was obtained before testing
began. Subjects completed a health history questionnaire;
all were apparently healthy and had no medical contraindi-
cations or history of any endocrine disorders that might
influence their responsiveness to an acute bout of resistance
exercise. No subject was taking any medication or nutri-
tional supplementation; all were non-steroid users and non-
smokers. Subjects were randomly allocated into one of four
groups: CHO group (n 8), EAA group (n 8), combined
CHO EAA group (n 8), or placebo (PLA) group (n
8). All experimentation was approved by the ethics in hu-
man research committee of the university.
Experimental design
To examine the influence of nutritive interventions on
acute hormonal responses to a single bout of heavy-
resistance exercise in untrained young men, a randomized,
double-blind, placebo-controlled design was used. Subjects
reported to the Exercise and Sport Sciences Laboratory on
three occasions separated by no more than 7 d. All sessions
were performed between 3:00 and 5:00 PM to minimize the
influence of diurnal variations on exercise performance and
hormonal response [25], with times held constant for each
subject. Subjects were required to refrain from all strenuous
activity, alcohol use, caffeine, and sexual activity and were
notified to maintain normal nocturnal sleep habits (i.e.,
approximately 8 h/night) throughout the experimental time-
line.
During the initial laboratory session (day 1), descriptive
data including height, weight, and body composition were
recorded. Height was measured to the nearest 0.1 cm by
using a stadiometer (Len Blayden, Lugarno, Australia) and
weight was measured to the nearest 0.01 kg by using an
electronic precision balance scale (HW-100KAI, GEC,
Avery Ltd., Miranda, Australia). Body composition was
determined by dual-energy x-ray absorptiometry with a to-
tal-body scanner (DPX-IQ, Prodigy, Lunar Corp., Madison,
 S.P. Bird et al. / Nutrition 22 (2006) 367375 369

Day 1 Table 1
Anthropometric measurements (height, w eight, body composition) and dietary instructions
Resistance exercise protocol
Exercises Sets Repetitions Equipment
Day 2 1. Leg press 3 810 Panatta Sports
Equipment familiarisation and one-repetition maximal strength testing 2. Leg curl 3 810 Panatta Sports
3. Leg extension 3 810 Panatta Sports
Day 7 4. Shoulder press 3 810 Panatta Sports
Pre-Ex Canula Pre-Ex Ex Ex Ex IP P15 P30
5. Lat pulldown 3 810 Panatta Sports
Rest insertion BD BD BD BD BD BD BD 6. Bench press 3 810 York Barbell
7. Barbell bicep curl 3 810 York Barbell
Pre-Ex Rest Period Resistance Exercise Bout Post-Ex Recovery 8. Supine tricep extension 3 810 York Barbell
Time
(min) -30 -15 0 15 30 45 60 75 90

Ingestion of 22.5-30.0 ml of nutritive Water intake

4-hour fast
intervention betw een each set
Fig. 1. Acute exercise bout protocol of blood sampling. BD, blood draw;
IP, immediately after exercise; P15, 15 min after exercise; P30, 30 min
after exercise; Post-Ex, after exercise; Pre-Ex, before exercise.
WI, USA). At the completion of the session, an accredited
practicing dietitian (Dietitians Association of Australia) in-
structed subjects with procedures for reporting detailed di-
etary intake and eating habits for a 3-d period. The second
session (day 2) consisted of equipment familiarization and
one-repetition maximal (1-RM) strength testing for all ex-
ercises in the resistance training protocol. During the final
session (day 7), subjects performed a single bout of heavy-
resistance exercise, during which they consumed one of four
nutritive interventions. Blood samples were collected every
15 min throughout the exercise bout and after the exercise
period. The experimental timeline is shown in Fig. 1.
Dietary control
Food intake diaries were used to control for 3 d before
the experimental period, to assess whether there were dif-
ferences in energy intake and macronutrient composition
between groups. Subjects were instructed to consume their
normal diet during this period. Nutritional analysis was
performed by an accredited practicing dietitian (Dietitians
Association of Australia) who used Serve 2.0 (Serve Nutri-
tion Management System, St. Ives, Australia). Each item
was entered onto a personal computer and the program
provided the total energy consumption and macronutrient
composition as an average over the 3 d. If a nutrient value
was missing, information from food tables or information
provided by food manufacturers was obtained. Subjects
were instructed to replicate their dietary patterns for the 3-d
period before the exercise bout.
Nutritive intervention
During the exercise bout, subjects consumed 6% CHO
(Gatorade, Quaker Oats Inc., Chicago, IL, USA), 6 g of
EAA (Musashi, Notting Hill, Australia), a CHO EAA
combination (6% CHO 6 g of EAA), or PLA (aspartame
ad libitum and citrus flavoring; Quaker Oats Inc.) dissolved in water at
a fluid volume of 8.5 mL/kg of body mass (average of 675
mL of solution). Total volume of fluid was divided by 25
servings, allowing for 22.5 to 30.0 mL, depending on body
size, to be ingested between each set of exercise. The EAA
composition consisted of histidine (0.65 g), isoleucine (0.60
g), leucine (1.12 g), lysine (0.93 g), methionine (0.19 g),
phenylalanine (0.93 g), threonine (0.88 g), and valine (0.70
g), which has been previously shown to enhance skeletal
muscle anabolism [8].
1-RM strength test
Maximal strength was assessed for each of the eight
selected exercises in the heavy-resistance protocol by
completing a 1-RM test (i.e., the heaviest load that could
be correctly performed once). Warm-up consisted of one
set of 5 to 10 repetitions at 40% to 60% of perceived
maximum. Subjects then rested for 1 min while perform-
ing light stretching. This was followed by three to five
repetitions performed at 60% to 80% of perceived max-
imum. Thereafter, three to four subsequent attempts were
made to determine the 1-RM strength, with the weight
increased progressively until the subject failed at the
given load. Three minutes of rest was allocated between
lifts. By definition, 1-RM is the maximum amount of
weight that can be lifted one time through a full range of
motion by using good form at a tempo of 2:0:2 (2 s
eccentric, 2 s concentric).
Resistance exercise protocol
The resistance exercise protocol (Table 1) used for this
investigation was that used by Bird and Tarpenning [23].
The subjects goal was to complete three sets of 8 to 10
repetitions at approximately 75% of that subjects 1-RM.
One minute of rest between each set and 2 min between
each exercise were allowed for recovery. Machine exercises
1 to 5 were performed on Panatta Sports Fit 2000 Line
weight machines (Panatta Sports, Apiro MC, Italy), and
exercises 6 to 8 were performed with an Olympic-style
barbell (York Barbell, York, PA, USA). The resistance
exercise session lasted approximately 60 min, was preceded
370 S.P. Bird et al. / Nutrition 22 (2006) 367375

Table 2
Physical characteristics, 1-RM performance data, and daily intake of dietary energy and nutrients*
PLA group (n 8) CHO group (n 8) EAA group (n 8) CHO EAA group (n 8)

Physical characteristics
Age (y) 20.3 0.8 22.3 1.2 21.3 0.7 20.6 0.5
Height (cm) 182.6 1.8 184.4 2.3 181.2 2.9 183.4 2.6
Weight (kg) 80.4 4.5 79.3 4.4 79.7 4.6 79.3 3.9
Fat mass (kg) 18.7 3.2 12.7 1.5 18.3 3.2 13.8 2.8
Fat-free mass (kg) 58.3 1.9 63.1 3.2 58.1 1.9 61.7 2.9
1-RM performance data
45 leg press (kg) 141.9 16.6 145.9 7.8 138.8 12.0 142.5 16.4
Bench press (kg) 58.1 6.7 67.8 5.2 65.6 3.1 63.1 6.0
Dietary intake
Energy (kcal) 2704.2 196.4 2895.8 225.6 2828.8 220.9 2744.0 253.1
Protein (g) 127.3 13.0 113.6 9.0 139.1 11.8 113.9 9.5
Carbohydrate (g) 299.7 20.9 343.6 22.9 298.1 30.0 338.3 33.8
Fat (g) 108.6 8.7 113.0 9.5 117.6 10.0 100.3 14.0

1-RM, one-repetition maximum; CHO, 6% carbohydrate solution; EAA, 6-g essential amino acid mixture; PLA, placebo
* Data are presented as mean standard error. No significant differences were found for any parameter.

by a 10-min warm-up, and concluded with a 10-min warm-
down period. Staff trained in the principles associated with
resistance training supervised all sessions.
Blood sampling and analysis
After a 4-h fast, subjects reported to the laboratory and
sat quietly for 15 min. A 20-gauge, 1.00-inch Teflon in-
dwelling cannula (Saf-T-Intima, Becton Dickinson Inc.,
Sandy, UT, USA) was inserted into an antecubital forearm
vein, after which subjects sat quietly for another 15-min
period before blood collection to minimize hormonal fluc-
tuations related to anticipatory responses [26]. Using a Va-
cutainer assembly and serum separator tubes (Monovette,
Sarstedt, Numbrecht, Germany), 5-mL blood samples were
drawn before exercise, every 15 min throughout the exercise
bout, immediately after exercise (IP), and 15 min (P15) and
30 min (P30) after exercise. To assist keeping the line clear
and prevent clotting, 1-mL of saline solution was injected
into the cannula line between each blood draw. Blood sam-
ples were gently inverted five times and allowed to stand at
room temperature for a minimum of 20 min. Samples were
then centrifuged for 10 min at 3000 rpm, with the superna-
tant removed, placed into plastic storage containers, and
frozen at 20C until analysis.
Before analysis the serum was allowed to reach room
temperature and then mixed gently by inversion. Glucose
was determined by an enzymatic spectrophotometric
method (Dimension Xpand, Dade Bearing Inc., Newark,
DE, USA). Total testosterone and cortisol concentrations
were determined by a competitive immunoassay technique
using chemiluminescent technology (VITROS ECi, Ortho-
Clinical Diagnostics Inc., Rochester, NY, USA) with detec-
tion limits lower than 0.03 nmol/L and lower than 3 nmol/L,
respectively. Insulin and growth hormone concentrations
were determined by a solid-phase, two-site chemilumines-
cent immunometric assay (Immulite 2000, Diagnostic Prod-
ucts Corp., Los Angeles, CA, USA), with detection limits of
2 IU/mL and 0.026 mIU/L, respectively. To avoid inter-
assay variations, all samples for each subject were assayed
in the same assay run. Analyses of glucose, total testoster-
one, cortisol, insulin, and growth hormone showed intraas-
say coefficients of variation of 5.9%, 2.6%, 2.7%, 3.1%, and
7.2%, respectively. Serum hormone concentrations were not
corrected for plasma volume shifts; thus, all statistical anal-
yses were performed on hormone values based on actual
measured circulating concentrations.
Statistical analysis
Descriptive data were generated for all variables and
expressed as mean standard error of the mean. Analysis
was performed with SPSS 11.5 (SPSS Inc., Chicago, IL,
USA). Statistical analysis involved a two-way analysis of
variance (group by time) with repeated measures. The
source of significant differences was located using Tukeys
Honestly Significant Difference post hoc procedure. Signif-
icant interactions were analyzed by simple main effects.
Statistical significance was set at P 0.05.
Results
Physical characteristics, 1-RM performance data, and
daily dietary intake
The physical characteristics, 1-RM performance data,
and daily dietary data of the research subjects are listed in
Table 2. The four groups did not significantly differ with
respect to physical characteristics and 1-RM performance
data. Thus, groups were matched for these variables. Nutri-
tional analysis of daily dietary intake showed that diets were
consistent across groups. The macronutrient composition of
dietary intake was also similar across groups.
 S.P. Bird et al. / Nutrition 22 (2006) 367375
a a
b 30
b c a b
6.5 d d b a
* * c b
b d d a
6.0 *
d *
*
Glucose (mM)
5.5
* 15
5.0
4.5
4.0
Exercise Recovery
3.5
Pre 15 30 45 IP P15 P30
Time (mins)
Fig. 2. Serum glucose concentrations for PLA (), CHO (
), EAA ( ),
and CHO EAA (). Significantly different from pre-exercise value (P
0.05): *CHO versus 30 min, 45 min, IP, P15, and P30; CHO EAA versus 30
min, 45 min, IP, P15, and P30. Significant difference between treatments (P
0.05): aCHO from PLA; bCHO EAA from PLA; cCHO from EAA;
d
CHO EAA from EAA. CHO, 6% carbohydrate solution; EAA, 6-g essential
amino acid mixture; IP, immediately after exercise; P15, 15 min after exercise;
P30, 30 min after exercise; PLA, placebo; Pre, before exercise.
Serum glucose
Figure 2 shows the serum glucose response during and
after a single bout of resistance exercise. The PLA group
showed no significant change in serum glucose concentra-
tion to the exercise bout from the pre-exercise value (4.6
0.1 mM) or up to P30 (4.2 0.1 mM). The EAA group
displayed a trend similar to that of the PLA group. Serum
glucose remained statistically unchanged from the pre-
exercise value (4.5 0.1 mM) throughout the exercise bout
and up to P30 (4.2 0.2 mM).
Conversely, CHO and CHO EAA ingestion during the
exercise bout resulted in significantly increased (P 0.05)
serum glucose levels of 31% and 40%, respectively, with all
time points significantly higher than pre-exercise levels.
The increase in glucose for the CHO group occurred within
15 min of commencing the exercise bout, peaked IP (5.6
0.2 mM), and remained significantly higher than the pre-
exercise value of 4.3 0.1 mM for at least P30 (5.1 0.1
mM, P 0.05). CHO ingestion also resulted in significantly
higher glucose levels compared with PLA at IP, P15, and
P30 and with EAA IP and P15 (P 0.05). The response of
the CHO EAA group mirrored that of the CHO group.
The CHO EAA group displayed a significant increase in
glucose within 15 min, peaked IP (6.1 0.1 mM), and
remained significantly higher than pre-exercise levels for at
least P30 (4.4 0.1 versus 5.5 0.4 mM, P 0.05). In
addition, the effect of CHO EAA ingestion compared
with that of PLA and EAA was pronounced, resulting in
significantly higher glucose levels at 30 min, 45 min, IP,
P15, and P30 (P 0.05).
371
a b
*
25 *
Insulin (lU/mL)
20
b
*
10
5
Exercise Recovery
0
Pre 15 30 45 IP P15 P30
Time (mins)
Fig. 3. Serum insulin concentrations for PLA (), CHO (
), EAA ( ),
and CHO EAA (). Significantly different from pre-exercise value (P
0.05): *CHO versus IP, P15, and P30; EAA versus P15 and P30; CHO EAA
versus IP, P15, and P30. Significant difference between treatments (P 0.05):
a
CHO from PLA; bCHO EAA from PLA. CHO, 6% carbohydrate solution;
EAA, 6-g essential amino acid mixture; IP, immediately after exercise; P15, 15
min after exercise; P30, 30 min after exercise; PLA, placebo; Pre, before
exercise.
Serum insulin
Figure 3 shows the serum insulin response during and
after a single bout of resistance exercise. The PLA group
showed no significant change in serum insulin concentration
to the exercise bout from the pre-exercise value (8.8 1.2
IU/mL), with a slight, but non-significant, decrease up to
P30 (8.1 0.8 IU/mL). The EAA group displayed sig-
nificant increases in insulin concentration relative to the
pre-exercise value (8.3 3.2 IU/mL) and at P15 and P30
(17.8 3.7 IU/mL, P 0.001; 14.0 2.1 IU/mL, P
0.05, respectively).
In contrast, serum insulin concentrations for the CHO
and CHO EAA groups reflected changes in glucose
concentrations. After the CHO intervention there was an
appreciable increase in insulin concentration after com-
mencement of the exercise bout relative to the pre-exercise
value (7.9 0.7 IU/mL), with significant increases noted
IP (16.0 1.6 IU/mL, P 0.01), followed by a peak
increase of 170% at P15 (21.2 3.5 IU/mL, P 0.001);
thereafter, insulin remained significantly higher for at least
P30 (20.3 2.5 IU/mL, P 0.001). CHO ingestion
resulted in significantly higher insulin levels than occurred
with PLA IP, P15, and P30 (P 0.05). The CHO EAA
treatment resulted in a pronounced increase in insulin
throughout the exercise bout compared with the pre-exer-
cise value of 7.6 0.6 IU/mL. Significant increases oc-
curred by the end of the exercise bout (IP; 17.3 0.8
IU/mL, P 0.01), with this response continuing into the
recovery period. Insulin remained significantly higher at
P15 (21.0 1.6 IU/mL, P 0.001), finishing 189%
above the pre-exercise value at P30 (22.0 4.5 IU/mL, P
 372 S.P. Bird et al. / Nutrition 22 (2006) 367375
a 20.0
b
700 a *
b *
*
a
a *
600 b
Cortisol (nmol/L)
b
* 15.0
* *
500
400
300
200
Exercise Recovery
100
Pre 15 30 45 IP P15 P30
Time (mins)
Fig. 4. Serum cortisol concentrations for PLA (), CHO ( ), EAA (),
and CHO EAA (). Significantly different from pre-exercise value (P
0.05): *PLA versus 30 min, 45 min, IP, P15, and P30; CHO versus P30;
CHO EAA versus P30. Significant difference between treatments (P 0.05):
a
CHO from PLA; bCHO EAA from PLA. CHO, 6% carbohydrate solution;
EAA, 6-g essential amino acid mixture; IP, immediately after exercise; P15, 15
min after exercise; P30, 30 min after exercise; PLA, placebo; Pre, before
exercise.
0.001). It is noteworthy that insulin appeared to still be
increasing after this time point. CHO EAA consumption
resulted in significantly higher insulin levels than did PLA
IP, P15, and P30 (P 0.05).
Serum cortisol
Figure 4 shows the serum cortisol response during and
after a single bout of resistance exercise. With ingestion of
the PLA beverage, cortisol increased immediately after
commencement of the exercise bout, with values for all time
points higher than the pre-exercise value (294.1 30.8
nmol/L). Significant increases in cortisol occurred within 30
min of initiating the exercise bout (473.6 52.1 nmol/L, P
0.01), with a peak increase of 105% IP (602.4 60.4
nmol/L, P 0.001). Cortisol remained 54% above the
pre-exercise value at P30 (452.3 69.1 nmol/L, P 0.05).
In contrast, the EAA group showed no significant change in
cortisol concentration to the exercise bout from the pre-
exercise value (294.4 33.4 nmol/L).
Consequent to the CHO and CHO EAA treatments,
the cortisol response was blunted, resulting in a significant
decrease before to after exercise. Both groups displayed a
noticeable decrease in cortisol concentration during the ex-
ercise bout compared with pre-exercise values (303.1
45.7 and 317.1 27.7 nmol/L, respectively). This decrease
continued during the recovery period, finishing 27% and
23%, respectively, below the pre-exercise value at P30
(221.5 45.8 nmol/L, P 0.01; 245.4.1 23.2 nmol/L, P
0.05, respectively). This blunted response was associated
with significantly lower cortisol levels compared with PLA
*
Testosterone (nmol/L)
17.5
*
12.5 *
10.0
7.5
Exercise Recovery
5.0
Pre 15 30 45 IP P15 P30
Time (min)
Fig. 5. Serum testosterone concentrations for PLA (), CHO (
), EAA
( ), and CHO EAA (). Significantly different from pre-exercise value (P
0.05): *PLA versus 15 min and 30 min; CHO versus 15 min and P30; EAA
versus 15 min, 30 min, 45 min, and IP; CHO EAA versus 15 min and 30 min.
CHO, 6% carbohydrate solution; EAA, 6-g essential amino acid mixture; IP,
immediately after exercise; P15, 15 min after exercise; P30, 30 min after
exercise; PLA, placebo; Pre, before exercise.
at 45 min, IP, P15, and P30, with the greatest difference
occurring IP (602.4 60.4 nmol/L versus 268.5 45.9 and
295.5 31.4 nmol/L, respectively).
Serum total testosterone
Figure 5 shows the serum total testosterone response
during and after a single bout of resistance exercise. All
groups displayed an acute increase in total testosterone
immediately after the initiation of the exercise bout;
opposing responses were noted for the PLA and EAA
groups versus the CHO and CHO EAA groups. The
PLA group demonstrated significant increases in total
testosterone compared with the pre-exercise value (10.1
1.2 nmol/L) and at P15 and P30 (12.2 1.8 nmol/L,
P 0.05; 12.5 1.9 nmol/L, P 0.01, respectively).
Thereafter, total testosterone remained statistically un-
changed from the pre-exercise value for all remaining
time points. The EAA group displayed a trend similar to
that of the PLA group. Total testosterone values in-
creased significantly at all time points during the exercise
bout compared with the pre-exercise value (9.1 1.0
nmol/L), with a peak increase of 32% at 45 min (12.3
1.8 nmol/L, P 0.001), and returned to the pre-exercise
value at P30 (9.2 1.2 nmol/L).
Subsequent to the CHO and CHO EAA treatments,
both groups demonstrated a significant increase in total
testosterone 15 min after the initiation of exercise (15.1
1.1, P 0.01; 15.9 2.4 nmol/L, P 0.01, respectively)
compared with pre-exercise values (12.9 1.2 and 12.9
1.8 nmol/L, respectively). CHO ingestion resulted in a sig-
nificant decrease before to after exercise (P30; 10.2 1.0
nmol/L, P 0.001). Total testosterone for the CHO EAA
 S.P. Bird et al. / Nutrition 22 (2006) 367375
75
Growth Hormone (mlU/L)
*
60 * monal milieu conducive to enhance skeletal muscle hyper-
* trophic adaptation to resistance training. The primary find-
45 * ings from this investigation were that performing a single
30
15
0 Exercise Recovery
Pre 15 30 45 IP P15 P30
Time (min)
Fig. 6. Serum growth hormone concentrations for PLA (), CHO (
),
EAA ( ), and CHO EAA (). Significantly different from pre-exercise
value (P 0.05): *PLA versus 30 min and 45 min; CHO versus 15 min, 30 min,
45 min, and IP; EAA versus 15 min, 30 min, 45 min, and IP; CHO EAA
versus 15 min, 30 min, 45 min, and IP. CHO, 6% carbohydrate solution; EAA,
6-g essential amino acid mixture; IP, immediately after exercise; P15, 15 min
after exercise; P30, 30 min after exercise; PLA, placebo; Pre, before exercise.
group remained statistically unchanged from the pre-
exercise value throughout this time. No group-by-time in-
teractions were present for total testosterone.
Serum growth hormone
Figure 6 shows the serum growth hormone response during
and after a single bout of resistance exercise. The resistance
exercise protocol resulted in a dramatic increase in growth
hormone above the pre-exercise value in all treatment groups
(PLA 1.0 0.4 mIU/L; EAA 1.3 0.3 mIU/L; CHO 1.4
0.5 mIU/L; CHO EAA 2.0 0.7 mIU/L). Significant
increases occurred within 15 min of initiation of the exercise
bout, with peak increases occurring at 30 min for the PLA,
EAA, and CHO EAA groups (36.1 11.1 mIU/L, P
0.01; 54.9 9.2 mIU/L, P 0.001; 40.7 9.1 mIU/L, P
0.001, respectively) and 45 min for the CHO group (44.2
13.7 mIU/L, P 0.001) compared with the pre-exercise value.
This increase remained significant IP for EAA (41.7 8.9
mIU/L, P 0.001), CHO (35.2 15.2 mIU/L, P 0.01), and
CHO EAA (30.4 8.3 mIU/L, P 0.01). Growth hormone
concentrations remained statistically unchanged from the pre-
exercise level during the recovery period. No group-by-time
interactions were present for growth hormone.
Discussion
The present investigation examined the influence of nu-
tritive interventions (a 6% CHO solution, a 6-g EAA mix-
ture, or a combined CHO EAA beverage) on acute
biochemical responses to a single bout of resistance exercise
in untrained young men. Specifically, the objective was to
373
determine whether exercise-induced cortisol release could
be attenuated by such interventions, thereby creating a hor-
bout of resistance exercise in conjunction with liquid CHO
ingestion resulted in significantly higher insulin concentra-
tions and a significant decrease in cortisol after exercise.
Moreover, such responses were amplified when the treat-
ments were combined (CHO EAA), thus creating a hor-
monal milieu associated with anabolism. These results are
in contrast to the non-significant change in insulin concen-
tration and significantly higher cortisol levels shown by the
PLA group.
Excessive levels of cortisol elicit several physiologic
responses, some of which may be detrimental to the positive
adaptations frequently attributed to resistance training [24],
namely improving nitrogen balance and increasing muscle
mass and strength. A primary function of increased cortisol
secretion has been reported to accelerate gluconeogenesis
[27]. Because of this glucose-regulatory action, it was hy-
pothesized that, if an individual ingested a CHO solution
while exercising, the exogenous glucose load would in-
crease blood glucose levels. In turn, the increased blood
glucose level would inhibit the stimulus for the adrenal
cortex to secrete cortisol to catabolize cellular protein for
gluconeogenic purposes. Moreover, this response may be
augmented by the addition of EAA, thus amplifying the
acute biochemical signals associated with resistance exer-
cise.
In the present experiment, the exercise-induced cortisol
response was somewhat antagonistic to that of glucose (i.e.,
when glucose levels were unchanged, cortisol concentration
increased, and when glucose levels were increased, cortisol
concentration decreased). The PLA group exhibited a peak
cortisol increase of 105% (P 0.001) IP, which corre-
sponded with no change in glucose levels (7%). Similar
increases in cortisol have been previously reported in un-
trained men [22,28,29]. Conversely, resistance exercise per-
formed in conjunction with CHO and CHO EAA con-
sumption resulted in a blunted cortisol response. Time
matched for PLA, the CHO and CHO EAA groups
showed decreases in cortisol of 11% and 7% IP, respec-
tively, with this decrease becoming significant at P30 (27%,
P 0.01; 23%, P 0.05, respectively). In addition, this
corresponded with significant increases (P 0.001) in
glucose levels of 31% and 40%, respectively. These data
provide evidence in support of the hypothesis by demon-
strating that the stimulatory effect of resistance exercise on
cortisol release can be overcome by CHO and/or
CHO EAA ingestion, with this response mediated, at
least to some extent, by higher blood glucose levels. Such
findings are in agreement with those reported by Tarpenning
et al. [10].
In contrast to the catabolic effects of cortisol, the ana-
bolic effects of insulin on muscle include stimulating amino
374 S.P. Bird et al. / Nutrition 22 (2006) 367375
acid transport and protein synthesis [18,30] and inhibiting
protein degradation [31]. According to Goldberg [32] a key
signaling event in the hypertrophic response of skeletal
muscle is an increased rate of amino acid transport, and this
directly corresponds to muscle contractile activity and in-
sulin concentrations. However, an acute bout of resistance
exercise is generally accompanied by a decrease in insulin
levels [11,33,34]. Moreover, it has been observed that in-
gestion of amino acid mixtures alone [35] or in combination
with CHO [36] can produce strong insulinotropic effects.
Such a response may increase the exercise-induced insulin
response after resistance exercise. In theory, increasing in-
sulin could influence the subsequent phase of recovery by
modulating anabolic and catabolic processes [37].
The insulin response in the present experiment mir-
rored changes in glucose kinetics (i.e., when glucose
levels were unchanged, insulin levels were unchanged,
and when glucose levels were increased, insulin levels
were increased). The PLA group showed a minor de-
crease in insulin (8%) and glucose (9%) concentrations
before to after exercise. However, ingestion of CHO
and/or EAA demonstrated a stimulatory effect on insulin
response, with the CHO and CHO EAA groups dis-
playing significantly increased insulin and glucose levels.
CHO consumption resulted in an approximately 2.5-fold
increase in insulin concentration. Further, addition of
EAA (CHO EAA) improved insulin responsiveness
approximately 3.0-fold. These findings are in agreement
with those of several reports [36 38], indicating that
inclusion of EAA with CHO acts in a synergistic fashion
to enhance the acute insulin response after resistance
exercise. It is noteworthy that the EAA group displayed
an approximately 2.0-fold increase in insulin concentra-
tion during the recovery period. Interestingly, this was
associated with a minor decrease in glucose (6%). Col-
lectively, these findings indicate that the inhibitory effect
of resistance exercise on insulin secretion can be over-
come by nutritive interventions. Although measurements
of protein turnover were not determined in this investi-
gation, it is possible that the modified hormonal response
associated with CHO EAA ingestion, that of increased
insulin and decreased cortisol, resulted in a hormonal
profile more favorable for positive nitrogen balance and
protein accretion.
In summary, the findings of the present investigation
suggest that CHO ingestion during the exercise bout can
suppress exercise-induced cortisol release, thereby altering
the balance between hormone-mediated anabolic and cata-
bolic activities. Further, such biochemical responses are
augmented with the addition of EAA (CHO EAA) toward
a profile more favorable for anabolism. However, the bio-
chemical events associated with resistance exercise and
nutrient status, and by which this phenomenon regulates
protein degradation, are yet to be elucidated.
Acknowledgments
The authors thank the dedicated group of subjects who
participated in this project. They also thank Brian Heffernan
and Matthew ONeal at the Central West Pathology Service,
Bathurst Base Hospital, NSW, Australia, and Dr. Gary Ma,
Trudi Jones, and Vicki Pitsiavas at the Institute of Clinical
Pathology and Medical Research, Laboratory of Endocri-
nology, Westmead Hospital, NSW, Australia, for technical
assistance with the immunoassay procedures. They also
thank Catherine Offner for nutritional support and dietary
analysis.
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