ArticlePDF AvailableLiterature Review

Abstract and Figures

Stress is now recognized as a universal premorbid factor associated with many risk factors of various chronic diseases. Acute stress may induce an individual's adaptive response to environmental demands. However, chronic, excessive stress causes cumulative, negatively impacts on health outcomes through "allostatic load". Thus monitoring of quantified levels of long-term stress mediators would provide a timely opportunity for prevention or earlier intervention of stress-related chronic illnesses. Although either acute or chronic stress could be quantified through measurement of changes in physiological parameters, such as heart rate, blood pressure, and levels of various metabolic hormones, it is still elusive to interpret whether changes in circulating levels of stress mediators, such as cortisol, reflect acute, chronic, or diurnal variations. Both serum and salivary cortisol levels reveal acute changes at a single point in time, but the overall long-term systemic cortisol exposure is difficult to evaluate because of circadian variations and its protein-binding capacity. Scalp hair has a fairy predictable growth rate of approximately 1 cm/month and the most 1 cm segment approximates the last month's cortisol production as the mean value. The analysis of cortisol in hair constitutes a highly promising technique for the retrospective assessment of chronic stress.
Content may be subject to copyright.
BMB
Re
p
orts
BMB Rep. 2015; 48(4): 209-216
www.bmbreports.org
*Corresponding authors. Eosu Kim, Tel: +82-2-2228-1620; Fax:
+82-2-318-0891; E-mail: kimeosu@yuhs.ac, Man Ho Choi, Tel:
+82-2-958-5081, Fax: +82-2-958-5059, E-mail: mh_choi@kist.re.kr
http://dx.doi.org/10.5483/BMBRep.2015.48.4.275
Received 17 October 2014
Key wor ds : Allostasis, Cortisol, Hair, Mass spectrometry, Metabolo-
mics, Stress
ISSN: 1976-670X (el ectronic edit ion)
Copyright 2015 by the The Korean Society for Biochemistry and Molecular Biology
This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/li-
censes/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Technical and clinical aspects of cortisol as a biochemical
marker of chronic stress
Do Yup Lee1, Eosu Kim2,* & Man Ho Choi3,*
1Department of Bio and Fermentation Convergence Technology, Kookmin University, Seoul 136-702, 2Department of Psychiatry, Institute
of Behavioral Science in Medicine, Brain Korea 21 Plus Project for Medical Science, Yonsei University College of Medicine, Seoul 120-752,
3Future Convergence Research Division, Korea Institute of Science and Technology, Seoul 136-791, Korea
Stress is now recognized as a universal premorbid factor
associated with many risk factors of various chronic diseases.
Acute stress may induce an individual’s adaptive response to
environmental demands. However, chronic, excessive stress
causes cumulative negative impacts on health outcomes
through “allostatic load”. Thus, monitoring the quantified
levels of long-term stress mediators would provide a timely
opportunity for prevention or earlier intervention of stress-
related chronic illnesses. Although either acute or chronic
stress could be quantified through measurement of changes in
physiological parameters such as heart rate, blood pressure,
and levels of various metabolic hormones, it is still elusive to
interpret whethe r the chan ge s in ci rculating levels o f stress
mediators such as cortisol can reflect the acute, chronic, or
diurnal variations. Both serum and salivary cortisol levels
reveal acute changes at a single point in time, but the overall
long-term systemic cortisol exposure is difficult to evaluate due
to circadian variations and its protein-binding capacity. Scalp
hair has a fairy predictable growth rate of approximately 1
cm/month, and the most 1 cm segment approximates the last
month’s cortisol production as the mean value. The analysis of
cortisol in hair is a highly promising technique for the retro-
spective assessment of chronic stress. [BMB Reports 2015;
48(4): 209-216]
INTRODUCTION
Stress can lead to both physical and psychological health
issues. Some stress can be beneficial at times by producing a
boost that provides the drive and energy to help people get
through situations like exams or work deadlines. However, an
extreme amount of stress can lead to negative consequences
and adversely affect the immune, cardiovascular, neuro-
endocrine, and central nervous systems (1). In particular, chron-
ic stress can have a serious impact due to sustained high levels
of the chemicals released in the “fight or flight” response, which
involves endocrine system releasing glucocorticoids (2, 3).
 Cortisol, which is synthesized from cholesterol, is the main
glucocorticoid in the zona fasciculate of human adrenal
cortex. Its secretion in response to biochemical stress contrib-
utes to the well-characterized suppression of the hypo-
thalamic-pituitary-adrenal (HPA) axis on health and cognition
events (4-6). Since the vast majority of cortisol actions rely on
binding to cytosolic receptors, only a small fraction of un-
bound, free cortisol is revealed to be biologically active. It
comes out of the mitochondrion, migrates out of the cell into
the extracellular space and into the bloodstream. Due to its
low molecular weight and lipophilic nature, unbound cortisol
enters the cells through passive diffusion, which makes it fea-
sible to measure the free cortisol in many body fluids (7).
 In general, cortisol levels in blood increase during the early
morning (highest at about 8 a.m.) and decrease slightly in the
evening and during the early phase of sleep (8). The timing of
blood sampling is therefore very important. While its assess-
ment in sweat or tears is only of theoretical importance and
urinary cortisol of decreasing interest, salivary cortisol may
have some advantages over the assessment of cortisol in blood
(9, 10). Since the hormone levels in biological fluids fluctuate
on a daily basis, cortisol extracted from the hair fiber has been
investigated (11-13). This review discusses on the methods in-
volved in mass spectrometry-based metabolomic studies for
identification of biomarkers in chronic stress, which is more
focused on hair cortisol. Comparative statistical analyses of
crucial aspects are also included to facilitate the understanding
of recent advances in the metabolic platform on mining
biomarkers.
STRESS AND THE ADRENAL GLAND
The two adrenal glands are located on top of the kidneys, and
these glands produce hormones in response to stress. Each
Invited Mini Revi ew
Metabolic biomarkers in chronic stress
Do Yup Lee, et al.
210 BMB Reports http://bmbreports.org
Fig. 1. The differential stress hor-
mones secreted by adrenal cortex and
medullar. Catecholamines cause gen-
eral physiological changes that pre-
pare the body for physical activity
(fight-or-flight response) in the short-
term response. Some typical effects in-
clude increases in heart rate, blood
pressure, and blood glucose levels,
and other general reactions of the
sympathetic nervous system. Corticoids
are involved in a wide range of phys-
iological processes including chronic
stress response, immune response, and
regulations of inflammation, carbohy-
drate metabolism, protein catabolism,
blood electrolyte levels, and behavior.
Data are taken from “https://www.
studyblue.com/notes/note/n/07-adrenal-
glands/deck/1109539”.
adrenal gland consists of a central area, called the medulla,
and an outer area of the cortex (Fig. 1). In case of the apparent
threat, the hypothalamus sends direct signals via the sym-
pathetic nervous system to the adrenal glands, causing them to
release a catecholamine and epinephrine (same as adrenaline).
It leads to an urgent action by stimulating faster breathing and
heart rates. The adrenal medullar also secrete another cat-
echolamine, norephinephrine, which works with epinephrine
to stimulate liver cells to release glucose to make more fuel
available for cellular respiration. These hormones have
short-term effects as the nerve impulses are sent from the
hypothalamus. Due to the short half-life of blood catechol-
amine, meticulous care must be taken to obtain blood samples
consistently vis-à-vis the stress immersion experience (14).
 Finding a “gold standard” biomarker for chronic stress has
been proven to be challenging, given its complex etiology and
highly individual manifestations, while the biomarkers of acute
stress have been well-defined and are primarily used to assess
the release of catecholamine. Hormones secreted by the adre-
nal cortex provide a slower, longer-acting (chronic) response
to the stress. In this event, the hypothalamus secretes a releas-
ing hormone which causes the anterior pituitary to secrete an
adrenal-stimulating hormone, adrenocorticotrophic hormone
(ACTH); and this signals the cells in the adrenal cortex to pro-
duce and secrete corticosteroids. Among them, mineralocorti-
coids, like aldosterone, can regulate water and sodium re-ab-
sorption in the kidneys. It also regulates the active secretion of
potassium in the principal cells of the cortical collecting tubule
and protons via proton ATPases in the luminal membrane of
the intercalated cells of the collecting tubule, which results in
an increase of blood pressure and blood volume. Glucocorti-
coids promote fat and protein breakdown and glucose syn-
thesis. Cortisol is the major glucocorticoids, and it regulates or
supports a variety of important cardiovascular, metabolic, im-
munologic, and homeostatic functions (1).
ALLOSTASIS AND ALLOSTATIC LOAD
The term, stress, was originally adopted from engineering
(measure of the internal forces induced by deformation of a
body), but it is now referred to ‘threats or anticipation of
threats to an organism’s homeostasis’ (15). Thus stress events
could be understood as any stimuli that cause alterations in
homeostasis for adaptation to the environment. These changes
in homeostasis are referred to as ‘allostasis’, which can be ex-
emplified by increased heart rate or blood pressure and en-
hanced systemic metabolism. In general, allostasis can be
adaptive or maladaptive depending on its degree or contextual
relevance; mediators of allostasis, such as metabolic hor-
mones, could contribute to healthy adaptation and pathophysi-
ology (16). The concept of ‘allostatic load’ indicates an altered,
‘new set point’ of homeostasis, resulting from cumulative ef-
fects of allostatic responses which are chronic, excessive, or
poorly regulated (15, 16). For example, the increased serum
glucose level is responsible for a single acute stressful event,
which can be called as ‘allostatic response’. Also, diabetes
(insulin resistance) resulting from repetitive chronic stress can
be understood as an ‘allostatic overload’, in which the base-
line fasting glucose level has been newly set to a higher level
than before (Fig. 2). Therefore, the biomarkers of allostatic
Metabolic biomarkers in chronic stress
Do Yup Lee, et al.
211http://bmbreports.org BMB Reports
Fig. 2. Stress, allostasis, and allostatic load. Stress is any stimulus
inducing either adaptive or maladaptive allostasis (changes in ho-
meostasis) of stress mediators, which constitute the autonomic
nervous system (blood pressure, catecholamines), metabolic hor-
mones (cortisol, insulin), and pro- & anti-inflammatory cytokines.
If stress stimuli are excessive and repetitive, recovery to the origi-
nal homeostatic levels may be incomplete (indicated by the sec-
ond blue arrow). As a result, chronic stress can make a body sys-
tem to anticipate, as if such a new (stressful) environment would
persist, demanding a newly defined set point for future adap-
tation. Thus, the difference between the new and old set points
can be understood as a ‘cumulative burden of adaptation to
stress’- i.e., allostatic load. Examples of allostatic load may be
found in the primary mediators (hypercortisolemia, increased infla-
mmatory cytokines), secondary outcomes (elevated blood pressure,
overweight, insulin resistance), or tertiary outcomes (hypertension,
diabetes, obesity, coronary heart disease, neurodegenerative dis-
orders). For more details, see reference # 6.
load, if available, could be used for measuring and predicting
the cumulative biological risks of impending illnesses, and
they are ideal indicators for detecting the ‘pre-phase’ of ill-
nesses (6, 17, 18).
 Mediators of allostasis constitute the autonomic nervous sys-
tem (catecholamines), metabolic hormones, and various cyto-
kines (15, 16). Among these, cortisol is the one that has been
paid great attention based on the concept of ‘glucocorticoid
cascade hypothesis’ (19). This hypothesis explains how and
why the cortisol actions could be related to pathophysiology
upon overload of stress. Psychological and physical stresses in-
crease the circulating cortisol levels. In the acute state, in-
creased cortisol induces adaptive responses via enhancing ca-
tabolic processes to supply more energy to the body. Normally,
increased cortisol levels will return to the basal levels by feed-
back inhibition mechanisms through the hypothalamus, pre-
frontal cortex, and most importantly, hippocampus (20). When
stressful stimuli are repeated chronically, circulating cortisol is
maintained at higher levels over a prolonged period. Chronically
elevated cortisol levels now causes damages on hippocampal
and cortical neurons (19), which are the main regions where
the feedback inhibition starts. As a result, even when stress
stimuli disappear, cortisol levels could be maintained at higher
levels beyond the physiologically normal range due to a vicious
cycle caused by the already damaged feedback mechanism.
 Consequently, a great number of studies have explored the
potential of utilizing cortisol as a biomarker for various chronic
illnesses or their pre-disease states (21, 22). Currently, it seems
that such role of cortisol, as one of biomarkers for the allostatic
load, may be largely accepted (6). However, there are still con-
troversial issues regarding the sampling and measurement
methods of these hormones that are fluctuated by environ-
mental contexts as well as by the circadian cycle. However,
the most important issues might be the possibility that the level
measured once at a current point could reflect the past history
of an individual’s stress load, just as HbA1c indicating the de-
gree of ‘glucose load’ for the past 3 months. The level of corti-
sol in scalp hair is now considered as a promising biomarker
for assessing the averaged level of one’s past stress burden dur-
ing a given period, and it will be further discussed in this
paper. Its utility has been demonstrated across diverse clinical
settings from neonates to old age, indicated by associations of
hair cortisol levels with babies’ stress in the neonatal intensive
care unit (23), children’s stress at school entry (24), and vari-
ous metabolic and neuropsychiatric disorders including acute
myocardial infarction (25), heart failure (26), metabolic syn-
drome (27), and post-traumatic stress disorder (28).
ADVANCES IN CORTISOL DETECTION IN
METABOLITE PROFILING
Because more than 90% of circulating cortisol in human se-
rum is protein-bound, changes in the binding proteins can al-
ter the levels serum total cortisol without influencing the free
concentrations of cortisol. Total cortisol could be misleadingly
lower than anticipated, resulting in the incorrect interpretation
that adrenal function is impaired. This is important because
the current standards for defining normal adrenal functions are
based on healthy people who have normal levels of binding
proteins. Measuring serum free cortisol levels in critical ill-
nesses may help to prevent the unnecessary use of glucocorti-
coid therapy (21). Although the free cortisol hypothesis has
been widely used, it has also been suggested that cortisol bind-
ing globulin (CBG)-bound cortisol may have physiological ef-
fects on target tissues (22).
 There is a high correlation between salivary cortisol levels
and unbound cortisol in plasma and serum, which remains
high during the circadian cycle and under different dynamic
test such as ACTH stimulation (22, 29). Since free cortisol rep-
resents the biologically active hormone fraction, salivary corti-
sol measures have early been considered as a better method
than serum cortisol for the evaluation of adrenocortical func-
tion (9). Recently, the late-night salivary cortisol has been
showing a superior diagnostic performance as the primary bio-
chemical diagnostic test for Cushing’s syndrome (10). Cortisol
in biological fluids has been extensively evaluated with corti-
sone, which may reveal the activity of 11-hydroxysteroid de-
hydrogenase (11-HSD) by mass spectrometry-based metabo-
lite profiling techniques (30-32). These analytical methods
have shown the acceptable analytical sensitivity and the se-
Metabolic biomarkers in chronic stress
Do Yup Lee, et al.
212 BMB Reports http://bmbreports.org
Fig. 3. A proposed mechanism of cortisol incorporation into hair
and retrospective reflection of its chronic secretion. Cortisol may
be incorporated into hair via passive diffusion from (A) blood ca-
pillary, (B) sweat, and (C) sebum, as well as from (D) external
sources. Data are taken from reference #12.
lectivity in trace amounts of biological samples such as urine
and serum. However, the methods have still hampered from
incorrect physiological levels of adrenal steroids, including
cortisol, due to the sampling problems with circadian
variation. In general, acute cortisol levels fluctuate markedly
depending on many physiological factors including circadian
rhythmicity, and it may provide a rather poor reflection of nor-
mal, chronic cortisol secretion (12, 13).
 In contrast to the biological fluids, hair can provide bio-
logical information of long-term exposure because its growth
rate is about 1 cm/month (Fig. 3). The hormones are mainly
delivered from the blood circulation to the capillaries of the
dermal papilla, which is located in the hair follicles (33). This
phenomenon enables retrospective examination of cortisol
production at the times when a stressor is most salient, without
needing to take a sample right at that time. Alternatively, it can
provide a baseline cortisol assessment for a time period during
which the stress has not yet occurred. In addition, its non-in-
vasive nature in sampling and easy storage at room temper-
ature has been highlighted as advantage in clinical applica-
tions (34-36). In the hair analysis, steroid hormones are ex-
tracted by solubilization or digestion of the hair matrix with al-
kaline hydrolysis for androgen and estrogens (37, 38). Also,
the acid hydrolysis or methanol extraction can enable an anal-
ysis of corticoids in hair due to its insufficient chemical stabil-
ity (11). When 62 biologically active steroids were analyzed
by an optimized extraction technique, and only 20 hair ste-
roids, including cortisol and cortisone, were quantitatively de-
tectable (11).
STRESS-MEDIATED HAIR CORTISOL IN ENDOCRINE
SYSTEM
Because more than 70% of diseases are believed to be
stress-related, prediction of chronic stress is an important step
in reducing the incidence of chronic illnesses. Hair cortisol
may provide an objective measurement of stress over time
rather than just ‘a day in life’. The symptoms of the metabolic
syndrome resemble those of Cushing’s syndrome, a disease
that is characterized by hypercortisolism. One of the questions
raised is whether chronically elevated cortisol concentrations
play a role in the development of obesity and the metabolic
syndromes. The increased hair cortisol levels are also asso-
ciated with children’s obesity caused by long-term activation
of HPA-axis (39), which is in accordance with the urinary lev-
els of cortisol in obese children (40). The risk of cardiovascular
disease (CVD) is associated with the increase in hair cortisol
levels by 2.7-times, which was similar to the risk associated with
hypertension or obesity. This suggests that high cortisol levels in
a long-term might be an important risk factor for CVD (41).
 Mitotane, an anti-neoplastic agent for adrenocortical cancer
(ACC), increases CBG and induces CYP3A4 activity, which
leads to high doses of hydrocortisone; however, there has
been no efficient biomarker to evaluate this therapy. Hair corti-
sol levels were higher in ACC patients compared to healthy in-
dividuals, and they were associated with body mass index
(42). However, there was no correlation between hair cortisol
levels and hydrocortisone doses. As a measure of long-term
cortisol exposure, the hair cortisol analysis in patients receiv-
ing glucocorticoid replacement therapy may be a useful tool.
Also, the hair cortisol content is correlated with hydro-
cortisone dose in the patients with adrenal insufficiency, who
had significantly higher subjective stress scores than the con-
trol subjects (43).
METABOLOMIC IDENTIFICATIONS IN CHRONIC
STRESS
The main benefit of metabolomic strategy is the high like-
lihood of identifying unpredicted changes in metabolic profiles
cued by abnormal conditions (44). In particular, the metab-
olomic information may offer novel diagnostic indicators and
therapeutic targets in clinical applications (45), as well as in
mechanistic studies heading to elucidate metabolic modules that
can regulate dysfunctional processes in disease statuses (46).
 Among dysfunctional heath conditions, chronic diseases are
mostly composed of subtle and long-term dysregulations of
cellular and physiological function that is often not measurable
even during disease onset (47). The stress-inducing abnormal
status is chronically developed with unpredictable combina-
tion of various types of etiological factors. Therefore, the mo-
Metabolic biomarkers in chronic stress
Do Yup Lee, et al.
213http://bmbreports.org BMB Reports
Fig. 4. Reconstructed metabolic net-
work for systematic screening of ther-
apeutic target point. The re-modeled
metabolic structure is composed o
f
metabolites, which consists of node
(e.g. metabolite) and edge (e.g. corre-
lation, structural similarity). The model
may be extended to protein infor-
mation via reaction pair that has al-
ready been built in the assembly,
which could unravel “hidden” or veil ed
metabolic modulation, particularly in
chronic disease. In this example, node
color and size indicate significant dif-
ference and fold change compared to
control (e.g. disease vs control), while
edge presents two layers of informa-
tion on structural similarity and re-
action likelihood, which leads to auto-
matic rearrangement as seen in the
figure indicating proximity of bioche-
mical module.
lecular characterization as well as clinical definition may not
be clear at specific disease course and it may not be homolo-
gous across individuals (48). The diversity of pathological traits
induced by chronic stress make it even harder to properly di-
agnose the abnormality and classify the progress stage. Most of
the studies are mainly focused on targeted single molecule and
one-point interaction, which may be insufficient to reflect the
dynamics and systematic effects of chronic stress (49). In this
context, metabolite profiling can be an effective tool for bio-
marker discovery and understanding of molecular mechanism,
particularly for the cases where a diagnostic/prognostic in-
dicator is unknown and molecular mechanism remains veiled.
Recently, a chronic unpredictable mild stress in animal model
showed aberrant profiling of amino acids that were grouped
into neurotransmitters and branched-chain amino acids (50).
The metabolic signatures under acute and chronic stress in rat
model and the biochemical cues are also closely associated
with behavior and physiological readouts (49).
DATA MINING IN METABOLOMICS
To capture disease-specific metabolic signatures, many studies,
including the case studies described above, primarily explored
on the multiple molecular constituents (metabolites), robustly
managed variability and heterogeneity of disease progress, and
defined the individuality of metabolic contents, which could
lead to well-defined diagnoses and prognoses. In this context,
multivariate statistics such as principal component analysis
(PCA) or partial least squared algorithm is a useful data mining
tool that can distinguish different groups with minimized loss
of information (51). By this nature, unidentified variation
source caused by transient (time-course) or individual specific-
ity can be handled within reconstructed statistical model,
which in turn offers more robust candidates for diagnostic/
prognostic information in clinical cases (Fig. 4).
 In many studies, metabolomic approach has been limitedly
applied to biomarker discovery with either single target or
group of metabolites. Yet, an ideal exploratory use of the tech-
nology would not only be able to capture the metabolite
changes associated with pathology, but it would also elucidate
the molecular mechanism responsible for the dysfunction and
propose the logical candidate for regulating the abnormality.
Pathway (enrichment) analysis, a systematic approach for data
mining from metabolite sets, can provide metabolic path-
way-wise information rather than the readouts of individual or
few metabolite contents (52). This bioinformatics tool, which
is rooted in gene ontology enrichment analysis, assigns groups
of metabolites at the level of metabolic pathway. The analysis
provides P value that presents significant difference at the lev-
els of metabolic pathways and information on pathway cen-
trality (pathway importance), which demonstrates how much
connectivity is formed surrounding the metabolites of interest
in the pathway (53). The output can be further investigated for
mechanistic understanding of disease, and it can be applied
for targeting and controlling the functional points of disease
process. However, although metabolic signature can be identi-
fied at the level of either single molecule, groups of metabo-
lites, or metabolic pathway, metabolite assessment cannot by
itself provide a complete understanding of disease process. To
gain comprehensive causality and mechanism in given ex-
perimental designs, integrative analysis should be accom-
panied by other molecular information such as mRNA ex-
Metabolic biomarkers in chronic stress
Do Yup Lee, et al.
214 BMB Reports http://bmbreports.org
pression levels and protein abundances (54). Since metab-
olomics joins systems with biology, mRNA has been the first
partner for integrative analysis mainly due to the well-estab-
lishment of technical platform and advancements in statistical
analysis. However, the expression level of mRNA showed a
low correlation with enzyme activity and protein expression
levels, which is a direct molecular companion with metabo-
lites, the substrate, and products. Thus protein information is
more suitable counterpart of metabolite readout (55).
 Most importantly, in order to have molecular information
(mRNA, protein, and metabolite) located at the cornerstone of
clinical application, various types of molecular levels should
be linked to physiological traits (e.g. clinical parameter) in a
statistically sound manner. This has been mainly done by
‘qualitative’ assessment in which the final readout of molec-
ular behaviors is linked to logical phenotypes of disease. For
example, aberrant glycolytic activity can be associated with tu-
mor metabolism, in which glycolytic intermediates and key
enzymes are overexpressed in tumor cells. But in the case of
chronic disease, we may not be able to detect “the stand-
ardized feature” like cancer metabolism case. Alternatively,
we can track and correlate the quantitative traits of pathology
with molecular dynamics, which can bear the variability
caused by individuality and differential disease progress. One
relevant statistical approach is canonical correlation analysis
(CCA), which has been often applied in psychological, cli-
mate, and ecological studies to enumerate the correlations be-
tween two distinct data sets measured on the identical ex-
perimental units (56-59). This statistics can analyze significant
relations between two different dataset (e.g. metabolite con-
tents and the associated clinical parameters). It is similar as
PCA, in the way that CCA seeks for linear combinations of the
variables to reduce the dimension of the data sets; but at the
same time, it explores to maximize the correlation between
the two variates (58).
CONCLUDING REMARKS
Because single cortisol assessments are strongly affected by the
acute context of the measurement situation (time of day, day
of the week, and other circumstances such as distress for
blood sampling), the assessment of long-term cortisol secretion
from the biological fluids, such as blood, urine, and saliva, is
highly labor-intensive and rather difficult to be implemented in
physical and psychological studies. Hair growth patterns also
vary across different regions of the scalp, and the precise
mechanisms of substances incorporating into hair are still in-
completely understood. But a growing observation supports
the notion that hair cortisol analysis provides a valid and reli-
able reflection of long-term cortisol secretion (12, 13, 27). We
mainly discussed about the advantages of hair analysis as an
index of chronic cortisol response. Due to their association
with various aspects of biochemical stress in adrenal steroids
and other steroid metabolites, advantages of hair analysis may
not only be restricted to cortisol (35). Further research on es-
tablishing the reference values of hair cortisol may be needed
to enhance the current knowledge on particular aspects of
chronic stress and other metabolic changes in many endocrine
diseases.
REFERENCES
1. Anderson NB (1998) Levels of analysis in health sciences:
A framework for integrating sociobehavioral and bio-
medical research. Ann N Y Acad Sci 840, 563-576
2. Edwards C (2012) Sixty tears after hench-Corticosteroids
and chronic inflammatory disease. J Clin Endocrinol
Metab 97, 1443-1451
3. Whirledge S and Cidlowski A (2013) A role for glucocorti-
coids in stress-impaired reproduction: Beyond the hypo-
thalamus and pituitary. Endocrinology 154, 4450-4468
4. Oswald LM, Zandi P, Nestadt G, Potash JB, Kalaydjian AE
and Wand GS (2006) Relationship between cortisol re-
sponse to stress and personality. Neuropsychopharmacology
31, 1583-1591
5. Staab CA and Maser E (2010) 11-Hydroxysteroid de-
hydrogenase type 1 is an important regulator at the inter-
face of obesity and inflammation. J Steroid Biochem Mol
Biol 119, 56-72
6. Juster RP, McEwen BS and Lupien SJ (2010) Allostatic load
biomarkers of chronic stress and impact on health and
cognition. Neurosci Biobehav Rev 35, 2-16
7. Miller WL (2008) Steroidogenic enzymes. Endocr Rev 13,
1-18
8. Weitzman ED, Fukushima D, Nogeire C, Roffwarg H,
Gallagher TF and Hellman L (1971) Twenty-four hour pat-
tern of the episodic secretion of cortisol in normal
subjects. J Clin Endocrinol Metab 33, 14-22
9. Hellhammer DH, Wüst S and Kudielka BM (2009)
Salivary cortisol as a biomarker in stress research. Psycho-
neuroendocrinology 34, 163-171
10. Elias PCL, Martinez EZ, Barone BFC, Mermejo LM, Castro
M and Moreira AC (2014) Late-night salivary cortisol has a
better performance than urinary free cortisol in the diag-
nosis of Cushing’s syndrome. J Clin Endocrinol Metab 99,
2045-2051
11. Jung HJ, Kim SJ, Lee WY, Chung BC and Choi MH (2011)
Gas chromatography/mass spectrometry based hair steroid
profiling may reveal pathogenesis in hair follicles of the
scalp. Rapid Commun Mass Spectrom 25, 1184-1192
12. Stalder T and Kirschbaum C (2012) Analysis of cortisol in
hair - State of the art and future directions. Brain Behav
Immun 26, 1019-1029
13. Russell E, Koren G, Rieder M and Uum SV (2012) Hair
cortisol as a biological marker of chronic stress: Current
status, future directions and unanswered questions. Psy-
choneuroendocrinology 37, 589-601
14. Dimsdale JE and Moss J (1980) Short-term catecholamine
response to psychological stress. Psychosom Med 42,
493-497
15. Karatsoreos IN and McEwen BS (2011) Psychobiological
allostasis: resistance, resilience and vulnerability. Tr Cognit
Sci 15, 576-584
Metabolic biomarkers in chronic stress
Do Yup Lee, et al.
215http://bmbreports.org BMB Reports
16. McEwen BS (1998) Protective and damaging effects of
stress mediators. N Engl J Med 338, 171-179
17. Seeman TE, McEwen BS, Rowe JW and Singer BH (2001)
Allostatic load as a marker of cumulative biological risk:
MacArthur studies of successful aging. Proc Natl Acad Sci
U S A 98, 4770-4775
18. Staufenbiel SM, Penninx BW, Spijker AT, Elzinga BM and
van Rossum EF (2013) Hair cortisol, stress exposure, and
mental health in humans: a systematic review. Psychoneu-
roendocrinology 38, 1220-1235
19. Sapolsky RM, Krey LC and McEwen BS (1986) The
Neuroendocrinology of Stress and Aging: The Glucocorti-
coid Cascade Hypothesis. Endocr Rev 7, 284-301
20. Mizoguchi K, Ishige A, Aburada M and Tabira T (2003)
Chronic stress attenuates glucocorticoid negative feed-
back: involvement of the prefrontal cortex and hippo-
campus. Neuroscience 119, 887-897
21. Hamrahian AH, Oseni TS and Arafah BM (2004) Measure-
ment of serum free cortisol in critically ill patients. N Engl
J Med 350, 1629-1638
22. Levine A, Zagoory-Sharon O, Feldman R, Lewis JG and
Weller A (2007) Measuring cortisol in human psycho-
biological studies. Physiol Behav 90, 43-53
23. Yamada J, Stevens B, de Silva N et al (2007) Hair cortisol
as a potential biologic marker of chronic stress in hospi-
talized neonates. Neonatology 92, 42-49
24. Groeneveld MG, Vermeer HJ, Linting M, Noppe G, van
Rossum EF and van IJzendoorn MH (2013) Children's hair
cortisol as a biomarker of stress at school entry. Stress 16,
711-715
25. Pereg Pereg D, Gow R and Mosseri M (2011) Hair cortisol
and the risk for acute myocardial infarction in adult men.
Stress 14, 73-81
26. Pereg D, Chan J, Russell E et al (2013) Cortisol and testos-
terone in hair as biological markers of systolic heart
failure. Psychoneuroendocrinology 38, 2875-2882
27. Stalder T, Kirschbaum C, Alexander N et al (2013) Cortisol
in hair and the metabolic syndrome. J Clin Endocrinol
Metab 98, 2573-2580
28. Steudte S, Kirschbaum C, Gao W et al (2013) Hair cortisol
as a biomarker of traumatization in healthy individuals
and posttraumatic stress disorder patients. Biol Psychiatry
74, 639-646
29. Vining RF, McGinley RA and Symons RG (1983) Hormones
in saliva: mode of entry and consequent implications for
clinical interpretation. Clin Chem 29, 1752-1756
30. Cho HJ, Kim JD, Lee WY, Chung BC and Choi MH (2009)
Quantitative metabolic profiling of 21 endogenous corti-
costeroids in urine by liquid chromatography-triple quad-
rupole-mass spectrometry. Anal Chim Acta 632, 101-108
31. Moon JY, Jung HJ, Moon MH, Chung BC and Choi MH
(2009) Heat-map visualization of gas chromatography-
mass spectrometry based quantitative signatures on ste-
roid metabolism. J Am Soc Mass Spectrom 20, 1626-1637
32. Maier B and Vogeser M (2013) Target analyte quantifica-
tion by isotope dilution LC-MS/MS directly referring to in-
ternal standard concentrations-validation for serum corti-
sol measurement. Clin Chem Lab Med 51, 833-837
33. Randall VA (1994) Androgens and human hair growth.
Clin Endocrinol (Oxf) 40, 439-457.
34. Choi MH, Kim KR, Kim IS, Lho DS and Chung BC (2001)
Increased hair polyamine levels in patients with
Alzheimer’s disease. Ann Neurol 50, 128
35. Cho SH, Choi MH, Sim WY, Lee WY and Chung BC
(2010) Metabolic alterations of DHEA and cholesterol sul-
phates in the hair of patients with acne measured by liq-
uid chromatography-mass spectrometry. Exp Dermatol 19,
694-696
36. Choi MH, Kim SJ, Lew BL, Sim WY and Chung BC (2013)
Hair steroid profiling reveals racial differences in male
pattern baldness between Korean and Caucasian popul-
ations. J Invest Dermatol 133, 822-824
37. Choi MH and Chung BC (1999) GC-MS determination of
steroids related to androgen biosynthesis in human hair
with pentafluorophenyldimethylsilyl-trimethylsilyl deriva-
tisation. Analyst 124, 1297-1300
38. Choi MH, Kim KR and Chung BC (2000) Determination of
estrone and 17-estradiol in human hair by gas chroma-
tography-mass spectrometry. Analyst 125, 711-714
39. Veldhorst MA, Noppe G, Jongejan MH et al (2014)
Increased scalp hair cortisol concentrations in obese
children. J Clin Endocrinol Metab 99, 285-290
40. Reinehr T, Kulle A, Wolters B et al (2014) Relationship be-
tween 24-hour urinary free cortisol concentrations and
metabolic syndrome in obese children. J Clin Endocrinol
Metab 99, 2391-2399
41. Manenschijn L, Schaap L, van Schoor NM et al (2013)
High long-term cortisol levels, measured in scalp hair, are
associated with a history of cardiovascular disease. J Clin
Endocrinol Metab 98, 2078-2083
42. Manenschijn L, Quinkler M and van Rossum EF (2014)
Hair cortisol measurement in mitotane-treated adrenocort-
ical cancer patients. Horm Metab Res 46, 299-304
43. Gow R1, Koren G, Rieder M and Van Uum S (2011) Hair
cortisol content in patients with adrenal insufficiency on
hydrocortisone replacement therapy. Clin Endocrinol
(Oxf) 74, 687-693
44. Lee do Y, Kind T, Yoon YR, Fiehn O and Liu KH (2014)
Comparative evaluation of extraction methods for simulta-
neous mass-spectrometric analysis of complex lipids and
primary metabolites from human blood plasma. Anal
Bioanal Chem 406, 7275-7286
45. Crews B, Wikoff WR, Patti GJ et al (2009) Variability anal-
ysis of human plasma and cerebral spinal fluid reveals
statistical significance of changes in mass spectrome-
try-based metabolomics data. Anal Chem 81, 8538-8544
46. Vinayavekhin N, Homan EA and Saghatelian A (2009)
Exploring disease through metabolomics. ACS Chem Biol
5, 91-103
47. Kim OY, Lee JH and Sweeney G (2013) Metabolomic
profiling as a useful tool for diagnosis and treatment of
chronic disease: focus on obesity, diabetes and cardio-
vascular diseases. Expert Rev Cardiovasc Ther 11, 61-68
48. Li ZY, Zheng XY, Gao XX et al (2010) Study of plasma
metabolic profiling and biomarkers of chronic un-
predictable mild stress rats based on gas chromatog-
raphy/mass spectrometry. Rapid Commun Mass Spectrom
24, 3539-3546
49. Wang X, Zhao T, Qiu Y et al (2009) Metabonomics ap-
proach to understanding acute and chronic stress in rat
Metabolic biomarkers in chronic stress
Do Yup Lee, et al.
216 BMB Reports http://bmbreports.org
models. J Proteome Res 8, 2511-2518
50. Ni Y, Su M, Lin J et al (2008) Metabolic profiling reveals
disorder of amino acid metabolism in four brain regions
from a rat model of chronic unpredictable mild stress.
FEBS Lett 582, 2627-2636
51. Broadhurst DI and Kell DB (2006) Statistical strategies for
avoiding false discoveries in metabolomics and related
experiments. Metabolomics 2, 171-196
52. Xia J and Wishart DS (2010) MetPA: a web-based metab-
olomics tool for pathway analysis and visualization.
Bioinformatics 26, 2342-2344
53. Xia J, Mandal R, Sinelnikov IV, Broadhurst D and Wishart
DS (2012) MetaboAnalyst 2.0-a comprehensive server for
metabolomic data analysis. Nucleic Acids Res 40, W127-
W133
54. Lee do Y, Park JJ, Barupal DK and Fiehn O (2012) System
response of metabolic networks in Chlamydomonas rein-
hardtii to total available ammonium. Mol Cell Proteomics
11, 973-988
55. Wienkoop S, Morgenthal K, Wolschin F, Scholz M, Selbig
J and Weckwerth W (2008) Integration of metabolomic
and proteomic phenotypes analysis of data covariance dis-
sects starch and RFO metabolism from low and high tem-
perature compensation response in arabidopsis thaliana.
Mol Cell Proteomics 7, 1725-1736
56. Meyer RC, Steinfath M, Lisec J et al (2007) The metabolic
signature related to high plant growth rate in Arabidopsis
thaliana. Proc Natl Acad Sci U S A 104, 4759-4764
57. Jozefczuk S, Klie S, Catchpole G et al (2010) Metabolomic
and transcriptomic stress response of Escherichia coli. Mol
Syst Biol 6, 364
58. González I, Déjean S, Martin PG, Gonçalves O, Besse P
and Baccini A (2009) Highlighting relationships between
heterogeneous biological data through graphical displays
based on regularized canonical correlation analysis. J Biol
Syst 17, 173-199
59. Song IS, Lee do Y, Shin MH et al (2012) Pharmacogene-
tics meets metabolomics: discovery of tryptophan as a
new endogenous oct2 substrate related to metformin
disposition. PLoS ONE 7, e36637
... Meditation, through all its forms, can significantly affect certain biochemical markers such as cortisol (associated with stress) (Lee et al., 2015) and oxytocin, dopamine, serotonin and endorphins (associated with sensation and perception of pleasure) (Dfarhud et al., 2014;Mitchell & Phillips, 2007). Cortisol is the main glucocorticoid produced in the adrenal cortex whose concentration in the blood increases upon awakening and in stressful situations, so its determination in intervention studies to treat chronic stress disorders and their effects on health is very common (Lee et al. 2015). ...
... Meditation, through all its forms, can significantly affect certain biochemical markers such as cortisol (associated with stress) (Lee et al., 2015) and oxytocin, dopamine, serotonin and endorphins (associated with sensation and perception of pleasure) (Dfarhud et al., 2014;Mitchell & Phillips, 2007). Cortisol is the main glucocorticoid produced in the adrenal cortex whose concentration in the blood increases upon awakening and in stressful situations, so its determination in intervention studies to treat chronic stress disorders and their effects on health is very common (Lee et al. 2015). On the other hand, molecules of happiness or pleasant sensations are associated with all pleasant physiological and emotional processes, negative emotions, those that promote interpersonal relationships and mediate events of stress and body pain (Dfarhud et al., 2014;Mitchell & Phillips, 2007;Pilozzi et al., 2021). ...
Article
Full-text available
Meditation, through its different practices such as yoga, mindfulness and transcendental meditation, can have beneficial effects on health, not only on a psychological level by reducing levels of stress and anxiety, but also physically by reducing blood pressure and heart rate. The objective of this systematic review is to know what are the effects of the different meditation practices on some biochemical markers such as cortisol, serotonin, dopamine, oxytocin and endorphins, which are associated with depression, anxiety, stress and happiness. The literature review was done according to the PRISMA methodology. The Science Direct, PubMed, Scopus and Web of Science databases were reviewed, narrowing the search between the years 2016-2020. After applying the inclusion/exclusion criteria, twenty-one articles were analyzed in depth. The different meditation practices have a positive effect by increasing the levels of oxytocin and endorphins, modulating the behavior of dopamine and serotonin in the central nervous system, and reducing the production of cortisol. These changes in the biologically active concentrations of these chemical markers are associated with improvements in the indicators of stress, anxiety and depression, and of the well-being perceived by the participants of the studies. The regular practice of the different forms of meditation improves the general well-being, having a positive effect on the indicators of happiness of the people. The systematization of the theoretical body of meditation, the development of standardized protocols and working with more people with active control groups, could improve the conclusions drawn from the research.
... (13) The adrenal gland produces steroid hormone namely cortisol which is a primary stress hormone and also helps in raising blood sugar level, intensify the use of sugar by the brain, and works as the finest stress biosignature. (14) (15) Its measures can be pomp by bodily stress, psychic stress, and ailments. In the event of an elevated level of stress, resulting in an elevation in serum cortisol level, consequently it is a well-founded standard in forbearing encountering stressful circumstances in life. ...
Article
The grounds of the contemporaneous examination is to evaluate the association in unite repercussions of mind-body intercession and serum cortisol measures and eggs attributes. The cognizance that psychical stress may avert females from accomplishing and keeping up a gestation has become vastly received. Mind-body intercession incorporates calmness, optimistic make-believe, biofeedback, balm, meditation, rumination, yoga, and more. All these acts work discreetly to help someone alleviate illness. Mind-body interventions (MBIs) may be an efficacious implement to assist females to cope with the ultimatum of infertility pinpointing and ministration Objective of this study is to sensitize all sub fertile females about the study, to investigate serum cortisol level of sub fertile females and to investigate the quality of oocyte of sub fertile females. This non-invasive methodology appends a set down of ministration history and the manifestations. Advice will be given to all the patients for research. Additional convention appends checking of oocytes quality after mind-body intercession in the second cycle of IVF after retrieving poor quality of oocytes in the first cycle and failure of the cycle due to it.
... Cortisol is known to be quickly and robustly induced by both psychological and physical stress and has a crucial role in stress adaptation and the restoration of homeostasis following acute stressors [53]. Acute, peak doses of psilocybin [54], and its serotonergic psychedelic analogs are known to induce cortisol release in humans [15], and thus may have a stress-mimicking effect via HPA-axis modulation [55]. Furthermore, cortisol and related glucocorticoids can rapidly and effectively suppress inflammation and have been used in the clinical therapy of both acute and chronic inflammatory diseases [56]. ...
Preprint
Full-text available
Patients characterized by stress-related disorders such as depression display elevated circulating concentrations of pro-inflammatory cytokines and a hyperactive HPA axis. Psychedelics are demonstrating promising results in treatment of such disorders, however the mechanisms of their therapeutic effects are still unknown. To date the evidence of acute and persisting effects of psychedelics on immune functioning, HPA axis activity in response to stress, and associated psychological outcomes is preliminary. To address this, we conducted a placebo-controlled, parallel group design comprising of 60 healthy participants who received either placebo (n=30) or 0.17 mg/kg psilocybin (n=30). Blood samples were taken to assess acute changes in immune status, and 7 days after drug administration. Seven days post-administration, participants in each treatment group were further subdivided: 15 underwent a stress induction protocol, and 15 underwent a control protocol. Ultra-high field magnetic resonance spectroscopy was used to assess whether acute changes in glutamate or glial activity were associated with changes in immune functioning. Finally, questionnaires assessed persisting self-report changes in mood and social behavior. Psilocybin immediately reduced concentrations of the pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-a), while other inflammatory markers (interleukin (IL)-1a, IL-1b, IL-6, and C-reactive protein (CRP)) remained unchanged. Seven days later, TNF-a concentrations returned to baseline, while IL-6 and CRP concentrations were persistently reduced in the psilocybin group. Changes in the immune profile were related to acute neurometabolic activity as acute reductions in TNF-a were linked to lower concentrations of glutamate in the hippocampus. Additionally, the more of a reduction in IL-6 and CRP seven days after psilocybin, the more persisting positive mood and social effects participants reported. Regarding the stress response, after a psychosocial stressor, psilocybin blunted the cortisol response compared to placebo. Such acute and persisting changes may contribute to the psychological and therapeutic effects of psilocybin demonstrated in ongoing patient trials.
... The trajectory of stress and related disorders has been widely investigated 1-3 , but there remains a gap in our understanding of the acute systemic effects of major stress 3,22,23 . Our aim was to characterize the metabolic fingerprint of major stress, in order to provide a comprehensive understanding of its acute physiological effects Figure 1. ...
Article
Full-text available
Major stress has systemic effects on the body that can have adverse consequences for physical and mental health. However, the molecular basis of these damaging effects remains incompletely understood. Here we use a longitudinal approach to characterise the acute systemic impact of major psychological stress in a pig model. We perform untargeted metabolomics on non-invasively obtained saliva samples from pigs before and 24 h after transfer to the novel physical and social environment of a slaughterhouse. The main molecular changes occurring include decreases in amino acids, B-vitamins, and amino acid-derived metabolites synthesized in B-vitamin-dependent reactions, as well as yet-unidentified metabolite features. Decreased levels of several of the identified metabolites are implicated in the pathology of human psychological disorders and neurodegenerative disease, suggesting a possible neuroprotective function. Our results provide a fingerprint of the acute effect of psychological stress on the metabolome and suggest candidate biomarkers with potential roles in stress-related disorders.
... However, most of available studies include cortisol. Testosterone to cortisol ratio (T/C) can be a determinant of various psychophysiological status [7]. T/C indicates biological balance and during stress conditions, moderates the automatic responses that are important for survival. ...
Article
Purpose Noise has become an integral part of human life. Noise stress affect various physiological indices. In the present study, the effects of acute noise stress on corticosterone and testosterone and testosterone to cortisol ratio (T/C) in male rats, trained with two types of high-intensity interval training (HIIT) and moderate-intensity continuous training (MCT) were evaluated. Methods: 42 male Wistar rats were divided randomly into seven groups, including the control group (C), control time (CT), exposure to acute noise stress (S), HIIT, MCT, HIIT with noise stress (HIIT+S), and MCT with noise stress (MCT+S). Exercise groups performed eight weeks of exercise training. One session of stress was induced in stress groups following the intervention (exercise or rest) period. Serum levels of corticosterone and T/C were measured through blood samples, taken 48 hours following the last session of exercise in the four exercise groups without noise stress and time control. Immediately after noise stress, blood samples were taken in 3 stress groups. Results: Serum level of corticosterone in the MCT group was significantly higher than CT and HIIT groups (P=0.001). Considering the effect of acute noise stress, corticosterone was significantly higher in HIIT+S and MCT+S, respectively, compared to the noise stress group (P<0.001). Testosterone level of the noise stress group was significantly lower than CT group (P<0.001). Testosterone level in the S group was significantly lower than other stress groups (MCT+S and HIIT+S) (P<0.001). T/C in HIIT+S group was significantly higher compared to S and MCT+S groups (P<0.001). Conclusion: HIIT and MCT, by priority, ameliorated the deteriorating effect of noise stress on testosterone and T/C; and it appears that the intensity and mode of previous exercise training affect the hormonal response to noise stress.
... The second group of indirect regulation of hormones by the GM includes cortisol (chief stress hormone in the hypothalamic-pituitary-adrenal axis (HPA) [129]) and the GI hormones ghrelin and leptin, which are also relevant mediators and modulators of pain. Cortisol is the principal stress hormone in humans [130]. While it does not directly affect pain [131], a chronic cortisol dysfunction can negatively impact the body's ability to cope with prolonged stress. ...
Article
Full-text available
Purpose Multiple diverse factors contribute to musculoskeletal pain, a major cause of physical dysfunction and health-related costs worldwide. Rapidly growing evidence demonstrates that the gut microbiome has overarching influences on human health and the body’s homeostasis and resilience to internal and external perturbations. This broad role of the gut microbiome is potentially relevant and connected to musculoskeletal pain, though the literature on the topic is limited. Thus, the literature on the topic of musculoskeletal pain and gut microbiome was explored. Methods This narrative review explores the vast array of reported metabolites associated with inflammation and immune-metabolic response, which are known contributors to musculoskeletal pain. Moreover, it covers known modifiable (e.g., diet, lifestyle choices, exposure to prescription drugs, pollutants, and chemicals) and non-modifiable factors (e.g., gut architecture, genetics, age, birth history, and early feeding patterns) that are known to contribute to changes to the gut microbiome. Particular attention is devoted to modifiable factors, as the ultimate goal of researching this topic is to implement gut microbiome health interventions into clinical practice. Results Overall, numerous associations exist in the literature that could converge on the gut microbiome’s pivotal role in musculoskeletal health. Particularly, a variety of metabolites that are either directly produced or indirectly modulated by the gut microbiome have been highlighted. Conclusion The review highlights noticeable connections between the gut and musculoskeletal health, thus warranting future research to focus on the gut microbiome’s role in musculoskeletal conditions.
... Nutrigenomic studies have shown that various dietary components can contribute to altered gene expressions which ineluctably affect metabolism and health (Kozul et al. 2008;Zhang and Kutateladze 2018). On the other hand, the homeostatic balance of the organism is directly related to stress exposure because it is well-known that stress condition affects numerous biological processes (neuroendocrine, physiological, and behavioural responses) through the hypothalamus-pituitary-adrenal cortex axis (Do Yup Lee and Choi 2015;Seyidoglu et al. 2021). In this context, many nutritional sources or additives (natural or synthetic), and their specific components have been long studied in vitro and in vivo to determine their anti-stress, therapeutic, and immunomodulatory potential, such as Spirulina platensis. ...
Article
Spirulina platensis has gradually gained more attention for its therapeutic, antioxidant, and anti-inflammatory potential worldwide. However, the current molecular knowledge about the effects of spirulina on stress-related genes is rather limited. The effects of dietary intake of spirulina on the HSP70 gene expression were assessed in a controlled in vivo experimental design. Moreover, alterations in serum corticosterone levels, IL-2, IL-4, IFN-γ, triglyceride, ALT, AST, relative gene expression values, and the correlations between them were evaluated. A total of 36 rats were divided into four groups: control group, stress-only group, spirulina group, and spirulina+stress group. To control the dose administration, S. platensis was applied by a gastric gavage in stress groups. Crowded environment stress and hosting alone stress were applied to the stress-only group and spirulina + stress group. RNA was extracted from brain samples using TRIpure and the relative gene expression assessment was performed using Roche-LightCycler-480-II real-time PCR-System. Gene expression values were remarkably different among the four experimental groups. The differences between stress-only and the spirulina groups were statistically significant (P<0.05). The correlation between the HSP70 gene expression and the IFN-γ was found to be statistically significant (P<0.05; r=0.50). Results indicate a novel effect of spirulina on the HSP70 expression related to the stress-response. Data presented in this study may be useful for further studies to define not only the molecular genetic aspects through dietary S. platensis but also the effects of spirulina on stress-response and animal welfare.Graphical abstract
Article
Background The chronic abnormal production of testosterone (T) is associated with many disorders in men. Fingernail clippings might be more suited for the diagnosis and medium-to-long term therapeutic monitoring for the T-related chronic disorders than the blood-derived specimens. The objective of this study was to characterize a thumbnail clipping as the specimen for assessing the several months-old T status. Methods Thumbnail clippings from various subjects were analyzed by liquid chromatography/electrospray ionization-tandem mass spectrometry to evaluate the gender difference, and changes caused by aging and androgen deprivation therapy (ADT) in the thumbnail T concentration. Results There was an evident gender difference in the thumbnail T concentrations [male; 2.55 ± 0.85 ng/g and female; 0.48 ± 0.29 ng/g, mean ± SD (n = 25 each), Welch t-test]. The thumbnail T concentrations significantly decreased with age in men (n = 268, Scheffé F-test), which was similar to those of the free or bioavailable T in serum/plasma. The thumbnail T concentrations sharply decreased by a 6-months ADT (especially the effect of the luteinizing hormone-releasing hormone agonist/antagonist) for patients with prostate cancer (n = 10). Conclusions The thumbnail clipping can be a specimen to retrospectively assess the average T production.
Article
Introduction: Breastfeeding is considered the best way to provide essential and necessary nutrients to the newborn, intervening in its growth and development. However, early abandonment of this method is quite common, due to various factors such as stress. Objectives: To determine whether the level of postpartum cortisol can serve as an indicator of maternal stress and whether there is a relationship between the level of cortisol and the rate of exclusive breastfeeding (EBF) at hospital discharge. Methodology: Systematic review of the literature under the PRISMA guidelines. PubMed, Web of Science, CINAHL, and Scopus databases were used. Original articles published from 2017 to 2022 in English, French, Portuguese, and Spanish were included. All study designs were eligible. Of the 3,712 records initially identified, 15 studies were included in this review. Results: Elevated cortisol levels, due to immediate postpartum stressors, have direct effects on the performance of the essential hormones in breast milk production. The EBF rates are negatively influenced by perceived maternal stress. Conclusion: Cortisol levels may be a good indicator of the level of stress to which the mother is subjected during the immediate postpartum period.
Article
Full-text available
Metabolomic results on human blood plasma largely depend on the sample preparation protocols employed for protein precipitation and metabolite extraction. Five different extraction methods were examined, which can be grouped into two categories, liquid-liquid extraction and protein precipitation methods, including long-standing protocols such as the Folch extraction and Bligh-Dyer extraction in comparison to modern methods such as the Matyash protocol and two global metabolite extraction methods. Extracts were subjected to analysis of blood plasma lipids and primary metabolites by using chip-based direct infusion nanoelectrospray tandem mass spectrometry and gas chromatography coupled to time-of-flight mass spectrometry, respectively. Optimal extraction schemes were evaluated based on the number of identified metabolites, extraction efficiency, compound diversity, reproducibility, and convenience for high-throughput sample preparations. Results showed that Folch and Matyash methods were equally valid and robust for lipidomic assessments while primary metabolites were better assessed by the protein precipitation methods with organic solvent mixtures. Graphical Abstract Schematic workflow of five extraction methods and subsequent mass spectrometry analysis using GC-TOF MS and nanoelectrospray direct-infusion ion trap MS/MSᅟ
Article
Full-text available
Context: Clinical features of Metabolic Syndrome (MetS) and Cushing's Syndrome are similar, suggesting a pathogenetic role of hypothalamus-pituitary-adrenal axis in MetS. Objective: The aim of the study was to determine whether MetS diagnosis and specific clusters of MetS components (waist circumference, dyslipidemia, hypertension, and impaired glucose metabolism) are associated with serum cortisol (SC) or 24-h urinary free cortisol (UFC) levels. Design and setting: We conducted cross-sectional analyses of data from our obesity cohort. We studied 264 obese children (age, 11.0 ± 2.8 years; male, 48%; BMI, 28.2 ± 5.4 kg/m(2)). We examined UFC, SC, homeostasis model assessment (HOMA), and features of MetS (waist circumference, blood pressure, fasting lipids, and glucose). Results: Slightly increased UFC concentrations were measured in 30.7% of the children. Obese children with MetS had significantly (P = .003) higher UFC levels compared with obese children without MetS. Girls demonstrated significantly higher UFC concentrations compared with boys independent of pubertal stage. UFC and SC levels were significantly related to features of MetS, but the associations were stronger for UFC. In multivariate analyses adjusted for age, sex, and body mass index, none of the features of MetS but HOMA index was correlated with UFC, whereas SC demonstrated no significant association to any parameter of MetS or HOMA. Conclusions: Our findings support the hypothesis that changes in the hypothalamus-pituitary-adrenal axis are related to MetS in obesity. UFC seems to be a suitable marker for this relationship. Norm values for UFC adapted to obese children may help to avoid unnecessary dexamethasone suppression tests.
Article
Full-text available
Context: The comparison of variability, reproducibility, and diagnostic performance of late-night salivary cortisol (LNSF) and urinary free cortisol (UFC) using concurrent and consecutive samples in Cushing's syndrome (CS) is lacking. Objectives, Patients, and Methods: In a prospective study, we evaluated 3 simultaneous and consecutive samples of LNSF by RIA and UFC by liquid chromatography associated with tandem mass spectrometry in Cushing's disease (CD) patients (n = 43), adrenal CS patients (n = 9), and obese subjects (n = 18) to compare their diagnostic performances. In CS patients, we also performed a modified CS severity index. Results: There was no difference in the coefficient of variation (percentage) between LNSF and UFC among the 3 samples obtained for each patient with Cushing's disease (35 ± 26 vs 31 ± 24), adrenal CS (28 ± 14 vs 22 ± 14), and obesity (39 ± 37 vs 48 ± 20). LNSF confirmed the diagnosis of hypercortisolism even in the presence of normal UFC in 17.3% of CS, whereas the inverse situation was not observed for UFC. The area under the receiver-operating characteristic curves for LNSF was 0.999 (95% credible interval [CI] 0.990-1.00) and for UFC was 0.928 (95% CI 0.809-0.987). The ratio between areas under the curve was 0.928 (95% CI 0.810-0.988), indicating better performance of LNSF than UFC in diagnosing CS. There was no association between the CS severity index and the degree of biochemical hypercortisolism. Conclusion: Our data show that despite similar variability between both methods, LNSF has a superior diagnostic performance than UFC and should be used as the primary biochemical diagnostic test for CS diagnosis.
Article
Full-text available
Context: Pathologically increased cortisol exposure induces obesity, but it is not known whether relatively high cortisol within the physiological range is related to childhood obesity. Objective: The aim of the study was to compare hair cortisol concentrations between obese and normal-weight children. Design: We performed an observational case-control study. Participants: Twenty obese children (body mass index-SD score [BMI-SDS]>2.3) and 20 age- and sex-matched normal-weight children (BMI-SDS<1.1) aged 8-12 years were recruited. Main outcome measures: Scalp hair samples from the posterior vertex were collected, and hair cortisol concentrations were measured using ELISA. Body weight, height, and waist circumference were measured. From the obese children, additional data on blood pressure and blood lipid concentrations were collected. Results: In both groups, five boys and 15 girls were included; their mean age was 10.8±1.3 vs 10.8±1.2 years (obese vs normal weight; not significant). Body weight, BMI, BMI-SDS, and waist circumference were higher in the obese children compared with the normal-weight children (69.8±17.2 vs 35.5±7.2 kg; 29.6±4.9 vs 16.4±1.6 kg/m2; 3.4±0.5 vs -0.2±0.8 SDS; 94±13 vs 62±6 cm; P<.001 all). Hair cortisol concentration was higher in obese than normal-weight children (median [interquartile range], 25 [17, 32] vs 17 [13, 21] pg/mg; P<.05). Conclusions: Hair cortisol concentration, a measure for long-term cortisol exposure, was higher in obese children than normal-weight children. This suggests long-term activation of the hypothalamus-pituitary-adrenal axis in obese children and may provide a novel target for treatment of obesity in children.
Over the past 5 yr, we have examined some of the sharpest edges of the pathology of aging. We have studied the capacity of aged organisms to respond appropriately to stress and the capacity of stress to cumulatively damage aging tissue. The idea of a relationship between stress and aging has permeated the gerontology literature in two forms. First, senescence has been thought of as a time of decreased adaptiveness to stress. This idea has been supported frequently, as many aged physiological systems function normally under basal conditions, yet do not adequately respond to a challenge. For example, aged and young humans have similar basal body temperatures, but the former are relatively impaired in thermoregulatory capacities when heat- or cold-challenged. A second theme in gerontology concerning stress is that chronic stress can accelerate the aging process. Selye and Tuchweber for example, postulated a finite "adaptational energy" in an organism, with prolonged stress prematurely depleting such reserves, thus accelerating the onset of senescence. This idea was derivative of earlier idea that the "rate of living" could be a pacemaker of aging. Experimentally, varied approaches have supported the notion that at least some biomarkers of age can be accelerated by stress. The above hypotheses led us to examine the adrenocortical axis, the endocrine axis which is among the most central to the stress response. Our findings support both of these concepts. We find that the aged male rat is impaired in terminating the secretion of adrenocortical stress hormones, glucocorticoids, at the end of stress. This hormonal excess may be due to degenerative changes in a region of the brain which normally inhibits glucocorticoid release; the degeneration, in turn, is caused by cumulative exposure to glucocorticoids. Together, these effects form a feed-forward cascade with potentially serious pathophysiological consequences in the aged subject. Reproduced by permission. Robert M. Sapolsky, Lewis C. Krey, Bruce S. McEwen, The Neuroendocrinology of Stress and Aging: The Glucocorticoid Cascade Hypothesis. Endocr. Rev. 7 , 284-301 (1986).
Article
An efficient procedure is described for the determination of estrone and 17 beta-estradiol in hair by gas chromatography-mass spectrometry (GC-MS). The method involves alkyloxycarbonylation with isobutyl chloroformate (isoBCF) of phenolic hydroxy groups after alkaline digestion of hair samples. The resulting isobutyloxycarbonyl derivatives of estrone and 17 beta-estradiol are extracted with hexane and subjected to chlorodifluoroacetyl derivatization in order to protect the remaining alcoholic hydroxy groups. When GC-MS with selected ion monitoring (SIM) was used, the quantitative ions were at m/z 270 and 384 in the electron ionization mass spectra for estrone and 17 beta-estradiol, respectively. The detection limits for SIM of the steroids were 1 and 2 pg, respectively, and the SIM responses were linear with correlation coefficients varying from 0.991 to 0.994 in the concentration range 0.2-4.0 ng g(-1) for the estrogens studied. The detection of estrone and 17 beta-estradiol in hair samples was possible in the concentration range of 0.24-1.30 ng g(-1). The concentrations of the two estrogens detected were different in male and female hair samples.
Article
The only approved drug for the treatment of adrenocortical cancer (ACC) is mitotane. Mitotane is adrenolytic and therefore, hydrocortisone replacement therapy is necessary. Since mitotane increases cortisol binding globulin (CBG) and induces CYP3A4 activity, high doses of hydrocortisone are thought to be required. Evaluation of hydrocortisone therapy in mitotane-treated patients has been difficult since there is no good marker to evaluate hydrocortisone therapy. Measurement of cortisol in scalp hair is a novel method that offers the opportunity to measure long-term cortisol levels. Our aim was to evaluate whether hair cortisol measurements could be useful in evaluating recent hydrocortisone treatment in mitotane-treated ACC patients. Hair cortisol levels were measured in 15 mitotane-treated ACC patients on hydrocortisone substitution and 96 healthy individuals. Cortisol levels were measured in 3 cm hair segments, corresponding to a period of 3 months. Hair cortisol levels were higher in ACC patients compared to healthy individuals (p<0.0001). Seven ACC patients (47%) had hair cortisol levels above the reference range. None of the patients had hair cortisol levels below normal. In contrast to hydrocortisone doses (β=0.03, p=0.93), hair cortisol levels were associated with BMI (β=0.53, p=0.042). There was no correlation between hair cortisol levels and hydrocortisone doses (β=0.41, p=0.13). Almost half of the ACC patients had high hair cortisol levels, suggesting long-term over-substitution of hydrocortisone in some of the patients, whereas none of the patients was under-substituted. Hair cortisol measurements might be useful in long-term monitoring hydrocortisone treatment in mitotane-treated ACC patients.
Article
In addition to the well-characterized role of the sex steroid receptors in regulating fertility and reproduction, reproductive events are also mediated by the hypothalamic-pituitary-adrenal axis in response to an individual's environment. Glucocorticoid secretion in response to stress contributes to the well characterized suppression of the hypothalamic-pituitary-gonadal axis through central actions in the hypothalamus and pituitary. However, both animal and in vitro studies indicate that other components of the reproductive system are also regulated by glucocorticoids. Furthermore, in the absence of stress, it appears that homeostatic glucocorticoid signaling plays a significant role in reproduction and fertility in all tissues comprising the hypothalamic-pituitary-gonadal axis. Indeed, as central regulators of the immune response, glucocorticoids are uniquely poised to integrate an individual's infectious, inflammatory, stress, nutritional, and metabolic status through glucocorticoid receptor signaling in target tissues. Endocrine signaling between tissues regulating the immune and stress response and those determining reproductive status provides an evolutionary advantage, facilitating the trade-off between reproductive investment and offspring fitness. This review focuses on the actions of glucocorticoids in tissues important for fertility and reproduction, highlighting recent studies which show glucocorticoid signaling plays a significant role throughout the hypothalamic-pituitary-gonadal axis, and characterizing these effects as permissive or inhibitory in terms of facilitating reproductive success.
Article
Congestive heart failure (CHF) is associated with increased stress and alterations in metabolism, favoring catabolism over anabolism. Hormonal profiles of patients with heart failure have been assessed using serum and saliva as matrices, which are only point measurements and do not provide long-term information. Scalp hair is a novel matrix that allows for measurement of hormones over a period of several months. We aimed to evaluate whether levels of cortisol and testosterone and their ratio (C/T) in hair correlate with severity of heart failure. We conducted a prospective study in ambulatory male patients with a left ventricular ejection fraction (LVEF)≤40%. Hormone levels were measured using immunoassays in the proximal 2cm of hair (representing approximately two months of systemic hormone exposure). Primary endpoints included the correlation of hair cortisol, testosterone, and C/T levels with the New York Heart Association (NYHA) class, LVEF, exercise capacity and NT-proBNP. The 44 CHF patients had a median hair level (range) of cortisol of 207 (117.7-1277.3)ng/g. Hair cortisol levels correlated positively with NYHA class (r=0.48, p=0.001) and negatively with treadmill stress test performance, (r=-0.37, p<0.05). The hair testosterone was 5.17 (2.39-24.64)ng/g and the C/T ratio was 39.89 (12.98-173.73). No associations were found between hair testosterone and C/T ratio and heart failure severity; however, the C/T ratio was higher in patients who required a CHF-related hospitalization than in patients who did not require this in the year following the inclusion in the study. Hair cortisol levels correlate with heart failure severity as assessed by the NYHA class and exercise capacity, while hair testosterone and C/T levels do not correlate with heart failure severity.