The role of deep breathing on stress

Abstract

The objective of this study was to verify, in a sample of university students, whether a relaxing technique called deep breathing (stress Intervention Functional IFA) is capable to improve the mood and to reduce the levels of stress. Thirty-eight adult healthy subjects (aged between 18 and 28 years) volunteered the study. They were randomly divided in two groups, the Experimental Group (N = 19) and the Control Group (N = 19). The subjects of the Experimental Group were submitted, once per week, to 10 treatment’s sessions of Anti-stress Protocol, each lasting 90 min, whereas subjects of the Control Group sat ten times for 90 min, once per week, without practicing any treatment. The psychological state of mood and stress was evaluated using Measurement of Psychological Stress (MSP) and Profile of Mood State (POMS), while the biological profile of the stress was detected by measuring the heart rate and the salivary cortisol. The results obtained from the present research support the possibility that deep breathing technique is capable to induce an effective improvement in mood and stress both in terms of self-reported evaluations (MPS and POMS) and of objective parameters, such as heart rate and salivary cortisol levels. No statistically significant difference was found between men and women.

Full text

ORIGINAL ARTICLE
The role of deep breathing on stress
Valentina Perciavalle
1
•Marta Blandini
2
•Paola Fecarotta
3
•Andrea Buscemi
2
•
Donatella Di Corrado
4
•Luana Bertolo
2
•Fulvia Fichera
2
•Marinella Coco
2
Received: 13 July 2016 / Accepted: 1 December 2016
ÓSpringer-Verlag Italia 2016
Abstract The objective of this study was to verify, in a
sample of university students, whether a relaxing technique
called deep breathing (stress Intervention Functional IFA)
is capable to improve the mood and to reduce the levels of
stress. Thirty-eight adult healthy subjects (aged between 18
and 28 years) volunteered the study. They were randomly
divided in two groups, the Experimental Group (N=19)
and the Control Group (N=19). The subjects of the
Experimental Group were submitted, once per week, to 10
treatment’s sessions of Anti-stress Protocol, each lasting
90 min, whereas subjects of the Control Group sat ten
times for 90 min, once per week, without practicing any
treatment. The psychological state of mood and stress was
evaluated using Measurement of Psychological Stress
(MSP) and Profile of Mood State (POMS), while the bio-
logical profile of the stress was detected by measuring the
heart rate and the salivary cortisol. The results obtained
from the present research support the possibility that deep
breathing technique is capable to induce an effective
improvement in mood and stress both in terms of self-
reported evaluations (MPS and POMS) and of objective
parameters, such as heart rate and salivary cortisol levels.
No statistically significant difference was found between
men and women.
Keywords Deep breathing Stress Cortisol Emotion
Introduction
Psychological stress is a major risk factor for the devel-
opment and progression of a number of diseases, including
cardiovascular disease, cancer, arthritis, and major
depression [1].
A fact known by the scientific community is that emo-
tions in general, and stress in particular, produce interre-
lated functional changes, mainly through the vegetative
nervous system, the endocrine system and the immune
system.
A protracted state of tension induces in the individual
psychological, physiological and behavioral effects that
over time can lead to harmful consequences [2].
Contemporary society maintains individuals in a con-
stant struggle for success without taking into account their
needs and how much all this costs them. For professional
sportsmen, mood and level of stress have been always
considered aspect they must learn to control. Moreover, it
has reported an increase of alteration of mood states and of
levels of stress in young students [3,4].
The objective of this research is to verify, in a sample of
university students, mood and stress levels, for evaluating
the efficacy of a particular relaxing technique, called deep
breathing (stress Intervention Functional IFA), for amelio-
rating the mood and for returning to normal levels of stress.
In Japan, deep breathing is widely used as a method of
reducing tension and mood. It is a fundamental technique
used in various relaxation methods and is also incorporated
&Marinella Coco
marinella.coco@unict.it
1
Department of Sciences of Formation, University of Catania,
Catania, Italy
2
Department of Biomedical and Biotechnological Sciences,
University of Catania, Via Santa Sofia 64, 95125 Catania,
Italy
3
Functional Psychotherapy Center of Catania, Catania, Italy
4
Faculty of Human and Social Sciences, University ‘‘Kore’’ of
Enna, Enna, Italy
123
Neurol Sci
DOI 10.1007/s10072-016-2790-8

in qigong, yoga and progressive muscle relaxation [5].
However, in that study the evaluation of the effects of the
different relaxation method on mood and stress was carried
out using self-reported evaluations.
In this study, we investigated the effects of deep
breathing on mood and stress not only with self-reported
evaluations but also measuring objective parameters, as
heart rate and salivary cortisol levels. The used self-re-
ported psychological tests were the Measurement of Psy-
chological Stress (MSP) and the Profile of Mood State
(POMS), while the biological profile of the stress was
evaluated by measuring heart rate and levels of salivary
cortisol [6]. This study is intended as a preliminary eval-
uation for a larger intervention study.
Materials and methods
Subjects
Thirty-eight adult healthy subjects, students of the degree
course in Psychology at the University of Catania, volun-
teered the study. Participants aged between 18 and 28 years
and were randomly divided in two groups, with 19 subjects
forming the Experimental Group and the other 19 repre-
senting the Control Group. Written informed consent was
obtained from the subjects.
Protocol
The subjects of the Experimental Group (three males and
16 females with a mean age 23.2 years ±2.74), were
subjected, once per week, to ten treatment sessions of Anti-
stress Protocol (see below), each lasting 90 min at the
Functional Psychotherapy Center of Catania. The treatment
took place between 9 and 11 a.m., and was always prac-
ticed by one of us (P.F.).
The subjects of the Control Group (four males and 15
females with a mean age 23.2 years ±2.46), summoned
ten times, once per week, at the Functional Psychotherapy
Center of Catania, where they sat between 9 and 11 a.m.
for 90 min without practicing any treatment.
At the beginning of the first session (I), fifth session (II)
and tenth session (III) to all 38 participants were admin-
istered MSP and POMS, and was measured heart rate and
salivary cortisone levels.
POMS test
To evaluate quantitatively the mood of the subjects they
had to complete the 30-item Profile of Mood States
(POMS) [7], which is a self-rating questionnaire consisting
of six mood dimensions: Tension–Anxiety (T–A),
Depression–Dejection (D), Anger–Hostility (A–H), Vigor
(V), Fatigue (F), and Confusion (C). The total mood dis-
turbance (TMD) score is calculated by subtracting the
Vscore from the sum of scores for the other dimensions.
MSP test
To describe the variables able to provide a global index of
the state of psychological stress, the Measurement of Psy-
chological Stress (MSP) Scale (Measure du Stress Test
Psychologique [8]) was used in the Italian translation and
adaptation of Di Nuovo and Rispoli [9]. The MSP is a
questionnaire consisting of 49 items based on different
aspects related to the perception that the individual has of its
condition (cognitive-affective, physiological, behavior). The
choice of responses is made of a scale (Likert-type) whose
possible answers are 1–4 (from ‘‘not at all’’ to ‘‘very’’).
Heart rate
Cacioppo and Berntson [10] demonstrated that an increase
in heart rate often associate with the stressors, so we
decided to use as a parameter for the evaluation of heart
rate stress. The heart rate was measured in the Experi-
mental as well as Control Group for 5 min at the beginning
of the first session (I), fifth session (II) and tenth session
(III). Measurement of heart rate started after that the sub-
ject sat for 15 min on a comfortable armchair.
Salivary cortisol
Levels of salivary cortisol were assessed using Radioim-
munoassay Cortisol Test (RIA CT from RADIM), as pre-
viously described [11]. The saliva samples were collected
three times in the same day (30 min after awakening and at
14:00 and 20:00 h; prior to breakfast, lunch and dinner,
respectively). Saliva samples were collected by adminis-
tering a tampon, chewed by the subject for about a minute;
the samples were immediately frozen at -70 °C.
Prior to testing, participants were instructed regarding
the purpose of the study and were asked to follow the
instructions for the collection of saliva samples: avoid
caffeine and acidic drinks, not brush teeth, eat or drink
anything for 15 min prior to a sample collection.
Anti-stress protocol based on the psychological
functional model
The anti-stress protocol is based on ten sessions and con-
sists in the use of techniques designed to adapt the Func-
tional Model proposed by Luciano Rispoli [12].
Table 1illustrates the guidelines for the deep breathing
technique that included.
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Statistical analysis
Data are presented as the mean ±standard deviation (SD).
The slope of the diurnal change in cortisol level was cal-
culated to estimate how each player fit the normal (i.e.,
descending) profile. A linear regression of the three cortisol
values on the sampling times was calculated, with data
pooled for each player. Steeper slopes represented smaller
bcoefficient values for the slope of the regression, indi-
cating that the decrease in cortisol was more rapid. Flatter
slopes (larger bcoefficient values) indicated slower
decreases or abnormally timed peaks. The average area
under the curve (AUC) was calculated by trapezoidal
estimation using cortisol values. The AUC ‘with respect to
ground’ (AUCg) was also calculated [11] which provides
information regarding the total hormonal output, and thus
the basal activity of the Hypothalamic–pituitary–adrenal
axis of the participants. Comparison were carried out using
the unpaired Student’s ttest or one-way repeated-measures
ANOVA (Friedman test) followed by Dunn’s multiple
comparison test. Correlation analysis was carried out using
one-tailed Pearson’s correlation analysis, and significance
was set at p\0.05. All analyses were performed using
GraphPad Prism version 6.03 for Windows (GraphPad
Software, San Diego, CA, USA) and were carried out
according to the guidelines for reporting statistics in jour-
nals published by the American Physiological Society [14].
Results
POMS test
The scores of the six subscales of the POMS separately
show significant differences in the group of deep breathing
(stress intervention functional IFA), in different parame-
ters, summarized by the total mood disturbance (TMD).
As can be seen in Fig. 1, the value of TMD shows a
significant reduction (p[0.05) in the Experimental Group
between first (I) and last (III) session of deep breathing
(stress Intervention Functional IFA). No significant chan-
ges were observed in the Control Group. No statistically
significant difference was found between men and women
in both Experimental and Control Group.
MSP test
As can be seen in Fig. 2, the values of MSP at the first
session test in both groups show a medium level of stress,
with values ranging between 90 and 100. The values
obtained after the fifth and tenth session were unchanged in
the control group (90 and 100). On the contrary, in the
Experimental Group the values of MPS show a significant
(p[0.05) reduction in the levels of stress between the first
and tenth session. No statistically significant difference was
found between men and women in both Experimental and
Control Group.
Heart rate
The heart rate were measured in the Experimental as well
as Control Group for 5 min at the beginning of the first
session (I), fifth session (II) and tenth session (III).
As can be observed in Fig. 3, the values obtained in the
Control Group do not show any significant change induced
by deep breathing, whereas in the Experimental Group the
level of heart rate exhibits a significant (p[0.05) reduc-
tion between the first and tenth session. No statistically
significant difference was found between men and women
in both Experimental and Control Group.
Table 1 Guidelines for the deep breathing technique For more details of each individual technique, see Rispoli [12] and Blandini et al. [13]
Session In orthostatic position, relaxing of the head and neck, rotating of shoulders, breathing deeply, yawning and imagining to follow the
alternation of the breath
Session Beat hands and feet, chilling down, staggering, imagining to do somersaults between the clouds, sharing
Session Feel parts of the body (standing and walking), contacting back to back the others of the group, remembering tenderness, sharing
Session In orthostatic position, moving the pelvis and modulating the voice, breathing as a butterfly, imagine smooth hills and a calm lake,
sharing
Session Contracting and releasing the body’s muscles, the shoulders to lift and leave, raising and lowering the shoulders, imagining to breathe
deeply, sharing
Session Grimacing, exhaling with eyes half-closed and mouth open, yawning, crawling in the direction of the head, sharing
Session Game with clothespins, breathing and moving the pelvis, imagining of getting lost, self-massaging, imagining to dive, sharing
Session Groping in the dark with eyes closed, producing tremors and convulsions, imagining to follow the alternation of the breath, sharing
Session Rolling on the others of the group, self-massaging the back, self-massaging the neck, imagining to be a leaf in the wind, sharing
Session In orthostatic position, relaxing of the head and neck, rotating the shoulders, breathing deeply, moving together body and arms,
imagining to fly, imagining sand dunes in the wind, sitting in a circle to greet
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Salivary cortisol
AscanbeseeninTable2and Fig. 4, the levels of
salivary cortisol at the beginning of the first session
show comparable values in Experimental and Control
Groups. The showed AUCg values are obtained by
averaging the levels measured 30 min after awakening,
at 14:00 and 20:00 h. As can be seen in both Table 2
and Fig. 4, in the Control Group the three measurements
did not exhibit significant differences. However, in the
Experimental Group there is a statistically significant
improvements (p[0.001) between the first (I) and the
last session (III), as well as between the fifth (II) and the
last session (III, p[0.05). No statistically significant
difference was found between men and women in both
Experimental and Control Group.
Discussion
It is generally believed that the deep breathing, practiced
using various techniques, such as qigong and yoga,
improves the sense of well being.
Scientific literature reports remarkable positive effects
induced by the oldest relaxing technique, yoga [15], as
improvement in motor abilities [16], heart rate, blood pressure
and body composition [17,18], cardiorespiratory function
Fig. 1 Mean values of total
mood disturbance (TMD) in the
Experimental as well as Control
Group, measured at the
beginning of the first session (I),
fifth session (II) and tenth
session (III). In the first row,
data of the entire sample are
presented, while in the second
and third row are shown
separately data for men and
women, respectively. Symbols
from ANOVA with Dunns’s
multiple comparison test:
*p\0.05
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[19], forced expiratory volume [18] and maximum vital
capacity [20]. Positive effects were found also on mood states
as anxiety [21] and perceived stress [22,23]. The study carried
out by Hayama and Inoue [5] showed a reduction of ‘tension
anxiety’ and fatigue through the use of deep breathing.
In this study, the use of the technique of deep breathing by a
group of university students showed a significant improve-
ment between the beginning and the end of the training.
The improvements occurred both on the examined
physiological effects (heart rate and salivary cortisol
levels) and on mood and perceived stress, improvements
not observed in the Control Group.
Comparable results were found by Paul et al. [24], who
observed in a longitudinal study, conducted on a sample of
students that the use of deep breathing is capable to reduce
stress, allowing an improvement in academic performance.
This has led the Authors to include in subsequent courses
of this method.
Recently, a systematic review on development of stress
in dental students, reported significant improvements with
deep breathing [25].
Plasma lactate concentrations increase during acute
psychosocial stress in humans [26] and in the last years it
has been observed that the excitability of the Primary
Motor Cortex is improved by high concentrations of blood
lactate, induced not only with a stressful condition as an
exhaustive exercise but also with an intravenous injection
[27–31]. It is worth noting that repetitive magnetic stimu-
lation of primary motor cortex is per se capable of
increasing blood lactate [32]. On the other hand, an
Fig. 2 Mean values of
Measurement of Psychological
Stress (MSP) in the
Experimental as well as Control
Group measured at the
beginning of the first session (I),
fifth session (II) and tenth
session (III). In the first row,
data of the entire sample are
presented, while in the second
and third row are shown
separately data for men and
women, respectively. Symbols
from ANOVA with Dunns’s
multiple comparison test:
*p\0.05
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exhaustive exercise induces a reduction of the excitability
of the Supplementary Motor Area [29] and a worsening of
attentional processes [33,34].
In conclusion, although this research have to be con-
sidered a pilot study, the use of deep breathing techniques
has lead to an effective improvement in the management of
stress in daily life, and therefore, could exert positive
influences on the stress conditions that the student must
face during the course of his/her studies.
Fig. 3 Mean values of heart
rate in the Experimental as well
as Control Group measured at
the beginning of the first session
(I), fifth session (II) and tenth
session (III). In the first row,
data of the entire sample are
presented, while in the second
and third row, are shown
separately data for men and
women, respectively. Symbols
from ANOVA with Dunns’s
multiple comparison test:
*p\0.05
Table 2 Mean levels (±standard deviation, SD) of salivary cortisol
(pg/ml) in the Experimental Group and Control Group
I II III
Experimental group
Mean values 9.62 9.53 7.34
SD 1.34 1.53 2.18
Control group
Mean values 9.55 9.53 9.58
SD 1.49 1.80 1.69
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Compliance with ethical standards
Conflict of interest The authors report no conflicts of interest. The
authors alone are responsible for the content and writing of the paper.
References
1. Muscatell KA, Eisenberger NI (2012) A social neuroscience
perspective on stress and health. Soc Pers Psychol Compass
6(12):890–904
2. Cooper CL, Dewe PJ, O’Driscoll MP (2001) Organizational
stress. A review and critique of theory, research, and applications.
SAGE publications, Thousand Oaks
3. Dweck CS (1990) Self-theories: their role in motivation, per-
sonality and development. Nebr Symp Motiv 38:199–235
4. Ford IW, Eklund RC, Gordon S (2000) An examination of psy-
chosocial variables moderating the relationship between life
stress and injury time-loss among athletes of a high standard.
J Sports Sci 18(5):301–312
5. Hayama Y, Inoue T (2012) The effects of deep breathing on
‘tensione–anxiety’ and fatigue in cancer patients undergoing
adjuvant chemotherapy. Complement Ther Clin Pract 18:94–98.
doi:10.1016/j.ctcp.2011.10.001
6. Vining RF, McGinley RA, Maksvytis JJ, Ho KY (1983) Salivary
cortisol: a better measure of adrenal cortical function than serum
cortisol. Ann Clin Biochem 20(Pt 6):329–335
7. McNair DM, Lorr M, Droppleman LM (1971) Manual for the
profile of mood states (POMS). Educational and Industrial
Testing Service, San Diego
8. Tessier R, Lemure L, Fillion L (1990) Mesure du Stress Psy-
chologique MSP. The Aviora, Brosard-Que
´bec
Fig. 4 Mean values of salivary
cortisol (pg/ml) in the
Experimental as well as Control
Group measured at the
beginning of the first session (I),
fifth session (II) and tenth
session (III). AUCg values of
salivary cortisol were obtained
by averaging the levels
measured 30 min after
awakening, at 14:00 and at
20:00 h. In the first row, data of
the entire sample are presented,
while in the second and third
row, are shown separately data
for men and women,
respectively. Symbols from
ANOVA with Dunns’s multiple
comparison test: *p\0.05;
***p\0.001
Neurol Sci
123

9. Di Nuovo S, Rispoli L (2000) Misurare lo stress. Linea test.
Editore Franco Angeli. ISBN: 9788846422521
10. Berntson GG, Cacioppo JT (2007) Heart rate variability: stress and
psychiatric conditions. In Malik M and Camm AJ (Eds) Dynamic
electrocardiography, Chapter 7. doi: 10.1002/9780470987483.ch7
11. Di Corrado D, Agostini T, Bonifazi M, Perciavalle V (2104)
Changes in mood states and salivary cortisol levels following two
months of training in elite female water polo players. Mol Med
Rep 9(6):2441–2446. doi:10.3892/mmr.2014.2115
12. Rispoli L (2011) Manuale delle tecniche Funzionali. Edizioni
S.E.F, Napoli
13. Blandini M, Fecarotta P, Buscemi B, Ramaci T, Buscemi A
(2015) An anti-stress protocol based on the psychological func-
tional model. Internat J Edu Res 3:459–468
14. Curran-Everett D, Benos DJ (2004) Guidelines for reporting
statistics in journals published by the American Physiological
Society. Am J Physiol 97:457–459
15. Cowen VS, Adams TB (2005) Physical and perceptual benefits of
yoga asana practice: results of a pilot study. J Bodywork Mov
Ther 9:211–219
16. Manjunath N, Telles S (1999) Factors influencing changes in
tweezer dexterity scores following yoga training. Indian J Physiol
Pharmacol 43(2):225–229
17. Murugesan R, Govindarajulu N, Bera T (2000) Effect of selected
yogic practices on the management of hypertension. Indian J
Physiol Pharmacol 44(2):207–210
18. Telles S, Nagarathna R, Nagendra HR, Desiraju T (1993) Phys-
iological changes in sports teachers following 3 months of
training in yoga. Indian J Med Sci 47(10):235–240
19. Raju PS, Prased KV, Venkata RY, Murthy KJ, Reddy MV (1997)
Influence of intensive yoga training on physiological changes in 6
adult women:a case report. J AlternComplement Med 3(3):291–295
20. Birkel DA, Edgren L (2000) Hatha yoga: improved vital capacity
of college students. Altern Ther Health Med 6(6):55–63
21. Malathi A, Damodaran A (1999) Stress due to exams in medical
students—role of yoga. Indian J Physiol Pharmacol 43(2):218–224
22. Berger BG, Owen DR (1988) Stress reduction and mood
enhancement in four exercise modes: swimming, body condi-
tioning, Hatha yoga, and fencing. Res Quart Exer Sport
59(2):148–159
23. Netz Y, Lidor R (2003) Mood alterations in mindful versus
aerobic exercise modes. J Psychol 137(5):405–419
24. Paul G, Elam B, Verhulst SJ (2007) A longitudinal study of
students’ perceptions of using deep breathing meditation to
reduce testing stresses. Teach Learn Med 19(3):287–292
25. Abdullah MA, Van der Molen HT, Alauja AH, De Boer BJ
(2014) Stress management in dental students: a systematic
review. Adv Med Educ Pract 5:176. doi:10.2147/AMEP.S46211
26. Kubera B, Hubold C, Otte S, Lindenberg AS, Zeiss I, Krause R,
Steinkamp M, Klement J, Entringer S, Pellerin L, Peters A (2012)
Rise in plasma lactate concentrations with psychosocial stress: a
possible sign of cerebral energy demand. Obes Facts
5(3):384–392. doi:10.1159/000339958
27. Coco M, Alagona G, Rapisarda G, Costanzo E, Calogero RA,
Perciavalle V, Perciavalle V (2009) Elevated blood lactate is
associated with increased motor cortex excitability. Somatosens
Motor Res 27(1):1–8. doi:10.3109/08990220.2010.507102
28. Coco M, Alagona G, Va Perciavalle, Cicirata V, Perciavalle V
(2011) Spinal cord excitability is not influenced by elevated
blood lactate levels. Somatosens Motor Res 28(1–2):19–24.
doi:10.3109/08990220.2011.598268
29. Coco M, Perciavalle V, Cavallari P, Va Perciavalle (2016)
Effects of an exhaustive exercise on motor skill learning and on
the excitability of primary motor cortex and supplementary motor
area. Medicine (Baltimore). 95(11):e2978. doi:10.1097/MD.
0000000000002978
30. Va Perciavalle, Coco M, Alagona G, Maci T, Perciavalle V
(2010) Gender differences in changes of motor cortex excitability
during elevated blood lactate levels. Somatosens Motor Res
27(3):106–110. doi:10.3109/08990220.2010.507102
31. Coco M, Caggia S, Musumeci G, Perciavalle V, Graziano AC,
Pannuzzo G, Cardile V (2013) Sodium L-lactate differently
affects brain-derived neurothrophic factor, inducible nitric oxide
synthase, and heat shock protein 70 kDa production in human
astrocytes and SH-SY5Y cultures. J Neurosci Res 91(2):313–320.
doi:10.1002/jnr.23154
32. Alagona G, Coco M, Rapisarda G, Costanzo E, Maci T, Restivo
D, Maugeri A, Perciavalle V (2009) Changes of blood lactate
levels after repetitive transcranial magnetic stimulation. Neurosci
Lett 450(2):111–113. doi:10.1016/j.neulet.2008.11.064
33. Coco M, Di Corrado D, Calogero RA, Va Perciavalle, Maci T,
Perciavalle V (2009) Attentional processes and blood lactate
levels. Brain Res 1302:205–211. doi:10.1016/j.brainres.2009.09.
032
34. Va Perciavalle, Di Corrado D, Scuto C, Perciavalle V, Coco M
(2014) Attention and blood lactate levels in equestrians per-
forming show jumping. Percept Mot Skills 118(3):733–745.
doi:10.2466/29.30.PMS.118k22w1
Neurol Sci
123