Ardalan Intensive Training Exercise 2014

Journal of Strength and Conditioning Research Publish Ahead of Print
DOI: 10.1519/JSC.0000000000000632
Intensive Resistance Exercise and Circadian Salivary Testosterone Concentrations among

Young Male Recreational lifters
Brief running head: Resistance Exercise and Salivary Testosterone
Laboratory: Dr. Shahrzad Baradaran laboratory, Isfahan, Iran
Ardalan Shariat1, 4, Mehdi Kargarfard2 , Mahmoud Danaee3, Shamsul Bahri Mohd Tamrin4
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1
Department of Sports Science, Faculty of Educational Studies, University Putra Malaysia,
Malaysia.
2
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Department of Exercise Physiology, Faculty of Physical Education and Sport Sciences,
University of Isfahan, Isfahan, Iran.
3
Department of Biotechnology, Faculty of Agriculture, Roudehen Branch, Islamic Azad
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University, Roudehen, Iran.

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4
Department of Occupational Health, Faculty of Medicine and Health Sciences, University Putra
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Malaysia, Malaysia.
Corresponding author: Ardalan Shariat, Department of Sports Science, Faculty of Educational
Studies, University Putra Malaysia, Serdang, Selangor 43400, Malaysia
Tel: +6173365494 E-mail: Ardalan_sh2002@yahoo.com
There was no financial support for this survey.
Copyright Ó Lippincott Williams & Wilkins. All rights reserved.

Resistance Exercise and Salivary Testosterone 3
ABSTRACT
Strength and morphological adaptations to resistance exercise are mediated in part by anabolic
hormones such as testosterone, yet the time course of variability in circadian hormone
concentrations is not well characterized. This study, investigated how the circadian rhythm of
salivary testosterone is altered by resistance exercise in young men. Twenty healthy young male
recreational lifters (age 18.0± 1.3) with two years of experience in weightlifting were recruited.
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A randomized controlled trial was conducted, and subjects were randomly assigned to either the
resistance exercise group (n=10), who completed a series of resistance exercise (3 times a week,
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in the afternoon, 6-7 repetitions, at 85% of 1RM for 3 weeks), or a control group (n=10), who did
not exercise during the 3 weeks. Before and after the study, an unstimulated saliva sample (2
mL) was taken every two hours for a maximum of 16 hours during each day. A significant
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decrease was observed in the resistance exercise (44.2%, P=0.001) and control group (46.1%,
P=0.001) for salivary testosterone at each time point compared to baseline (P=0.001). There was
also no significant difference between the exercise and resting conditions in both groups for
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salivary testosterone (P>0.05), except a significantly higher increase by 38.4% vs. -0.02%
(P=0.001), at 17:30 during exercise sessions in the resistance exercise group compared to the
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control group. Resistance exercise has no noteworthy effect on circadian secretion of salivary
testosterone throughout the 16 waking hours. These results indicate that athletes can undertake
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resistance exercise in either the morning or afternoon with the knowledge that a similar
testosterone response can be expected regardless of the time of day.
Key words: androgens, chronobiology, saliva samples

INTRODUCTION
The testes secrete testosterone at a rate which varies in a circadian way and exerts a powerful
anabolic impact on muscles (32). Recent evidence suggests that the amount of exercise time and
the level of hormones, especially with regard to the circadian rhythm, have some effects on
physical and psychological performance (18,31). Currently, there are many unanswered
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questions about the responses of both endocrine hormone secretion mechanisms and salivary
testosterone to intensive resistance exercise and their subsequent effects on the waking circadian
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scale in men, particularly in a population of young recreational lifters. Most of these questions
are in relation to the following variables: the time of exercise and sampling, the variety, duration,
and intensity of exercise, and the basic fitness level of the subjects, followed by the circadian
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rhythm (20). Investigations on the endocrine hormone levels with anabolic characteristics could
be performed as a clinical method for analysing and monitoring the physical and psychological
attributes of the athletes. Also, it would inform coaches, athletes and strength researchers in
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prescribing resistance exercise workouts. Therefore, the role of testosterone as an anabolic
hormone must be considered.

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It is generally accepted that testosterone is responsive to physical exercises and other forms of
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stress. Testosterone is an anabolic hormone which promotes protein synthesis and has a major
role in the growth and preservation of the muscles and tissues (2). The resulting effects on
skeletal muscle anabolism are often due to a spike in the rate of testosterone during the healing
stages (27). Until now, a large number of studies have used blood samples to assess the levels of
hormones (7,11,12,15). Some of these studies (7,11,12) have shown how resistance exercise can

create fluctuations in the levels of cortisol and testosterone. However, there is a lack of
information which specially addresses the effects of intensive resistance exercise on the circadian
concentrations of salivary testosterone in young male recreational lifters.
Examining testosterone levels in saliva is a convenient, non-invasive way to determine the
concentrations of testosterone in the body. Previous studies (6,8,16,23) have indicated that
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salivary testosterone levels provide a reliable reflection of gonad function and, in particular, the
circadian rhythm of the hormone, when frequent sampling is performed (5,23). A circadian
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rhythm has also been observed in salivary testosterone, with lower concentrations detected in the
afternoon than in the morning (8,22,33). Previous research has clarified the effect of an intensive
resistance exercise session performed in the early morning on the circadian rhythm of salivary
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testosterone in body builders (23); however, it is important to establish an understanding of the
circadian variation, which could be further enhanced by the implementation of a morning
resistance exercise routine (11).

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The practical question is whether athletes are better off undertaking their resistance exercise in
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the morning or afternoon. We determined whether any significant differences in hormonal
response were likely between the exercise training in the morning and in the afternoon.
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As this study is essentially a variation of a similar study performed by Kraemer et al. (23), in
which testing was performed solely in the morning, we will compare our results, which were
based on exercise performed in the afternoon and with a slight modification in the intensity, with
the findings of the aforementioned study.

We hypothesized that the circadian rhythm of testosterone concentrations is under circadian
patterns, using the modified protocol (increasing the intensity) which was reported by Kraemer et
al. (23), and also that altering the time of exercise from early morning to the afternoon would not
change the outcome. Therefore, the purpose of this investigation was to determine the effects of
intensive resistance exercise performed in the afternoon on circadian salivary testosterone
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concentrations among young male recreational lifters.
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METHODS
Experimental Approach to the Problem
In this study, a parallel group design was used to investigate the effects of intensive resistance
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exercise (as the independent variable) on circadian salivary testosterone concentrations (as the
dependent variable) among young male recreational lifters. Subjects were either exposed to a
series of resistance exercise sessions or functioned as a control group, in which they performed
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no exercise. The salivary testosterone concentrations of control and resistance exercise groups
were examined on 2 occasions 3 weeks apart. Unstimulated saliva samples were collected at
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various time intervals (from 06:00 until 22:00) in order to establish a measure of the circadian
rhythm of testosterone secretion. Due to the literature which believes that the resistance training
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athletes have higher performance in the afternoon (14), this study was required to focus on
different angles to investigate the variables of time and employing higher intensity in the
exercise session.
Subjects

Twenty healthy young, recreational lifter men (age 18.0± 1.3 years, body mass 75.2 ± 3.2 kg,
height 1.79 ± .03 m, body fat % 8.1 ± 1.9 (mean ± SD)) who had two years of experience in
weightlifting participated in the study. Subjects were randomly selected to be part of either the
resistance exercise (n= 10) or control (n= 10) group. All subjects were active members of weight
lifting gyms in the city of Isfahan, Iran when the study was conducted in April of 2012.
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Before undergoing their physical examinations by the physician, all subjects filled out a
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questionnaire (23), and those who had any chronic medical condition which could create an
unnecessary risk during their exercise testing were excluded from the study.
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The resistance exercise group performed regular exercise sessions (3 times per week at 16:05)
for three weeks, and the control group completed all of the required tests with the absence of
physical exercises. The subjects included in this research did not consume dietary supplements in
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the form of carbohydrates, proteins (protein and/or carbohydrate intake impacts androgen
receptors and testosterone concentrations (35)) or amino acids (amino acids affect hormonal
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responses (8)), nor were any of them taking anabolic steroids either prior to or while
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participating in this research.
The subjects’ schedule of daily activity and diets were carefully evaluated by a dietician 2 days
beforehand and during implementation of the experiment, until the final sample of saliva had
been collected. The dietician instructed them on their food intake in 3-5 daily meals, which
contained an approximate fat percentage of 30%, carbohydrate percentage of 50% and protein

percentage of 20%, as this combination has no demonstrable effect on the circadian rhythm of
testosterone concentrations (23, 31, 37). A diet containing more than 44% fat is known to
influence the level of testosterone (37,8).
All test subjects were given instruction on how to remain properly hydrated in order to avoid the
possibility of hypohydration impacting their performance during the study (21) (the subjects’
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water consumption was recorded and found to be 3.8±0.2 L daily).
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The subjects were prohibited from consuming drinks containing alcohol, caffeine, or any other
stimulants. They were also asked not to engage in sexual or other strenuous physical activity
within 24 hours of the days in which saliva collection was to occur. The recruited subjects were
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not involved in other physical exercises, and both groups were advised to continue with a normal
sleep pattern of approximately 8 hours per night for the duration of the experimental study
period.
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Ethical approval for the study was obtained from the University of Isfahan, Faculty of Physical
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Education and Sport Sciences, in Iran. The subjects were given a clear explanation of the
objectives of the study, as well as the potential risks involved, and consent forms (parental
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consent forms for those below 18 years of age) were obtained for all subjects.
Procedures
A modified protocol, introduced by Kraemer and his colleagues (23), (Table 1) was used in this
study. Exercises 3-8 were done with proper machines (Techno Gym Equipment’s, UK, 2008) and

the other exercise (except sit-ups) was done with an Olympic-style barbell (Iron Grip Barbell
Company, USA).
Table 1 about here
Replication of the Study
In the present study, the intensity and time of the exercise was adapted and altered from Kraemer
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et al. (2001) (23). After 10-15 minutes of general warm up exercises and stretching, the
resistance exercise group subjects started the testing session at 16:05. They performed 3 sets of
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each exercise, with 6-7 repetitions and 2 minutes of rest in between each set as the exercise
protocol, with a daily exercise session duration of around 1 hour and 30 minutes. The
experimental process took place over a period of three weeks. It should be noted that in each
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session, the heart rate was found to be more than 180 beats per minute (bpm). For the purposes
of monitoring the intensity of the exercise, the heart rates of 7 random athletes were determined
based on Karvonen’s method of Heart Rate Calculating (19).The environmental factors, such as
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the noise level (82 dB), temperature (19.0 ± 1.0°C), humidity (40-50 %), and comfort, were
strictly controlled. The subjects were permitted to drink 1.0 ± 0.2 L of cool water (12 °C) during
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their exercise sessions.

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Assessment of the Body Composition
Anthropometric characteristics, including height, weight, and body mass, were measured by the
same investigator using standard procedure before the beginning of the study for all participants.
For weight, a physician’s beam digital scale (± 0.10kg), with the participants barefoot and
without heavy clothing, was used, followed by a barefoot measurement using a height rod (±

.005 m) (SECA700, CA, USA). The Body Mass Index (BMI) was calculated as weight in
kilograms divided by squared height in meters, and the body composition was assessed by means
of a Lange sector for the cutaneous fold, which was based on the Jackson and Pollock protocol
(17). A Lange skin-fold caliper was used to determine the fat in skin folds of the triceps,
abdomen and upper iliac areas of the test subjects. This process was repeated three times to
validate the accuracy (3) and the thickness of skin folds was used to evaluate the percentage of
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body fat (BF %) (25).
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1RM Measurement
Once the resistance exercise group had a full understanding of the equipment and were taught the
necessary techniques by the CSCS trainer, they participated in the 1RM test. Determination of
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1RM was done for bench press, leg press and shoulder press. The participants initially performed
two warm-up sets of 2-5 repetitions. The weight for these was set at approximately 50% and 80%
of their perceived 1-repetition maximum (1-RM), respectively. These were followed by 3 to 4

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sets of steadily increasing weight, which included rest intervals of 3 to 5 minutes, until a 1-RM
weight was established for each individual. The same investigator was used to monitor all of the
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1-RM tests, ensuring that all participants performed the exercises accurately using a complete
range of motion (22).

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Hormonal Assay
The unstimulated saliva sampling (2 mL) pre-test was done on a day when no exercise session
took place. The post-test sampling was done 3 weeks later, while the resistance exercise group
did the exercise and the control group refrained from physical exertion. Samples were collected

from both groups in 10 separate intervals, beginning at 06:00 and repeated every 2 hours until
16:00; further samples were collected at 17:30, 18:00, 20:00 and 22:00. The samples were
obtained while the subjects were leaning forward in a seated position and collected by means of
the passive drool method. They were asked to wash their mouths with water, and after waiting
for one minute, deposit the unstimulated saliva (2 mL) into the receptacles (9,23). In order to
avoid the circadian influences on their performance, sampling was conducted at the same time
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during the day (9,29). The subjects were asked to avoid any oral contact with objects such as
floss or toothbrushes before and during the testing session (23).
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The process of sample collection was monitored carefully, and the specimens were immediately
transferred to the professional medical and pathology laboratory and stored at -80ºC (1). In order
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to avoid inter-assay variation, the collected samples were all examined in the same assessment.
RADIM kit (SEAC Company, Italy) and the quantitative ELISA protocol (ELISA Technologies,
Inc., USA) were used to determine the concentrations of testosterone. All of the samples for each
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subject were analyzed on the same micro plate (23).

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For quality control, a few subjects were randomly selected from both groups and the experiments
were repeated for a second time, and the results were 99% consistent with the original findings.
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Statistical Analyses
The sample size for the study was calculated using G power software, and all of the variables
were subjected to the normality test. The result revealed that all variables were distributed
normally in order to determine the means and the standard deviations, and descriptive statistics

were used in reporting the data. The saliva testosterone concentrations were analyzed at ten
points throughout the day (from 06:00 until 22:00).
Regarding the statistical method, the two-way repeated measure ANOVA analysis following
Bonferroni post-hoc test was used for within group comparison, and all related assumptions,
including the normal distribution of dependent variables and sphericity (homogeneity of variance
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and co-variance), were met. Using Pearson product-moment correlation coefficient the linearity
and correlations of the dependent variable among all of the three or more repeated measures
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were equal. A retest correlation was used to quantify the reliability of the correlation, which was
0.94, and represents a very high level of reliability.

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The separate resistance exercise and control groups’ circadian rhythm data was analyzed using
repeated measure ANOVA methods. Additionally, two-way repeated measure ANOVA analysis
was performed, including two-between factors (exercise, control) and two-within factors (pre-
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and post-test) at the 10 sampling time periods (06:00, 08:00, 10:00, 12:00, 14:00, 16:00,17.30,
18:00, 20:00, 22:00 hours) to evaluate testosterone changes. SPSS version 19 (SPSS, Inc.,
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Chicago, IL.) was used for analyzing the data. The differences for all analyses were considered
significant if P<0.05.
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RESULTS
With reference to Figure 1, one can observe the activity of testosterone levels in the condition of
normal daily rest of the two different groups, namely exercise and control, based on the outcome

of salivary samplings taken every two hours. The results obtained in the control group showed
that peak salivary testosterone levels (607.62 ± 19.76 pmol/L) occurred in early morning (06:00),
with a corresponding reduction throughout the day until an evening nadir (22:00) (323.18 ±
4.914pmol/L). Similarly, in the RE group the testosterone level within saliva (611.78 ±
16.406 pmol/L) was highest in early morning (06:00), with a gradual reduction throughout the
day until an evening nadir (22:00) (321.88 ± 5.018 pmol/L). The data showed that the
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concentrations of salivary testosterone among our subjects were in the normal range reported by
Teo et al., (33) (normal range: 693±243 pmol/L in the early morning and 451± 173 pmol/L in the
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evening) and it was also consistent with the salivary testosterone concentrations reported in
similar research performed by Kraemer et.al (23).
Figure 1 about here:

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Both the control and resistance exercise groups had their salivary testosterone concentrations
examined on 2 separate occasions 3 weeks apart. The resistance exercise group performed 3
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weeks of prescribed resistance exercise and had their circadian rhythm of salivary testosterone
concentrations examined to measure what difference could be observed after 3 weeks of exercise
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training. On the first occasion, this was done on a rest day (on which they performed no
exercise), while on the second occasion, 3 weeks later; the measurement was taken on a day
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when they performed the resistance exercise session at 16:00. As expected, for the control group
there was no change in the circadian pattern of salivary testosterone concentrations after 3 weeks
without exercise (a within group comparison) (F=21.9, P= 0.164). For the resistance exercise
group, there was a temporary rise in salivary testosterone immediately after the training session
at 17:30 (F=458. 665, P=0.001), but no difference at any other time point compared with the

salivary testosterone concentrations on the rest day (a within group comparison) (F= 37.4, P=
0.216). Subsequent escalation of testosterone levels was also observed in the hours after the
exercise, as shown in Figure 2. The intra- and inter-assay variance for testosterone was recorded
at 2.3-6.1% and 7.7-8.1%, respectively.
Figure 2 about here:
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With reference to the current study, resting and exercise conditions exhibited no apparent
differences, despite the fact that the previous time period was actually much higher than the
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ensuing one. However, there was a significant group × time during the workout interaction for
the testosterone levels (F= 11.965, P< 0.05). Bonferroni post-hoc analyses indicated that only the
males who performed resistance exercise significantly increased salivary testosterone, from
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371.28 ± 7.228 pmol/L to 513.76 ± 20.332 pmol/L, respectively, (P= 0.001) (Figure 2) but that
the change was temporary, and appeared not to have any lasting effects on the circadian levels of
testosterone. Therefore, the comparison in the mean of testosterone secretion in pre- and post-
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test, between resistance exercise and control groups showed that intensive resistance exercise
had no significant effect on circadian secretion of salivary testosterone in 16 hours of waking

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time (Figure 2).

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DISCUSSION
In 2001, Kraemer investigated the effect of resistance exercise in the early morning on the
circadian rhythm of testosterone concentrations, and the result showed that the circadian rhythm
did not change significantly (23). However, a couple of key points set this study apart from our
own. Firstly, the time when exercise training was undertaken was moved to the afternoon rather

than the morning. The reason for this decision was that the afternoon is typically the time during
which most lifters perform their routines (9). Secondly, the previous research (23) was done
without a control group with which to compare results. In this study, however, a control group
who did not train was used to show the differences more clearly in comparison with the
resistance exercise group. Thirdly, the previous research was done with 10 repetitions (performed
at roughly 75% 1RM) for each set of each exercise. Here we have increased the intensity (to
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approximately 85% 1 RM) by decreasing the number of repetitions (from 10 to 6-7 repetitions)
without changing the duration of rest between sets. The reason for this decision was to evoke a
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larger effect on hormonal responses in comparison with Kraemer’s 2001 study.
What has not been answered yet is whether intensive resistance exercise in the afternoon has
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different effects on the circadian rhythm of testosterone concentrations throughout a complete
day in comparison with exercise strictly in the morning. The main finding of the present study
was that an intense period of intensive resistance-oriented sport activities provides a nominal
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temporary effect upon the testosterone circadian rhythm that diminishes an hour after the
workout, after which testosterone levels return to normal. The findings obtained through both
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resistance exercise and control groups obviously manifest a circadian pattern in which there is a
presence of higher concentrations during the morning and lower concentrations during the
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evening and it is under circadian patterns. These results are in line with previous studies
(9,15,22,23).
This rhythm was consistent with the hypothesized rhythm for the secretion of testosterone. The
measurements taken on both occasions 3 weeks apart resulted in levels which were within a

normal range as reported by Teo et al. (33), and these data were in line with the results of
Kraemer et al. (23), providing further evidence of stability in the daily pattern of salivary
testosterone concentrations during the waking hours.
The results of the sampling at various times over the course of a day showed that there was a
remarkable increase in the level of testosterone immediately after the intensive resistance
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exercise at 17:30 and this seems to imply an acute alteration rather than a change in the circadian
rhythm of testosterone secretion in response to intensive resistance exercise. The research
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performed by Kraemer et.al (23) likewise showed a significant increase in salivary testosterone
concentrations immediately after the resistance exercise, but given the time frame in which the
research was conducted, this increase was observed in the early morning, with no corresponding
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data regarding the afternoon. The rhythm returned to its normal level a few hours after
concluding the exercise. However, the precise physiological mechanism inducing the increase in
testosterone resulting from exercise remains unclear. In resting conditions, the testes were
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primarily under the influence of the gonadotropins (LH, FSH). However, no relationship has
been found between the levels of the gonadotropins and the increase of testosterone with regard
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to exercise (30,31). Meanwhile, gonadotropins work as regulators and based on the results of this
study, intensive exercise (in the afternoon or morning) may not affect their performance. An
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increase in the level of free testosterone due to alterations in the binding affinity of the sex
hormone binding globulin (SHGB) has also been discounted by Fahrner (11). It has been
suggested that the increase in testosterone is due to a reduced metabolic clearance rate (MCR),
and alternatively, Tsigos et al.(34) postulated that the increase may be mediated by sympathetic
stimulation of the testes via the adrenal gland.

The results of several studies (2,4,6,8,14,28) indicate that the way in which exercise is
performed, with respect to the following variables, is critical in hormonal responses: intensity
and duration, type of the exercise; fitness level and gender of the subjects; rest intervals between
the exercises, and the conditions under which the recovery of the muscles takes place . Certain
factors, such as sleep or waking periods, nutrition, meal time, physical exercises, hormones and
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stress, can influence and change the pattern of the circadian rhythm (10,27,28). Intensive
resistance exercise without sufficient rest intervals and the stress caused by competitive sports
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can result in changes in the physiological, immunological, psychological, and functional status of
the athletes in the long term (12,15,16). Physical exercises affect the hormonal responses, so
considering the pattern of the circadian rhythm might lead to achieving the ideal metabolism and
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allow us to maximize the effect of resistance exercise on the skeletal muscles (30,38). It has been
well established that intensive resistance exercise is able to dramatically boost the serum levels
of testosterone (24), but the results of this study showed that this effect is temporary, and after
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each session of exercise will return to the normal level. It should be noted that free testosterone is
rigidly controlled (e.g., by mechanisms of nitric oxide and blood flow), and these mechanisms
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are important factors in terms of the body’s response to resistance exercise and the
concentrations of testosterone (23).

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Utilization of salivary testosterone proved to be trustworthy in comparison to the blood serum
and accurately reflected the circadian rhythms (16), especially in frequent sampling (26).
In a report by Beaven et al. (4), it was shown that the increase in serum testosterone coincides
with an increase in salivary testosterone for the first hour. Likewise, in another study, Vittek et al.

(36) investigated the relation between serum and salivary testosterone. The results showed that
both free and total testosterone were highly correlated (r = 0.97 and r = 0.70 – 0.87
respectively) and denoted that they are statistically significant. This data, in combination with the
results of the present research, can be deemed as a good indication that salivary testosterone is
indicative of fluctuations in free testosterone (12,22).
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It is plausible that after a period of exercise, regulatory components are consistently involved due
to the quick transition experienced in salivary testosterone levels and the normal circadian
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rhythm (13,15,16,28,33).
In the previous research by Kreamer et al. (23), they referred to their exercise protocol as a heavy
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protocol, but their results showed that it did not significantly affect the circadian concentrations
of salivary testosterone. Therefore, we decided to make the protocol in our study more intensive
by decreasing the number of repetition to 6-7 and increasing the weight, though we did not
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change the rest time between the sets. But, with regard to the results of Kraemer et al., we
hypothesised that the circadian rhythm of testosterone concentrations, using the modified
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protocol (increasing the intensity) which was reported by Kraemer et al. (23), and also that
altering the time of exercise from early morning to the afternoon would not change the outcome
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in comparison with Kraemer et al. (23) research. This hypothesis was based on an individual
period of resultantly intensive exercise within the length of the afternoon, which could account
for the slight alterations within the circadian rhythm of waking hours. Therefore, the present
hypothesis was built upon previously-done studies within which long sampling times, i.e. 6-8
hours, have been applied (8). Such data denotes that homeostatic processes become swiftly

involved in bringing back the concentrations level of serum testosterone to usual circadian
quantities (23). The findings of this study provide further support for such a conclusion, and
illustrate a tight adjustment of the concentration levels of salivary testosterone subsequent to
physical stress.
Kraemer et al. (23) showed that heavy resistance exercise has no significant effect on the
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circadian rhythm of salivary testosterone among male body builders, and that it may instead be
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the circadian variation which could be enhanced through a repeated morning resistance exercise
routine. Changing the time of exercise (from early morning to the afternoon), and increasing the
intensity of the exercise in this study did not significantly affect the circadian rhythm of
testosterone concentrations and it had merely a temporary effect immediately after the exercise.
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Based on these data, it can be said that the signals for androgen receptors were directly linked to
the time in which exercise training was performed, and that the decrease was most likely related
to not only circadian values but also the increased activity of the exercise group, as we did not
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see the same pattern within the control group.

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According to the aforementioned information, it seems that the results of the present study, with
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regard to testosterone, are in agreement with the findings of most of the previously published
studies, especially with Kraemer et al. (2001) (23), which was only based on morning exercise
training. These studies both showed that resistance exercise in the morning or afternoon did not
substantially affect the circadian rhythm of testosterone concentrations and that free testosterone
appears to be regulated by the body's homeostatic systems.

PRACTICAL APPLICATIONS
The evaluation and measurement of testosterone in the saliva can be regarded as a highly
practical and helpful method in primary research studies and clinical environments. The results
of the current study showed that intensive resistance exercise has no noteworthy effect on
circadian secretion of salivary testosterone throughout the 16 waking hours. Therefore, this
information would be useful for athletes, coaches and researchers and athletes can undertake
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resistance exercise in the morning or afternoon with the knowledge that both will result in
similar testosterone responses. It seems that there are other potential factors, such as diet, which
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can affect the circadian secretion of salivary testosterone. It is suggested for further researchers
to investigate the circadian secretion of salivary testosterone in response to the concurrent effects
of aerobic and resistance exercise.

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ACKNOWLEDGEMENT
We would like to have special thanks to University Putra Malaysia, Department of Sport
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Sciences and University of Isfahan (Iran) departments of Exercise Physiology also, to all the
subjects who helped us in this project. We also would like to express our special thanks to Prof.
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David Pyne for reviewing the manuscript in spite of his very busy schedule. Last but not least,
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our sincere gratitude to Matthew Ballard who helped us a lot in finalizing this text within the
limited time frame. The results of the present study do not constitute endorsement by the authors
or the National Strength and Conditioning Association (NSCA). The authors did not have any
conflicts of interests.

References
1. Allgrove, JE, Gomes, E, Hough, J and Gleeson, M. Effect of exercise intensity on salivary antimicrobial
proteins and markers of stress in active men. J Sports Sci 26: 653– 661, 2008.
2. Auyeung, TW, Lee, JSW, Kwok, T, Leung, J, Ohlsson, C, Vandenput, L, Leung, PC and Woo, J.
Testosterone but not estradiol level is positively related to muscle strength and physical performance
independent of muscle mass: a cross-sectional study in 1489 older men. Eur JEndocrinol 164: 811– 817,
2011.
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3. Barker, JL, Olsen, LW and Sorensen, TIA. Childhood body-mass index and the risk of coronary heart
disease in adulthood. N Engl J Med 357: 2329– 2337, 2007.
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4. Beaven, CM, Gill, ND, Ingram, JR and Hopkins, WG. Acute salivary hormone responses to complex
exercise bouts. J Strength Cond Res 25: 1072– 1078, 2011.
5. Bribiescas, RG and Hill, KR. Circadian variation in salivary testosterone across age classes in Ache
Amerindian males of Paraguay. Am J Hum Biol 22: 216– 220, 2010.

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6. Cadoux-Hudson, TA, Few, JD and Imms, FJ. The effect of exercise on the production and clearance of
testosterone in well trained young men. Eur J Appl Physiol Occup Physiol 54: 321– 325, 1985.
7. Cefalu, W, Pardridge, WM, Chaudhuri, G and Judd, HL. Serum bioavailability and tissue metabolism of
C
testosterone and estradiol in rat salivary gland.J Clin Endocrinol Metab 63: 20– 28, 1986.
8. Chandler, RM, Byrne, HK, Patterson, JG and Ivy, JL. Dietary supplements affect the anabolic hormones
after weight-training exercise. J Appl Physiol 76: 839– 845, 1994.

C
9. Chtourou, H and Souissi, N. The effect of training at a specific time of day: a review. J Strength Cond Res
26: 1984– 2005, 2012.

A
10. Duclos, M, Gouarne, C and Bonnemaison, D. Acute and chronic effects of exercise on tissue sensitivity to
glucocorticoids. J Appl Physiol 94: 869– 875, 2003.
11. Fahrner, CL and Hackney, AC. Effects of endurance exercise on free testosterone concentration and the
binding affinity of sex hormone binding globulin (SHBG). Int J Sports Med 19: 12– 15, 1998.
12. Fry, AC, Kraemer, WJ, Stone, MH, Koziris, Lp, Thrush, JT and Fleck, SJ. Relationships Between Serum
Testosterone, Cortisol, and Weightlifting Performance. J Strength Cond Res 14: 338– 343, 2000.

13. Garber, CE, Blissmer, B, Deschenes, MR, Franklin, BA, Lamonte, MJ, Lee, IM, Nieman, DC and Swain,
DP. American college of sports Medicine position stand. Quantity and quality of exercise for developing and
maintaining cardiorespiratory, musculoskeletal, and neuromotor fitness in apparently healthy adults:
guidance for prescribing exercise. Med Sci Sports Exerc 43:1334– 1359, 2011.
14. Hayes, LD, Bickerstaff, GF and Baker, JS. Interactions of cortisol, testosterone, and resistance training:
influence of circadian rhythms.Chronobiol Int 27: 675– 705, 2010.
15. Hough, J, Corney, R, Kouris, A and Gleeson, M. Salivary cortisol and testosterone responses to high-
D
intensity cycling before and after an 11-day intensified training period. J Sports Sci 31:1– 10, 2013.
16. Huang, W, Ramsey, KM, Marcheva, B and Bass, J. Circadian rhythms, sleep, and metabolism. J Clin Invest
TE
121: 21– 33, 2011.
17. Jackson, AS and Pollock, ML. Generalized equations for predicting body density of men.1978. Br J Nutr
91: 161– 168, 2004.
18. Jankowski, CM, Gozansky, WS, MacLean, PS, Shulman, B, Wolfe, P, Schwartz, RS and Cohrt, WM. N-
EP
acetyl-4-aminophenol and musculoskeletal adaptations to resistance exercise training. Eur J Appl
Physiol 113: 1127– 1136, 2013.
19. Karvonen, J and Vuorimaa, T. Heart rate and exercise intensity during sports activities. Sports Med 5: 303–
311, 1988.
C
20. Kraemer, RR, Acevedo, EO, Dzewaltowski, D, Kilgore, JL, Kraemer, GR and Castracane, VD. Effects of
low-volume resistive exercise on beta-endorphin and cortisol concentrations. Int J Sports Med 17: 12– 16,
C
2007.
21. Kraemer, WJ, Flanagan, SD, Comstock, BA, Fragala, MS, Earp, JE, Dunn-Lewis, C, Ho, JY, Thomas,
A
GA, Solomon-Hill, G, Penwell, ZR, Powell, MD, Wolf, MR, Volek, JS, Denegar, CR and Maresh, CM.
Effects of a whole body compression garment on markers of recovery after a heavy resistance workout in
men and women. J Strength Cond Res 24: 804– 814, 2010.
22. Kraemer, WJ, Gordon, SE, Fleck, SJ, Marchitelli, LJ, Mello, R, Dziados, JE, Friedl, K, Harman, E, Maresh,
C and Fry, AC. Endogenous anabolic hormonal and growth factor responses to heavy resistance exercise in
males and females. Int J Sports Med 12: 228– 235, 1991.

23. Kraemer, WJ, Loebel, CC, Volek, JS, Ratamess, NA, Newton, RU, Wickham, RB, Gotshalk, LA, Duncan,
ND, Mazzetti, SA, Gómez, AL, Rubin, MR, Nindl, BC and Häkkinen, K. The effect of heavy resistance
exercise on the circadian rhythm of salivary testosterone in men. Eur J Appl Physiol 84: 13– 18, 2001.
24. Kraemer, WJ, Solomon-Hill, G, Volk, BM, Kupchak, BR, Looney, DP, Dunn-Lewis, C, Comstock,
BA, Szivak, TK, Hooper, DR, Flanagan, SD, Maresh, CM andVolek, JS. The effects of soy and whey protein
supplementation on acute hormonal responses to resistance exercise in men. J Am Coll Nutr 32: 66– 74,
2013.
D
25. Lambert, BS, Oliver, JM, Katts, GR, Green, JS, Martin, SE and Crouse, SF. DEXA or BMI: Clinical
Considerations for Evaluating Obesity in Collegiate Division IA American Football Athletes. Clin J Sport
TE
Med 22: 436– 438, 2012.
26. Leymarie, P, Roger, M, Castainer, M and Scholler, R. Circadian variations of plasma testosterone and
estrogens in normal men. J Steroid Biochem 5: 167– 171, 1974.
27. McMurray, RG, Eubank, TK and Hackney, AC. Nocturnal hormonal responses to resistance exercise. Eur J
EP
Appl Physiol Occup Physiol 72: 121– 126, 1995.
28. Naclerio, F, Faigenbaum, AD, Larumbe-Zabala, E, Perez-Bibao, T, Kang, J, Ratamess, NA and Triplett, NT.
Effects of different resistance training volumes on strength and power in team sport athletes.
J Strength Cond Res 27: 1832– 1840, 2013.

C
29. Pulford, DJ, Mosteller, M, Briley, JD, Johansson, KW and Nelsen, AJ. Saliva sampling in global clinical
studies: the impact of low sampling volume on performance of DNA in downstream genotyping
C
experiments. BMC Med Genomics 6: 20– 27, 2013.
30. Raastad, T, Bjøro, T and Hallen, J. Hormonal responses to high-and moderate-intensity strength exercise.
A
Eur J Appl Physiol 82: 121– 128, 2000.
31. Ruige, JB, Mahmoud, AM, De Bacquer, D and Kaufman, JM. Endogenous testosterone and cardiovascular
disease in healthy men: a meta-analysis. Heart 97: 870– 875, 2011.
32. Srinivas-Shankar, U, Roberts, SA, Connolly MJ, O’Connell MDL, Adams JE, Oldham JA and Wu, FC.
Effects of testosterone on muscle strength, physical function, body composition, and quality of life in
intermediate-frail and frail elderly men: a randomized, double-blind, placebo-controlled study. J Clin
Endocrinol Metab 95: 639– 650, 2010.

33. Teo, W, McGuigan, MR and Newton, MJ. The effects of circadian rhythmicity of salivary cortisol and
testosterone on maximal isometric force, maximal dynamic force, and power output. J Strength Cond Res
25: 1538– 1545, 2011.
34. Tsigos, C and Chrousos, GP. Hypothalamic–pituitary–adrenal axis, neuroendocrine factors and stress. J
Psychosom Res 53: 865– 871, 2002.
35. Vingren, JL, Kraemer, WJ, Ratamess, NA, Anderson, JM, Volek, JS and Maresh, CM. Testosterone
physiology in resistance exercise and training. Sports Med 40: 1037–1053, 2010.
D
36. Vittek, J, L`Hommedieu, DG, Gordon, GG, Rappaport, SC and Southren, Al. Direct radioimmunoassay
(RIA) of salivary testosterone:correlation with free and total serum testosterone. Life Sci 37: 711– 716,
TE
1985.
37. Volek, JS, Kraemer, WJ, Bush, JA, Incledon, T and Boetes, M. Testosterone and cortisol in relationship to
dietary nutrients and resistance exercise. J Appl Physiol 82: 49– 54, 1997.
38. Zambon, AC, McDearmon, EL, Salomonis, N, Vranizan, KM, Johansen, KL, Adey, D, Takahashi,
EP
JS, Schambelan, M and Conklin, BR. Time-and exercise-dependent gene regulation in human skeletal
muscle. Genome Biol 4: R61, 2003.

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Table1. The protocol of Intensive Resistance Exercise
Exercise order Sets× RM
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1- Hang pulls 3×6-7 RM
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2- Bench press 3×6-7 RM
3- Leg press 3×6-7 RM
4- Seated row 3×6-7 RM

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5- Leg curl 3×6-7 RM
6- Shoulder press 3×6-7 RM
7- Lat pulls 3×6-7 RM

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8- Knee extensions 3×6-7 RM
9- Arm curls 3×6-7 RM

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10- Sit-ups 3×20 RM

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Figure legends:
Figure1. Circadian rhythm of salivary testosterone in resting condition for both resistance
exercise and control groups (different sampling time).
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Figure2. Circadian rhythm of salivary testosterone in exercise condition for both resistance
exercise and control groups (different sampling time).
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Figure 1:
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Figure 2:
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Journal of Strength and Conditioning Research Publish Ahead of Print

Intensive Resistance Exercise and Circadian Salivary Testosterone Concentrations among

Brief running head: Resistance Exercise and Salivary Testosterone

Laboratory: Dr. Shahrzad Baradaran laboratory, Isfahan, Iran

University of Isfahan, Isfahan, Iran.

University, Roudehen, Iran.

Corresponding author: Ardalan Shariat, Department of Sports Science, Faculty of Educational

Studies, University Putra Malaysia, Serdang, Selangor 43400, Malaysia

Tel: +6173365494 E-mail: Ardalan_sh2002@yahoo.com

There was no financial support for this survey.

Copyright Ó Lippincott Williams & Wilkins. All rights reserved.

testosterone response can be expected regardless of the time of day.

Key words: androgens, chronobiology, saliva samples

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prescribing resistance exercise workouts. Therefore, the role of testosterone as an anabolic

hormone must be considered.

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concentrations of salivary testosterone in young male recreational lifters.

Examining testosterone levels in saliva is a convenient, non-invasive way to determine the

circadian variation, which could be further enhanced by the implementation of a morning

resistance exercise routine (11).

the morning or afternoon. We determined whether any significant differences in hormonal

the findings of the aforementioned study.

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We hypothesized that the circadian rhythm of testosterone concentrations is under circadian

intensive resistance exercise performed in the afternoon on circadian salivary testosterone

Experimental Approach to the Problem

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participating in this research.

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influence the level of testosterone (37,8).

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Table 1 about here

Replication of the Study

their exercise sessions.

Assessment of the Body Composition

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of their perceived 1-repetition maximum (1-RM), respectively. These were followed by 3 to 4

range of motion (22).

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floss or toothbrushes before and during the testing session (23).

subject were analyzed on the same micro plate (23).

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points throughout the day (from 06:00 until 22:00).

0.94, and represents a very high level of reliability.

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similar research performed by Kraemer et.al (23).

Figure 1 about here:

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at 2.3-6.1% and 7.7-8.1%, respectively.

Figure 2 about here:

had no significant effect on circadian secretion of salivary testosterone in 16 hours of waking

time (Figure 2).

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Copyright Ó Lippincott Williams & Wilkins. All rights reserved.

testosterone concentrations during the waking hours.

rhythm of testosterone secretion in response to intensive resistance exercise. The research

stimulation of the testes via the adrenal gland.

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concentrations of testosterone (23).

Utilization of salivary testosterone proved to be trustworthy in comparison to the blood serum

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indicative of fluctuations in free testosterone (12,22).