P.

Paccotti1
M. Minetto1
M. Terzolo1
M. Ventura1
G. P. Ganzit2
Effects of High-Intensity Isokinetic Exercise on Salivary
P. Borrione1
A. Termine1 Cortisol in Athletes with Different Training Schedules:
A. Angeli1 Relationships to Serum Cortisol and Lactate

Training & Testing

Abstract
Physical exercise is associated with increases of serum and sali-
vary levels of cortisol. The concomitant increase in serum lactate
has been implicated as one of the mechanisms responsible for
adrenocortical activation. We evaluated the responses of serum
lactate and serum and salivary cortisol to an acute bout of high-
intensity isokinetic exercise in eleven non-competitive and
twenty competitive athletes (NCA and CA, respectively). The lat-
ter group was composed of endurance- and power-trained ath-
letes (EA and PA, respectively). Aims of the study were to deter-
mine interindividual differences in the lactate and cortisol re-
sponses as a function of type and intensity of training and to
search for relationships both between lactate and cortisol pro-
duction and between serum and salivary cortisol levels.
The isokinetic exercise test elicited significant cortisol and lac-
tate responses. No difference was evident in the lactate re-
sponses between NCA and CA, while the PA showed a higher re-
sponse during and after the exercise in comparison to EA (peak
levels immediately after the exercise: PA 15.0 ± 1.5 mmol/l vs. EA
11.1 ± 2.6 mmol/l, p < 0.01). Serum cortisol was higher in the CA
in comparison to the NCA group at 30 and 120 minutes after the
termination of the exercise, while no differential response was
evident between EA and PA groups. Salivary cortisol response
was higher in the CA group in comparison to NCA immediately
after the exercise and at 90 and 120 minutes after the termina-
tion and was higher in PA in comparison to EA at 60, 90, and
120 minutes after the termination (peak levels at 60 minutes:
PA 51.2 ± 18.5 nmol/l vs. EA 27.5 ± 20.8 nmol/l, p < 0.05). No sig-
nificant correlations were found between serum or salivary cor-
tisol and lactate levels. The relationship between serum and sali-
vary cortisol was markedly non-linear, the slope of the serum-
saliva regression line being lower for serum cortisol concentra-
tions over 500 nmol/l than for concentrations below that value
(0.019 and 0.037, respectively, p < 0.01).
We have confirmed in this particular setting the existence of an
important adrenocortical response that can be reliably and non
invasively assessed by a serial saliva sampling and have sup-
ported the concept that cortisol and lactate responses to a high-
intensity isokinetic exercise are independent. The interindivid-
ual differences in cortisol changes are likely to be related to the
training status and mode as well as to the correspondence be- 747
tween the evaluation protocol and the discipline individually
performed.
Key words
Hypothalamic-pituitary-adrenal (HPA) axis · strength exercise ·
saliva

Introduction paraventricular nucleus in the hypothalamus, which secrete cor-

ticotrophin-releasing hormone and vasopressin, hence activate
Stress is a term defining a physiological response to events per- the hypothalamic-pituitary-adrenal (HPA) axis and eventually
ceived as potentially or actually threatening the body’s integrity. increase the cortisol output from the adrenal glands. Strenuous
Among the major effectors of this response are the neurons of the exercise is a prototype of physical stressor. Acute intense aerobic

Affiliation
1
Clinica Medica Generale, Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, Italy
2
Istituto di Medicina dello Sport di Torino, Torino, Italy

Correspondence
P. Paccotti · Clinica Medica Generale · ASO San Luigi, Regione Gonzole 10 · 10043 Orbassano (TO) · Italy ·
Phone: + 39 0119 02 6514 · Fax: + 39 0119 03 86 55 · E-mail: piero.paccotti@unito.it

Accepted after revision: September 30, 2004

Bibliography
Int J Sports Med 2005; 26: 747 – 755 © Georg Thieme Verlag KG · Stuttgart · New York ·
DOI 10.1055/s-2004-830449 · Published online February 2, 2005 ·
ISSN 0172-4622
 exercise that exceeds 60% of the maximum aerobic power
[9,10, 25] and anaerobic exercise above the maximum oxygen
uptake as well [4] admittedly enhance the blood levels of corti-
sol. Several factors have been identified to influence the adreno-
cortical response, notably type and timing of exercise. In fact,
anaerobic exercise has been shown to induce a greater increase
in serum cortisol than aerobic exercise of comparable total work
output [13]. With regard to resistance exercise, the duration and
number of repetitions in each set and the duration of the interset
rest period were found to be the primary determinants of the
cortisol response [14,17, 21]. Moreover, peak cortisol concentra-
tions during exercise are higher when it is done in the morning
than in the evening or night [18]. The mechanisms by which
physical exercise activates the HPA axis have not been fully elu-
Training & Testing
cidated. Increases in serum lactate have been implicated as one
of the mechanisms responsible for the HPA activation during ex-
ercise [25] but remains unclear at which level of HPA axis lactate
could be functionally active. Petrides et al. have provided exper-
imental evidence that lactate does not play a direct role at the pi-
tuitary level [30].
Serum cortisol is 90 to 97 percent bound to plasma proteins,
mostly to corticosteroid-binding globulin (CBG), which binds
cortisol specifically and with high affinity. Salivary cortisol was
found to reflect the unbound biologically active fraction of serum
cortisol both in normal individuals and in women with elevated
CBG [39]. The correlation between salivary and serum “free” cor-
tisol was excellent in dynamic tests for the assessment of HPA
axis both in healthy subjects and in patients with adrenal insuf-
ficiency [24, 29, 39] as well as in randomly collected samples at
different phases of the circadian cycle [1, 29, 32]. Clearly, the rela-
tionship between salivary and “total” serum cortisol levels is
748 non-linear, with a more rapid increase in salivary concentrations
once the serum CBG is saturated [1, 26, 39].
In the last decade, several publications have appeared concern-
ing the use of salivary cortisol for assessing the adrenocortical re-
sponse to exercise. These studies have examined the hormonal
changes in response to exercise testing on treadmill [33, 35, 37]
or cycle ergometer [2, 6,16, 23, 27, 31, 33, 35] or kayak ergometer
[35, 36] (incremental tests to volitional exhaustion or constant
intensity exercises or short-duration anaerobic tests). To our
knowledge, no studies have provided evidence of a significant
correlation between saliva and serum in the cortisol response to
an acute bout of high-intensity strength exercise. Moreover, no
data about such correlation are available in relation to the isoki-
netic muscle performance. This type of muscle contraction has
become a popular method to assess dynamic muscle function
with the interfacing of an isokinetic dynamometer. This device
provides constant velocity with accommodating resistance
throughout a joint’s range of motion. We focused our interest on
this type of exercise in order to obtain objective measures of
muscle function on variables related to torque and mechanical
fatigue in different groups of athletes and in order to accurately
verify the clustering between the different groups, based on the
information of the discipline individually performed. Aims of the
present study were to determine interindividual differences in
the lactate and cortisol responses to an acute bout of high-inten-
sity isokinetic exercise as a function of type and intensity of
training; to evaluate the relationship between cortisol and lac-
Paccotti P et al. Effects of High-Intensity Isokinetic Exercise … Int J Sports Med 2005; 26: 747 – 755
tate responses; to assess the correlation between serum and sali-
vary cortisol responses.
Materials and Methods
Subjects and exercise protocol
Twenty competitive male athletes (13 endurance-trained and 7
power-trained) and eleven non-competitive male athletes (Table
1 summarizes their characteristics) participated in the study.
The detail of the disciplines for the former group is the following:
three marathon runners; one long-distance cyclist; one triath-
lete; seven rowers; one cross-country skier; three alpine skiers;
three body builders; one volleyball player. The athletes were all
at the national or international level and they were tested during
the training out-of-competition period of their season.
The detail of the disciplines for the latter group is the following:
three runners; two body builders; two volleyball players; one
basketball player; three soccer players.
None of the subjects was a current smoker, or was receiving any
medication. They were instructed to abstain from caffeine and
alcohol consumption and to refrain from any strenuous physical
activity for 24 hours before the study. Testing was always per-
formed in the afternoon (1600 h) after 3 hours from a standard-
ized meal. A complete medical examination was performed on
each subject; the probands were thoroughly informed about the
purposes and procedures of the study before giving a written
consent. The study conformed with the guidelines in the Decla-
ration of Helsinki and was approved by the Regional Ethics Com-
mittee.
We submitted the athletes to an isokinetic exercise test on a Cy-
bex 6000 device (Cybex, Division of Lumex Inc., Ronkonkoma,
USA). After the completion of a warm-up exercise on a bicycle er-
gometer (consisting of 15 minutes cycling at 75 – 100 W and
stretching of the muscle groups subsequently involved in exer-
cise), they fulfilled for each leg four sets of twenty maximal con-
tractions of the knee flexor and extensor muscle groups at 1808/
Table 1 Mean ± SD of age, weight, height, training history and vol-
ume (over the last three years) in the two groups of com-
petitive athletes (CA: 13 endurance-trained and 7 power-
trained, EA and PA, respectively) and non-competitive ath-
letes (NCA)
CA NCA (n = 11)
EA (n = 13) PA (n = 7)
Age (years) 24.9 ± 7.6 33.0 ± 6.0 25.3 ± 5.4
Weight (kg) 76.6 ± 8.1 85.0 ± 7.1 74.1 ± 6.7
Height (m) 1.83 ± 0.10 1.85 ± 0.11 1.79 ± 0.10
Training history 6.8 ± 5.2 11.4 ± 3.9 7.9 ± 3.1
(years)
Training volume 14.8 ± 2.6 13.1 ± 2.7 4.5 ± 1.9
(hours/week)
Fig. 1 Schematic representation of the isokinetic exercise. Four sets
of twenty maximal contractions of the knee flexor and extensor
muscle groups were performed for each leg. The rates of change of
the regression line (dotted line) fitting the A1, A2, A3, and A4 points
sec angular velocity throughout a constant range of motion
(1008), starting from the dominant leg. There was a 30-second
rest period between each set and a 3-minute rest period between
the two legs (Int). The complete exercise routine (a total of 160
maximal contractions) was completed in about 15 minutes. An
antecubital venous catheter was placed 30 minutes before start-
ing the warm-up. Blood for hormonal evaluation was sampled 15
and 5 minutes before the warm-up (Pre), immediately after ter-
mination of the test (Post), and at + 7, + 15, + 30, + 45, + 60, + 90,
+ 120 minutes thereafter. Saliva was collected at the same time,
by using cotton salivettes (Sarstedt, Nümbrecht, Germany).
Blood for lactate determination was sampled 15 and 5 minutes
before the warm-up (Pre), during the test (Int), immediately after
termination of the test (Post), and at + 30, + 60, + 120 minutes
thereafter.
The average of the initial two blood and salivary samples for each
subject was taken to establish baseline pre-exercise serum and
salivary cortisol and lactate levels.
Saliva was also collected at 0800 h, 1600 h, and 2400 h in a large
group of healthy individuals (n = 110; men 58, women 52; age
35.3 ± 12.5 yr and 38.2 ± 12.7 yr; BMI 24.7 ± 3.2 kg/m2 and
22.5 ± 3.5 kg/m2, respectively) in order to establish reference
ranges for cortisol in saliva and to compare the pre-exercise sali-
vary cortisol levels in the athletes with the afternoon salivary
cortisol levels in the control group.
Assays
Serum lactate concentrations were measured spectrophotomet-
rically by an Aeroset analyser (Abbott Laboratories, Abbott Park,
IL, USA). Serum cortisol was measured by radioimmunoassay
(Sorin Biomedica, Saluggia, Italy), salivary cortisol was measured
by radioimmunoassay (Radim SpA, Pomezia, Italy), on a 1272
CliniGamma gamma counter (LKB Wallac, Turku, Finland). All
the samples of any subject were processed in duplicate in the
same assay session. In order to separate serum and to extract sa-
liva, blood samples and salivettes were centrifuged at 1000 g for
15 minutes at room temperature and then stored at – 20 8C until
assayed. All salivary samples were subjected to a single freeze–
thaw cycle and on the day of the assay were centrifuged at
1000 g for 15 min to remove mucopolysaccharides that can inter-
fere with pipetting [40].
Paccotti P et al. Effects of High-Intensity Isokinetic Exercise … Int J Sports Med 2005; 26: 747 – 755
(i.e. the intercepts of the regression lines – solid lines – within each of
the four sets) were calculated for the dominant leg. SLOPEISK unit is de-
fined as Nm/set. MVCISK is defined as the point A1 and the correspond-
ing unit is Nm.
Training & Testing
Calculated sensitivities of the assays were 13.8 nmol/l for serum
cortisol, 2.7 nmol/l for salivary cortisol, and 0.9 mmol/l for lac-
tate. Intra- and inter-assay coefficients of variation for all the
above-mentioned assays were below 6 % and 10%, respectively.
Data management and statistical analysis
The studied mechanical variables were: 1) the peak extension
torque produced during the isokinetic exercise (MVCISK) by the
dominant leg, and 2) the rate of change (SLOPEISK) of the maximal
voluntary extension torque produced during the whole isoki-
netic exercise by the dominant leg. The intercept of the regres-
sion over the twenty isokinetic contractions during the first set
of the dominant leg was selected as MVCISK (point A1 in Fig. 1).
For each of the four isokinetic sets, performed with the dominant
leg, a regression among the four extension peak torque values
during the test was done and the intercept calculated (A1, A2, 749
A3, and A4). SLOPEISK was obtained as the slope of the regression
line fitted over the four intercepts, hence providing a description
of the whole fatiguing exercise (Fig. 1).
Normal distribution of the data was tested by the Shapiro-Wilk
test. Because data were not normally distributed the non para-
metric Wilcoxon paired test was used to determine whether
there were any significant differences within each group over
time among the samples. The differences between the groups
(Competitive Athletes [CA] vs. Non-Competitive Athletes [NCA];
Endurance-trained Athletes [EA] vs. Power-trained Athletes [PA])
were analyzed at each time point by means of the Mann-Whit-
ney U test. The area under the response curve (AUC) was calcu-
lated according to the trapezoid method and the comparisons
between the groups were made by means of the Mann-Whitney
U test. The relationship between serum and salivary cortisol val-
ues was evaluated by means of Spearman rank correlation anal-
ysis, applied both to raw data and after logarithmic transforma-
tion. Also the relationships between cortisol and lactate levels
and between mechanical variables and lactate response were
evaluated by means of Spearman rank correlation analysis. The
Kruskal-Wallis test followed by the Dunn’s post hoc test was
used to compare salivary cortisol levels from healthy controls in
the three different time-points and Mann-Whitney U test was
used to compare the pre-exercise salivary cortisol values in the
athletes and the afternoon salivary cortisol levels in the controls.
 Table 2 Cortisol and lactate responses to isokinetic exercise in competitive athletes (CA, n = 20) and non-competitive athletes (NCA, n = 11).
Serum and salivary cortisol AUCs units: nmol/l · 150 minutes; Lactate AUC units: mmol/l · 150 minutes. The last columns report the
significances obtained in the comparison of each value with respect to pre-exercise levels and those obtained in the comparison of
the two groups

Competitive athletes Non-competitive athletes Within group p Between group p

Mean SD Mean SD CA NCA CA vs. NCA

Serum cortisol (nmol/l)

Pre 526.9 228.5 443.5 257.8 – – ns
Post 673.7 313.8 522.5 361.5 < 0.05 ns ns
+7 819.2 398.2 611.2 386.3 < 0.001 ns ns
+ 15 807.3 288.2 671.3 366.0 < 0.001 < 0.05 ns
Training & Testing

+ 30 893.8 342.2 606.6 392.8 < 0.001 ns < 0.05

+ 45 856.4 390.5 548.4 393.7 < 0.01 ns ns
+ 60 740.6 323.9 484.0 375.2 < 0.01 ns ns
+ 90 714.8 408.7 424.8 354.0 ns ns ns
+ 120 567.8 242.3 338.6 303.8 ns ns < 0.05
AUC 41779.5 25769.1 33199.0 21630.6 ns
Salivary cortisol (nmol/l)
Pre 12.5 9.1 10.2 9.8 – – ns
Post 22.1 16.4 10.7 5.9 < 0.001 ns < 0.05
+7 22.9 15.9 13.9 8.2 < 0.001 ns ns
+ 15 26.5 16.7 17.9 10.8 < 0.01 ns ns
+ 30 31.1 19.9 22.4 16.9 < 0.001 < 0.05 ns
+ 45 33.4 19.9 21.8 19.4 < 0.001 ns ns
+ 60 35.8 22.7 22.8 25.6 < 0.001 ns ns
+ 90 29.2 19.8 17.1 20.0 < 0.001 ns < 0.05
+ 120 22.2 16.2 11.7 12.1 < 0.01 ns < 0.05
AUC 2308.4 1665.6 1617.9 1851.0 ns
750 Lactate (mmol/l)
Pre 1.2 0.4 1.3 0.6 – – ns
Int 11.7 3.4 11.0 4.2 < 0.001 < 0. 01 ns
Post 12.4 3.0 12.2 2.6 < 0.001 < 0.01 ns
+ 30 6.8 2.7 6.4 4.0 < 0.001 < 0.01 ns
+ 60 3.7 1.4 3.2 1.4 < 0.001 < 0.01 ns
+ 120 2.0 0.6 2.0 0.8 < 0.001 < 0.05 ns
AUC 638.7 238.9 579.8 271.9 ns

Statistical analysis was performed with the Statistica 6 for Win-
dows (Statsoft Inc., Tulsa, OK, USA) software package. All values
are reported as mean ± SD. Statistical significance was set at
p < 0.05.
Results
The isokinetic exercise test was completed by the whole group of
athletes and elicited significant cortisol and lactate responses
both in the CA and in the NCA groups (see Tables 2 and 3 for de-
tails). Serum cortisol was found higher in the CA group at + 30
minutes and + 120 minutes post-exercise in comparison to the
NCA group (Fig. 2), while no differential response was evident
between EA and PA groups (Fig. 3). Salivary cortisol response
was higher in the CA group at Post, + 90 minutes, and + 120 min-
utes post-exercise in comparison to NCA (Fig. 2), and was higher
in the PA group at + 60 minutes, + 90 minutes, and + 120 minutes
in comparison to EA (Fig. 3). No difference was evident in the
lactate responses between CA and NCA (Fig. 2), while the PA
showed a higher response at each time-point in comparison to
EA (Fig. 3). No difference was evident between CA and NCA both
in the SLOPEISK (CA: – 11.96 ± 11.88 Nm/set vs. NCA: – 14.25 ±
9.82 Nm/set, p = ns) and in the MVCISK (CA: 159.2 ± 35.4 Nm vs.
NCA: 140.8 ± 40.7 Nm, p = ns), while the PA showed a lower
SLOPEISK value (PA: – 28.92 ± 4.06 Nm/set vs. EA: – 6.31 ±
6.96 Nm/set, p < 0.01) and a higher MVCISK value (PA: 191.4 ±
43.4 Nm vs. EA: 143.1 ± 15.7 Nm, p < 0.05) in comparison to EA.
The correlation analysis showed significant relationships be-
tween the SLOPEISK and both the lactate peak levels (r = – 0.68,
p < 0.01) and the peak-pre-exercise delta (r = – 0.66, p < 0.01), as

Paccotti P et al. Effects of High-Intensity Isokinetic Exercise … Int J Sports Med 2005; 26: 747 – 755
 Table 3 Cortisol and lactate responses to isokinetic exercise in endurance-trained competitive athletes (EA, n = 13) and power-trained com-
petitive athletes (PA, n = 7). Serum and salivary cortisol AUCs units: nmol/l · 150 minutes; lactate AUC units: mmol/l · 150 minutes. The
last columns report the significances obtained in the comparison of each value with respect to pre-exercise levels and those obtained
in the comparison of the two groups

Endurance athletes Power athletes Within group p Between group p

Mean SD Mean SD EA PA EA vs. PA

Serum cortisol (nmol/l)

Pre 573.3 226.2 437.1 220.2 – – ns
Post 714.0 343.9 598.9 255.3 ns < 0.05 ns
+7 857.7 466.8 747.7 238.8 < 0.05 < 0.05 ns
+ 15 775.4 331.4 866.4 192.4 < 0.05 < 0.05 ns

Training & Testing

+ 30 808.0 374.1 1053.0 213.2 < 0.05 < 0.05 ns
+ 45 800.8 443.1 959.6 266.9 ns < 0.05 ns
+ 60 684.3 327.2 845.1 313.8 ns < 0.05 ns
+ 90 702.9 463.3 736.9 314.2 ns < 0.05 ns
+ 120 516.3 237.2 663.4 238.8 ns ns ns
AUC 35952.5 27369.3 52600.3 19897.9 ns
Salivary cortisol (nmol/l)
Pre 10.7 5.0 16.0 13.8 – – ns
Post 20.7 14.6 24.5 20.5 < 0.01 < 0.05 ns
+7 22.0 14.5 24.6 19.4 < 0.01 < 0.05 ns
+ 15 25.4 16.3 28.5 18.5 < 0.01 ns ns
+ 30 28.8 19.2 35.4 21.9 < 0.01 < 0.05 ns
+ 45 29.5 21.3 40.6 15.7 < 0.01 < 0.05 ns
+ 60 27.5 20.8 51.2 18.5 < 0.01 < 0.05 < 0.05
+ 90 20.9 13.2 44.4 21.6 < 0.01 < 0.05 < 0.05
+ 120 15.6 10.7 34.6 18.3 < 0.05 < 0.05 < 0.05
AUC 1907.3 1457.0 2798.2 1182.8 0.055
Lactate (mmol/l) 751
Pre 1.2 0.5 1.1 0.2 – – ns
Int 10.4 2.8 14.3 3.0 < 0.01 < 0.05 < 0.05
Post 11.1 2.6 15.0 1.5 < 0.01 < 0.05 < 0.01
+ 30 5.6 2.4 9.1 1.7 < 0.01 < 0.05 < 0.01
+ 60 3.2 1.2 4.5 1.2 < 0.01 < 0.05 < 0.05
+ 120 1.7 0.5 2.5 0.6 < 0.01 < 0.05 < 0.01
AUC 528.5 202.8 843.4 152.4 < 0.01

well as between the MVCISK and both the lactate peak levels
(r = 0.64, p = 0.01) and the peak-pre-exercise delta (r = 0.63,
p = 0.01), while no correlations were found between serum or
salivary cortisol and lactate levels at each of the nine sampling
points.
The relationship between serum and salivary cortisol was mark-
edly non-linear, the slope of the serum-saliva regression line
being lower for serum cortisol concentrations over 500 nmol/l
than for concentrations below that value (0.019 and 0.037, re-
spectively, p < 0.01). After logarithmic transformation of raw data
a significant positive correlation was apparent (r = 0.62, p < 0.001,
Fig. 4 a): this linear relationship between the logarithmic values
involves the existence of an exponential relationship between
serum and salivary cortisol raw data (Fig. 4 b). Moreover, when
analysing each of the nine sampling points significant correla-
tions were observed between serum and salivary cortisol, except
for the + 30 minutes recovery time: the r values for these corre-
lations ranged between 0.40 and 0.71.
The salivary mean (± SD) cortisol levels evaluated in the control
group were as follows: 20.4 ± 7.7 nmol/l at 0800 h (range, the
central 95 %: 18.9 – 21.9 nmol/l); 9.2 ± 4.3 nmol/l at 1600 h (8.4 –
10.0 nmol/l); 5.5 ± 3.1 nmol/l at 2400 h (4.8 – 6.1 nmol/l). Signifi-
cant differences (p < 0.001) were observed between these three
values, while no difference was found between the pre-exercise
salivary cortisol values in the athletes and the afternoon salivary
cortisol levels in the controls.

Paccotti P et al. Effects of High-Intensity Isokinetic Exercise … Int J Sports Med 2005; 26: 747 – 755

Training & Testing
752
Discussion
In our athletes the isokinetic exercise elicited significant lactate
and cortisol responses. The lack of correlation between these two
biochemical markers suggests that the adrenocortical activation
was in this particular setting at least partly independent from
the muscular lactate production. In this respect, our data are
compatible with previous experimental observations [30], but
the following limitations deserve attention. The use of venous
blood appears suitable for determining the parameters for lac-
tate disappearance [28], but venous concentrations are less accu-
rate for assessing lactate production [12]. Moreover, blood sam-
pling was made at the times usually chosen for evaluating the
cortisol response to exercise. Also for this reason we could have
underestimated the peak levels of this chemical. Notwithstand-
ing this, the expected differences in peak and AUC levels between
athletes with different muscle performances were found, as dis-
cussed beyond. Finally, we must underlie the limits of a correla-
tion analysis made to study the relationship between two analy-
tes with different dynamics of response to the exercise: in fact,
the peak levels of cortisol were reached at + 15 minutes and + 30
minutes post-exercise for the serum levels and at + 60 minutes
Paccotti P et al. Effects of High-Intensity Isokinetic Exercise … Int J Sports Med 2005; 26: 747 – 755
Fig. 2 Serum and salivary cortisol and lac-
tate responses (and relative AUCs) to isoki-
netic exercise in competitive athletes (CA,
n = 20) and non-competitive athletes (NCA,
n = 11). The black stripe indicates the exer-
cise’s duration. Differences between groups:
* p < 0.05.
post-exercise for the salivary levels, at a time when lactate was
back almost completely.
We did not find differences in the lactate response between NCA
and CA groups: this could be explained by a comparable effort
raised by the exercise and by the heterogeneity of sports disci-
plines in both groups, conceivably requiring different muscular
adaptations. Serum and salivary cortisol responses resulted
higher in CA than in NCA: these data support the view that HPA
function has different degrees of adaptation to a high-volume
and high-intensity training schedule. It was suggested that adap-
tation to continuous or repeated exposure to stressful stimuli is
stressor-specific [15,19]. Kraemer et al. found that the HPA acti-
vation in relation to an acute bout of maximal endurance exer-
cise demonstrates a differential response to different training
programs (strength vs. endurance training) [22]. Also Tremblay
et al. have recently reported effects of training status and exer-
cise mode on the hormonal responses to various exercise tests
[38].
When analysing the lactate response in competitive athletes we
found higher lactate levels in PA than EA, negative correlations
between SLOPEISK and lactate and positive correlations between

MVCISK and lactate. Interestingly enough, it is agreed that in ath-
letes the muscle composition is also related to the mode and in-
tensity of training. Type I (“red”) and II (“white”) fibres have dif-
ferent properties: it is well-known that in all age groups a higher
oxidative and a lower glycolytic capacity is found in type I than in
type II fibres [3]. A higher torque expression, torque decrease
rate, and lactate production in PA is likely to be the consequence
of a higher proportion of type II fibres [3, 7, 8, 34]. The response of
salivary cortisol resulted higher in PA (individuals more accus-
tomed to strength exercise) in comparison to EA, while no differ-
ence was apparent in serum cortisol. Saturation of CBG sites at
serum cortisol levels above 500 – 600 nmol/l implies relatively
higher rises of the free, unbound, fraction. As a consequence,
slight differences of “total” cortisol concentrations in the blood-
stream could be faced by appreciable differences in saliva.
To the best of our knowledge, this is the first study that assesses
the relationship between saliva and serum in the cortisol re-
sponse to high-intensity isokinetic exercise: our finding of non-
linear relationship between salivary and serum cortisol confirms
and expands previous data, providing evidence of a clean-cut
slope variation of the serum-saliva regression line for serum cor-
tisol concentrations above 500 nmol/l, in agreement with Aardal
Paccotti P et al. Effects of High-Intensity Isokinetic Exercise … Int J Sports Med 2005; 26: 747 – 755
Fig. 3 Serum and salivary cortisol and lac-
tate responses (and relative AUCs) to isoki-
netic exercise in endurance-trained compet-
itive athletes (EA, n = 13) and power-trained
competitive athletes (PA, n = 7). The black
stripe indicates the exercise’s duration.
Differences between groups: * p < 0.05;
** p < 0.01.
Training & Testing
753
and Holm who reported the variation threshold for serum corti-
sol concentrations above 450 nmol/l [1].
Our data on morning, afternoon, and nighttime salivary cortisol
in healthy controls are in agreement with the previously report-
ed normality ranges by Laudat et al. (0800 h: 10.2 – 27.3 nmol/l;
2000 h: 2.2 – 4.1 nmol/l) [24], Aardal et al. (0800 h: 3.5 –
27.0 nmol/l; 2200 h: < 6.0 nmol/l) [1], Raff et al. (0700 h: 4.7 –
32.6 nmol/l; 2300 h: 0.4 – 3.6 nmol/l) [32].
In the present study serum cortisol and lactate concentrations
were not adjusted for changes in plasma volume. It is well-
known that exercise in humans is accompanied by a loss of plas-
ma fluid [5] and by a redistribution of regional and organ blood
volume with increased blood flow to the exercising muscles [11].
The former adaptation prevails in the case of submaximal pro-
longed exercises, the latter in maximal short-duration efforts.
For this reason the estimation of changes in plasma volume was
not made. Moreover, the observed changes in salivary cortisol
levels were not corrected for any exercise-induced variations in
salivary secretion rate and/or composition. It was ascertained,
in fact, that neither maximal stimulation of saliva flow nor mini-
mal secretion of saliva following medication with anticholinergic

Training & Testing
Fig. 4 a and b a Linear correlation between serum and salivary corti-
sol levels after logarithmic transformation of raw data in the whole
group of 31 athletes submitted to isokinetic exercise (r = 0.62,
p < 0.001). b Exponential correlation between serum and salivary cor-
tisol raw data in the whole group of 31 athletes (the dotted line is the
linear regression for serum cortisol concentrations below 500 nmol/l,
slope 0.037, r = 0.61, p < 0.001).
754
drugs (“dry mouth”) influence the concentration of cortisol in
saliva [20]. We made hormonal evaluations in absence of a rest-
ing control session; however, we performed the exercise session
under well standardized conditions at the chosen time of the cir-
cadian cycle, when serum cortisol levels are physiologically low
and relatively stable. In fact, no difference was evident between
the pre-exercise salivary cortisol levels of the athletes and the
corresponding levels at 1600 h of the controls.
In conclusion, we have demonstrated that the adrenocortical re-
sponse of athletes to a maximal exercise can be measured in a re-
liable and non invasive way by serial saliva sampling. The collec-
tion procedure is easy and applicable under a variety of field set-
tings, while activation of HPA axis, hence the increase of gluco-
corticoid secretion, is related to the sport discipline and training
status, as demonstrated in the present series by the higher re-
sponses of PA and CA in comparison to EA and NCA, respectively.
On the contrary, this HPA axis activation was found independent
on lactate production; the latter appears in relation to power
training and could be a function of muscle fibers composition.
Paccotti P et al. Effects of High-Intensity Isokinetic Exercise … Int J Sports Med 2005; 26: 747 – 755
Acknowledgements
The authors are indebted to Dott. Tiziana Armano, Dipartimento
di Matematica, Università di Torino, for her excellent assistance
with mathematical and statistical analysis.
This work was supported by grants from the University of
Turin (“ex-60%”) and by the MIUR Project “ENERME”
(2 003 062 904_002).
References
1
Aardal E, Holm AC. Cortisol in saliva–reference ranges and relation to
cortisol in serum. Eur J Clin Chem Clin Biochem 1995; 33: 927 – 932
2
Ben-Aryeh H, Roll N, Lahav M, Dlin R, Hanne-Paparo N, Szargel R,
Shein-Orr C, Laufer D. Effect of exercise on salivary composition and
cortisol in serum and saliva in man. J Dent Res 1989; 68: 1495 – 1497
3
Borges O, Essen-Gustavsson B. Enzyme activities in type I and II
muscle fibres of human skeletal muscle in relation to age and torque
development. Acta Physiol Scand 1989; 136: 29 – 36
4
Buono MJ, Yeager JE, Hodgdon JA. Plasma adrenocorticotropin and
cortisol responses to brief high-intensity exercise in humans. J Appl
Physiol 1986; 61: 1337 – 1339
5
Convertino VA, Keil LC, Bernauer EM, Greenleaf JE. Plasma volume, os-
molality, vasopressin, and renin activity during graded exercise in
man. J Appl Physiol 1981; 50: 123 – 128
6
del Corral P, Mahon AD, Duncan GE, Howe CA, Craig BW. The effect of
exercise on serum and salivary cortisol in male children. Med Sci
Sports Exerc 1994; 26: 1297 – 1301
7
Essen B, Haggmark T. Lactate concentration in type I and II muscle fi-
bres during muscular contraction in man. Acta Physiol Scand 1975;
95: 344 – 346
8
Essen B, Jansson E, Henriksson J, Taylor AW, Saltin B. Metabolic char-
acteristics of fibre types in human skeletal muscle. Acta Physiol Scand
1975; 95: 153 – 165
9
Farrell PA, Garthwaite TL, Gustafson AB. Plasma adrenocorticotropin
and cortisol responses to submaximal and exhaustive exercise. J Appl
Physiol 1983; 55: 1441 – 1444
10
Few JD. Effect of exercise on the secretion and metabolism of cortisol
in man. J Endocrinol 1974; 62: 341 – 353
11
Flamm SD, Taki J, Moore R, Lewis SF, Keech F, Maltais F, Ahmad M,
Callahan R, Dragotakes S, Alpert N. Redistribution of regional and or-
gan blood volume and effect on cardiac function in relation to upright
exercise intensity in healthy human subjects. Circulation 1990; 81:
1550 – 1559
12
Grassi B, Quaresima V, Marconi C, Ferrari M, Cerretelli P. Blood lactate
accumulation and muscle deoxygenation during incremental exer-
cise. J Appl Physiol 1999; 87: 348 – 355
13
Hackney AC, Premo MC, McMurray RG. Influence of aerobic versus
anaerobic exercise on the relationship between reproductive hor-
mones in men. J Sports Sci 1995; 13: 305 – 311
14
Hakkinen K, Pakarinen A. Acute hormonal responses to two different
fatiguing heavy-resistance protocols in male athletes. J Appl Physiol
1993; 74: 882 – 887
15
Hashimoto K, Suemaru S, Takao T, Sugawara M, Makino S, Ota Z. Cor-
ticotropin-releasing hormone and pituitary-adrenocortical responses
in chronically stressed rats. Regul Pept 1988; 23: 117 – 126
16
Jacks DE, Sowash J, Anning J, McGloughlin T, Andres F. Effect of exer-
cise at three exercise intensities on salivary cortisol. J Strength Cond
Res 2002; 16: 286 – 289
17
Jurimae T, Karelson K, Smirnova T, Viru A. The effect of a single-circuit
weight-training session on the blood biochemistry of untrained uni-
versity students. Eur J Appl Physiol Occup Physiol 1990; 61: 344 – 348
18
Kanaley JA, Weltman JY, Pieper KS, Weltman A, Hartman ML. Cortisol
and growth hormone responses to exercise at different times of day. J
Clin Endocrinol Metab 2001; 86: 2881 – 2889
19
Kant GJ, Eggleston T, Landman-Roberts L, Kenion CC, Driver GC,
Meyerhoff JL. Habituation to repeated stress is stressor specific. Phar-
macol Biochem Behav 1985; 22: 631 – 634
20
Kirschbaum C, Hellhammer DH. Salivary cortisol in psychobiological
research: an overview. Neuropsychobiology 1989; 22: 150 – 169
21
Kraemer WJ, Dziados JE, Marchitelli LJ, Gordon SE, Harman EA, Mello
R, Fleck SJ, Frykman PN, Triplett NT. Effects of different heavy-resist-
ance exercise protocols on plasma beta-endorphin concentrations. J
Appl Physiol 1993; 74: 450 – 459
22
Kraemer WJ, Patton JF, Gordon SE, Harman EA, Deschenes MR, Rey-
nolds K, Newton RU, Triplett NT, Dziados JE. Compatibility of high-in-
tensity strength and endurance training on hormonal and skeletal
muscle adaptations. J Appl Physiol 1995; 78: 976 – 989
23
Lac G, Marquet P, Chassain AP, Galen FX. Dexamethasone in resting
and exercising men. II. Effects on adrenocortical hormones. J Appl
Physiol 1999; 87: 183 – 188
24
Laudat MH, Cerdas S, Fournier C, Guiban D, Guilhaume B, Luton JP.
Salivary cortisol measurement: a practical approach to assess pitui-
tary-adrenal function. J Clin Endocrinol Metab 1988; 66: 343 – 348
25
Luger A, Deuster PA, Kyle SB, Gallucci WT, Montgomery LC, Gold PW,
Loriaux DL, Chrousos GP. Acute hypothalamic-pituitary-adrenal re-
sponses to the stress of treadmill exercise. Physiologic adaptations to
physical training. N Engl J Med 1987; 21: 1309 – 1315
26
Obminski Z, Stupnicki R. Comparison of the testosterone-to-cortisol
ratio values obtained from hormonal assays in saliva and serum. J
Sports Med Phys Fitness 1997; 37: 50 – 55
27
O’Connor PJ, Corrigan DL. Influence of short-term cycling on salivary
cortisol levels. Med Sci Sports Exerc 1987; 19: 224 – 228
28
Oyono-Enguelle S, Gartner M, Marbach J, Heitz A, Ott C, Freund H.
Comparison of arterial and venous blood lactate kinetics after short
exercise. Int J Sports Med 1989; 10: 16 – 24
29
Peters JR, Walker RF, Riad-Fahmy D, Hall R. Salivary cortisol assays for
assessing pituitary-adrenal reserve. Clin Endocrinol (Oxf) 1982; 17:
583 – 592
Paccotti P et al. Effects of High-Intensity Isokinetic Exercise … Int J Sports Med 2005; 26: 747 – 755
30
Petrides JS, Deuster PA, Mueller GP. Lactic acid does not directly acti-
vate hypothalamic-pituitary corticotroph function. Proc Soc Exp Biol
Med 1999; 220: 100 – 105
31
Port K. Serum and saliva cortisol responses and blood lactate accumu-
lation during incremental exercise testing. Int J Sports Med 1991; 12:
490 – 494
32
Raff H, Raff JL, Findling JW. Late-night salivary cortisol as a screening
test for Cushing’s syndrome. J Clin Endocrinol Metab 1998; 83: 2681 –
2686
33
Rowbottom DG, Keast D, Garcia-Webb P, Morton AR. Serum free corti-
sol responses to a standard exercise test among elite triathletes. Aust J
Sci Med Sport 1995; 27: 103 – 107
34
Stallknecht B, Vissing J, Galbo H. Lactate production and clearance in
exercise. Effects of training. A mini-review. Scand J Med Sci Sports
1998; 8: 127 – 131
35
Stupnicki R, Obminski Z. Glucocorticoid response to exercise as mea-
sured by serum and salivary cortisol. Eur J Appl Physiol Occup Physiol
Training & Testing
1992; 65: 546 – 549
36
Stupnicki R, Obminski Z, Klusiewicz A, Viru A. Pre-exercise serum cor-
tisol concentration and responses to laboratory exercise. Eur J Appl
Physiol Occup Physiol 1995; 71: 439 – 443
37
Thuma JR, Gilders R, Verdun M, Loucks AB. Circadian rhythm of corti-
sol confounds cortisol responses to exercise: implications for future
research. J Appl Physiol 1995; 78: 1657 – 1664
38
Tremblay MS, Copeland JL, Van Helder W. Effect of training status and
exercise mode on endogenous steroid hormones in males. J Appl
Physiol 2004; 96: 531 – 539
39
Vining RF, McGinley RA, Maksvytis JJ, Ho KY. Salivary cortisol: a better
measure of adrenal cortical function than serum cortisol. Ann Clin
Biochem 1983; 20: 329 – 335
40
Worthman CM, Stallings JF, Hofman LF. Sensitive salivary estradiol as-
say for monitoring ovarian function. Clin Chem 1990; 36: 1769 – 1773
755
Copyright of International Journal of Sports Medicine is the property of Georg Thieme Verlag Stuttgart and its
content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's
express written permission. However, users may print, download, or email articles for individual use.