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Twice-daily sessions result in a greater muscle strength and a similar muscle

hypertrophy compared to once-daily session in resistance-trained men
Article in The Journal of sports medicine and physical fitness · February 2021

DOI: 10.23736/S0022-4707.21.12118-8
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See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/349657740
Twice-daily sessions result in a greater muscle strength and a similar muscle

hypertrophy compared to once-daily session in resistance-trained men
Article in The Journal of Sports Medicine and Physical Fitness · February 2021

DOI: 10.23736/S0022-4707.21.12118-8
CITATIONS READS
0 12
7 authors, including:
Felipe Brigatto Tiago Volpi Braz

Universidade Metodista de Piracicaba (Unimep) Faculdade de Americana
Marcelo Saldanha Aoki Paulo Marchetti

University of São Paulo California State University, Northridge
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Page 34 of 65
Twice-daily sessions result in a greater muscle strength and a similar muscle hypertrophy compared to
once-daily session in resistance-trained men
1
2
3
4
5
6
7
8 Daniel
A. CORRÊA 1, Felipe A. BRIGATTO 1 *, Tiago V. BRAZ 1, Júlio B. B. DE CARMARGO 1,
9
Marcelo S. AOKI 2, Paulo H. MARCHETTI 3, Charles R. LOPES 1,4
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11
12
13
14
15 1
Methodist University of Piracicaba, Human Performance Research Laboratory, Piracicaba, São Paulo,
16
17 Brazil.

18 2
School of Arts, Sciences and Humanities, University of São Paulo, São Paulo, Brazil.
19
20 3
Department of Kinesiology. California State University Northridge, CA, USA.
21
22 4
Adventist Faculty of Hortolândia, Hortolândia, São Paulo, Brazil.
23
24
25
26
27
*Corresponding author: Felipe A. Brigatto, Human Performance Research Laboratory, Human Movement
28
Sciences Postgraduate Program, Methodist University of Piracicaba, Campus Taquaral. Rodovia do Açúcar,
29 km 156. Piracicaba (SP) Brazil. Zip code: 13400-911. E-mail: filephi@gmail.com
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Page 35 of 65
ABSTRACT
1
2
3 BACKGROUNDː
The present study investigated the mid-term effects of training muscle groups once- versus
4
twice-daily on morphofunctional adaptations in trained men.
5
6 METHODSː
Participants were randomly assigned to 1 of 2 experimental groups: 1 daily session per muscle
7
8 group

(1S, n = 11), where every muscle group was trained once a day or 2 daily sessions per muscle group
9 n = 12), where every muscle group was trained twice. Testing was conducted before intervention and
(2S,
10
11 after
8 weeks for maximal strength (1RM) and muscular endurance (60%1RM) for bench press and parallel
12
back squat exercises, and muscle thickness (MT) of the biceps brachii, triceps brachii, vastus lateralis, anterior
13
14
quadriceps and pectoralis major.
15
16 RESULTSː
The major findings were as follows: (a) the increase in 1RM back squat was significantly greater
17 in 2S (∆=16.1%) compared to 1S (∆=7.8%) (p<0.05) and (b) both groups significantly increased bench press
18
19 1RM
(1S: ∆=4.6%; 2S: ∆=6.8%), back squat 60% 1RM (1S: ∆= 19.0%; 2S: ∆= 24.3%), bench press 60%
20
1RM (1S: ∆= 15.4%; 2S: ∆= 24.0%) and all MT outcomes (p< 0.05 for all), with no differences between
21
22
experimental groups (1S and 2S).
23
24 CONCLUSIONSː
This study provides evidence that a twice-daily resistance training augments lower-body
25
muscular strength; however, the daily frequency does not seem to have any additive effect on upper-body
26
27
muscular strength, muscular endurance, and muscle hypertrophy in trained men.
28
29
30 words: resistance training; training frequency; muscle thickness; volume load.
Key
31
32
33
34
35
36
37
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39
40
41
42
43
44
45
46
47
48
49
50
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55

Page 36 of 65
Introduction
1
It has been well known that progressive resistance training (RT) promotes neuromuscular adaptations,
2
3 such
as strength gain and muscle hypertrophy 1,2. The magnitude of such muscular adaptations is believed to
4
5 be maximized by the manipulation of RT variables 1. Training frequency is, therefore, considered a
6
7
determinant variable in the hypertrophic response to a given RT program 3. On a general level, RT frequency
8
9
refers to the number of sessions performed during a specific period, usually described on a weekly basis.
10
11
Considering muscle hypertrophy as a primary goal, frequency can be characterized as the number of sessions
12
13
per week (sessions·wk-1) in which the same muscle group is trained 4.
14
15
16 In this context, a higher training frequency may help to achieve a greater volume of training, which may
17
18 in turn enhance the hypertrophic response 5–7. However, systematic reviews with meta-analysis showed no
19
20
significant difference between a higher and a lower frequency on a volume-equated basis for both muscle
21
22 8
23 strength

and hypertrophy 6. Therefore, a higher weekly RT frequency seems to exert an influence on muscle
24
25 strength
and hypertrophy gains only when such protocol results in a higher total load lifted (TLL − sets ×
26
27 repetitions
× load [kg]) compared to a low RT frequency 6,7.
28
29
Although it is well established that the weekly RT frequency does not influence the magnitude of
30
31
32 strength
and hypertrophy gains on a volume-equated basis, there is a paucity of evidence on the effects of
33
34 increased
daily training frequency on morphofunctional adaptations. Anecdotal evidence suggests that there
35
36 1
both physiological and performance benefits associated with a twice-daily training session , as many high-
are
37
38
level RT athletes from the bodybuilding, powerlifting, and weightlifting use multiple daily training sessions
39
40 to increase performance, especially during periods of a high training load 9,10. Previous studies have shown
41
42 weightlifters can perform 2 training sessions on the same day, with no injuries or decrements in
that
43
44 11–18
45 performance

. Moreover, given sufficient recovery after a twice-daily training period, an increase in
46
47 performance
has also been observed 16,17.
48
49 Häkkinen and Kallinen16 investigated neuromuscular adaptations in 10 female athletes during an
50
51
intensive RT period for 3 weeks and during a separate second 3-week training period, when the same TLL
52
53
54 was
distributed into 2 daily sessions. The authors concluded that isometric strength, muscle cross-sectional
55

Page 37 of 65
area, and muscle activation increased during the twice-daily training period, but not in the first training period
1 with

one daily session.
2
3 Additionally, Hartman et al.18 compared the physiological responses in 10 nationally competitive male
4
5
weightlifters during 3 weeks of once- versus twice-daily RT sessions with a similar training volume. There
6
7
was no significant group difference for any of the dependent variables, but the twice-daily training showed an
8
9
10 increase
in: a) percentage change in isometric knee-extension strength (+5.1% vs +3.2%), b) neuromuscular
11
12 activation
(+20.3% vs +9.1%), c) testosterone concentration (+10.5% vs +6.4%), and a decrease in d)
13
14
testosterone:cortisol ratio (–10.5% vs +1.3%) as compared to once-daily training group 18.
15
16
17
Although the aforementioned studies 16,18 suggest that 2 daily sessions can be an effective stimulus to
18
19 maximize
morphofunctional adaptations, both studies were conducted in a high-level competitive
20
21 weightlifters,
thus, the findings cannot necessarily be generalized to other populations such as resistance-
22
23
training practitioners. Moreover, the intervention period lasted only 3 weeks in both studies 16,18. Indeed, to
24
25 best of the authors’ knowledge, no previous study has investigated the effect of increased daily training
the
26
27
frequency on strength and hypertrophy after a longer time frame (e.g., 8 weeks) in resistance-trained subjects.
28
29
30 In light of this scenario, the aim of the present study was to investigate the mid-term effects of training
31
32
muscle groups once- versus twice-daily (where the number of sets per muscle group was equated: 32 weekly
33
34 per muscle group) on neuromuscular performance and morphological adaptations in trained men. It was
sets
35
36
hypothesized that training muscle groups with twice-daily training with 8 sets per muscle per session would
37
38
39 promote

a greater neuromuscular adaptation compared to a once-daily session with 16 sets per muscle. It is
40
41 reasonable to expect that the twice-daily training scheme may result in a greater accumulation of TLL which,
42
43 in turn, has a dose-response relationship with muscle strength 19 and hypertrophy 20,21.
44
45
46
47 Materials and methods
48
49
50
51
Subjects
52
53
54 Twenty-three healthy men (27 ± 5 years [range 20-32 years]; height 175 ± 6 cm; total body mass 80 ±
55
11 kg; RT experience 36 ± 35 months [range 12-60 months]); RT frequency 5 ± 1 sessions·wk-1) volunteered

Page 38 of 65
to participate in this study. The sample size was justified by a priori power analysis based on a pilot study 4,
1 where

the vastus lateralis muscle thickness (MT) was assessed as the outcome measure with a target effect
2
3 size
(ES) difference of 0.75, an alpha level of 0.05, and a power (1 − 𝛽) of 0.80 . Accordingly, the sample
22
4
5 required to achieve 80% power was 9 subjects for each group. All participants were resistance-trained;
size
6
7 performing
RT on a minimum of 3 days per week for at least 1 year at the university’s RT gym. All participants
8
9
regularly performed (minimum frequency of once a week) all exercises used in the training intervention and
10
11 in the strength tests for at least 1 year before entering the study. Participants were free from any existing
12
13
musculoskeletal disorders, history of injury with residual symptoms (pain, “giving-away” sensations) in the
14
15
16 trunk,
upper limbs, and lower limbs within the last year and stated that they had not taken anabolic steroids or
17
18 any
other illegal agents known to increase muscle size at the time or during the previous year. Moreover, the
19
20
participation in the study required a negative answer to all questions on the Physical Activity Readiness
21
22
Questionnaire and had a minimum 1RM parallel back squat of 1.25x total body mass and a 1RM bench press
23
24 of at least equal to total body mass 23. All participants read and signed an informed consent document approved
25
26 by the University Research Ethics Committee (#1.749.141).
27
28
29
30
31 Experimental
procedures
32
33 Participants were pair-matched according to baseline muscle thickness and strength and then randomly
34
35
assigned to 1 of 2 experimental groups: 1 daily session per muscle group (1S, n = 11), which every muscle
36
37
group was trained once a day with 16 sets or 2 daily sessions per muscle group (2S, n = 12), which every
38
39
muscle group was trained twice a day with 8 sets per session. After baseline testing, 4 participants (2 from
40
41
each experimental group) dropped out from the study due to personal reasons not related to the current study.
42
43
44 Additionally,
1 participant of the 2S group reported a knee pain during the training intervention, resulting in
45
46 additional
drop out. Therefore, data from 18 participants were included in the statistical analysis (Table 1).
47
48 All other RT variables (e.g., exercise performed, exercise order, range of repetitions, rest interval
49
50
between sets, and exercises, etc.) were maintained constant. The experimental period lasted 11 weeks: 1st
51
52
week— familiarization period; 2nd week—pre-intervention period (baseline); 3rd-10th week—training
53
54
intervention period; and 11th week—post-intervention period. The training intervention period lasted 8 weeks,
55

Page 39 of 65
and the TLL was calculated for every RT session to compare the accumulated external training load among
1 experimental

groups, across the intervention period.
2
3 Testing was conducted before intervention (pre) and after the 8-week (post-8) period for maximal
4
5 voluntary
muscle strength (1 repetition maximum test [1RM] for bench press and parallel back squat
6
7
exercises), muscular endurance (maximum repetitions at 60% of 1RM test [60%1RM] for bench press and
8
9
parallel back squat exercises), and MT of the biceps brachii, triceps brachii, vastus lateralis, anterior
10
11
quadriceps (rectus femoris and vastus intermedius), and pectoralis major. In the first week, participants
12
13
attended 2 familiarization sessions in the laboratory. The participants refrained from all physical exercises
14
15
16 other
than activities of daily living for at least 48 hours before the first familiarization session. In the first
17
18 session,
participants were familiarized with the 1RM and 60%1RM tests. The following day (24 hours after),
19
20
participants were familiarized with standard procedures adopted in all RT exercises, such as body position,
21
22
cadence, range of motion, rest, etc. In addition, participants were trained and instructed to record their dietary
23
24
intake.
25
26
27
28
29
*** Table 1 about here ***
30
31
32
33 Resistance
training program
34
35 The RT protocol consisted of 9 exercises targeting each of the major muscle groups. Participants were
36
37
instructed to refrain from any additional resistance-type training for the duration of the study. The specific
38
39
protocols for experimental groups are outlined in Table 2. The exercises were chosen based on their common
40
41
inclusion in bodybuilding and strength- type RT programs 24.
42
43
44 The weekly training protocol for all groups consisted of 2 split routines targeting specific muscle groups:
45
46 split
routine A (Arout) and split routine B (Brout). The S1 weekly training consisted of 4 days of RT (Arout +
47
48 B rout + Arout + Brout) with a once-daily session of 16 sets per muscle group, whereas S2 weekly training
49
50
consisted of 4 days of RT with twice-daily sessions of 8 sets per muscle group (Arout [AM/PM] + Brout
51
52
[AM/PM] + Arout [AM/PM] + Brout [AM/PM]). Thus, both experimental groups performed 16 daily sets and
53
54 32 weekly sets per muscle group, comprising 16 sets of multi-joint exercises and 16 sets of single-joint
55
exercises, except for hamstrings muscles that were stimulated with 32 weekly sets of single-joint exercise

Page 40 of 65
(machine lying leg curl). In this context, both experimental groups stimulated each muscle group 2 days per
1 week,

but the RT frequency was 2 and four sessions·wk-1 per muscle group for 1S and 2S, respectively.
2
3 In an effort to help ensure that the participants of the S1 group were not exposed to a new stimulus
4
5 concerning
the time of day-specific RT, they held their sessions in the period of the day when they usually
6
7
trained before the study. Moreover, this procedure was adopted due to the logistical difficulty in getting all
8
9
participants to train in the same period of the day. Four participants performed all sessions in the morning (9-
10
11 11 AM), 2 in the afternoon (4-5 PM), and 2 in the night (8-9 PM). For the S2 group, the first session of the
12
13
day was standardized between 6-8 AM. The second session of the day was performed according to the
14
15
16 availability
of the participants, respecting an interval of at least 8 hours for the first session. Therefore, the
17
18 second
session of all participants was always between 5-7 PM.
19
20 Each set involved 8–10 maximum repetitions (8-10RM) with 90 seconds of rest between sets and 180
21
22
seconds rest between exercises. All sets were conducted to the point of momentary concentric muscular
23
24
failure, operationally defined as the inability to perform another concentric repetition while maintaining
25
26
adequate technique. The external load (kg) was adjusted for each exercise, as needed, on successive sets to
27
28
29 ensure

that participants achieved failure in the target repetition range. The cadence of repetitions was
30
31 conducted
in a controlled fashion, with concentric and eccentric actions of approximately 1.5 seconds, for
32
33 total
repetition duration of approximately 3 seconds. All RT sessions were preceded by a specific warm-up
34
35
consisting of 2 sets of 10 repetitions with 50% of the load used in the first set of all exercises of the session.
36
37 participants reported a rating of perceived exertion (RPE) based on the RPE/repetitions in reserve scale 25
All
38
39 of 9.5–10 for all sets and exercises in RT sessions.
40
41
All routines were directly supervised by the research assistants to ensure the correct performance of the
42
43
44 respective
routines. Before the training intervention period, all participants underwent 10RM testing 24 to
45
46 determine
individual initial training loads for each exercise. Attempts were made to progressively increase the
47
48
load lifted each week while maintaining the target repetition range. No injuries were reported and the
49
50
adherence to the program was 100% for all groups.
51
52
53
54 *** Table 2 about here ***
55
Estimate of food intake

Page 41 of 65
To avoid potential dietary confounding of results, participants were advised to maintain their customary
1 nutritional

regimen and to avoid taking any supplements during the study period. Dietary nutrient intake was
2
3 assessed
by a 24-hour food diary on 2 nonconsecutive weekdays and 1 day of the weekend. The participants
4
5 were
instructed to record in detail the time of consumption and types and quantity of food preparations
6
7
consumed during 24 hours. The quantity of food was recorded in cooking units (spoons, cups, and glasses)
8
9 transformed into grams. The estimation of energy intake (macronutrients) was analyzed by NutWin
and
10
11
software (UNIFESP, Sao Paulo, Brazil). The estimated food intake was assessed during weeks 1, 4, and 8 of
12
13
the training intervention period.
14
15
16
17
19
20
21
22
Maximum Dynamic Strength (1RM)
23
24 Maximum dynamic strength was assessed through 1RM testing using the bench press (1RMBENCH), and
25
26
parallel back squat (1RMSQUAT) exercises (Nakagym, SP, Brazil). The testing protocol followed previous
27
28
29 recommendations

by Haff and Triplett10. The test-retest intraclass correlation coefficient (ICC), coefficient of
30
31 variation
(CV), and the standard error of the measurement (SEM) from our laboratory for 1RMBENCH were
32
33 0.989,
0.8%, and 2.05 kg, respectively. The ICC, CV, and SEM for 1RM leg press were 0.990, 0.7%, and 1.95
34
35 respectively.
kg,
36
37
38
Muscle endurance (ME)
39
40 For assessments of muscle endurance (ME), participants performed as many repetitions as possible to
41
42
43 muscular

failure with proper form at 60% of 1RM load 26 on both the bench press (60%1RMBENCH) and parallel
44
45 back
squat (60%1RMSQUAT). The test-retest ICC, CV, and TEM calculated through the data collected during
46
47 the
familiarization period and the pre-intervention period (5 days between the test-retest) for 60%1RMBENCH
48
49
were 0.926, 2.7%, and 1.1 repetitions, respectively. The ICC, CV, and TEM for 60%1RM SQUAT were 0.903,
50
51
3.6%, and 1.6 repetitions, respectively.
52
53
54
55
Muscle thickness (MT)

Page 42 of 65
Ultrasound imaging was used to obtain the assessment of MT. A trained technician performed all testing
1 using

an A-mode ultrasound imaging unit (Bodymetrix Pro System; Intelametrix, Inc., Livermore, CA, USA).
2
3 After
a generous application of a water-soluble transmission gel (Mercur S.A. – Body Care, Santa Cruz do
4
5 Sul,
RS, Brazil) to the site to be evaluated, a 2.5-MHz linear probe was placed perpendicular to the tissue
6
7
interface without depressing the skin. Equipment settings were optimized for image quality, according to the
8
9
manufacturer’s user manual, and held constant among testing sessions. When the quality of the image was
10
11
deemed to be satisfactory, the image was saved to the hard drive, and MT dimensions were obtained by
12
13
measuring the distance from the subcutaneous adipose tissue–muscle interface to the muscle-bone interface,
14
15
16 as per methods used by Abe et al.27.
17
18 Measurements were taken on the right side of the body at 5 sites: biceps brachii (MTBB), triceps brachii
19
20 TB), vastus lateralis (MTVL), anterior quadriceps (MTAQ) and pectoralis major (MTPM). Upper arm
(MT
21
22
measurements were conducted while participants were standing. Afterwards, participants laid supine on an
23
24
examination table for measurements of the thigh muscles. For the anterior and posterior upper arm,
25
26
measurements were taken 60% distal between the lateral epicondyle of the humerus and the acromion process
27
28
29 of the scapula; for the thigh muscle, measurements were taken at 50% of the distance between the lateral
30
31 condyle
of the femur and greater trochanter. For chest, measurements were taken at the site between third and
32
33 fourth
of costa (third intercostal space) under the clavicle midpoint.
34
35 For each measurement, the examined limb was secured to minimize unwanted movement. To maintain
36
37
consistency between pre- and post-intervention testing, each site was marked with henna ink (reinforced every
38
39
week). In an effort to help ensure that swelling in the muscles from training did not obscure results, images
40
41
were obtained 72 h before commencement of the study and after the final training session. This is consistent
42
43
44 with
research showing that an acute increase in muscle thickness returns to baseline within 48 h following a
45
28
46 RT session .
47
48 To further ensure the accuracy of measurements, at least 3 images were obtained for each site. If
49
50
measurements were within 1mm of one another the figures were averaged to obtain a final value. If
51
52
measurements were more than 1mm of one another, a fourth image was obtained, and the closest 3
53
54
measurements were then averaged. All images were performed by the experienced researcher who was blind
55
to the RT protocol performed. The test-retest ICC for MTBB, MTTB, MTVL, MTAQ, and MTPM were 0.972,

Page 43 of 65
0.981, 0.993, 0.996 and 0.984, respectively. The CV for these measures were 1.7, 1.2, 0.9, 0.8%, 1.1%,
1 respectively.

The TEM for these measurements were 0.44, 0.34, 0.52, 0.43, 0.32 mm, respectively.
2
3
4
5 Total
load lifted (TLL)
6
7 Total load lifted (TLL − sets × repetitions × load (kg)] was calculated from training logs filled out by
8
9
research assistants for every RT session. The accumulated TLL (ATLL) was the sum of all RT weeks. Only
10
11
repetitions performed through a full range of motion were included for analysis. The data were expressed in
12
13
kilograms (kg). The ∆TLL described the difference in the TLL between weeks 8 and 1 (e.g., TLL in the week
14
15
16 8 minus the TLL in the week 1).
17
18
19
20
Overall well-being status (WB)
21
22 The well-being questionnaire (WB) assessed participants’ fatigue, sleep quality, general muscle
23
24
soreness, stress levels, and mood on a five-point scale (scores of 1 to 5). Well-being status was then determined
25
26 by summing the five scores 29. The WB questionnaire was completed 10-min before each RT session’ warm-
27
28
29 up.

The total WB status was the sum of all weeks of RT. The data were expressed in arbitrary units (AU).
30
31 Moreover,
the weekly coefficient of variation (CV) of the WB over the 8 weeks of the intervention training
32
33 was
calculated.
34
35
36
37
38 Statistical
analysis
39
40 The normality and homogeneity of the variances were verified using the Shapiro-Wilk and Levene tests,
41
42
respectively. A paired t-test was used to compare mean values of the baseline descriptive variables, well-being
43
44
status (WB), weekly coefficient of variation (CV%) of the WB, ATLL, and ∆TLL (week 8 – week 1) between-
45
46
groups (1S vs. 2S). A 2-way repeated-measures ANOVA was used to compare 1RMBENCH, 1RMSQUAT,
47
48
60%1RM BENCH, 60%1RMSQUAT, MTBB, MTTB, MTVL, MTAQ, and MTPM, time effect (pre vs post week 8) x 2
49
50
51 groups

(1S vs. 2S). The absolute changes (i.e., post 8 weeks – pre) in 1RM, 60%1RM, and MT measures were
52
53 compared
between the protocols using a t-test. A 2-way repeated-measures ANOVA (3x2) (pre vs post week 4
54
55 vs post week 8) x 2 groups (1S vs. 2S) was used to compare the food intake variables (Total kcal, protein,
carbohydrate, and lipids). Post hoc comparisons were performed with the Bonferroni correction. Effect sizes

Page 44 of 65
(ES) in absolute differences (post-8 week - pre) in raw values of the variables using the standardized difference
1 based

on Cohen’s d units by means (d value) 30.
2
3 The ES results were qualitatively interpreted using the following thresholds: < 0.2, trivial; 0.2 - 0.6, small;
4
5 0.6
-1.2, moderate; 1.2 - 2.0, large; 2.0 - 4.0; very large and > 4.0, extremely large. The smallest worthwhile
6
7
change (SWC) for the dependent variables was calculated by the formula SWC = typical error of measurement
8
9
(TEM) x 2. The SWC for 1RMBENCH, 1RMSQUAT, 60%1RMBENCH, 60%1RMSQUAT, MTBB, MTTB, MTVL, MTAQ,
10
11 MTPE for these measures are 4.56 kg, 4.06kg, 2.2 repetitions, 3.2 repetitions, 0.88 mm, 0.68 mm, 1.04 mm,
and
12
13
0.86 mm, and 0.64 mm, respectively. SWC was used as a trivial area (gray bar) of the smallest true individual
14
15
16 change
of subjects 31. We defined an individual as “responding” positively to training with a response greater
17
18 than
+1SWC from zero for increases in the dependent variables; if not, he was considered as a non-responder.
19
20 percentage of participants exceeding the SWC was calculated for all dependent variables. All analyses were
The
21
22
conducted in SPSS-22.0 software (IBM Corp., Armonk, NY, USA). The adopted significance was p ≤ 0.05.
23
24
25
26 Results
27
28
29
No significant difference was noted between groups in any baseline measurements (all p > 0.05 [Table
30
31 1]).
There was no significant difference in any dietary intake variables (total kcal, proteins, carbohydrate, and
32
33 lipids
grams) either within- or between-groups over the course of the study (all p > 0.05 [Table 3]).
34
35 A significant main effect of time (p = 0.001), but not group x time interaction (p = 0.188), was observed
36
37 1RMBENCH. There was a significant main effect of time (p = 0.001) and group x time interaction (p =
for
38
39
0.043) for 1RMSQUAT (Table 4). The absolute change analysis (Figure 1A) revealed a significant difference
40
41
between groups for the 1RMSQUAT (p = 0.014, ES = 1.30, 90%CI = 1.05 to 1.55) but not for 1RMBENCH (p =
42
43
44 0.182,
ES = 0.66, 90%CI = 0.33 to 0.99). The individual analyses showed that 44.4% and 66.6% of the
45
46 participants
in 1S and 2S groups increased the 1RMBENCH. However, 77.7% (1S) and 100% (2S) of sample
47
48
increased in 1RMSQUAT after training period (Figure 1A).
49
50 A significant main effect of time (all p = 0.001), but not group x time interaction (all p > 0.05), was
51
52
observed for 60%1RMBENCH and 60%1RMSQUAT, respectively (Table 4). The absolute change analysis (Figure
53
54 did not reveal any significant difference between protocols for the 60%1RM test (all p > 0.05). The effect
1A)
55
size (in absolute differences post 8 weeks – pre) was moderate between 1S vs 2S in 60%1RMBENCH (ES = 0.75,

Page 45 of 65
90%CI = 0.48 to 1.02) and small in 60%1RMSQUAT (ES = 0.28, 90%CI = -0.18 to 0.74) (Figure 1A). The
1 individual

analyses showed that 60%1RMBENCH increased in 33.3% of the participants in 1S group and 77.7%
2
3 of the participants in group 2S. Similarly, 55.5% of the participants in 1S group and 77.7% of the participants
4
5 in group 2S increased the 60%1RMSQUAT (Figure 1A).
6
7
8
9
10
12
13
14
15
16 A significant main effect of time (all p < 0.05), but not group x time interaction (all p > 0.05) was
17
18 observed
for MTBB, MTTB, MTVL, MTAQ, and MTPM (Table 5). The absolute change analysis (Figure 1B) did
19
20 reveal any significant differences between protocols for the MT increases (all p > 0.05). The effect size (in
not
21
22
absolute differences post 8 weeks – pre) was small between 1S vs 2S in MTBB (ES = 0.10, 90%CI = -0.12 to
23
24
0.32), MTTB (ES = 0.37, 90%CI = 0.17 to 0.57), MTVL (ES = 0.09, 90%CI = -0.13 to 0.21), MTAQ (ES = 0.13,
25
26
90%CI = -0.01 to 0.27) and MTPM (ES = 0.29, 90%CI = 0.03 to 0.65) (Figure 1B). The percentual of
27
28
29 participants

responding positively to training between groups in the muscle thickness was similar (>88% in
30
31 MT BB, >77% in MTTB and MTVL, >66% in MTAQ, and > 55% in MTPM) (Figure 1B).
32
33
34
36
37 *** Figure 1 about here ***
38
39
40
41
A significant between-group difference was observed for ATLL (p = 0.0002, mean difference = 116,221
42
43
44 CI95% = 61,427 to 170,816 kg) (Figure 2A). For ∆TLL, no significant difference was observed between-
kg,
45
46 group
(p = 0.412; 1S = 15.3% vs 2S = 18.5%) (Figure 2B).
47
48
49
51
52
53
54
55

Page 46 of 65
No significant between-group difference was observed for total WB status (p = 0.872, mean difference
1 = 1.7 AU, CI95% = -2.1 to 2.4 AU) (Figure 3A). No significant between-group difference was observed for
2
3 the
CV% of WB (p = 0.787; 1S = 9.8 ± 4.7% vs 2S = 9.2 ± 4.7%) (Figure 3B).
4
5
6
8
9
10
11 Discussion
12
13
This study investigated the mid-term effects of training muscle groups once- versus twice-daily on
14
15
16 neuromuscular
performance and morphological adaptations in trained men. The primary and novel finding of
17
18 this
study was that training a muscle group only once a day is as efficient as training twice a day to promote
19
20
increase in upper-body maximal strength, muscular endurance, and muscle size. Alternatively, the increase in
21
22
lower-body maximal strength seems to be more pronounced when this portion of the body is stimulated with
23
24
twice-daily sessions.
25
26 Regarding maximal dynamic strength, previous research has demonstrated a possible neuromuscular
27
28
29 benefit

from dividing the same daily training load into 2 separate sessions. Häkkinen and Kallinen16
30
31 investigated
neuromuscular adaptations in 10 female athletes during an intensive strength-training period for
32
33 3 weeks and during a separate second 3-week training period, when the same total training load was distributed
34
35 2 daily sessions. The authors concluded that isometric strength and muscle activation increased during
into
36
37 twice-daily training period, but not in the first training period with 1 daily session 16. Likewise, Häkkinen
the
38
39 Pakarinen13 reported that male strength athletes performing 2 weeks of twice-daily training experienced
and
40
41
a significant increase in isometric knee-extension strength compared with the same daily training load
42
43
44 performed
in once-daily sessions. Moreover, Hartman et al.18 compared the physiological responses in 10
45
46 nationally
competitive male weightlifters during 3 weeks of once- versus twice-daily RT sessions with similar
47
48
training volumes. There were no significant group differences for any of the dependent variables, but the
49
50
twice-daily training group had a greater percentage change in isometric knee-extension strength (+5.1% vs
51
52
+3.2%), clean and jerk performance (+1.9% vs +0.3%), and neuromuscular activation (+20.3% vs +9.1%).
53
54
Therefore, the authors concluded that there were no additional benefits from increased daily training
55

Page 47 of 65
frequency, but the increase in isometric strength and neuromuscular activation for the twice-daily group might
1 provide

some rationale for dividing training load in an attempt to reduce the risk of overtraining 18.
2
3 Although the aforementioned studies 13,16,18 suggest that 2 daily sessions can be an effective stimulus to
4
5 maximize
muscle strength, these studies were conducted in high-level competitive weightlifters. Additionally,
6
7 intervention period lasted only 2-3 weeks. Thus, it is difficult to draw conclusions and/or compare the
the
8
9
findings of Häkkinen and Pakarinen13, Häkkinen and Kallinen16 and, Hartman et al.18 with the results of the
10
11
present study. Interestingly, in the present study, the magnitude of increase for training with 2 sessions was
12
13
higher than 1 session per day for 1RMSQUAT but not for 1RMBENCH.
14
15
16 The motor learning theory dictates that practicing a given exercise more frequently over time leads to
17
18 better
skill acquisition, conceivably through neural enhancements 32. Compared to the bench press, the back
19
20
squat exercise involves a greater amount of muscle mass, requires more stabilization and balance, which may
21
22
result in greater neural demand. In this context, it is plausible to assume that due to the greater coordinative
23
24 neural demand of this exercise, the more frequently practice of the squat in the 2S group led to better
and
25
26
neural enhancements and, consequently, a greater strength development. Another possible explanation for this
27
28
29 result

is that, even though not significantly different from each group, the training frequency that the
30
31 participants
reported to adopt previously to the experiment was lower for the lower limbs muscles compared
32
33 to the upper ones (Table 1), which might actually be observed in the higher percentage increases in 1RMSQUAT
34
35
(7.8% and 16.1% for 1S and 2S, respectively) compared to 1RMBENCH (4.6 and 6.8 for 1S and 2S, respectively)
36
37 in both groups. In this sense, it could be speculated that different muscle groups may present distinct curve-
38
39
responses to increased training frequency.
40
41
Additionally, the higher TLL accumulated in the back squat exercise (128570 ± 18211 kg vs 106289
42
43
44 ± 10344 kg [p = 0.005] for 2S and 1S groups, respectively) might also suggest that the 2S group was able to
45
46 maintain
a higher intensity (more external load per set) during the experimental period, explaining the larger
47
48
magnitude-increase observed in 1RMSQUAT compared to the 1S group. While 1S group performed 8 sets of
49
50
squat exercise over the same session with only 90 seconds of rest between sets, 2S group distributed the 8 sets
51
52 in 2 blocks of 4 sets, with and interval of 8 hours between blocks. Therefore, this 8-hour recovery between the
53
54
fourth and fifth sets, possibly resulted in less accumulated fatigue along the sets and, consequently,
55
maintenance of intensity (load) compared to the 1S. In fact, the findings of Schoenfeld et al. 33 indicate that

Page 48 of 65
maximal strength benefits are obtained from the use of heavy loads. Conversely, the TLL accumulated in the
1 bench

press exercise did not differ between the groups (73287 ± 10046 kg vs 78004 ± 8333 kg [p = 0.342] for
2
3 1S
and 2S groups, respectively).
4
5 Another interesting aspect of the present findings is that although a significant within-group difference
6
7 observed in the 1S for 1RMBENCH, the absolute difference from pre- to post-8 weeks (4.1 kg) was below
was
8
9 SWC for this measurement (4.6 kg). Therefore, it is possible that the change observed for 1RM BENCH in
the
10
11 1S might be explained by the expected variation of this outcome, instead of due to the RT intervention.
the
12
13
Conversely, the current findings indicate that daily frequency RT does not influence the magnitude of
14
15
16 muscular
endurance. Although no significant difference between groups was observed for measures of upper-
17
18 and
lower-body muscular endurance, on a percentage basis, an advantage was observed for 2S compared to
19
20 1S with respect to the increase in 60%1RMBENCH (24.0% vs. 15.4%, respectively) and 60%1RMSQUAT (24.3%
21
22 19.0%, respectively). Moreover, the effect size between conditions in absolute difference (post - pre) was
vs.
23
24
small to moderate and favorable to 2S (ES = 0.75 and 0.28 for 60%1RMBENCH and 60%1RMSQUAT,
25
26
respectively), suggesting a meaningful difference in results.
27
28
29
When comparing 1 vs 2 sessions per muscle group per week, Brigatto et al. 4 observed that only the 2
30
31 sessions/week
intervention resulted in a significant increase in 60%1RMBENCH, suggesting a possible benefit
32
33 of a higher RT frequency for the development of muscular endurance 4. The authors suggest that the results
34
35
observed for 60%1RM outcome can be explained by the greater TLL accumulated by 2 compared to 1
36
37
session/week intervention, which implies that distributing the weekly RT volume in 2 sessions results in a
38
39
higher weekly TLL per muscle group 4. Alternatively, it is possible that spreading out the TLL over more daily
40
41
sessions enhances buffering capacity to a greater extent than performing a higher per-session volume less
42
43
44 frequently,
thereby increasing fatigue resistance. Further research is needed to determine the causal effects of
45
46 this
phenomenon.
47
48 Regarding the measurement of MT, while both groups demonstrated increased MTBB, MTTB, MTVL,
49
50 MT AQ, and MTPM no significant between-groups difference was observed. Thus, the present findings
51
52
essentially reflected recent meta-analytic data showed no significant difference between a higher and a lower
53
54
weekly RT frequency for muscle hypertrophy 6.
55

Page 49 of 65
Häkkinen and Kallinen16 investigated morphological adaptations in 10 female athletes during an
1 intensive

RT period for 3 weeks and during a separate second 3-week training period when the same TLL was
2
3 distributed
into 2 daily sessions. The authors concluded that muscle cross-sectional area of the quadriceps
4
5 femoris
increased during the twice-daily training period, but not in the first training period with 1 daily session.
6
7
They suggested that twice-daily training sessions period was more conducive to muscle hypertrophy and might
8
9
offer a more effective neuromuscular training stimulus compared to once-daily training session period 16. The
10
11
results observed in the present study did not corroborate the assumption proposed by Häkkinen and Kallinen16,
12
13
possibly due to the methodological differences (sample size and gender, duration of intervention, etc.) between
14
15
16 the
studies.
17
18 The present study expands on previous findings by providing a robust evidence of the greater TLL
19
20
increase with a distribution of RT daily sets into smaller units, such as 2 daily sessions, compared to the same
21
22
23 number

of sets applied in a single daily session. This is important, as the increment in muscle strength and
24
25 mass
is strongly dependent on TLL of RT. In fact, a clear dose-response relationship has been reported
26
27 between
TLL and both muscle strength 19 and hypertrophy 20,21. Therefore, it is plausible to hypothesize that
28
29 greater TLL achieved through 2 daily sessions protocol, if maintained for a longer time frame (more than
the
30
31 8 weeks), might culminate in a significantly greater increase in hypertrophy compared to an once-daily session
32
33
protocol. This hypothesis requires further investigation.
34
35
36 Additionally, it is possible that the high RT volume applied in both experimental groups was the main
37
38
factor for similar hypertrophy gains observed between groups. Thirty-two weekly sets were performed per
39
40
muscle group in both 1S and 2S. The authors employed a high RT volume due to the dose-response
41
42
relationship previously reported by Brigatto et al. (2019b) in which 32 weekly sets resulted in a greater
43
44
45 increase

in MT compared to 16 and 24 sets in resistance-trained men. Thus, even the 2S group accumulating
46
47 16.4
% more TLL than the 1S group, it is plausible to hypothesize that, when performing 32 weekly sets per
48
49 muscle
group, a training volume plateau is reached and no further increase in muscle hypertrophy would be
50
51
observed. In other words, it is possible that when weekly RT sets employed is high enough, there is a limited
52
53
advantage of a higher training frequency and a greater TLL to maximize muscle hypertrophy. This hypothesis
54
55
warrants further investigation, since Schoenfeld et al. (2019a) reported a greater increase in MT of rectus

Page 50 of 65
femoris and vastus lateralis with 45 weekly sets per muscle group versus 9 and 27 weekly sets for 8 weeks in
1 resistance-trained

men. In addition, it is interesting to note that the increment in lower limbs muscles thickness
2 35
3 was
not influenced by the larger increase observed in 1RMSQUAT in the 2S group. Indeed, Dankel et al.
4
5 suggest
that training induced an increase in muscle size appear to play little if any role with respect to training-
6
7
induced increase in muscle strength.
8
9
10 It might be speculated that the difference observed in ATLL between both groups was not sufficient to
11
12 induce
a significant difference on morphological adaptations. Indeed, the training dose required for increasing
13
14
muscle strength seems to be smaller than the one needed for increasing muscle size 33. In this sense, individuals
15
16 large RT-experience as the ones of the current study might need to accumulate a greater training load or
with
17
18 be submitted to a longer training intervention period in order to enhance morphological adaptations.
19
20
21 On the other hand, the ΔTLL was similar between conditions (1S = 15.3% vs 2S = 18.5%). Thus, these
22
23
findings provide a straight evidence that both daily RT frequencies result in a similar capacity to accumulate
24
25
TLL over the weeks. This may help to explain why the muscle hypertrophy was similar between groups even
26
27
with 2S resulting in a higher ATLL. Therefore, the TLL progression could be an important variable to
28
29
maximize the muscle hypertrophy responses in resistance-trained men at least when determined at the group
30
31 36
32 level
. In this context, the relative analysis of the TLL (ΔTLL) can be a complementary and more appropriate
33
34 tool
for comparing the accumulation (progression) of external load among interventions performed by
35
36
different groups of participants.
37
38
39
Participants’ fatigue, sleep quality, general muscle soreness, stress levels, and mood 29 were assessed
40
41 immediately before performing each RT session’ warm-up. Regardless of the number of daily sessions, the
42
43 WB status was similar during the 8-weeks intervention period as well as the weekly percentage variation
44
45
between sessions. That is, the number of daily sessions did not influence the psychobiological stress imposed
46
47 on the 2 experimental groups during the intervention. This finding is quite relevant from a practical application
48
49
standpoint, since those aforementioned variables, especially psychological stress, may negatively influence
50
51
isometric peak power 37. Psychological questionnaires provide a simple and cost-effective monitoring tool for
52
53
54 coaches
and can be implemented regularly throughout a RT periodization. Future studies adopting such a
55
refined methodology are encouraged.

Page 51 of 65
Some limitations of the current investigation should be acknowledged. First, the study period lasted only
1 8 weeks. Although this duration was sufficient to achieve a significant increase in muscular strength and
2
3 hypertrophy
for both groups, it is conceivable that results between groups would have diverged over a longer
4
5 time
frame. Second, the small sample size affected statistical power. As is the case in most longitudinal RT
6
7
studies, a high degree of interindividual variability was noted among participants, which limited the ability to
8
9
detect a significant difference in several outcome measures. Despite this limitation, the analysis of the effect
10
11 provides a good basis for drawing inferential conclusions from the results. The findings of the current
size
12
13
study are specific to resistance-trained men and, therefore, cannot necessarily be generalized to other
14
15
16 populations
including adolescents, women, and the elderly. It is possible that a higher RT daily frequency and
17
18 volume
may not be as well tolerated in these individuals and perhaps could hasten the onset of overtraining,
19
20
when combined with a high intensity of effort and longer training periods. Future research is required to
21
22
determine the response related to daily sessions frequency across different populations. Furthermore, the
23
24
results may have been influenced by the novelty of changing RT programs. Pre-study interviews revealed that
25
26 all participants regularly trained with a frequency of 1-2 sessions per muscle group per week and once per
27
28
29 day.

Therefore, the participants who were randomized to the 2S group were exposed to a new stimulus
30
31 concerning
the weekly frequency (2 times per muscle group per day), whereas the 1S group trained with their
32
33 usual
frequency (1 time per muscle group per day). Finally, although each participant performed the training
34
35
sessions at the same period of the day throughout the intervention, it was not possible to standardize the time
36
37 of day at which the RT was performed among the participants. This may have exerted some possible influence
38
39 on the neuromuscular outcomes, since it is well established that human exercise performance varies according
40
41
to the time of day, due to factors such as chronobiology and circadian rhythm 38. However, a systematic review
42
43
44 with
meta-analysis regarding the effects of time of day-specific RT on morphofunctional adaptations showed
45
46 that
the increase in strength and muscle hypertrophy is similar irrespective of the time of day at which the
47
48
training is performed 39. This topic warrants further investigation.
49
50
51
52
53 Conclusions
54
55 In conclusion, this study provides evidence that a twice-daily RT augments lower-body muscular
strength; however, the RT daily frequency does not seem to have any additive effect on upper-body muscular

Page 52 of 65
strength, muscular endurance, and muscle hypertrophy. Given that training the same muscle group into smaller
1 units,

such as 2 daily sessions, is thought to be less energetically taxing compared to condensing the daily
2
3 volume
in a single session, dividing the muscle group RT volume in 2 sessions/day provides a practical means
4
5 to perform a higher TLL per muscle group, while maintaining the intensity of effort and providing adequate
6
7
recovery between sessions. Alternatively, 1S may be more time-efficient for those with limited time for RT,
8
9 as it requires only 1 training session/day while producing a similar improvement in most outcome measures.
10
11
12
13
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30. Dankel SJ, Mouser JG, Mattocks KT, Counts BR, Jessee MB, Buckner SL, et al. The widespread
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29 misuse of effect sizes. J Sci Med Sport [Internet]. 2017 May;20(5):446–50. Available from:
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31 http://dx.doi.org/10.1016/j.jsams.2016.10.003
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31. Hopkins WG, Marshall SW, Batterham AM, Hanin J. Progressive statistics for studies in sports
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34 medicine and exercise science. Med Sci Sports Exerc. 2009;41(1):3–12.
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36 32.

Shea CH, Lai Q, Black C, Park JH. Spacing practice sessions across days benefits the learning of
37 motor skills. Hum Mov Sci. 2000;19(5):737–60.
38
39 33.
Schoenfeld BJ, Grgic J, Ogborn D, Krieger JW. Strength and hypertrophy adaptations between low-
40 vs. High-load resistance training: A systematic review and meta-analysis. J Strength Cond Res.
41
42 2017;31(12):3508–23.
43
44 34.
Schoenfeld BJ, Contreras B, Krieger J, Grgic J, Delcastillo K, Belliard R, et al. Resistance Training
45 Volume Enhances Muscle Hypertrophy but Not Strength in Trained Men. Med Sci Sports Exerc.
46
47 2019;51(1):94–103.
48
35. Dankel SJ, Buckner SL, Jessee MB, Grant Mouser J, Mattocks KT, Abe T, et al. Correlations Do Not
49
50 Show Cause and Effect: Not Even for Changes in Muscle Size and Strength. Sport Med. 2018;48(1).
51
52 36.
Damas F, Angleri V, Phillips SM, Witard OC, Ugrinowitsch C, Santanielo N, et al. Myofibrillar
53 protein synthesis and muscle hypertrophy individualized responses to systematically changing
54
55 resistance training variables in trained young men. J Appl Physiol. 2019;127(3):806–15.
37. Stults-Kolehmainen MA, Bartholomew JB. Psychological stress impairs short-term muscular

Page 55 of 65
recovery from resistance exercise. Med Sci Sports Exerc. 2012;44(11):2220–7.

38. Drust B, Waterhouse J, Atkinson G, Edwards B, Reilly T. Circadian rhythms in sports performance -
1
2 An update. Chronobiol Int. 2005;22(1):21–44.
3
39. Grgic J, Lazinica B, Garofolini A, Schoenfeld BJ, Saner NJ, Mikulic P. The effects of time of day-
4
5 specific resistance training on adaptations in skeletal muscle hypertrophy and muscle strength: A
6
systematic review and meta-analysis. Chronobiol Int [Internet]. 2019;36(4):449–60. Available from:
7
8 https://doi.org/10.1080/07420528.2019.1567524
9
10
11
12
13 NOTES
14
15
16
17
Conflicts of interest. The authors certify that there is no conflict of interest with any financial organization
18
regarding the material discussed in the manuscript.
19
20
21 Authors’
contributions. DAC, FAB, TVB, PHM and CRL conceived and designed research. DAC, FAB, TVB
22
23 and

JBBDC conducted experiments. FAB, TVB and MSA analyzed data. DAC, FAB, TVP, JBBDC, MSA,
24
PHM and CRL wrote the manuscript. All authors interpreted the results of experiments, edited, revised, and
25
26 approved
the manuscript.
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47 TABLES
48
49
50 Table 1. Baseline descriptive statistics (mean ± SD).
51
52 Variables 1S (n=9) 2S (n=9) P-value
53
54
55 Age (years) 26.3 ± 6.2 24.9 ± 4.2 0.573

Page 56 of 65
Total Body Mass (kg) 79.7 ± 17.1 81.4 ± 6.3 0.774
1 Height (cm) 174 ± 6.6 176 ± 5.5 0.446

2
3 RT Experience (months) 36.8 ± 35 36.3 ± 34 0.978
4
5 RT Frequency (sessions·wk-1) 4.8 ± 0.8 4.9 ± 0.9 0.792
6
7 RT Frequency UL (sessions·wk-1) 3.6 ± 0.9 3.6 ± 0.7 0.992
8
9 RT Frequency LL (sessions·wk-1) 1.2 ± 0.4 1.3 ± 0.5 0.623
10
11 1RMBENCH (kg) 94 ± 14 101 ± 14 0.292
12
13 1RMSQUAT (kg) 138 ± 14 126 ± 23 0.221
14
15 Legend: 1S = one session per day; 2S = two sessions per day; RT = resistance training; UL =
16 upper limbs; LM = lower limbs; 1RMBENCH = one maximal repetition test in bench press
17 exercise; 1RMSQUAT = one maximal repetition test in parallel back squat exercise.
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Page 57 of 65
1 2.
Table Training protocols for 1S and 2S.
2
3 Period Monday Tuesday Wednesday Thursday Friday
4
5 Arout Brout
6
7 Bench press 8x8-10RM Lat pulldown 8x8-10RM Bench press 8x8-10RM Lat pulldown 8x8-10RM
8 Straight-arm pulldown 8x8- Dumbbell flat fly 8x8- Straight-arm pulldown
Dumbbell flat fly 8x8-10RM
9 10RM
10 Cable triceps 8x8-10RM 10RM XXXX Cable triceps 8x8-10RM 8x8-10RM
11 1S Biceps curl 8x8-10RM Biceps curl 8x8-10RM
(n=9)
12 Parallel back squat 8x8-
Parallel back squat 8x8-10RM
13 Lying leg curl 16x8-10RM 10RM Lying leg curl 16x8-10RM
14
15 Leg extension 8x8-
Leg extension 8x8-10RM
16 10RM
17
18 Arout Brout Arout Brout
19
21 Dumbbell flat fly 4x8-10RM
Straight-arm pulldown 4x8- Dumbbell flat fly 4x8- Straight-arm pulldown
22 10RM
XXXX
10RM 4x8-10RM
23 AM Cable triceps 4x8-10RM Biceps curl 4x8-10RM Cable triceps 4x8-10RM Biceps curl 4x8-10RM
24 Parallel back squat 4x8-
25 Parallel back squat 4x8-10RM Lying leg curl 8x8-10RM
10RM
Lying leg curl 8x8-10RM
26 Leg extension 4x8-
27 Leg extension 4x8-10RM
10RM
28 2S
29
(n=9)
31 PM Straight-arm pulldown 4x8- Dumbbell flat fly 4x8- Straight-arm pulldown
Dumbbell flat fly 4x8-10RM
32 10RM 10RM 4x8-10RM
33 Cable triceps 4x8-10RM
34 Biceps curl 4x8-10RM Cable triceps 4x8-10RM Biceps curl 4x8-10RM
35 Parallel back squat 4x8-10RM Parallel back squat 4x8-
Lying leg curl 8x8-10RM Lying leg curl 8x8-10RM
36 10RM
37 Leg extension 4x8-10RM Leg extension 4x8-
38 10RM

Page 58 of 65
Legend: 1S = one session per day; 2S = two sessions per day; Arout = split routine A; Brout = split routine B; RM = repetition maximum; AM
= ante
1 meridiem;

PM = post meridiem.
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3
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Page 59 of 65
Table 3. Estimated dietary nutrient intake for 1S and 2S (mean ±SD).
1 ANOVA 3x2
2
3 Variables
Pre-test Post-4 weeks Post-8 weeks Time Time X Group
4 Effect Interaction
5 P-value P-value
6
7 Total

(kcal)
8 1S 2,566 ± 650 2,559 ± 491 2,495 ± 482 0.522
9 2S 0.196
2,697 ± 527 2,552 ± 343 2,709 ± 487 0.313
10
11 Protein

(g)
12 1S 157 ± 79 152 ± 73 172 ± 55 0.280
13 2S 0.097
146 ± 43 142 ± 61 172 ± 68 0.332
14
15 Carbohydrate

(g)
16 1S 298 ± 54 312 ± 48 302 ± 62 0.402
0.145
17 2S 372 ± 78 342 ± 40 345 ± 54 0.496
18
Lipids (g)
19
20 1S 77 ± 22 72 ± 23 63 ± 21 0.188
21 2S 0.264
70 ± 24 70 ± 17 72 ± 23 0.429
22
23 Legend:
1S = one session per day; 2S = two sessions per day; Total (Kcal) = total kilocalories intake (3
24 recorded
days’ average); g = grams
25
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Page 60 of 65
Table 4. Pre- vs. Post-8 weeks Muscle Strength measures (mean ±SD).
ANOVA 2x2
Post- MD
1 Variables
Pre-test ∆% time time*group
2 8 weeks [95% CI]
P-value P-value η p
2
3
4 1RM
BENCH (kg)
5
6 1S 94 ± 14 98 ± 14a 4 [2 - 6] 4.6
0.001 0.188 0.206
7 2S 101 ± 15 108 ± 15a 7 [4 - 10] 6.8
8
9 1RM
SQUAT (kg)
10 1S 138 ± 14 149 ± 16a 11 [6 - 12] 7.8
11 0.001 0.043 0.420
12 2S 126 ± 23 146 ± 27a 20 [12 - 28] 16.1
13
14 60%1RM
BENCH (rep)
15 1S 14 ± 3 17 ± 3a 3 [2 - 4] 15.4
16 0.001 0.161 0.229
17 2S 17 ± 4 21 ± 3a 4 [2 - 6] 24.0
18 60%1RM

SQUAT (rep)
19
20 1S 17 ± 4 20 ± 3a 3 [1 - 5] 19.0
21 a
0.001 0.602 0.036
22 2S 17 ± 4 21 ± 3 4 [1 - 7] 24.3
23
24 Legend:
1S = one session per day; 2S = two sessions per day; 1RMBENCH = one maximal
25 repetition
test in bench press exercise; 1RMBENCH = one maximal repetition test in
26
parallel back squat exercise; 60% 1RMBENCH = 60% of 1RM test in bench press exercise;
SQUAT = 60% of 1RM test in parallel back squat exercise; rep = repetitions η p
2
27
60% 1RM
28 = partial eta squared; MD [95% CI] = mean difference and 95% confidence interval.
29 a Significantly greater than the corresponding pre-intervention value (p < 0.05).
30
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Page 61 of 65
1
2
3 Table 5. Pre and Post 8 weeks- Muscle Morphology measures (mean ±SD).
4 ANOVA 2x2
5 Post- MD
Variables Pre-test ∆% time time*group
6 8 weeks [95% CI]
P-value P-value η p
2
7
8 MTBB (mm)
9
10 1S 28.1 ± 5.5 31.7 ± 4.0a 3.6 [1.1 - 5.1] 12.9
0.001 0.790 0.009
11 2S 28.6 ± 3.4 32.5 ± 4.2a 3.9 [1.2 - 5.6] 13.8
12
13 MTTB (mm)
14 1S 44.2 ± 4.9 47.8 ± 4.0a 3.6 [1.8 - 5.4] 8.1
15 0.001 0.381 0.097
16 2S 44.0 ± 6.8 46.3 ± 9.2a 2.3 [0.5 - 4.1] 5.2
17 MTVL (mm)
18
19 1S 43.6 ± 7.2 48.3 ± 7.7a 4.7 [2.2 - 7.2] 10.8
20 0.001 0.875 0.003
21 2S 43.5 ± 4.2 47.8 ± 5.6a 4.3 [3.3 - 5.3] 9.9
22 MTAQ (mm)
23
24 1S 43.7 ± 7.7 47.2 ± 7.6a 3.5 [1.0 - 5.0] 8.0
0.001 0.795 0.009
25 2S 44.0 ± 4.0 48.3 ± 4.8a 4.3 [1.4 - 6.3] 9.8
26
27 MTPM (mm)
28 1S 33.1 ± 3.6 36.4 ± 6.5a 3.3 [1.6 - 5.0] 9.8
29 0.001 0.587 0.039
30 2S 36.4 ± 7.5 38.6 ± 7.7a 2.2 [0.2 - 4.2] 6.0
31
32 Legend: 1S = one session per day; 2S = two sessions per day; MTTB = muscle thickness
33 of the triceps brachii muscle; MTEF = muscle thickness of the elbow flexors muscles;
34 MTVL = muscle thickness of the vastus lateralis muscle; MTAQ = muscle thickness of the
35 anterior quadriceps muscle; MTPM = muscle thickness of pectoralis major; η2p = partial
36 eta squared; MD [95% CI] = mean difference and 95% confidence interval.
37
38 a Significantly greater than the corresponding pre-intervention value (p < 0.05).
39
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28

Page 62 of 65
1
2 TITLES OF FIGURES
3
4 Figure 1. Univariate scatterplot comparison of groups 1S (one session per day) and 2S
5 (two sessions per day) in 1RMBENCH, 1RMSQUAT, 60%1RMBENCH and 60%1RMSQUAT
6 (Figure 1A), muscle thickness of the triceps brachii (MTTB), biceps brachii (MTBB), vastus
7 lateralis (MTVL), anterior quadriceps (MTAQ) and pectoral muscles (MTPM) (Figure 1B).
8 Trivial gray areas were the smallest worthwhile change (SWC) (see methods). # = p <
9 0.05 between 1S vs 2S.
10
11 Figure 2. The weekly accumulated total load lifted (ATLL) of subjects during the 8-
12 weeks of intervention training (2A). Individual delta (∆%) relative differences of the TLL
13 (kg) in the week 8 minus week 1 (2B). # Significantly greater than 1S (p < 0.05)
14
15
16 Figure 3. Well-being (WB) status between groups 1S (one session per day) and 2S (two
17 sessions per day) during the 8-weeks of intervention training (3A). Weekly coefficient of
18 variation (CV) of the WB over the 8 weeks of the intervention training (3B).
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Twice-daily sessions result in a greater muscle strength and a similar muscle

Article in The Journal of sports medicine and physical fitness · February 2021

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Twice-daily sessions result in a greater muscle strength and a similar muscle

Article in The Journal of Sports Medicine and Physical Fitness · February 2021

Felipe Brigatto Tiago Volpi Braz

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Häkkinen and Kallinen16 investigated morphological adaptations in 10 female athletes during an

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recovery from resistance exercise. Med Sci Sports Exerc. 2012;44(11):2220–7.

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Total Body Mass (kg) 79.7 ± 17.1 81.4 ± 6.3 0.774

1 Height (cm) 174 ± 6.6 176 ± 5.5 0.446

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Table 3. Estimated dietary nutrient intake for 1S and 2S (mean ±SD).

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