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Introduction:

 Cortisol,
a glucocorticoid and a stress hormone (1),
an important Biomarker, has diverse effects on inflammation and protein synthesis. As Cortisol is under the influence of Hypothalamo-Pituitary-Adrenal (HPA) axis, it is secreted in response to stress whether-physical, chemical or psychological. Various
researches indicate that Salivary Cortisol is nearly as accurate as Serum Cortisol (2), and even preferred over Serum Cortisol in cases of Child
(3) and Maternal care
(4),  adrenal disease
(5,6),  neurological disease.
 The method of collection of Salivary Cortisol is a simpler,
non-invasive approach, causing minimal-to no- discomfort to the patient (7). It reduces the risk of false values related to needle-stress (7). For this
purpose, Salivary Cortisol is routinely used in
western clinical setups (6, 8). It is also used as a Biomarker in Stress Research
(9, 10). Altered diurnal variation in Cortisol secretion is
indicated in various disease states (11). However, In
the
Indian Clinical scenario Serum Cortisol is being used for diagnostic and prognostic purposes,
there are fewer studies done on salivary cortisol. We aim to comparatively evaluate and correlate the salivary and Serum Cortisol levels
in order to establish preference of Salivary Cortisol over Serum Cortisol and to use Salivary Cortisol as a substitute to Serum Cortisol Assay in Indian Population.

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Objectives of the Study:

 

1    To
estimate the levels of Salivary and Serum Cortisol in Healthy Adults.

2.   To correlate the morning Salivary and Serum Cortisol
levels.

3.    To study the mean
difference between morning and evening Salivary Cortisol levels.

 

Material and Methods:

 

 Research Approach:

 Cross sectional study.
This study will be conducted among apparently normal Individuals.

 

Study Design:

 

 

 

 

 

 

 

 

 

 

 

Study Technique:

·     
2mL un-stimulated whole saliva sample will be collected 5
min after rinsing mouth repeatedly with distilled water, centrifuged at 3000rpm for 15 min, supernatant will be used for Salivary Cortisol Assay (12).

·     
2mL venous blood will be drawn in
a vaccutainer, centrifuged at 3000rpm for 15 min; supernatant will be used for Serum cortisol Assay.

·     
Serum
and salivary samples
are collected between 7-10am. And again between 4-8pm,
only saliva
will be collected.

 

Study participants:

To study the correlation between saliva and serum,
n=25 will be chosen, for the power of 0.9, number of predictor -1, with alpha
significance of 0.05 and effect size f2-0.35.

To study the mean difference between the morning
and evening Salivary Cortisol groups- n=25 will be chosen for the power of 0.8,
0.05 alpha significance, large effect size-0.35.

From 25 participants morning
and evening saliva samples and morning serum sample will be collected.

 

Study Tool:

Cortisol will be estimated by ECLIA method in both saliva and serum samples.

 

Inclusion Criteria:

 

·        
Age group: 18-40years.

·        
Sex: Both males and females.

·        
Healthy individuals with Sound Sleep before collecting samples.

 

 

Exclusion Criteria:

 

·        
Age: < 18 years and > 40years.

·        
Pregnancy

·        
Chronic illness: liver disease, acute or chronic renal failure,
hypertension, diabetes,
infectious
disease, Haematopoeitic disease.

·        
Autoimmune Disorders: Rheumatoid Arthritis, Sjögren’s Syndrome, SLE.

·        
Pre-operative
Head and Neck Surgery, Periodontitis.

·        
Acute illness

·        
Neuropsychological disorders:  Stress, Depression.

·        
Chronic smokers,
alcoholics, tobacco chewers.

·        
Malnutrition

 

 

 

 

 

 

 

Statistical Analysis:

Pearson’s
Correlation Test
will be done to study the correlation between saliva and serum cortisol levels.
To determine the significance of mean difference between the morning and
evening saliva levels groups, Mann- Whitney U test will be used.

 

Implications of the Study:

 

For estimating night Serum Cortisol level in
paediatric age group or even in adults, the stress caused by needle stick prick
and waking the patient in the night will falsely elevate the cortisol levels causing
loss of diurnal variation and will affect the report interpretation. 

For improved patient care and better prognosis, this study is directed toward the measurement of Salivary Cortisol, which
is a simple and non-invasive method,
and is especially required in pediatric age group, where collecting
Serum is cumbersome and painful.

Our study reinforces the use
of Serum Cortisol Assay as better, simple and non-invasive method.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Reference:

 

(1)  Lee DY, Kim E, Choi MH. Technical and clinical
aspects of cortisol as a biochemical
marker of chronic stress.
BMB Reports. 2015 Apr;48(4):209-216. Available from:
https://pubmed/25560699 Accessed on 17th December 2017

 

(2)  Tara
Kidd, Livia A. Carvalho, Andrew Steptoe. The relationship between cortisol responses
to laboratory stress and cortisol profiles in
daily life. Biological
Psychology. 2014 May;99:34–40.
Available from: https://doi.org/10.1016/j.biopsycho.2014.02.010

 

(3) Moore TA, Schmid KK, French JA. Comparison of cortisol samples
in the first two weeks of life in preterm infants. Journal of  Pediatric 
Endocrinology & Metabolism.
2015 Mar;28(3-4):415-420. doi: 10.1515/jpem-2014-0246.

 

(4) Lindsay F Hofman.
Human Saliva as a Diagnostic Specimen. In: Dennis M. Bier(ed.), D’Ann Finley(ed.). Innovative Non- or
Minimally-Invasive Technologies for Monitoring Health and Nutritional Status in
Mothers and Young Children. Children’s Nutrition Research Center, Baylor College of Medicine, Houston, TX. The Journal of Nutrition. 2001;131:p.1621S–1625S.

 

 (5) Ross F
Vining, Robynne A McGinley , JosephJ Maksvytis, Kian Y Ho. Salivary Cortisol: a better measure for Adrenal cortical function than Serum Cortisol. Annals of  Clinical Biochemistry:International
Journal of Laboratory Medicine. 1983;20(6):329-335. Available from: http://
doi/abs/10.1177/000456328302000601.

 

(6) Cinzia  Carrozza, Salvatore Maria Corsello, Rosa Maria Paragliola, Francesca Ingraudo, Sara Palumbo, Pietro Locantore et al. Clinical accuracy
of midnight salivary cortisol
measured by automated electrochemiluminescence
immunoassay method in Cushing’s Syndrome. Annals of  Clinical   Biochemistry:International
Journal of Laboratory Medicine. 2010;47:228-232. Available from:doi:10.1258/acb.2010.010020.

 

(7)  Kirschbaum C., Hellhammer D.H.
Salivary Cortisol in Psychobiological Research: An

       
Overview.
Neuropsychobiology. 1989;22:150-169. Available from: doi: 10.1159/issn.0302-282X

 

 (8) Jung C,
Greco S, Nguyen HH, Ho JT, Lewis JG, Torpy
DJ
et al. Plasma ,Saliva and Urinary
Cortisol levels following psychological stress doses of hydrocortisone in
Normal Volunteers. BMC Endocrine Disorder.2014
Nov;14:91. Available from: https://doi.org/10.1186/1472-6823-14-91

 

(9) Dirk H. Hellhammer, Stefan Wu¨st, Brigitte M. Kudielka. Salivary Cortisol as a Biomarker in Stress Research. Psychoneuroendocrinology. 2009
Feb;34(2):163-71. Available from: doi: 10.1016/j.psyneuen.2008.10.026.

 

 (10)
Isabelle E. Widmer, Jardena J. Puder, Caroline Konig, Hans Pargger,
Hans Reinhard Zerkowski Ju?rg Girard,
Beat Muller. Cortisol Response in
Relation to the Severity of Stress and Illness. The Journal of Clinical Endocrinology & Metabolism. 2005 Aug;90(8): 4579–4586. Available from:  https://doi.org/10.1210/jc.2005-0354

 

 (11)
Mantagos S, Moustogiannis A, Vagenakis AG. Diurnal variation of plasma cortisol
levels in infancy. Journal of  Pediatric 
Endocrinology & Metabolism. 1998 Jul-Aug;11(4):549-53. Available
from: https://www.ncbi.nlm.nih.gov/pubmed/9777576Acessed on 27th Dec2017

 

 (12)  Mahvash Navazesh,
Satish K.S. Kumar. Measuring salivary flow-Challenges and opportunities. The Journal of American Dental Association. 2008 May;139:35S-40S.
Available from:http://dx.doi.org/10.14219/jada.archive.2008.0353

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